Monsanto Company 10-SY-210U 1 of 721 

Petition for the Determination of Nonregulated Status for Dicamba-Tolerant Soybean 

MON 87708 

The undersigned submits this petition under 7 CFR §340.6 to request that the Administrator 

make a determination that the article should not be regulated under 7 CFR Part 340 

July 6, 2010 

(Revised on October 10, 2011, March 1, 2012 and March 26, 2012) 

OECD Unique Identifier: MON-877Ø8-9 

Monsanto Petition Number: 10-SY-210U 

Submitted by:

Rhonda M. Mannion, B.S. 

Monsanto Company 

800 North Lindbergh Blvd. 

St. Louis, MO 63167 

Phone: (314) 694-8785 

Fax: (314) 694-3080 

E-mail: rhonda.m.mannion@monsanto.com 

Prepared by:

L.A. Burzio, Ph.D., B.A. Comstock, B.S., A. Deffenbaugh, B.S., K.S. Giddings, Ph.D., R.L. Girault M.S., 

G.G. Harrigan, Ph.D., J.L. Honegger, Ph.D., D.L. Kendrick, M.S., R.M. Mannion B.S., 

M.C. McCann, M.S., B. Morrison, M.S., K.E. Niemeyer B.S., G.J. Rogan, M.S., R.W. Schneider, B.S., 

J.K. Soteres Ph.D., C. Wang, Ph.D., G. Watson, Ph.D., J. Wright, Ph.D. 

Contributors and/or Principal Investigators:

C. Arnevik, M.S., J. Berry, M.S., M. Bleeke, Ph.D., C.R. Brown, M.S., K. Carr, M.S., D. Carson, Ph.D., 

C.M. Dalton, B.A., A.J. Evans, M.P.H., R.A. Heeren, B.S., S. Hill, M.S., K.M. Huizinga, Ph.D., 

J. Kronenberg, Ph.D., T.C. Laufer, M.S., K.D. Lawry, M.S., V. Leopold, M.S., F. Lloyd, M.S., S.L. 

Phillips, M.S., J.F. Rice, Ph.D., Z. Song, M.S., R.S. Thoma, M.S., Q. Tian, Ph.D. M. Miller, Ph.D., M. 

Starke, Ph.D., J. Whitsel, M.S.
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RELEASE OF INFORMATION

Monsanto is submitting the information in this petition for review by the USDA as part of the 

regulatory process. By submitting this information, Monsanto does not authorize its release to 

any third party. In the event the USDA receives a Freedom of Information Act request, pursuant 

to 5 U.S.C., § 552, and 7 CFR § 1, covering all or some of this information, Monsanto expects 

that, in advance of the release of the document(s), USDA will provide Monsanto with a copy of 

the material proposed to be released and the opportunity to object to the release of any 

information based on appropriate legal grounds, e.g., responsiveness, confidentiality, and/or 

competitive concerns. Monsanto understands that a copy of this information may be made 

available to the public in a reading room and upon individual request as part of a public comment 

period. Except in accordance with the foregoing, Monsanto does not authorize the release, 

publication or other distribution of this information (including website posting) without 

Monsanto's prior notice and consent. 

©2012 Monsanto Company. All Rights Reserved. 

This document is protected under copyright law. This document is for use only by the regulatory 

authority to which it has been submitted by Monsanto Company and only in support of actions 

requested by Monsanto Company. Any other use of this material, without prior written consent 

of Monsanto, is strictly prohibited. By submitting this document, Monsanto does not grant any 

party or entity any right to license or to use the information or intellectual property described in 

this document. 

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EXECUTIVE SUMMARY 

The Animal and Plant Health Inspection Service (APHIS) of the United States (U.S.) 

Department of Agriculture (USDA) has responsibility, under the Plant Protection Act (Title IV 

Pub. L. 106-224, 114 Stat. 438, 7 U.S.C. § 7701-7772) to prevent the introduction and 

dissemination of plant pests into the U.S. APHIS regulation 7 CFR § 340.6 provides that an 

applicant may petition APHIS to evaluate submitted data to determine that a particular regulated 

article does not present a plant pest risk and should no longer be regulated. If APHIS determines 

that the regulated article does not present a plant pest risk, the petition is granted, thereby 

allowing unrestricted introduction of the article. 

Monsanto Company is submitting this request to APHIS for a determination of nonregulated 

status in whole for the new biotechnology-derived soybean product, MON 87708, any progeny 

derived from crosses between MON 87708 and conventional soybean, and any progeny derived 

from crosses of MON 87708 with other biotechnology-derived soybean that has been granted 

nonregulated status under 7 CFR Part 340. 

Product Description

Monsanto Company has developed biotechnology-derived soybean MON 87708 that is tolerant 

to dicamba (3,6-dichloro-2-methoxybenzoic acid) herbicide. MON 87708 offers growers an 

expanded use of dicamba in soybean production from the current preplant and preharvest labeled 

uses. The tolerance of MON 87708 to dicamba facilitates a wider window of application in 

soybean, allowing preemergence application up to the day of crop emergence and in-crop 

postemergence applications through the early reproductive (R1/R2) growth stage. Dicamba 

provides effective control of over 95 annual and biennial weed species, and suppression of over 

100 perennial broadleaf and woody plant species. Dicamba is efficacious on broadleaf weeds 

that are hard-to-control with glyphosate, such as common lambsquarters, hemp sesbania, 

morning glory species, nightshade, Pennsylvania smartweed, prickly sida, velvetleaf, waterhemp 

and wild buckwheat. Hard-to-control weeds generally require a higher rate and/or application at 

a smaller growth stage in order to consistently achieve commercially acceptable control. Refer 

to the Roundup WeatherMax label (U.S. EPA Reg. No. 524-537) for a listing of these weeds. 

Additionally, dicamba provides effective control of herbicide-resistant broadleaf weeds, 

including glyphosate-resistant weeds such as marestail, common ragweed, giant ragweed, palmer 

pigweed, and waterhemp. Herbicide resistant weeds are those listed on the International Survey 

of Resistant Weeds website (www.weedscience.org). 

MON 87708 will be combined with MON 89788 (Roundup Ready 2 Yield® soybean) utilizing 

traditional breeding techniques. Dicamba is an effective broadleaf herbicide and the potential 

use of dicamba and glyphosate herbicides at the same time in mixtures for weed control will 

provide growers greater application flexibility prior to planting as well as in-crop for greater 

consistency of control in both conventional and conservation tillage situations. Use of dicamba, 

in addition to glyphosate and the other herbicide options currently labeled for use on soybean, 

provides more options to implement diversified weed management programs to control a broad 

 

® Roundup Ready 2 Yield and Roundup Ready are registered trademarks of Monsanto Technology LLC. 
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spectrum of grass and broadleaf weed species. Successful adoption of the dicamba tolerance 

trait, into the Roundup Ready® soybean system, will provide: 1) growers with an opportunity for 

an efficient, effective weed management system; 2) an option to delay or prevent further 

resistance to glyphosate and other critically important soybean herbicides, in particular, 

herbicides in the ALS and PPO class of chemistry; 3) excellent crop safety, and 4) continue to 

provide soybean growers with effective weed control systems necessary for production yields to 

meet the growing needs of the food, feed, and industrial markets. The combination of dicamba 

and glyphosate tolerance in soybeans will also provide the basis for delaying or preventing the 

evolution of further weed resistance to glyphosate, dicamba, and herbicides in general, because 

of the ability to use these two modes of action in mixtures and sequences. 

MON 87708 contains a gene from Stenotrophomonas maltophilia that expresses a monooxygenase enzyme that rapidly demethylates dicamba rendering it inactive, thereby conferring 

tolerance to dicamba. The demethylation of dicamba produces 3,6-dichlorosalicylic acid 

(DCSA), a known soybean, soil, and livestock metabolite whose safety has been evaluated by the 

Environmental Protection Agency (U.S. EPA). DCSA, in addition to dicamba, is included in the 

current 10 ppm pesticide residue tolerance for soybean seed that supports the existing uses of 

dicamba on commercial soybean (40 CFR § 180.227). Even with the expanded use of dicamba 

on MON 87708, compared to commercial soybean uses, the rapid metabolism of dicamba results 

in residues in dicamba-treated MON 87708 seed, including the DCSA metabolite, that are well 

below the established 10 ppm tolerance, and therefore no modification to the existing soybean 

seed tolerance is needed. Consequently, only approval for the expanded use pattern of dicamba 

on MON 87708 has been requested of EPA. 

Data and Information Presented Confirm the Lack of Plant Pest Potential of MON 87708 

Compared to Conventional Soybean

The data and information presented in this petition demonstrate MON 87708 is agronomically, 

phenotypically, and compositionally comparable to conventional soybean with the exception of 

its tolerance to dicamba. Moreover, the data presented demonstrate MON 87708 is unlikely to 

pose an increased plant pest risk, including weediness or adverse environmental impact, 

compared to conventional soybean. The food, feed, and environmental safety of MON 87708 

was confirmed based on multiple, well-established lines of evidence: 

1. Soybean is a familiar crop that does not possess any of the attributes commonly 

associated with weeds and has a history of safe consumption. 

2. A detailed molecular characterization of the inserted DNA demonstrated a single, intact 

copy of the T-DNA insert in a single locus within the soybean genome. 

3. Data confirmed that the dicamba mono-oxygenase (DMO) in MON 87708 (MON 87708 

DMO) is unlikely to be a toxin or allergen based on extensive information collected. 

4. A compositional assessment of seed and forage confirmed that MON 87708 is 

compositionally equivalent to conventional soybean. 
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5. An extensive evaluation on phenotypic and agronomic characteristics and environmental 

interactions of MON 87708 demonstrated no increased plant pest potential compared to 

conventional soybean. 

6. An assessment of potential impact on non-target organisms (NTOs) and endangered 

species indicated that, under normal agricultural conditions, MON 87708 is unlikely to 

have adverse effects on these organisms compared to conventional soybean. 

7. Evaluation of MON 87708 using current cultivation and management practices for 

soybean concluded that deregulation of MON 87708 will not significantly impact 

soybean agronomic practices or land use, with the exception of the expanded window of 

dicamba application. 

Soybean is a Familiar Crop Lacking Weedy Characteristics 

There is a longstanding history of safe use and consumption of conventional soybean and 

processed products. Soybean is grown as a commercial crop in over 35 countries. Domestication 

occurred as early as 1000 B.C. and is now the most widely grown oilseed crop in the world, with 

approximately 211 million metric tons of harvested seed produced in 2008, which represented 

56% of world oilseed seed production that year. 

The commercial soybean species in the U.S. (Glycine max L. Merr.) does not exhibit weedy 

characteristics, does not invade established ecosystems, and does not outcross to weedy relatives. 

Soybean is not listed as a weed in major weed references, nor is it present on the lists of noxious 

weed species distributed by the federal government (7 CFR Part 360). During 2004 to 2008, U.S. 

growers planted between 64.7 and 75.7 million acres of soybean. Soybean does not possess any 

of the attributes commonly associated with weeds, such as long persistence of the seed in the soil, 

ability to disperse, invade, or become a dominant species in new or diverse landscapes, or the 

ability to compete well with native vegetation. However, due to a pronounced lack of dormancy 

it is known that soybean seed can germinate quickly under adequate temperature and moisture 

conditions, and can potentially grow as a volunteer plant. However, a volunteer soybean plant 

likely would be killed by frost during the autumn or winter of the year it germinated. 

Furthermore, if a volunteer plant were to survive, it would not compete well with the succeeding 

crop, and would be controlled readily via mechanical or other chemical means. Twenty 

commonly used agricultural herbicides, representing eight modes-of-action (i.e., ALS-inhibitor, 

chloroacetamide, EPSPS, PPO inhibitor, PSI disruption, PSII inhibitor, synthetic auxin, and 

tubulin inhibitor classes) were tested as potential substrates for MON 87708 DMO. None of the 

herbicides tested were found to effect the tolerance of MON 87708 at commercial application 

rates, therefore, herbicides effective for control of volunteer soybean can still be used to control 

MON 87708 volunteers. Finally, since wild populations of Glycine species are not known to 

exist in the U.S., there is no potential for MON 87708 to outcross to wild or weedy relatives. 

Conventional Soybean A3525 is an Appropriate Comparator to MON 87708

Soybean variety A3525 is the near isogenic line to MON 87708 and was used as the 

conventional soybean comparator to support the safety assessment of MON 87708. MON 87708 

and the near isogenic conventional soybean control A3525 have similar genetic backgrounds 
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with the exception of the dmo expression cassette, thus, the effect of the dmo expression cassette 

and the expressed MON 87708 DMO could be assessed in an unbiased manner. 

Molecular Characterization Verifies the Integrity and Stability of the Inserted DNA in 

MON 87708

MON 87708 was developed through Agrobacterium-mediated transformation of conventional 

soybean A3525 meristem tissue with the 2T-DNA plasmid vector PV-GMHT4355. 

PV-GMHT4355 contains two separate T-DNAs that are each delineated by Left and Right 

Border sequences. The first T-DNA, designated as T-DNA I, contains the dmo expression 

cassette regulated by the peanut chlorotic streak virus (PC1SV) promoter and the pea E9 3′ nontranslated region. The second T-DNA, designated as T-DNA II, contains the cp4 epsps

expression cassette under the regulation of the figwort mosaic virus (FMV) promoter and the pea 

E9 3′ non-translated region. During transformation, both T-DNAs were inserted into the 

soybean genome, where T-DNA II, containing the cp4 epsps expression cassette, functioned as a 

marker gene for the selection of transformed plantlets. Subsequently, conventional selfpollination breeding methods and segregation were used to isolate a plant containing the dmo

expression cassette but not containing the cp4 epsps expression cassette, resulting in the 

production of marker-free, dicamba-tolerant soybean MON 87708. 

Molecular characterization by Southern blot analyses determined that MON 87708 contains one 

copy of the T-DNA I at a single integration locus and all expression elements are present. These 

data also demonstrated that MON 87708 does not contain detectable backbone sequences from 

the plasmid vector or T-DNA II sequences. The complete DNA sequence of the insert and 

adjacent genomic DNA sequence in MON 87708 confirmed the integrity of the inserted dmo

expression cassette within the inserted sequences and identified the 5′ and 3′ insert-to-genomic 

DNA junctions. Furthermore, Southern blot analysis demonstrated that the insert in MON 87708 

has been maintained through at least five generations of breeding, thereby confirming the 

stability of the insert over multiple generations. 

Data Confirm MON 87708 DMO Safety

MON 87708 contains a dmo expression cassette that results in two forms of the DMO protein; 

referred to as DMO and DMO+27 (Section V.A.). The active form of these proteins, necessary 

to confer dicamba tolerance, is a trimer comprised of three DMO monomers. In MON 87708, 

the trimer can be comprised of DMO, DMO+27, or a combination of both. Therefore, this 

document will refer to both forms of the protein and all forms of the trimer as MON 87708 

DMO. 

A multistep approach was used to characterize MON 87708 DMO. This detailed 

characterization and assessment confirmed that MON 87708 DMO is safe for human and animal 

consumption. The assessment involved: 1) characterization of the physicochemical and 

functional properties of MON 87708 DMO; 2) quantification of MON 87708 DMO levels in 

plant tissues; 3) comparison of the amino acid sequence of MON 87708 DMO to known 

allergens, gliadins, glutenins, toxins, and other biologically active proteins known to have 

adverse effects on mammals; 4) evaluation of the digestibility of MON 87708 DMO in simulated 

gastric and intestinal fluids; 5) endogenous and exogenous substrate specificity of DMO; 6) 
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documentation of the history of safe consumption of mono-oxygenases (the class of enzymes to 

which MON 87708 DMO belongs); and 7) investigation of the potential mammalian toxicity 

through an oral gavage assay. 

DMO was found to be specific to dicamba when tested using structurally similar endogenous 

substrates and exogenous herbicide substrates representing a wide range of modes-of-action. 

MON 87708 DMO has no relevant amino acid sequence similarities with known allergens, 

gliadins, glutenins, toxins, and other biologically active proteins that may have adverse effects on 

mammals. MON 87708 DMO was rapidly degraded in simulated gastric and intestinal fluids 

and a high dose of this protein in a mouse acute oral toxicity evaluation demonstrated that it is 

not acutely toxic, and does not cause any adverse effect. The safety assessment supports the 

conclusion that exposure to MON 87708 DMO poses no meaningful risk to the environment, or 

human and animal health. 

MON 87708 is Compositionally Equivalent to Conventional Soybean

Detailed compositional analyses in accordance with OECD guidelines were conducted to assess 

whether levels of key nutrients and anti-nutrients in MON 87708 were comparable to levels 

present in the aforementioned near isogenic conventional soybean control A3525 and several 

commercial reference soybean varieties. Seed and forage were harvested from five individual 

sites in which MON 87708 (both treated with dicamba herbicide at the V2-V3 growth stage and 

not treated with dicamba herbicide), the conventional control, and a range of commercial 

reference varieties were grown concurrently in the same field trial. The commercial reference 

varieties used to establish a range of natural variability for the key nutrients and anti-nutrients in 

commercial soybean varieties have a history of safe consumption. Nutrients assessed in this 

analysis included proximates (ash, carbohydrates by calculation, moisture, protein, and fat), fiber, 

amino acids (18 components), fatty acids (FA, C8-C22), and vitamin E (α-tochopherol) in seed, 

and proximates (ash, carbohydrates by calculation, moisture, protein, and fat) and fiber in forage. 

The anti-nutrients assessed in seed included raffinose, stachyose, lectin, phytic acid, trypsin 

inhibitors, and isoflavones (daidzein, genistein, and glycitein). 

The combined-site analysis was conducted to determine statistically significant differences (5% 

level of significance) between MON 87708 and the near isogenic conventional control A3525. 

The results from the combined-site data were reviewed using considerations relevant to food and 

feed safety and nutritional quality. These considerations included assessments of: 1) the relative 

magnitudes of the difference in the mean values of nutrient and anti-nutrient components of 

MON 87708 and the conventional control, 2) whether the MON 87708 component mean value 

was within the range of natural variability of that component as represented by the 99% tolerance 

interval of the commercial reference varieties grown concurrently in the same field trial, 3) 

analyses of the reproducibility of the statistically significant combined-site component 

differences at individual sites, and 4) assessing the differences within the context of natural 

variability of commercial soybean composition published in the scientific literature and in the 

International Life Sciences Institute (ILSI) Crop Composition Database. 

Assessment of the analytical results confirmed that the differences observed in the combined-site 

analysis were not meaningful to food and feed safety or the nutritional quality of MON 87708 

soybean. In addition, the levels of assessed components in MON 87708 were compositionally
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 equivalent to the conventional control and within the range of variability of the commercial 

reference varieties that were grown concurrently in the same field trial. 

MON 87708 Does Not Change Soybean Plant Pest Potential or Environmental Interactions

Assessing the plant pest potential of a biotechnology-derived crop includes the concept of 

familiarity that the USDA recognizes as an important consideration. Familiarity is based upon 

the fact that the new biotechnology-derived plant is developed from a conventional plant variety 

whose biological properties and plant pest potential are well known. Familiarity considers the 

biology of the plant, the introduced trait, the receiving environment, and the interactions among 

these factors that provides a basis for comparative risk assessment between a biotechnologyderived plant and the conventional control. Following this concept, the phenotypic, agronomic, 

and environmental interaction assessment of MON 87708 included the near isogenic 

conventional soybean control A3525 and the commercial reference varieties. Characteristics 

assessed included: seed dormancy and germination, pollen morphology, and symbiont 

interactions conducted in the laboratory and greenhouse; and plant phenotypic and agronomic 

evaluations and environmental interaction observations conducted in the field. The commercial 

soybean reference varieties grown concurrently were used to establish a range of natural 

variability for each assessed characteristic in soybean. The phenotypic, agronomic, and 

environmental interaction assessment demonstrated that MON 87708 is equivalent to the 

conventional control. Thus, MON 87708 is unlikely to have a changed plant pest potential 

compared to conventional soybean. 

Seed dormancy and germination characterization demonstrated that MON 87708 seed had 

germination characteristics similar to seed of the conventional control. In particular, the lack of 

hard seed, a well-accepted characteristic of weediness affecting seed germination, supports a 

conclusion of no increased weediness of MON 87708 when compared to the conventional 

control. For pollen characteristics and symbiont interactions, there were no statistically 

significant differences (5% level of significance) observed between MON 87708 and the 

conventional control for any of the parameters measured, including pollen viability and diameter, 

nodule number and dry weight, shoot total nitrogen, and shoot and root dry weight. Collectively, 

these results support the conclusion that MON 87708 is not likely to exhibit increased plant pest 

potential compared to conventional soybean. 

The field evaluation of phenotypic, agronomic, and environmental interaction characteristics of 

MON 87708 also support the conclusion that MON 87708 is not likely to have an increased plant 

pest potential compared to conventional soybean. The evaluations were conducted at 18 

replicated field sites across North American soybean production regions. These assessments 

included plant growth and development characteristics, as well as observations for plant 

responses to abiotic stressors and plant-disease and plant-arthropod interactions. The observed 

phenotypic characteristics were similar between MON 87708 and the conventional control. 

In a combined-site analysis, data show no statistically significant differences (5% level of 

significance) between MON 87708 and the conventional control for early stand count, seedling 

vigor, days to 50% flowering, lodging, pod shattering, final stand count, seed moisture, seed test 

weight, or yield. Two statistically significant differences were detected between MON 87708 
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and the conventional control for plant height and 100 seed weight. MON 87708 was slightly 

taller and had a lower 100 seed weight than the conventional control. However, both differences 

were small in magnitude. Additionally, MON 87708 and the conventional control were within 

the same range of plant growth stages for 131 out of the 132 growth stage observations among 

the sites. Except for the differences in plant height, 100 seed weight, and a single growth stage 

observation at one site, all values for MON 87708 fell within the range of the commercial 

reference varieties grown concurrently. None of these differences were considered biologically 

meaningful in terms of increased plant pest potential of MON 87708 compared to conventional 

soybean. 

In an individual-site assessment of abiotic stress response and disease damage, no differences 

were observed between MON 87708 and the conventional control for 193 out of 194 

comparisons for the assessed abiotic stressors or for any of the 215 comparisons for the assessed 

diseases among all observations at the 18 sites. One difference was observed between 

MON 87708 and the conventional control for wind damage during a single observation at one 

site. The damage rating for MON 87708 (slight damage) was outside the range of the 

commercial reference varieties (no damage); however, the difference was not observed during 

any of the other 29 wind damage observations among the sites. Thus, the slight difference in 

wind damage rating was not indicative of a consistent plant response associated with 

MON 87708 and is not considered biologically meaningful in terms of increased plant pest 

potential or an altered environmental impact from MON 87708 compared to conventional 

soybean. 

In an assessment of arthropod-related damage, no statistically significant differences (5% level 

of significance) were detected between MON 87708 and the conventional control for 89 out of 

95 comparisons for the assessed arthropods. Lack of variability in the data precluded statistical 

comparisons between MON 87708 and the conventional control for 121 additional comparisons; 

however, the means for MON 87708 and the conventional control were the same value for these 

comparisons, indicating no biological differences. For each of the six statistically significant 

differences between MON 87708 and the conventional control, the severity of arthropod-related 

damage to MON 87708 was within or slightly outside the range of the commercial reference 

varieties. The differences between MON 87708 and the conventional control were small in 

magnitude and were not consistent across observations or sites. Thus, the differences in 

arthropod-related damage are not indicative of a consistent plant response associated with 

MON 87708 and are not considered biologically meaningful in terms of increased plant pest 

potential or an altered environmental impact from MON 87708 compared to conventional 

soybean. 

In an assessment of pest and beneficial arthropod abundance, no statistically significant 

differences (5% level of significance) were detected between MON 87708 and the conventional 

control for 142 out of 151 comparisons (including 74 arthropod pest and 77 beneficial arthropod 

comparisons) among the multiple collections conducted during the season at four sites. For the 

nine detected differences in arthropod abundance, seven were arthropod pests (green cloverworm, 

Japanese beetles, and stink bugs) and two were beneficial arthropods (spiders and Nabis spp). 

The differences detected in pest and beneficial arthropod abundance were small in magnitude 

and were not consistent with other collection times at the individual sites or across the sites. 

Consequently, it is concluded that the differences in pest and beneficial arthropod abundance are 
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not indicative of a consistent plant response associated with MON 87708 and are not biologically 

meaningful in terms of increased plant pest potential or an altered environmental impact from 

MON 87708 compared to conventional soybean. 

Field evaluations of phenotypic, agronomic, and environmental interaction characteristics of 

MON 87708 treated with dicamba herbicide were also conducted. Data were collected from 

field trials conducted at eight sites within the U.S. soybean producing regions. These 

assessments included plant growth and development characteristics, as well as observations for 

plant responses to abiotic stressors, plant-disease and plant-arthropod interactions. The 

phenotypic, agronomic, and environmental interaction assessment demonstrated that treated 

MON 87708 is equivalent to the conventional control. Thus, MON 87708 is unlikely to have an 

altered plant pest potential compared to conventional soybean. 

The observed phenotypic characteristics were similar between the dicamba-treated MON 87708 

and the conventional control. In a combined-site assessment, no statistically significant 

differences were detected between treated MON 87708 and the conventional control for early 

stand count, seedling vigor, days to 50% flowering, plant height, lodging, pod shattering, final 

stand count, seed moisture, or yield. One statistically significant difference was detected 

between treated MON 87708 and the control, for 100 seed weight. The difference in 100 seed 

weight was relatively small in magnitude and the mean 100 seed weight of treated MON 87708 

was slightly below the reference range. It is unlikely that this small difference in 100 seed 

weight would contribute to increased weed potential of MON 87708 when treated with dicamba 

compared to conventional soybean. Additionally, treated MON 87708 and the control were 

within the same range of plant growth stages for all growth stage observations among the sites. 

None of these differences were considered biologically meaningful in terms of increased plant 

pest potential of treated MON 87708 compared to conventional soybean. 

In an assessment of plant response to abiotic stressors and disease damage, no differences were 

observed between treated MON 87708 and the conventional control for 181 of 182 comparisons 

among all observations at the eight sites. One difference was observed between treated 

MON 87708 and the control for white mold during a single observation (slight vs. none). The 

damage rating for treated MON 87708 was outside of the reference range (no damage was 

observed in the references). This difference was not observed in any of the other two white mold 

evaluations across the sites and is not considered biologically meaningful in terms of increased 

plant pest potential or an altered environmental impact from treated MON 87708 compared to 

conventional soybean. 

In an assessment of arthropod-related damage, there were no statistically significant differences 

detected between treated MON 87708 and the control for 56 out of 59 comparisons. Lack of 

variability in the data precluded statistical comparisons between treated MON 87708 and the 

conventional control for 34 additional comparisons. The mean damage ratings for bean leaf 

beetle and grasshopper damage was outside the reference range however the response was not 

consistent across observations or sites. Thus, the results are not considered biologically 

meaningful in terms of adverse environmental impacts of treated MON 87708 compared to the 

conventional soybean. 
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In summary, the phenotypic, agronomic, and environmental interaction data were collected to 

provide a detailed characterization of MON 87708 and to assess whether the introduction of the 

dicamba tolerance trait in MON 87708 and the associated application of dicamba herbicide alters 

the plant pest potential compared to conventional soybean. The analysis considered the 

comparisons of MON 87708 to the conventional control, the reproducibility, magnitude, and 

direction of detected differences (trends), and comparison to the range of the commercial 

reference varieties. Results from the phenotypic, agronomic, and environmental interactions 

assessment indicated that MON 87708 does not possess weedy characteristics, increased 

susceptibility or tolerance to specific abiotic stress, diseases, or arthropods, or characteristics that 

would confer a plant pest risk or a significant environmental impact compared to conventional 

soybean. 

MON 87708 Will Not Adversely Affect NTOs or Threatened and Endangered Species

Evaluation of the impacts of a biotechnology-derived crop on Non-Target Organisms (NTOs) 

and threatened and endangered species is a component of the plant pest risk assessment. Since 

MON 87708 does not possess pesticidal activity, all organisms that interact with MON 87708 are 

considered to be NTOs. The environmental assessment demonstrated that the presence of the 

dicamba tolerance trait in MON 87708 and the associated application of dicamba did not alter 

plant-arthropod interactions, including beneficial arthropods, or alter disease susceptibility 

compared to the conventional control. 

The biochemical information and experimental data for evaluation of MON 87708 included 

molecular characterization, MON 87708 DMO safety assessments, the history of environmental 

exposure to mono-oxygenases (the class of enzymes to which MON 87708 DMO belongs), 

information from the environmental interaction assessment, demonstration of compositional 

equivalence to conventional soybean, and demonstration of agronomic and phenotypic 

equivalence to conventional soybean. Taken together, these data support the conclusion that 

MON 87708 has no reasonable mechanism for harm to NTOs, or to pose an additional risk to 

threatened and endangered species compared to the cultivation of conventional soybean. 

The potential for outcrossing and gene introgression from MON 87708 to sexually-compatible 

species in the U.S. is unlikely since no known wild Glycine species related to cultivated soybean 

are known to be present in North America. Furthermore, should cross-pollination occur, 

MON 87708 and its progeny are not expected to exhibit a significant environmental impact 

because, as described above, evaluations have shown that the presence of the dicamba tolerance 

trait is not likely to enhance weediness or plant-pest potential. Therefore, the environmental 

consequence of pollen transfer from MON 87708 to other Glycine species is considered 

negligible. 

Deregulation of MON 87708 Will Not Significantly Impact Soybean Agronomic Practices 

or Land Use

Soybean fields are typically highly managed agricultural areas that are dedicated to crop 

production for many years. Cultivation of MON 87708 would not be expected to differ from 

typical soybean cultivation, with the sole exception of an expanded window of dicamba 

applications due to the presence of the dicamba tolerance trait in MON 87708. MON 87708 
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likely would be used in common rotations on land currently used for agricultural purposes. As 

demonstrated, MON 87708 is similar to conventional soybean in its agronomic, phenotypic, 

ecological, and compositional characteristics and has comparable levels of resistance to insects 

and diseases as compared to commercial soybean. Therefore, the introduction of MON 87708 

into the Roundup Ready soybean system is not expected to have a significant impact on current 

cultivation and management practices for soybean. The adoption of MON 87708 into the 

Roundup Ready soybean system will provide growers with another herbicide mode-of-action and 

the means to control broadleaf weeds, including hard-to-control and herbicide-resistant broadleaf 

weeds, and will help preserve conservation tillage practices by providing growers with an 

additional weed management tool. Based on these considerations, there is no apparent potential 

for significant impacts on agronomic practices or land use, with the exception of the expanded 

application window of dicamba. 

Conclusion

Based on the data and information presented in this petition, it is concluded that MON 87708 is 

not likely to be a plant pest. Therefore, Monsanto Company requests a determination from 

APHIS that MON 87708 and any progeny derived from crosses between MON 87708 and 

conventional soybean or previously deregulated biotechnology-derived soybean, be granted 

nonregulated status under 7 CFR Part 340. 
Monsanto Company 10-SY-210U 14 of 721 

TABLE OF CONTENTS 

RELEASE OF INFORMATION .........................................................................................2

CERTIFICATION ...............................................................................................................3

EXECUTIVE SUMMARY .................................................................................................4

TABLE OF CONTENTS ...................................................................................................14

LIST OF TABLES .............................................................................................................19

LIST OF FIGURES ...........................................................................................................26

ABBREVIATION AND DEFINITIONS ..........................................................................28

I. RATIONALE FOR THE DEVELOPMENT OF MON 87708 .....................................32

I.A. Basis for the Request for a Determination of Nonregulated Status under 

7 CFR § 340.6 ...............................................................................................................32

I.B. Rationale for the Development of Dicamba-Tolerant Soybean 

MON 87708 ...................................................................................................................32

I.C. Submissions to Other Regulatory Agencies ..........................................................34

I.C.1. Submission to FDA .....................................................................................34

I.C.2. Submission to EPA .....................................................................................35

I.C.3. Submissions to Foreign Government Agencies ..........................................35

II. THE BIOLOGY OF SOYBEAN .................................................................................36

II.A. Soybean as a Crop ................................................................................................36

II.B. Characteristics of the Recipient Plant ..................................................................36

II.C. Soybean as a Test System in Product Safety Assessment ...................................37

III. DESCRIPTION OF THE GENETIC MODIFICATION ...........................................38

III.A. The Plasmid Vector PV-GMHT4355 .................................................................38

III.B. Description of the Transformation System .........................................................43

III.C. The dmo Coding Sequence and MON 87708 DMO (T-DNA I) ........................45

III.D. The cp4 epsps Coding Sequence and the CP4 EPSPS Protein 

(T-DNA II) ....................................................................................................................45

III.E. Regulatory Sequences .........................................................................................45

III.F. T-DNA Borders ...................................................................................................46

III.G. Genetic Elements Outside of the T-DNA Borders .............................................46

IV. CHARACTERIZATION OF THE GENETIC MODIFICATION .............................47

IV.A. Insert and Copy Number of T-DNA I in MON 87708 .......................................52

IV.A.1. Probe 8 .....................................................................................................53

IV.A.2. Probe 9 .....................................................................................................53
Monsanto Company 10-SY-210U 15 of 721 

IV.A.3. Probe 10 ...................................................................................................54

IV.B. Southern Blot Analysis to Determine the Presence or Absence of 

T-DNA II Sequences in MON 87708 ............................................................................58

IV.B.1. Probe 4 .....................................................................................................58

IV.B.2. Probe 5 .....................................................................................................58

IV.B.3. Probe 6 .....................................................................................................59

IV.C. Southern Blot Analysis to Determine the Presence or Absence of 

PV-GMHT4355 Backbone Sequences in MON 87708 ................................................63

IV.C.1. Plasmid Vector Backbone Probes 1, 2, 3, and 7 ......................................63

IV.D. Organization and Sequence of the Insert and Adjacent DNA in 

MON 87708 ...................................................................................................................65

IV.E. PCR and DNA Sequence Analyses to Examine the MON 87708 

Insertion Site ..................................................................................................................65

IV.F. Southern Blot Analysis to Examine Insert Stability in Multiple 

Generations of MON 87708 ..........................................................................................65

IV.F.1. Probe 9 ......................................................................................................66

IV.G. Inheritance of the Genetic Insert in MON 87708 ...............................................69

IV.H. Genetic Modification Characterization Conclusion ...........................................73

V. CHARACTERIZATION AND SAFETY ASSESSMENT OF THE 

MON 87708 DMO .............................................................................................................74

V.A. Function of DMO and MON 87708 DMO ..........................................................74

V.A.1. Formation of MON 87708 DMO ..............................................................75

V.A.2. Specificity of MON 87708 DMO .............................................................75

V.B. Characterization of MON 87708 DMO ...............................................................77

V.C. Expression Levels of MON 87708 DMO ............................................................78

V.D. Assessment of Potential Allergenicity of MON 87708 DMO .............................81

V.E. Safety Assessment Summary of MON 87708 DMO ...........................................81

V.F. MON 87708 DMO Characterization and Safety Conclusion ...............................84

VI. COMPOSITIONAL ASSESSMENT OF MON 87708 ..............................................86

VI.A. Compositional Equivalence of MON 87708 Seed and Forage to 

Conventional Soybean ...................................................................................................86

VI.A.1. Composition of Soybean Seed and Forage (Treated) ..............................88

 VI.A.2. Composition of Soybean Seed and Forage (Untreated)..........................93

VI.B. Compositional Assessment Conclusion ...........................................................144

VII. PHENOTYPIC, AGRONOMIC, AND ENVIRONMENTAL 

INTERACTIONS ASSESSMENT ..................................................................................146
Monsanto Company 10-SY-210U 16 of 721 

VII.A. Characteristics Measured for Assessment ......................................................146

VII.B. Interpretation of Phenotypic and Environmental Interaction Data .................150

VII.B.1. Interpretation of Detected Differences Criteria ....................................150

VII.C. Comparative Assessments of the Phenotypic, Agronomic, and 

Environmental Interaction Characteristics of MON 87708 ........................................153

VII.C.1. Seed Dormancy and Germination Characteristics ................................153

VII.C.2. Phenotypic, Agronomic, and Environmental Interaction 

Characteristics Evaluated under Field Conditions ...............................................156

VII.C.2.1. Phenotypic, Agronomic, and Environmental Interaction 

Characteristics for Untreated MON 87708 Evaluated under 2008 Field 

Conditions ............................................................................................................156

VII.C.2.2. Phenotypic, Agronomic, and Environmental Interaction 

Characteristics for Dicamba-Treated Evaluated under 2009 Field 

Conditions ............................................................................................................157

VII.C.2.3. Field Phenotypic and Agronomic Characteristics .............................158

VII.C.2.4. Environmental Interaction Characteristics ........................................162

VII.C.3. Pollen Characteristics ...........................................................................166

VII.C.4. Symbiont Interactions ...........................................................................167

VII.D. Phenotypic, Agronomic, and Environmental Interactions Assessment 

Conclusion ...................................................................................................................169

VIII. U.S. AGRONOMIC PRACTICES ........................................................................170

VIII.A. Introduction ...................................................................................................170

VIII.B. Overview of U.S. Soybean Production ..........................................................171

VIII.B.1. Soybean Production .............................................................................171

VIII.B.2. Soybean Seed Production ....................................................................176

VIII.C. Production Management Considerations .......................................................179

VIII.C.1. Pre-Season ...........................................................................................179

VIII.C.2. Planting and Early Season ...................................................................180

VIII.C.3. Mid to Late Season ..............................................................................182

VIII.C.4. Harvest Season ....................................................................................183

VIII.D. Management of Insects ..................................................................................183

VIII.E. Management of Diseases and Other Pests .....................................................184

VIII.F. Weed Management .........................................................................................186

VIII.F.1. Methods of Weed Control in Soybean .................................................189

VIII.G. Dicamba Herbicide Use in the U.S. ...............................................................197

VIII.G.1. Dicamba Application Timing for Labeled Crops ................................200
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VIII.G.2. Distribution of Dicamba Use in the U.S. ............................................202

VIII.G.3. Potential Impacts to Adjacent Crops ...................................................207

VIII.H. Dicamba-Tolerant Soybean MON 87708 ......................................................208

VIII.H.1. Potential Impact of Dicamba Application Timing to 

MON 87708 .........................................................................................................217

VIII.H.2. Impact on Dicamba Use in U.S. Soybean Production Following 

Integration of MON 87708 into the Roundup Ready Soybean System ...............221

VIII.H.3. Conclusions on Dicamba-Tolerant Soybean MON 87708 ..................224

VIII.I. Crop Rotation Practices in Soybean ................................................................224

VIII.J. Soybean Volunteer Management ....................................................................238

VIII.K. Weed Resistance to Dicamba Herbicide........................................................241

VIII.L. Stewardship of MON 87708 ..........................................................................241

VIII.M. Impact of the Introduction of MON 87708 on Agricultural Practices ..........243

IX. ENVIRONMENTAL CONSEQUENCES ...............................................................245

IX.A. Introduction ......................................................................................................245

IX.B. Plant Pest Assessment of the MON 87708 Insert and Expressed Protein ........246

IX.B.1. Characteristics of the Genetic Insert and the Expressed Protein ...........246

IX.B.2. Compositional Characteristics ...............................................................247

IX.B 3. Phenotypic and Agronomic and Environmental Interaction 

Characteristics ......................................................................................................248

IX.C. Weediness Potential of MON 87708 ................................................................253

IX.D. Potential for Pollen Mediated Gene Flow ........................................................254

IX.D.1. Hybridization with Cultivated Soybean .................................................255

IX.D.2. Hybridization with Wild Annual Species within Subgenus Soja ..........255

IX.D.3. Hybridization with the Wild Perennial Species of Glycine 

Subgenus ..............................................................................................................256

IX.D.4. Transfer of Genetic Information to Species with Which Soybean 

Cannot Interbreed (Horizontal Gene Flow) .........................................................256

IX.E. Potential Impact on Soybean Agronomic Practices ..........................................257

IX.F. Summary of Plant Pest Assessments ................................................................257

X. ADVERSE CONSEQUENCES OF INTRODUCTION ...........................................260

REFERENCES ................................................................................................................261

APPENDICES .................................................................................................................282

Appendix A: USDA Notifications ..................................................................................283

Appendix B: Materials, Methods, and Results for Molecular Analyses of 

MON 87708 .....................................................................................................................287
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Appendix C: Materials, Methods, and Results for the Characterization of 

MON 87708 DMO and Substrate Specificity of DMO ...................................................294

Appendix D: Materials and Methods Used for the Analysis of the Levels of 

MON 87708 DMO ...........................................................................................................323

Appendix E: Materials, Methods, and Individual-Site Results for Compositional 

Analysis of MON 87708 Soybean Seed and Forage .......................................................326

Appendix F: Materials, Methods, and Individual Site Results for the Seed 

Dormancy and Germination Assessment of MON 87708 ...............................................437

Appendix G: Materials, Methods, and Individual-Site Results from the 

Phenotypic, Agronomic, and Environmental Interaction Assessment of 

MON 87708 under Field Conditions ...............................................................................443

Appendix H: Materials and Methods for Pollen Morphology and Viability 

Assessment .......................................................................................................................486

Appendix I: Materials and Methods for Symbiont Assessment .....................................489

Appendix J: Petitioner’s Environmental Report .............................................................492

Appendix K: Herbicide Resistance .................................................................................588

Appendix L: Comparative Analysis of Dicamba and Alternative Soybean 

Herbicides ........................................................................................................................606

Appendix M: Potential Impact of Dicamba on Human Health and the 

Environment .....................................................................................................................676

Appendix N: Overview of Evaluation of Potential Exposure and Biological 

Effects on Endangered Species for Dicamba Use in MON 87708 ..................................703
Monsanto Company 10-SY-210U 19 of 721 

LIST OF TABLES 

Table III-1. Summary of Genetic Elements in the Plasmid Vector 

PV-GMHT4355 .................................................................................................................40

Table IV-1. Summary Chart of the Expected DNA Segments Based on 

Hybridizing Probes and Restriction Enzymes Used in MON 87708 Analysis ..................50

Table IV-2. Summary of Genetic Elements in MON 87708 ............................................51

Table IV-3. Segregation of the dmo Expression Cassette During the Development 

of MON 87708 ...................................................................................................................72

Table V-1. Herbicides Applied to MON 87708 and Conventional Control .....................77

Table V-2. Tissues Collected and Analyzed for MON 87708 DMO ...............................79

Table V-3. Summary of the Levels of MON 87708 DMO in Leaf, Root, Forage, 

and Seed from MON 87708 Grown in 2008 U.S. Field Trials ..........................................80

Table VI-1. Summary of Differences (α=0.05) for the Comparison of Soybean 

Component Levels for MON 87708 (Treated) vs. Conventional Control .........................98

Table VI-2. Statistical Summary of Combined-Site Soybean Seed Nutrients for 

MON 87708 (Treated) vs. Conventional Control ............................................................112

Table VI-3. Statistical Summary of Combined-Site Soybean Seed Anti-Nutrients 

for MON 87708 (Treated) vs. Conventional Control ......................................................118

Table VI-4. Statistical Summary of Combined-Site Soybean Forage Nutrients for 

MON 87708 (Treated) vs. Conventional Control ............................................................120

Table VI-5. Summary of Differences (α=0.05) for the Comparison of Soybean C

omponent Levels for MON 87708 (Untreated) vs. Conventional Control ......................122

Table VI-6. Statistical Summary of CombinedSite Soybean Seed Nutrients for MON 87708 (Untreated) vs. Conventional 

Control .............................................................................................................................132

Table VI-7. Statistical Summary of Combined-Site Soybean Seed AntiNutrients for MON 87708 (Untreated) vs. Conventional Control ...................................138

Table VI-8. Statistical Summary of CombinedSite Soybean Forage Nutrients for MON 87708 (Untreated) Conventional Control ......140

Table VI-9. Literature and ILSI Database Ranges for Components in Soybean 

Seed and Forage ...............................................................................................................142

Table VII-1. Phenotypic, Agronomic, and Environmental Interaction 

Characteristics Evaluated in U.S. Field Trials, Laboratory, or Greenhouse Tests ..........148

Table VII-2. Combined-Site Comparison of MON 87708 to Conventional 

Control for Seed Dormancy and Germination Characteristics ........................................155

Table VII-3. 2008 Field Phenotypic Evaluation Sites for Untreated MON 87708 ........157
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Table VII-4. 2009 Field Phenotypic Evaluation Sites for Dicamba Treated 

MON 87708 .....................................................................................................................158

Table VII-5. Combined-Site Comparison of Untreated MON 87708 to 

Conventional Control during 2008 for Phenotypic and Agronomic Characteristics .......159

Table VII-6. Combined-Site Comparison of Dicamba Treated MON 87708 to 

Conventional Control during 2009 for Phenotypic and Agronomic Characteristics .......161

Table VII-7. Pollen Characteristics of MON 87708 Compared to Conventional 

Control .............................................................................................................................167

Table VII-8. Symbiont Interaction Assessment of MON 87708 and Conventional 

Control .............................................................................................................................169

Table VIII-1. Soybean Production in the U.S., 1999 – 20081 ........................................173

Table VIII-2. U.S. Soybean Production by Region and State in 20081 ..........................174

Table VIII-3. U.S. Soybean Production Costs and Returns in 20081 .............................175

Table VIII-4. Common Weeds in Soybean Production: Midwest Region ....................188

Table VIII-5. Common Weeds in Soybean Production: Southeast Region ...................189

Table VIII-6. Common Weeds in Soybean Production: Eastern Coastal Region .........189

Table VIII-7. Herbicide Use in Soybean in the U.S. from 1995 through 20011 .............192

Table VIII-8. Agricultural Chemical Applications Registered for Soybean Use in 

AR, IA, IL, IN, KS, KY, LA, MI, MN, MS, MO, NE, NC, ND, OH, SD, TN, VA, 

and WI in 20061

 ...............................................................................................................193

Table VIII-9. Crop Tolerance and Common Grass Weed Responses to Herbicides 

Applied in Soybean Production .......................................................................................195

Table VIII-10. Common Broadleaf Weed Responses to Herbicides Applied in 

Soybean Production .........................................................................................................196

Table VIII-11. Dicamba Use in All Labeled Crops from 1990 to 20081 ........................198

Table VIII-12. Dicamba-Treated Acres and Amounts Applied to Labeled Crops 

and Uses in 20081

.............................................................................................................199

Table VIII-13. Dicamba Applications – Average Number and Rates to Labeled 

Crops1 ...............................................................................................................................199

Table VIII-14. Dicamba-Treated Acres (000) by Application Timing and Crop in 

20081

 ................................................................................................................................201

Table VIII-15. Dicamba-Treated Acres by State and Labeled Crop in 20081 ................203

Table VIII-17. Common Broadleaf Weed Responses to Preplant Burndown 

Herbicides ........................................................................................................................213

Table VIII-18. Common Broadleaf Weed Responses to Dicamba Compared to 

Labeled Postemergence Herbicides in Soybean Production ............................................214

Table VIII-19. Summary of Comparative Analysis of Dicamba and Alternative 

Herbicides ........................................................................................................................215
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Table VIII-20. Dates and Days to Reach Various Growth Stages in Corn and 

Soybean in Central Illinois ...............................................................................................218

Table VIII-21. Dates and Days to Reach Various Growth Stages in Corn and 

Soybean in Western Tennessee ........................................................................................220

Table VIII-22. Average Daily Temperatures in Major Soybean Producing States1 .......220

Table VIII-23. Average Relative Humidity (1971-2000) in Major Soybean 

Producing States1

 .............................................................................................................221

Table VIII-24. Rotational Practices in the U.S. Following Soybean Production ...........227

Table VIII-25. Rotational Practices Following Soybean Production in the 

Midwest Region ...............................................................................................................228

Table VIII-26. Rotational Practices Following Soybean Production in the 

Southeast Region .............................................................................................................233

Table VIII-27. Rotational Practices Following Soybean Production in the East 

Coastal Region .................................................................................................................236

Table VIII-28. Ratings for Postemergence Control of Volunteer Soybean in 

Labeled Rotational Crops1 ...............................................................................................240

Table IX-1. Summary of Published Literature on Soybean Cross Pollination ...............259

Table A-1. USDA Notifications Approved for MON 87708 and Status of Trials 

Conducted under These Notifications and Permits ..........................................................284

Table B-1. Hybridization Conditions of Utilized Probes ................................................289

Table C-1. Molecular Weight and Purity of the MON 87708-Produced DMO 

Proteins ............................................................................................................................302

Table C-2. Summary of the Tryptic Masses Identified for the MON 87708 DMO 

Protein Using MALDI-TOF MS ......................................................................................306

Table C-3. Summary of the Tryptic Masses Identified for the MON 87708 

DMO+27 Protein Using MALDI-TOF MS .....................................................................307

Table C-4. MON 87708 DMO Functional Activity Assay .............................................312

Table C-5. Herbicides Tested in Exogenous Specificity Herbicide Tolerance 

Greenhouse Trials ............................................................................................................313

Table C-6. Compounds Used in Specificity In Vitro Experiments .................................314

Table C-7. Herbicide Tolerance Trials Injury Ratings ...................................................317

Table E-1. Commercial Reference Varieties ..................................................................326

Table E-2. Re-Expression Formulas for Statistical Analysis of Composition Data .......332

Table E-3. Component with Observations Below the Assay Limit of Quantitation 

Not Excluded from Statistical Analysis ...........................................................................332

Table E-4. Statistical Summary of Site IARL Soybean Seed Nutrients for 

MON 87708 (Treated) vs. Conventional Control ............................................................334
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Table E-5. Statistical Summary of Site IARL Soybean Seed Anti-nutrients for 

MON 87708 (Treated) vs. Conventional Control ............................................................340

Table E-6. Statistical Summary of Site IARL Soybean Forage Nutrients for 

MON 87708 (Treated) vs. Conventional Control ............................................................342

Table E-7. Statistical Summary of Site ILCY Soybean Seed Nutrients for 

MON 87708 (Treated) vs. Conventional Control ............................................................344

Table E-8. Statistical Summary of Site ILCY Soybean Seed Anti-nutrients for 

MON 87708 (Treated) vs. Conventional Control ............................................................350

Table E-9. Statistical Summary of Site ILCY Soybean Forage Nutrients for 

MON 87708 (Treated) vs. Conventional Control ............................................................352

Table E-10. Statistical Summary of Site ILWY Soybean Seed Nutrients for 

MON 87708 (Treated) vs. Conventional Control ...........................................................354

Table E-11. Statistical Summary of Site ILWY Soybean Seed Anti-nutrients for 

MON 87708 (Treated) vs. Conventional Control ............................................................360

Table E-12. Statistical Summary of Site ILWY Soybean Forage Nutrients for 

MON 87708 (Treated) vs. Conventional Control ............................................................362

Table E-13. Statistical Summary of Site INRC Soybean Seed Nutrients for 

MON 87708 (Treated) vs. Conventional Control ............................................................364

Table E-14. Statistical Summary of Site INRC Soybean Seed Anti-nutrients for 

MON 87708 (Treated) vs. Conventional Control ............................................................370

Table E-15. Statistical Summary of Site INRC Soybean Forage Nutrients for 

MON 87708 (Treated) vs. Conventional Control ............................................................372

Table E-16. Statistical Summary of Site PAHM Soybean Seed Nutrients for 

MON 87708 (Treated) vs. Conventional Control ............................................................374

Table E-17. Statistical Summary of Site PAHM Soybean Seed Anti-nutrients for 

MON 87708 (Treated) vs. Conventional Control ............................................................380

Table E-18. Statistical Summary of Site PAHM Soybean Forage Nutrients for 

MON 87708 (Treated) vs. Conventional Control ............................................................382

Table E-19. Statistical Summary of Site IARL Soybean Seed Nutrients for 

MON 87708 (Untreated) vs. Conventional Control ........................................................384

Table E-20. Statistical Summary of Site IARL Soybean Seed Anti-Nutrients for 

MON 87708 (Untreated) vs. Conventional Control ........................................................390

Table E-21. Statistical Summary of Site IARL Soybean Forage Nutrients for 

MON 87708 (Untreated) vs. Conventional Control ........................................................392

Table E-22. Statistical Summary of Site ILCY Soybean Seed Nutrients for 

MON 87708 (Untreated) vs. Conventional Control ........................................................394

Table E-23. Statistical Summary of Site ILCY Soybean Seed Anti-Nutrients for 

MON 87708 (Untreated) vs. Conventional Control ........................................................400

Table E-24. Statistical Summary of Site ILCY Soybean Forage Nutrients for 

MON 87708 (Untreated) vs. Conventional Control ........................................................402
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Table E-25. Statistical Summary of Site ILWY Soybean Seed Nutrients for 

MON 87708 (Untreated) vs. Conventional Control ........................................................404

Table E-26. Statistical Summary of Site ILWY Soybean Seed Anti-Nutrients for 

MON 87708 (Untreated) vs. Conventional Control ........................................................410

Table E-27. Statistical Summary of Site ILWY Soybean Forage Nutrients for 

MON 87708 (Untreated) vs. Conventional Control ........................................................412

Table E-28. Statistical Summary of Site INRC Soybean Seed Nutrients for 

MON 87708 (Untreated) vs. Conventional Control ........................................................414

Table E-29. Statistical Summary of Site INRC Soybean Seed Anti-Nutrients for 

MON 87708 (Untreated) vs. Conventional Control ........................................................420

Table E-30. Statistical Summary of Site INRC Soybean Forage Nutrients for 

MON 87708 (Untreated) vs. Conventional Control ........................................................422

Table E-31. Statistical Summary of Site PAHM Soybean Seed Nutrients for 

MON 87708 (Untreated) vs. Conventional Control ........................................................424

Table E-32. Statistical Summary of Site PAHM Soybean Seed Anti-Nutrients for 

MON 87708 (Untreated) vs. Conventional Control ........................................................430

Table E-33. Statistical Summary of Site PAHM Soybean Forage Nutrients for 

MON 87708 (Untreated) vs. Conventional Control ........................................................432

Table F-1. Starting Seed of MON 87708, Conventional Control, and Commercial 

Reference Varieties Used in the Dormancy and Germination Assessment .....................437

Table F-2. Dormancy and Germination Characteristics of MON 87708 and 

Conventional Control Seed Produced at each of Three Field Sites .................................440

Table G-1. Starting Seed for the Phenotypic, Agronomic, and Environmental 

Interaction Assessment for 2008 Field Trials ..................................................................445

Table G-2. Starting Seed for the Phenotypic, Agronomic, and Environmental 

Interaction Assessment for 2009 Field Trials ..................................................................446

Table G-3. Field and Planting Information for 2008 Field Trials...................................448

Table G-4. Field and Planting Information for 2009 Field Trials...................................450

Table G-5. 2008 Individual-Site Phenotypic Comparison of Untreated 

MON 87708 to Conventional Control .............................................................................457

Table G-6. 2009 Individual-Site Phenotypic Comparison of Dicamba Treated 

MON 87708 to Conventional Control .............................................................................460

Table G-7. Growth Stage Monitoring of Untreated MON 87708, Conventional 

Control, and the Commercial Reference Varieties from 2008 Field Trials .....................463

Table G-8. Growth Stage Monitoring of Dicamba Treated MON 87708, 

Conventional Control, and the Commercial Reference Varieties from 2009 Field 

Trials ................................................................................................................................467

Table G-9. Abiotic Stress Response Evaluations of Untreated MON 87708 and 

Conventional Control from 2008 Field Trials Using an Observational Severity 

Scale .................................................................................................................................469
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Table G-10. Disease Damage Evaluations of Untreated MON 87708 and 

Conventional Control from 2008 Field Trials Using an Observational Severity 

Scale .................................................................................................................................470

Table G-11. Arthropod–Related Damage Evaluations of Untreated MON 87708 

and Conventional Control from 2008 Field Trials Using an Observational Severity 

Scale .................................................................................................................................471

Table G-12. Abundance of Pest Arthropods in Beat Sheet Samples Collected 

from Untreated MON 87708, Conventional Control, and the Commercial 

Reference Varieties from 2008 Field Trials .....................................................................473

Table G-13. Abundance of Beneficial Arthropods in Beat Sheet Samples 

Collected from Untreated MON 87708, Conventional Control, and the 

Commercial Reference Varieties from 2008 Field Trials ................................................477

Table G-14. Abiotic Stress Response Evaluations of Dicamba-Treated 

MON 87708 and Conventional Control from 2009 Field Trials Using an 

Observational Severity Scale ...........................................................................................481

Table G-15. Disease Damage Evaluations of Dicamba-Treated MON 87708 and 

Conventional Control from 2009 Field Trials Using an Observational Severity 

Scale .................................................................................................................................482

Table G-16. Arthropod–Related Damage Evaluations of Dicamba-Treated 

MON 87708 and Conventional Control from 2009 Field Trials Using an 

Observational Severity Scale ...........................................................................................483

Table I-1. Starting Seed of MON 87708, Conventional Control, and Commercial 

Reference Varieties Used in the Symbiont Assessment ..................................................489

Table J-1. Summary of Dicamba Uses on Soybean ........................................................493

Table J-2. 2008 Soybean Productivity by Region ..........................................................496

Table J-3. Known Weed Resistance in the Southern U.S. 1 ............................................508

Table J-4. Known Weed Resistance in the Midwest U.S. 1 ............................................509

Table J-5. Deregulated or Submitted Biotechnology-derived Soybean Products ...........514

Table J-6. Organic and Conventional Soybean Seed Sources .........................................544

Table K-1. Management Recommendations for Control of Dicamba- and Other 

Synthetic Auxin-Resistant Weeds....................................................................................596

Table L-1. Dicamba Acid Acute Toxicity Study Findings .............................................611

Table L-2. Dicamba Acid Reproductive, Developmental, Mutagenic, and 

Neurotoxicologic Findings...............................................................................................612

Table L-3. Dicamba Acid Subchronic, Chronic and Cancer Findings ...........................613

Table L-4. Summary of Toxicological Findings from Testing of DCSA .......................615

Table L-5. Dicamba Acid Ecotoxicity Findings on Mammals, Birds and Fish 

(U.S. EPA. 2005a) ...........................................................................................................616
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Table L-6. Dicamba Acid Ecotoxicity Findings on Aquatic and Terrestrial 

Invertebrates (U.S. EPA. 2005a) ......................................................................................617

Table L-7. Dicamba Acid Ecotoxicity Results on Aquatic and Terrestrial Plants 

(U.S. EPA. 2005a) ...........................................................................................................617

Table L-8. Dicamba, Diglycolamine (DGA) Salt Ecotoxicity Findings (U.S. 

EPA. 2005a) .....................................................................................................................618

Table L-9. Environmental Fate and Physical Properties of Dicamba Acid ....................619

Table L-10. Ten Most Widely Used Alternative Herbicides in U.S. Soybean 

Production ........................................................................................................................623

Table L-11. Alternative Registered Soybean Herbicides ...............................................624

Table L-12. Active Ingredients Contained in Alternative Herbicides ............................629

Table L-13. Human Health Risk Parameters for Alternative Herbicides .......................632

Table L-14. Aquatic Toxicity Parameters for Fish and Aquatic Invertebrates for 

Alternative Herbicides .....................................................................................................637

Table L-15. Aquatic Toxicity Parameters for Aquatic Plants for Alternate 

Herbicide Active Ingredients ...........................................................................................641

Table L-16. Herbicide Efficacy Comparison: Herbicide Resistant Weeds and 

Hard-to-Control Weeds in Soybean .................................................................................646

Table L-17. Summary of Comparative Analysis of Dicamba and Alternative 

Herbicides ........................................................................................................................653

Table L-18. Planting Restrictions (months) for Alternative Herbicide Products ...........659

Table M-1. Summary of Dietary Exposure and Risk for Dicamba: Food and 

Water ................................................................................................................................682

Table M-2. Aggregate (Short-term) Exposure Assessment for Dicamba .......................683

Table M-3. Residues of Dicamba, DCSA, 5-Hydroxydicamba and DCGA in 

Dicamba-Tolerant Soybean Forage, Hay and Seed .........................................................684

Table M-4. Dicamba Concentrations (μg/L) in Groundwater in Major Soybean 

Growing States .................................................................................................................688

Table M-5. Dicamba Concentrations (μg/L) in Surface Water in Major Soybean 

Growing States .................................................................................................................690

Table N-1. EPA Levels of Concern for Threatened and Endangered Species ...............705

Table N-2. Comparison of Default Kenaga Residues and Dicamba-specific 

Residues in Food-Items....................................................................................................707

Table N-3. Risk Quotients for Chronic Exposure a for Mammalian Species Based 

on Dicamba-Specific Residue Values and DT50. .............................................................709

Table N-4. Risk Quotients for Acute Exposure to Birds Using Measured 

Dicamba Residue and Residue Decline Values ...............................................................710
Monsanto Company 10-SY-210U 26 of 721 

LIST OF FIGURES 

Figure III-1. Circular Map of Plasmid Vector PV-GMHT4355 Showing Probes 1-

10........................................................................................................................................39

Figure III-2. Schematic of the Development of MON 87708 ...........................................44

Figure III-3. Deduced Amino Acid Sequence of the RbcS Targeting Sequence 

and MON 87708 DMO ......................................................................................................45

Figure IV-1. Schematic Representation of the Insert and DNA Flanking 

Sequences in MON 87708 .................................................................................................49

Figure IV-2. Southern Blot Analysis to Determine Insert and Copy Number of 

T-DNA I in MON 87708: Probe 8 ....................................................................................55

Figure IV-3. Southern Blot Analysis to Determine Insert and Copy Number of 

T-DNA I in MON 87708: Probe 9 ....................................................................................56

Figure IV-4. Southern Blot Analysis to Determine Insert and Copy Number of 

T-DNA I in MON 87708: Probe 10 ..................................................................................57

Figure IV-5. Southern Blot Analysis to Detect the Presence or Absence of 

T-DNA II Sequences in MON 87708: Probe 4 .................................................................60

Figure IV-6. Southern Blot Analysis to Detect the Presence or Absence of 

T-DNA II Sequences in MON 87708: Probe 5 .................................................................61

Figure IV-7. Southern Blot Analysis to Detect the Presence or Absence of 

T-DNA II Sequences in MON 87708: Probe 6 .................................................................62

Figure IV-8. Southern Blot Analysis to Determine the Presence or Absence of 

PV-GMHT4355 Backbone Sequences in MON 87708: Probes 1, 2, 3, and 7 .................64

Figure IV-9. Breeding History of MON 87708 ................................................................67

Figure IV-10. Southern Blot Analysis to Examine Insert Stability in Multiple 

Generations of MON 87708: Probe 9 ...............................................................................68

Figure IV-11. Breeding Path for Generating Segregation Data for MON 87708 .............71

Figure V-1. Three Components of the DMO Oxygenase System ....................................74

Figure V-2. Dicamba and Set of Potential Endogenous Substrates Tested in in 

vitro Experiments with DMO ............................................................................................76

Figure VII-1. Schematic Diagram of Agronomic and Phenotypic Data 

Interpretation Methods .....................................................................................................151

Figure VIII-1. Mean Annual Dicamba Use for Agricultural Uses in the U.S. 

During 1999-20041...........................................................................................................206

Figure VIII-2. Planted Soybean Acreage by County in the U.S. in 20091 .....................206

Figure B-1. Overlapping PCR Analysis Across the Insert in MON 87708 ....................292

Figure B-2. PCR Amplification of the MON 87708 Insertion Site ................................293
Monsanto Company 10-SY-210U 27 of 721 

Figure C-1. Molecular Weight and Purity Analysis of MON 87708 DMO 

Proteins ............................................................................................................................303

Figure C-2. Immunoblot Analysis of MON 87708 DMO Proteins ................................304

Figure C-3. MALDI-TOF MS Coverage Map of the MON 87708 DMO Protein .........308

Figure C-4. MALDI-TOF MS Coverage Map of the MON 87708 DMO+27 

Protein ..............................................................................................................................308

Figure C-5. N-Terminal Sequence of the MON 87708 DMO Protein ...........................310

Figure C-6. N-Terminal Sequence of the MON 87708 DMO+27 Protein .....................310

Figure C-7. Glycosylation Analysis of the MON 87708 DMO Proteins ........................311

Figure C-8. UPLC separation of dicamba (DCB) and DCSA ........................................319

Figure C-9. E. coli-produced DMO Conversion of Dicamba (DCB) to DCSA and 

2,4-D to 2,4-Dichlorophenol (2,4-DCP) ..........................................................................320

Figure C-10. E. coli-produced DMO Conversion of Endogenous Substrates ................321

Figure K-1. Weed Resistance to Various Herbicide Families1 .......................................592

Figure L-1. Structure of Dicamba ...................................................................................609

Figure L-2. Structure of DCSA .......................................................................................614

Figure M-1. Ground Water Sampling Sites for Dicamba (Top: Detects only; 

Bottom: detects and non-detects) .....................................................................................689

Figure M-2. Surface Water Sampling Sites for Dicamba (Top: Detects only; 

Bottom: detects and non-detects) .....................................................................................691

Figure N-1. Identification of Taxa Exceeding Endangered Species Levels of 

Concern ............................................................................................................................708

Figure N-2. County-Level Analysis Process ..................................................................712

Figure N-3. Counties with Soybean Farms and Adjacent Counties ...............................713

Figure N-4. Counties with Soybean Farms (or Adjacent Counties) with Listed 

Species in the Taxa of Interest .........................................................................................715
Monsanto Company 10-SY-210U 28 of 721 

ABBREVIATION AND DEFINITIONS

~ approximately 

aadA Bacterial promoter and coding sequence for an aminoglycosidemodifying enzyme, 3’(9)-O-nucleotidyltransferase from the 

transposon Tn7 that confers spectinomycin and streptomycin 

resistance 

AAPSE American Association of Pesticide Safety Educators 

ADF acid detergent fiber 

ALS acetolactate synthase 

AOSA Association of Official Seed Analysts 

aPAD acute population adjusted dose 

APHIS Animal and Plant Health Inspection Service 

B. japonicum Bradyrhizobium japonicum 

BIO Biotechnology Industry Organization 

bp base pair 

BRS Biotechnology Regulatory Services 

BSA bovine serum albumin 

bu/A bushels per acre 

CEQ Council on Environmental Quality 

CES Cooperative Extension Service 

CFR Code of Federal Regulations 

CHT ceramic hydroxyapatite column 

cp4 epsps codon modified coding sequence of the aroA gene from Agrobacterium 

species strain CP4 encoding CP4 EPSPS 

CP4 EPSPS 5-Enolpyruvylshikimate-3-phosphate synthase protein 

cPAD chronic population adjusted dose 

CTAB Hexadecyltrimethylammonium bromide 

CTP2 Sequences encoding the chloroplast transit peptide region of 

Arabidopsis thaliana EPSPS used to direct proteins into chloroplasts 

CV column volume 

Da Dalton 

DAP days after planting 

dATP Deoxyadenosine triphosphate 

DCSA 3,6-dichlorosalicylic acid also known as 

3,6-dichloro-2-hydroxybenzoic acid 

DCGA 3,6-dichlorogentisic acid 

dCTP Deoxycytidine triphosphate 

DDI daily dietary intake 

DEEM dietary exposure evaluation model 

DGA Diglycolamine 

DHB dihydroxybenzoic acid 

dicamba 3,6-dichloro-2-methoxybenzoic acid 

dmo Coding sequence of the dicamba mono-oxygenase gene from 

Stenotrophomonas maltophilia encoding DMO 

DMO full-length dicamba mono-oxygenase protein 

DMO+27 full-length dicamba mono-oxygenase protein with an additional 24 

 

Note: Standard abbreviations, e.g., units of measure, are used according to the format described in 

‘Instructions to Authors’ in the Journal of Biological Chemistry. 
Monsanto Company 10-SY-210U 29 of 721 

amino acids from the Rubisco small subunit and 3 amino acids from 

an intervening sequence 

DnaK 5′ non-translated leader sequence from the Petunia hybrid Hsp70 gene 

that is involved in regulating gene expression 

dNTP deoxynucleoside triphosphate 

DTT dithiothreitol 

DWCF dry weight conversion factor 

dwt dry weight of tissue 

E9 3′ non-translated region of the pea RbcS2 gene that functions to direct 

polyadenylation of the mRNA 

E. coli Escherichia coli 

EEC estimated environmental exposure concentration 

EFED Environmental Fate and Effects Division 

ELISA enzyme-linked immunosorbent assay 

EPA U.S. Environmental Protection Agency 

ERS Economic Research Service 

ETS Excellence Through Stewardship 

Exo exonuclease I 

FA fatty acid 

FASTA algorithm used to find local high scoring alignments between a pair of 

protein or nucleotide sequences 

FDA U.S. Food and Drug Administration 

FFDCA Federal Food, Drug, and Cosmetic Act 

FIFRA Federal Insecticide, Fungicide, and Rodenticide Act 

FLt full-length transcript 

FMV Promoter for the figwort mosaic virus 35S RNA 

FQPA Food Quality Protection Act 

fwt fresh weight of tissue 

GLP good laboratory practice 

H.U. hemagglutinating unit

HAL health advisory level 

HED Health Effects Division 

HPPD 4-hydroxyphenylpyruvate dioxygenase 

HRP horseradish peroxidase 

IAA indole acetic acid 

ILSI International Life Sciences Institute 

IPM integrated pest management 

kb kilobase 

kDa kiloDalton 

LB loading buffer 

lb/bu pounds per bushel 

LC lethal concentration 

LD lethal dose 

Left Border DNA region from Agrobacterium tumefaciens containing the left 

border sequence used for transfer of the T-DNA 

LOAEL lowest dosing level that produced an observable effect 

LOC level of concern 

LOD limit of detection 

LOEC lowest observed effect concentration 

LOQ limit of quantitation 

MALDI-TOF MS matrix-assisted laser desorption/ionization time-of-flight mass 
Monsanto Company 10-SY-210U 30 of 721 

spectrometry 

MMT million metric tons 

MOE margin of exposure 

MON 87708 DMO The active form of DMO, a trimer comprised of three monmers. The 

DMO trimer can be comprised of DMO, DMO+27 or a combination 

of both forms. 

MSTA Monsanto Technology Stewardship Agreement 

MW molecular weight 

MWCO molecular weight cut off 

N/A not applicable 

NADH nicotinamide adenine dinucleotide 

NASS National Agricultural Statistics Service, a branch of the USDA 

NAWQA National Water-Quality Assessment 

NDF neutral detergent fiber 

NEPA National Environmental Policy Act 

NFDM non-fat dried milk 

NOAEL no observable adverse effect level 

NOEC no observable effect concentration 

NTOs non-target organisms 

OECD Organization of Economic Co-Operation and Development 

OPP Office of Pesticide Programs 

ORETF Outdoor Residential Exposure Task Force 

ori-pBR322 Origin of replication from pBR322 necessary for maintenance of 

plasmid in E. coli 

ori V Origin of replication from the broad host range plasmid RK2 necessary 

for maintenance of plasmid in Agrobacterium 

OSL over-season leaf 

PAD population adjusted dose 

PBS phosphate buffered saline solution 

PBST phosphate buffered saline solution containing 0.05% (v/v) Tween-20 

PC1SV Promoter for the full length transcript of peanut chlorotic streak virus 

that directs transcription in plant cells 

PCR polymerase chain reaction 

pea Pisum sativum 

PHED Pesticide Handler Exposure Database

RED Reregistration Eligibility Decision

RfD reference dose 

PMSF phenylmethylsulfonyl fluoride 

PPA Plant Protection Act 

PPO protoporphyrinogen oxidase 

PRZM/EXAMS Pesticide Root Zone Model/Exposure Analysis Modeling System 

PVDF polyvinylidene difluoride 

RbcS Sequences encoding the transit peptide region of the ribulose-1,5-

bisphosphate carboxylase oxygenase gene from Pisum sativum

Right Border DNA region from Agrobacterium tumefaciens containing the right 

border sequence used for transfer of the T-DNA 

rop Coding sequence for repressor of primer protein used for maintenance 

of plasmid copy number in E. coli 

RQ risk quotient 

RT room temperature 

SAP shrimp alkaline phosphate 
Monsanto Company 10-SY-210U 31 of 721 

SCI-GROW Screening Concentration in Ground Water 

SCN soybean cyst nematode 

SD standard deviation 

SDS-PAGE sodium dodecyl sulfate polyacrylamide gel electrophoresis

S.E. standard error 

SGF simulated gastric fluid 

SIF simulated intestinal fluid 

S. maltophilia Stenotrophomonas maltophilia

soybean Glycine max (L.) Merr. 

T-DNA transfer(ed) DNA 

TBS tris buffered saline 

TES threatened and endangered species 

TEV 5′ non-translated region from the Tobacco Etch RNA virus genome that 

is involved in regulating gene expression 

TFA trifluoroacetic acid 

TIU trypsin inhibitor units 

Tm melting temperature 

TMRC theoretical maximum resiude concentration 

TMB 3,3',5,5' tetramethyl-benzidine 

TRED tolerance reregistration eligibility decision 

TSSP tissue-specific pool 

TUG Technology Use Guide 

U.S. United States of America 

USDA U.S. Department of Agriculture 

v/v volume per volume 

VMD volume median diameter 

w/v weight per volume 

WAT weeks after treatment 

WPS Worker Protection standard 

WSSA Weed Science Society of America 
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I. RATIONALE FOR THE DEVELOPMENT OF MON 87708 

I.A. Basis for the Request for a Determination of Nonregulated Status under 

7 CFR § 340.6 

The Animal and Plant Health Inspection Service (APHIS) of the United States (U.S.) 

Department of Agriculture (USDA) has responsibility, under the Plant Protection Act 

(Title IV Pub. L. 106-224, 114 Stat. 438, 7 U.S.C. § 7701-7772), to prevent the 

introduction and dissemination of plant pests into the U.S. APHIS regulation 

7 CFR § 340.6 provides that an applicant may petition APHIS to evaluate submitted data 

to determine that a particular regulated article does not present a plant pest risk and no 

longer should be regulated. If APHIS determines that the regulated article does not 

present a plant pest risk, the petition is granted, thereby allowing unrestricted introduction 

of the article. A listing of all regulated MON 87708 field trials conducted under USDA 

notification can be found in Appendix A. 

Monsanto Company is submitting this request to APHIS for a determination of 

nonregulated status for new biotechnology-derived soybean product, MON 87708, any 

progeny derived from crosses between MON 87708 and conventional soybean, and any 

progeny derived from crosses of MON 87708 with biotechnology-derived soybean that 

have previously been granted nonregulated status under 7 CFR Part 340. 

I.B. Rationale for the Development of Dicamba-Tolerant Soybean MON 87708 

The Roundup Ready® soybean system permits over-the-top application of Roundup®

agricultural herbicides containing the active ingredient glyphosate for effective weed 

control. The value of the Roundup Ready soybean system has been demonstrated by the 

significant growth in the number of planted acres since its introduction in 1996. Today 

more than 90% of all soybean acres grown in the U.S. are Roundup Ready (USDA-NASS, 

2009c). The Roundup Ready soybean system delivers effective broad spectrum weed 

control, provides flexibility of application timing, has facilitated increased adoption of 

reduced tillage practices, and has resulted in increased grower income (Carpenter and 

Gianessi, 2001; Bonny, 2008; Hurley et al., 2009). Additionally, the Roundup Ready 

soybean system provides incremental environmental benefits (Bonny, 2008; Brookes and 

Barfoot 2009), and glyphosate, as concluded by the U.S. EPA (1993), has a favorable 

safety profile. Continued use of the Roundup Ready soybean system will maintain 

effective and familiar weed control management practices that are fully compatible with 

all current tillage and land management systems including conservation tillage practices. 

Growth of conservation tillage in the U.S. was greatly accelerated with the introduction 

of glyphosate-tolerant crops in large part because of the broad spectrum postemergence 

control offered by glyphosate (Price et al. 2011). 

As with all herbicides used in agriculture, there is potential for weeds to develop 

resistance to the herbicide over time.1

 If unmanaged, herbicide resistance can become a 

limiting factor in crop production. Glyphosate has had few cases of weed resistance, 

 

1

 http://www.weedscience.org/In.asp 
Monsanto Company 10-SY-210U 33 of 721 

particularly in relation to other herbicides. In the U.S., while there have been thirteen 

confirmed glyphosate-resistant weeds (Heap, 2011), glyphosate still controls more than 

160 weed species (Roundup WeatherMax herbicide label, EPA Reg. No.524-537) and 

remains an extremely valuable tool for U.S, soybean crop production. Herbicide resistant 

weeds are those listed on the International Survey of Resistant Weeds website 

(www.weedscience.org). Additionally, studies have shown that resistance can be 

postponed, contained and managed through good management practices. One of the 

management practices most often recommended by University/Cooperative Extension 

Service and industry is the use of multiple herbicide modes-of-action. Simultaneously 

using two herbicides with different modes-of-action significantly reduces the probability 

of weeds developing resistance to either or both herbicides (Powles et al., 1996; Beckie 

and Reboud, 2009). Other recommended management practices to manage herbicide 

resistance includes the use of multiple herbicide modes-of-action in sequence, and/or the 

inclusion of mechanical or cultural practices in addition to the use of an herbicide. 

Monsanto Company has developed biotechnology-derived soybean MON 87708 that is 

tolerant to dicamba (3,6-dichloro-2-methoxybenzoic acid) herbicide. Dicamba is a 

synthetic auxin herbicide that kills plants by mimicking naturally-occurring plant growth 

hormones called auxins, thereby destroying tissue through uncontrolled cell division and 

growth (Ahrens, 1994). Dicamba’s mode-of-action is different from glyphosate, and it 

provides efficacious control of broadleaf weeds and is complementary to glyphosate on 

hard-to-control weeds such as common lambsquarters, hemp sesbania, morning glory 

species, nightshade, Pennsylvania smartweed, prickly sida, velvetleaf, waterhemp, and 

wild buckwheat (Johnson et al., 2010). Additionally, dicamba provides effective control 

of herbicide-resistant broadleaf weeds, including glyphosate-resistant weeds such as 

marestail, common ragweed, giant ragweed, palmer pigweed, and waterhemp (Johnson et 

al., 2010). Hard-to-control weeds generally require a higher rate and/or application at a 

smaller growth stage in order to consistently achieve commercially acceptable control. 

Refer to the Roundup WeatherMax label (U.S. EPA Reg. No. 524-537) for a listing of 

these weeds. Herbicide resistant weeds are those listed on the International Survey of 

Resistant Weeds website (www.weedscience.org). Since its introduction in 1967, only 

four species with known dicamba-resistant biotypes have been identified in North 

America (Heap, 2011). 

MON 87708 will be combined with MON 89788 (Roundup Ready 2 Yield soybean) 

utilizing traditional breeding techniques. Dicamba is an effective broadleaf herbicide and 

the potential use of dicamba and glyphosate herbicides at the same time in mixtures for 

weed control will provide growers greater application flexibility prior to planting as well 

as in-crop for greater consistency of control in both conventional and conservation tillage 

situations (Johnson et al., 2010). Use of dicamba, in addition to glyphosate and the other 

herbicide options currently labeled for use on soybean, provides more options to 

implement diversified weed management programs to control a broad spectrum of grass 

and broadleaf weed species (Johnson et al, 2010). Successful adoption of the dicamba 

tolerance trait, into the Roundup Ready soybean system, will provide: 1) growers with an 

opportunity for an efficient, effective weed management system; 2) an effective tool for 

the management of glyphosate resistant weeds that will help to conserve reduced tillage 

practices; 3) an option to delay or prevent further resistance to glyphosate and other 
Monsanto Company 10-SY-210U 34 of 721 

critically important soybean herbicides, in particular herbicides in the ALS and PPO class 

of chemistry; 4) excellent crop safety; and 5) soybean growers with effective weed 

control systems necessary for production yields to meet the growing needs of the food, 

feed, and industrial markets. 

MON 87708 expresses a mono-oxygenase enzyme that rapidly demethylates dicamba 

rendering it inactive, thereby conferring tolerance to dicamba. The demethylation of 

dicamba produces 3,6-dichlorosalicylic acid (DCSA), a known soybean, soil, and 

livestock metabolite whose safety has been evaluated by the Environmental Protection 

Agency (U.S. EPA). DCSA, in addition to dicamba, is included in the current 10 ppm 

pesticide residue tolerance for soybean seed that supports the existing uses of dicamba on 

conventional soybean (40 CFR § 180.227). Even with the expanded use of dicamba on 

MON 87708, compared to conventional soybean uses, the rapid metabolism of dicamba 

results in residues in dicamba-treated MON 87708 seed, including the DCSA metabolite, 

that are well below the established 10 ppm tolerance, and therefore no modification to the 

existing soybean seed tolerance is needed. Consequently, only approval for the expanded 

use pattern of dicamba on MON 87708 has been requested of EPA. Furthermore, the 

U.S., Canada and the EU have recently completed reviews of dicamba where the safety 

of dicamba has been confirmed (U.S. EPA, 2009; PMRA, 2008; European Commission, 

2008). The proposed use pattern of dicamba in MON 87708 falls within the use pattern 

criteria (rates and methods) evaluated and approved by EPA in association with existing 

dicamba agricultural uses. 

I.C. Submissions to Other Regulatory Agencies 

Under the Coordinated Framework for Regulation of Biotechnology, the responsibility 

for regulatory oversight of biotechnology-derived crops that do not include 

plant-incorporated protectants falls on two federal agencies: U.S. Food and Drug 

Administration (FDA) and United States Department of Agriculture (USDA). 

Deregulation of MON 87708 by USDA constitutes only one component of the overall 

regulatory oversight and review of this product. As a practical matter, MON 87708 

cannot be released and marketed until FDA and USDA have completed their reviews and 

assessments under their respective jurisdictions. Additionally, EPA must complete its 

review and assessments prior to approving the use of dicamba on MON 87708. 

I.C.1. Submission to FDA

MON 87708 falls within the scope of the 1992 FDA policy statement concerning 

regulation of products derived from new plant varieties, including those developed 

through biotechnology (U.S. FDA, 1992). In compliance with this policy, Monsanto has 

initiated a consultation with the FDA (BNF No. 125) on the food and feed safety and 

compositional assessment of MON 87708. A safety and nutritional assessment summary 

document for MON 87708 (BNF No. 125) was submitted to FDA on November 9, 2010. 

FDA completed the consultation process for MON 87708 on October 11, 2011.2

 

 

2

 http://www.accessdata.fda.gov/scripts/fcn/fcnDetailNavigation.cfm?rpt=bioListing&id=86 
Monsanto Company 10-SY-210U 35 of 721 

I.C.2. Submission to EPA

The EPA has authority over the use of pesticidal substances under the Federal Insecticide, 

Fungicide and Rodenticide Act (FIFRA), as amended (7 U.S.C. § 136 et seq.). Monsanto 

has submitted to the EPA an application to amend Registration Number 524-582 to 

register a new use pattern for dicamba on MON 87708. The new use pattern facilitates a 

wider window of application of dicamba on MON 87708, allowing dicamba to be applied 

preemergence through crop emergence (cracking) and in-crop postemergence through the 

early R1/R2 reproductive phase. EPA has reviewed the safety of dicamba and DCSA, the 

primary metabolite in MON 87708, during the reregistration of dicamba in 2006. EPA 

concluded in the 2006 dicamba Reregistration Eligibility Decision (RED) document that 

risks to human health and the environment associated with exposure to dicamba and its 

metabolites, including DCSA, were below the Agency’s level of concern for all 

registered uses of dicamba including conventional soybean (U.S. EPA, 2009). Dicamba 

residues on soybean seed (less than 0.07 ppm average residue and less than 0.5 ppm 

maximum residue) resulting from its application on MON 87708 at the maximum labeled 

use rate are well below the established 10 ppm soybean seed pesticide residue tolerance. 

Therefore, a change to the current soybean seed tolerance is not needed to support the use 

of dicamba on MON 87708. However Monsanto has requested the establishment of new 

tolerances for soybean forage and hay, which will allow for the feeding of forage and hay 

to livestock. No other revisions to dicamba pesticide residue tolerances are needed 

including animal products such as meat or milk. Furthermore, the use of dicamba on 

MON 87708 does not present any new environmental exposure scenarios not previously 

evaluated and deemed acceptable by EPA. Additional details regarding dicamba and its 

use on MON 87708 are available in Appendix M. 

I.C.3. Submissions to Foreign Government Agencies 

To support commercial introduction of MON 87708 in the U.S., regulatory submissions 

will be made to countries that will eventually commercialize or import significant 

quantities of soybean or its processed fractions from the U.S. These will include 

submissions to a number of foreign government regulatory authorities, including: 

Ministry of Agriculture, People’s Republic of China; Japan’s Ministry of Agriculture, 

Forestry, and Fisheries, Ministry of Environment, and the Ministry of Health, Labor, and 

Welfare; the Canadian Food Inspection Agency and Health Canada; the Intersectoral 

Commission for Biosafety of Genetically Modified Organisms, Mexico; the European 

Food Safety Authority, as well as to regulatory authorities in other soybean importing 

countries with functioning regulatory systems. As appropriate, notifications of 

importation will be made to importing countries that do not have a formal approval 

process. 
Monsanto Company 10-SY-210U 36 of 721 

II. THE BIOLOGY OF SOYBEAN 

The Organization for Economic Co-Operation and Development (OECD) Consensus 

Document (OECD, 2000) on the biology of soybean provides key information on: 

 a general description of soybean biology, including taxonomy and morphology as 

well as soybean use as a crop plant 

 agronomic practices in soybean cultivation 

 geographic centers of origin 

 reproductive biology 

 cultivated soybean as a volunteer weed 

 inter-species/genus introgression into relatives and interactions with other 

organisms, and 

 a summary of the ecology of soybean 

The taxonomic information for soybean is available in the USDA’s PLANTS Profile 

(USDA-NRCS, 2010). 

To support the evaluation of the plant pest potential of MON 87708 relative to 

conventional soybean, additional information regarding several aspects of soybean 

biology can be found elsewhere in this petition. This includes: agronomic practices for 

soybean in Section VIII; volunteer management of soybean in Section VIII.J; and interspecies/genus introgression potential in Section VII.C.3. 

II.A. Soybean as a Crop 

Soybean is the most widely grown oilseed in the world, with approximately 211 million 

metric tons of harvested seed produced in 2008. This represents 56% of world oilseed 

seed production that year. Soybean is grown as a commercial crop in over 35 countries. 

The major producers are the U.S., Brazil, Argentina, China, India, and Paraguay, 

accounting for approximately 94% of the global soybean production in 2008. 

Approximately one-third of the 2008 world soybean production was in the U.S. 

(Soyatech, 2010). The U.S. was also the largest soybean seed exporting country in 2008 

(ASA, 2009). 

Soybean has a long history of planting and production in North America. Soybean was 

originally introduced into North America from China in 1765 and has since been 

reintroduced several times by scientists, seed dealers, merchants, military expeditions, 

and various individuals (Singh and Hymowitz, 1999). Conventional plant breeding is 

based on the interplay and combination of genes present in the particular crop genome, 

and soybean is limited with regard to genetic diversity (Chung and Singh, 2008). 

II.B. Characteristics of the Recipient Plant 

The conventional soybean variety A3525, used as the recipient for the dmo expression 

cassette insertion that produced MON 87708, was developed by Asgrow Seed Company. 

A3525 is a mid−maturity group III soybean variety with very high yield potential. 
Monsanto Company 10-SY-210U 37 of 721 

A3525 has superior yields relative to varieties of similar maturity and has excellent 

agronomic characteristics (Steffen, 2004). 

II.C. Soybean as a Test System in Product Safety Assessment 

Soybean variety A3525 is the near isogenic line to MON 87708 and was used as the 

conventional soybean comparator (hereafter referred to as the conventional control) in the 

safety assessment of MON 87708. MON 87708 and the conventional control have 

similar genetic backgrounds with the exception of the dmo expression cassette, thus, the 

effect of the dmo expression cassette and the expressed MON 87708 DMO could 

therefore be assessed in an unbiased manner. In addition, commercial soybean varieties 

that were derived through conventional methods and Roundup Ready soybean varieties 

(hereafter referred to as commercial reference varieties) were used as reference materials 

to establish ranges of natural variability or responses representative of commercial 

soybean varieties. The commercial reference varieties used at each location were 

selected based on their availability and agronomic fit for the respective geographic region. 
Monsanto Company 10-SY-210U 38 of 721 

III. DESCRIPTION OF THE GENETIC MODIFICATION 

MON 87708 was developed through Agrobacterium tumefaciens-mediated 

transformation of conventional soybean A3525 meristem tissue utilizing transformation 

plasmid vector PV-GMHT4355. This section describes the plasmid vector, the donor 

genes, and the regulatory elements used in the development of MON 87708 and the 

deduced amino acid sequence of the MON 87708 DMO. In this section, transfer DNA 

(T-DNA) refers to DNA that is transferred to the plant during transformation. An 

expression cassette is comprised of sequences to be transcribed and the regulatory 

elements necessary for the expression of those sequences. 

III.A. The Plasmid Vector PV-GMHT4355 

PV-GMHT4355 was used for the transformation of conventional soybean to produce 

MON 87708 and is shown in Figure III-1; PV-GMHT4355 is approximately 11.4 kb and 

contains two T-DNAs, each delineated by Left and Right Border sequences to facilitate 

transformation. The first T-DNA, designated as T-DNA I, contains the dmo coding 

sequence under regulation of the peanut chlorotic streak virus (PC1SV) promoter and the 

pea E9 3′ non-translated region. The second T-DNA, designated as T-DNA II, contains 

the cp4 epsps coding sequence under the regulation of the figwort mosaic virus (FMV) 

promoter and the pea E9 3′ non-translated region. During transformation, both T-DNAs 

were inserted into the soybean genome (Section III.B) where T-DNA II, containing the 

cp4 epsps expression cassette, functioned as a marker gene for the selection of 

transformed plantlets. Subsequently, conventional self-pollinated breeding methods and 

segregation, along with a combination of analytical techniques, were used to isolate those 

plants that contain the dmo expression cassette (T-DNA I) and did not contain the cp4 

epsps expression cassette (T-DNA II). 

The backbone region of PV-GMHT4355 that is outside both of the T-DNAs contains two 

origins of replication for maintenance of the plasmid vector in bacteria (ori V, oripBR322), a bacterial selectable marker gene (aadA), and a coding sequence for repressor 

of primer (rop) protein which is necessary for the maintenance of the plasmid vector copy 

number in E. coli. A description of the genetic elements and their prefixes (e.g., P-, L-, I-, 

TS-, OR-, B-, CS-, and T-) in PV-GMHT4355 is provided in Table III-1. 
Monsanto Company 10-SY-210U 39 of 721 

Probe 

Number 

Probe 

Type 

Probe 

Name 

Start Position 

(bp) 

End Position 

(bp) 

Total Length 

(bp) 

1 Backbone Probe B1 443 1328 886 

2 Backbone probe B2 1250 2754 1505 

3 Backbone Probe B3 2625 4384 1760 

4 T-DNA II Probe TII-1 4796 5637 842 

5 T-DNA II Probe TII-2 5575 7021 1447 

6 T-DNA II Probe TII-3 6937 7761 825 

7 Backbone Probe B4 8119 8289 171 

8 T-DNA I Probe TI-1 8290 9523 1234 

9 T-DNA I Probe TI-2 9448 10668 1221 

10 T-DNA I Probe TI-3 10610 442 1185 

Figure III-1. Circular Map of Plasmid Vector PV-GMHT4355 Showing Probes 1-10 

The plasmid vector PV-GMHT4355 containing the T-DNAs used in Agrobacterium-mediated 

transformation to produce MON 87708. Genetic elements and restriction sites for enzymes used in the 

Southern blot analyses (with positions relative to the size of the plasmid vector) are shown on the exterior 

of the map. The probes used in the Southern blot analyses (labeled 1-10 on the interior of the map) are 

detailed in the accompanying table above. 

*The Left and Right Border sequences of T-DNA II share 100% identity to those of T-DNA I, which were 

covered by probes 8 and 10 and thus not included in the T-DNA II probes. 

PV-GMHT4355

11352 bp

Aat II 6042

Bsp1286 I 386

Bsp1286 I 554

Bsp1286 I 830

Bsp1286 I 1217

Bsp1286I 2041

Bsp1286 I 2539

Bsp1286 I 3370

Bsp1286 I 3720

Bsp1286 I 4444

Bsp1286 I 5867

Bsp1286 I 7733

Kpn I 6322

Kpn I 10278

Nde I 2044

Pvu II 1814

Pvu II 5637

Pvu II 5823

Pvu II 7397

Pvu II 10380

B-Left Border

OR-ori V

CS-rop

OR-ori-pBR322

aadA

B-Left Border

T-E9

CS-cp4 epsps

TS-CTP2

L-DnaK

P-FMV

B-Right Border

B-Right Border

P-PC1SV

L-TEV

TS-RbcS

CS-dmo

T-E9

1

2

3

4 5

6

7

8

9

10

*

*
Monsanto Company 10-SY-210U 40 of 721 

Table III-1. Summary of Genetic Elements in the Plasmid Vector PV-GMHT4355 

Genetic Element 

Location in 

Plasmid (bp) Function (Reference) 

T-DNA I (Present in MON 87708) 

B1-Right Border 8290-8646 

DNA region from Agrobacterium tumefaciens 

containing the Right Border sequence used for 

transfer of the T-DNA (Depicker et al., 1982; 

Zambryski et al., 1982) 

Intervening sequence 8647-8691 Sequence used in DNA cloning 

P2-PC1SV 8692-9124 

Promoter for the Full-Length Transcript (FLt) 

of peanut chlorotic streak caulimovirus (Maiti 

and Shepherd, 1998) that directs transcription 

in plant cells 

Intervening sequence 9125-9144 Sequence used in DNA cloning 

L3-TEV 9145-9276 

5′ non-translated region from the Tobacco Etch 

virus genome (Niepel and Gallie, 1999) that is 

involved in regulating gene expression 

Intervening sequence 9277 Sequence used in DNA cloning 

TS4-RbcS 9278-9520 

Sequences encoding the transit peptide and the 

first 24 amino acids of the mature protein of the 

RbcS gene from Pisum sativum (pea) (Fluhr et 

al., 1986) that directs transport to the DMO 

precursor protein of the chloroplast 

Intervening Sequence 9521-9529 Sequence used in DNA cloning 

CS5-dmo 9530-10552 

Coding sequence for the dicamba 

mono-oxygenase from Stenotrophomonas 

maltophilia (Herman et al., 2005; Wang et al., 

1997) 

Intervening Sequence 10553-10620 Sequence used in DNA cloning 

T6-E9 10621-11263

3′ non-translated region from the RbcS2 gene of 

Pisum sativum (pea) encoding the Rubisco 

small subunit, which functions to direct 

polyadenylation of the mRNA (Coruzzi et al., 

1984) 

Intervening Sequence 11264-11352 Sequence used in DNA cloning 

B-Left Border 1-442 

DNA region from Agrobacterium tumefaciens

containing the Left Border sequence used for 

transfer of the T-DNA (Barker et al., 1983) 

 
Monsanto Company 10-SY-210U 41 of 721 

Table III-1 (continued). Summary of Genetic Elements in the Plasmid Vector 

PV-GMHT4355 

Genetic Element 

Location in 

Plasmid (bp) Function (Reference) 

Plasmid Vector Backbone (Not present in MON 87708) 

Intervening Sequence 443-528 Sequence used in DNA cloning 

OR7-ori V 529-925 

Origin of replication from the broad host range 

plasmid RK2 for maintenance of plasmid in 

Agrobacterium (Stalker et al., 1981) 

Intervening Sequence 926-1662 Sequence used in DNA cloning 

CS-rop 1663-1854 

Coding sequence for repressor of primer 

protein derived from the ColE1 plasmid for 

maintenance of plasmid copy number in E. coli

(Giza and Huang, 1989) 

Intervening Sequence 1855-2281 Sequence used in DNA cloning 

OR-ori-pBR322 2282-2870 

Origin of replication from pBR322 for 

maintenance of plasmid in E. coli (Sutcliffe, 

1979) 

Intervening Sequence 2871-3400 Sequence used in DNA cloning 

aadA 3401-4289 

Bacterial promoter, coding and 3′ UTR 

sequences for an aminoglycoside-modifying 

enzyme, 3′′ (9)-O-nucleotidyltransferase from 

transposon Tn7 (Fling et al., 1985) that confers 

spectinomycin and streptomycin resistance 

Intervening Sequence 4290-4384 Sequence used in DNA cloning 

T-DNA II (Not present in MON 87708)

B-Left Border 4385-4795 

DNA region from Agrobacterium tumefaciens

containing the Left Border sequence used for 

transfer of the T-DNA (Barker et al., 1983) 

Intervening Sequence 4796-4809 Sequence used in DNA cloning 

T-E9 4810-5452 

3′ non-translated sequence from RbcS2 gene of 

Pisum sativum (pea) encoding the Rubisco 

small subunit, which functions to direct 

polyadenylation of the mRNA (Coruzzi et al., 

1984) 

Intervening Sequence 5453-5458 Sequence used in DNA cloning 

CS-cp4 epsps 5459-6826 

Codon optimized coding sequence of the aroA

gene from Agrobacterium spp. strain CP4 

encoding CP4 EPSPS (Barry et al., 1997; 

Padgette et al., 1996a) 

 
Monsanto Company 10-SY-210U 42 of 721 

Table III-1 (continued). Summary of Genetic Elements in the Plasmid Vector 

PV-GMHT4355 

Genetic Element 

Location in 

Plasmid 

(bp) Function (Reference) 

TS-CTP2 6827-7054 

Sequences encoding the chloroplast transit 

peptide region from the shkG gene of Arabidopsis 

thaliana encoding EPSPS (Herrmann, 1995; Klee 

et al., 1987) that directs transport of the CP4 

EPSPS precursor protein to the chloroplast 

Intervening Sequence 7055-7063 Sequence used in DNA cloning 

L-DnaK 7064-7159 

5′ non-translated leader sequence from the 

Petunia hybrida Hsp70 gene (Rensing and Maier, 

1994) that is involved in regulating gene 

expression 

Intervening Sequence 7160-7162 Sequence used in DNA cloning 

P-FMV 7163-7714 

Promoter for the 35S RNA from figwort mosaic 

virus (Rogers, 2000) that directs transcription in 

plant cells 

Intervening Sequence 7715-7761 Sequence used in DNA cloning 

B-Right Border 7762-8118 

DNA region from Agrobacterium tumefaciens 

containing the Right Border sequence used for 

transfer of the T-DNA (Depicker et al., 1982; 

Zambryski et al., 1982) 

Intervening sequence 8119-8289 Sequence used in DNA cloning 

1

B -border. 

2

P-promoter. 

3

L- leader. 

4

TS- targeting sequence. 

5

CS-coding sequence. 

6

T- 3' non-translated transcriptional termination sequence and polyadenylation signal sequences. 

7

OR-origin of replication. 
Monsanto Company 10-SY-210U 43 of 721 

III.B. Description of the Transformation System 

The Agrobacterium-mediated soybean transformation used to produce MON 87708 was 

based on the method described by Martinell et al. (2002), which allows for the generation 

of transformed plants without utilization of callus. Briefly, meristem tissues were excised 

from the embryos of germinated conventional seed. After co-culturing with the 

Agrobacterium carrying the vector, the meristems were placed on selection medium 

containing glyphosate, carbenicillin, cefotaxime, and ticarcillin/clavulanate acid mixture, 

to inhibit the growth of untransformed plant cells and excess Agrobacterium. The 

meristems were then placed in media conducive to shoot and root development. Rooted 

plants with normal phenotypic characteristics were selected and transferred to soil for 

growth and further assessment. 

The R0 plants generated through this transformation were self-pollinated to produce R1

plants, and the unlinked insertions of T-DNA I and T-DNA II were segregated. A 

non-lethal dose of glyphosate was applied to R1 plants and those plants with minor 

herbicide injury were selected for further analyses, whereas plants showing no injury, 

indicating that they contained the cp4 epsps coding sequence from T-DNA II, were 

eliminated from further development. Subsequently, plants that were homozygous for 

T-DNA I were identified by quantitative polymerase chain reaction (PCR) analysis. 

MON 87708 was selected as the lead event based on superior phenotypic characteristics, 

dicamba tolerance, and its molecular profile. The major development steps of 

MON 87708 are depicted in Figure III-2. The result of this process was the production of 

marker-free, dicamba-tolerant soybean MON 87708. 
Monsanto Company 10-SY-210U 44 of 721 



Figure III-2. Schematic of the Development of MON 87708 

Transformed meristem tissue from A3525 with PV-GMHT4355 in 

Agrobacterium tumefaciens

Selected transformants and generated rooted shoots from the 

transformed meristem tissues 

Screening of transformed plants for the presence of T-DNA I (dmo

expression cassette) and absence of the T-DNA II 

(cp4 epsps expression cassette) 

Identified MON 87708 as lead candidate based on analysis of the 

genomic insert and evaluation of progeny generations in laboratory and 

field assessments 

Assembled Agrobacterium binary plasmid vector PV-GMHT4355 and 

transferred to Agrobacterium tumefaciens strain ABI 
Monsanto Company 10-SY-210U 45 of 721 

III.C. The dmo Coding Sequence and MON 87708 DMO (T-DNA I) 

The dmo expression cassette (T-DNA I) present in MON 87708 encodes MON 87708 

DMO (Figure III-3). The dmo expression cassette contains the coding region for the 

DMO from Stenotrophomonas maltophilia (Herman et al., 2005; Wang et al., 1997). The 

presence of MON 87708 DMO confers tolerance to dicamba (refer to Section V.A for 

more details). 

1 MASMISSSAV TTVSRASRGQ SAAMAPFGGL KSMTGFPVRK VNTDITSITS NGGRVKCMQV

61 WPPIGKKKFE TLSYLPPLTR DSRAMATFVR NAWYVAALPE ELSEKPLGRT ILDTPLALYR 

121 QPDGVVAALL DICPHRFAPL SDGILVNGHL QCPYHGLEFD GGGQCVHNPH GNGARPASLN 

181 VRSFPVVERD ALIWICPGDP ALADPGAIPD FGCRVDPAYR TVGGYGHVDC NYKLLVDNLM 

241 DLGHAQYVHR ANAQTDAFDR LEREVIVGDG EIQALMKIPG GTPSVLMAKF LRGANTPVDA 

301 WNDIRWNKVS AMLNFIAVAP EGTPKEQSIH SRGTHILTPE TEASCHYFFG SSRNFGIDDP 

361 EMDGVLRSWQ AQALVKEDKV VVEAIERRRA YVEANGIRPA MLSCDEAAVR VSREIEKLEQ 

421 LEAA

Figure III-3. Deduced Amino Acid Sequence of the RbcS Targeting Sequence and 

MON 87708 DMO 

The transit peptide and the first 24 amino acids of the mature protein of the RbcS gene are 

underlined. Accumulation of MON 87708 DMO is targeted to the chloroplasts using the RbcS

transit peptide. (see Section V.A for more detail). 

III.D. The cp4 epsps Coding Sequence and the CP4 EPSPS Protein (T-DNA II) 

The cp4 epsps expression cassette (T-DNA II), that is not present in MON 87708, 

encoded a 47.6 kDa CP4 EPSPS protein, consisting of a single polypeptide of 455 amino 

acids (Padgette et al., 1996b). The cp4 epsps coding sequence is the codon optimized 

coding sequence of the aroA gene from Agrobacterium spp. strain CP4 encoding CP4 

EPSPS (Barry et al., 1997; Padgette et al., 1996a). CP4 EPSPS confers tolerance to 

glyphosate and was used as a selectable marker during the transformation selection 

process. Through conventional self-pollinated breeding methods and segregation, along 

with a combination of analytical techniques, plants that did not contain the cp4 epsps

expression cassette were isolated. 

III.E. Regulatory Sequences 

The dmo coding sequence in T-DNA I is under the regulation of the PC1SV promoter, 

TEV leader, the RbcS targeting sequence, and the E9 3′ non-translated region. The 

PC1SV promoter is the promoter for the Full-Length Transcript (FLt) of peanut chlorotic 

streak caulimovirus (Maiti and Shepherd, 1998) that directs transcription in plant cells. 

The TEV leader is the 5′ non-translated region from the Tobacco Etch virus (Niepel and 

Gallie, 1999) and is involved in regulating gene expression. The RbcS targeting sequence 

is the sequence encoding the chloroplast transit peptide and the first 24 amino acids of the 

mature protein of the ribulose-1,5-bisphosphate carboxylase oxygenase gene from pea 

(Pisum sativum) (Fluhr et al., 1986) that directs transport of the DMO precursor protein 

to the chloroplast. The E9 3′ non-translated region is the 3′ non-translated region from 
Monsanto Company 10-SY-210U 46 of 721 

the RbcS2 gene of pea encoding the Rubisco small subunit, which functions to direct 

polyadenylation of the mRNA (Coruzzi et al., 1984). 

T-DNA II contains the cp4 epsps coding sequence under the regulation of the FMV

promoter, DnaK leader, the CTP2 targeting sequence, and the E9 3′ non-translated region. 

The FMV promoter is the promoter for the 35S RNA from figwort mosaic virus (Rogers, 

2000) that directs transcription in plant cells. The DnaK leader is the 5′ non-translated 

leader sequence from the Petunia hybrida Hsp70 gene (Rensing and Maier, 1994) that is 

involved in regulating gene expression. The CTP2 targeting sequence is the sequence 

encoding the chloroplast transit peptide region from the shkG gene of Arabidopsis

thaliana encoding EPSPS (Herrmann, 1995; Klee et al., 1987) that directs transport of the 

CP4 EPSPS precursor protein to the chloroplast. The E9 3′ non-translated region is the 3′

non-translated region from the RbcS2 gene of pea encoding the Rubisco small subunit, 

which functions to direct polyadenylation of the mRNA (Coruzzi et al., 1984). 

III.F. T-DNA Borders 

PV-GMHT4355 contains Right and Left Border regions (Figure III-1 and Table III-1) 

that were derived from Agrobacterium tumefaciens (Barker et al., 1983; Depicker et al., 

1982; Zambryski et al., 1982). The border regions each contain a 24-25 bp nick site that 

is the site of DNA exchange during transformation. The border regions separate the 

T-DNA from the backbone region and are involved in their efficient transfer into the 

soybean genome. Because PV-GMHT4355 is a 2T-DNA vector, it contains two Right 

Border regions and two Left Border regions, where one set flanks T-DNA I and the other 

set flanks T-DNA II. 

III.G. Genetic Elements Outside of the T-DNA Borders 

Genetic elements that exist outside of the T-DNA borders are those that are essential for 

the maintenance or selection of PV-GMHT4355 in bacteria and are referred to as the 

plasmid backbone. The ori V, derived from the broad host plasmid RK2, is required for 

the maintenance of the plasmid vector in Agrobacterium (Stalker et al., 1981), whereas 

the ori-pBR322, derived from the plasmid vector pBR322, is required for the 

maintenance of the plasmid vector in E. coli (Sutcliffe, 1979). The rop is necessary for 

the maintenance of plasmid vector copy number in E. coli (Giza and Huang, 1989). The 

aadA is a bacterial promoter and coding sequence for an enzyme from transposon Tn7 

that confers spectinomycin and streptomycin resistance (Fling et al., 1985) in E. coli and 

Agrobacterium during molecular cloning. Because these elements are outside the border 

regions, they were not expected to be transferred into the soybean genome. The absence 

of the backbone sequence in MON 87708 was confirmed by Southern blot analyses (see 

Section IV.C). 
Monsanto Company 10-SY-210U 47 of 721 

IV. CHARACTERIZATION OF THE GENETIC MODIFICATION 

A multi-faceted approach was taken to characterize the genetic modification that 

produced MON 87708. The results confirmed that MON 87708 contains a single copy of 

the dmo expression cassette (T-DNA I) that is stably integrated at a single locus and is 

inherited according to Mendelian principles over multiple generations (Section IV.G). 

The results confirmed that no T-DNA II or plasmid vector backbone sequences are 

detected in MON 87708. These conclusions are based on several lines of evidence: 

1) Southern blot analyses to assay the entire soybean genome for the presence of DNA 

derived from PV-GMHT4355, and to confirm that a single copy of T-DNA I was inserted 

at a single site and that the insert is stably inherited; 2) DNA sequencing analyses to 

determine the exact sequence of the inserted DNA and allowed a comparison to the 

T-DNA I sequence in PV-GMHT4355 to confirm that only the expected sequences were 

integrated; and 3) a comparison of the DNA flanking T-DNA I to the sequence of the 

insertion site in conventional soybean to identify any rearrangements that occurred at the 

insertion site during transformation. Taken together, the characterization of the genetic 

modification demonstrates that a single copy of the T-DNA I was inserted at a single 

locus of the genome. 

Southern blot analyses were used to determine the number of copies and the insertion 

sites of T-DNA I as well as the presence or absence of T-DNA II and plasmid vector 

backbone sequences. The Southern blot strategy was designed to ensure that all potential 

inserted segments would be identified. The entire soybean genome was assayed with 

probes that spanned the complete plasmid vector PV-GMHT4355 to detect the presence 

of T-DNA I as well as confirm the lack of any detectable T-DNA II and plasmid vector 

backbone sequences. This was accomplished by using probes that were less than 2 kb in 

length, ensuring a high level of sensitivity. This high level of sensitivity was 

demonstrated for each blot by detection of a positive control added at 0.1 copies per 

genome equivalent. Two restriction enzyme sets were specifically chosen to fully 

characterize T-DNA I and look for any potential fragments of T-DNA I. This two 

enzyme set design also maximizes the possibility of detecting an insertion elsewhere in 

the genome that could be overlooked if that band comigrated with an expected band. 

Additionally, the restriction enzyme sets were chosen such that at least one enzyme from 

each set resides in the known 5′ or 3′ flanking sequence and that together the enzyme sets 

result in overlapping segments covering the entire insert. Therefore, at least one segment 

for each flank is of a predictable size and overlaps with another predictable size segment. 

This overlapping strategy confirms that the entire insert sequence is identified in a 

predictable hybridization pattern. 

To determine the number of copies and the insertion sites of T-DNA I, and the presence 

or absence of T-DNA II and the plasmid vector backbone sequences, duplicated samples 

that consisted of equal amounts of digested DNA were run on the agarose gel. One set of 

samples was run for a longer period of time (long run) than a second set (short run). The 

long run allows for greater resolution of large molecular weight DNA, whereas the short 

run allows the detection of small molecular weight DNA. The molecular weight markers 

on the left of the figures were used to estimate the sizes of the bands present in the long 

run lanes of the Southern blots, and the molecular weight markers on the right of the 
Monsanto Company 10-SY-210U 48 of 721 

figures were used to estimate the sizes of bands present in the short run lanes of the 

Southern blots. 

The DNA sequencing analyses complement the Southern blot analyses. Southern blot 

results demonstrated that MON 87708 contains a single copy of T-DNA I at a single 

insertion site. Sequencing of the insert and the flanking DNA confirmed the organization 

of the elements within the insert, determined the 5′ and 3′ insert-to-plant junctions, as 

well as the complete DNA sequence of the insert and adjacent DNA. In addition, DNA 

sequencing analyses confirmed that each genetic element in the insert is intact and the 

sequence of the insert matches the corresponding sequence in PV-GMHT4355. 

Furthermore, genomic organization at the insertion site was assessed by comparing the 

insert and flanking sequence to the insertion site in conventional soybean. 

The stability of the T-DNA I present in MON 87708 across multiple generations (R2-R6) 

was demonstrated by Southern blot analysis. Genomic DNA from five generations of 

MON 87708 was digested with one of the enzyme sets used for the insert and copy 

number analysis and was hybridized with a probe that detects restriction segments that 

encompass the entire T-DNA I. This fingerprint strategy consists of two border segments 

that assess not only the stability of T-DNA I, but also the stability of genomic DNA 

directly adjacent to T-DNA I. 

The results of these analyses for MON 87708 demonstrated that a single copy of the 

T-DNA I was inserted at a single locus of the genome. Generational stability analysis 

demonstrated that an expected Southern blot fingerprint of MON 87708 was maintained 

through five generations of the breeding history, thereby confirming the stability of 

T-DNA I in MON 87708. Results from segregation analyses showed heritability and 

stability of the insert occurred as expected across multiple generations, which 

corroborates the molecular insert stability analysis and establishes the genetic behavior of 

the T-DNA I at a single chromosomal locus. 

The Southern blot analysis confirmed that T-DNA I reported in Figure IV-1 represents 

the only detectable insert in MON 87708. Figure IV-1 is a linear map depicting 

restriction sites within the insert as well as within the known soybean genomic DNA 

immediately flanking the insert in MON 87708. The circular map of PV-GMHT4355 

annotated with the probes used in the Southern blot analysis is presented in Figure III-1. 

Based on the linear map of the insert and the plasmid map, a table summarizing the 

expected DNA segments for Southern analyses is presented in Table IV-1. The genetic 

elements integrated in MON 87708 are summarized in Table IV-2. The generations used 

are depicted in the breeding history shown in Figure IV-9. Materials and methods used 

for characterization of T-DNA I in MON 87708 are found in Appendix B. 
Monsanto Company

Figure IV-1. Schematic Representation of the Insert and A linear map of the insert and genomic DNA flanking the insert in MONinsert, as well as restriction sites with positions relative to the size of the linear map for enzymesand locations of the T-DNA I probes, which are described in Figure IIImap are the expected sizes of the DNA segments after digestion insert and the beginning of the genomic DNA sequence flanking the 5the elements in MON 87708. 

10-SY-210U

1. Schematic Representation of the Insert and DNA Flanking Sequences in MON 87708 

A linear map of the insert and genomic DNA flanking the insert in MON 87708 is shown. Identified on the map are geninsert, as well as restriction sites with positions relative to the size of the linear map for enzymes used in the Southern ana

I probes, which are described in Figure III-1, are shown on the middle portion. Shown on 

map are the expected sizes of the DNA segments after digestion with respective restriction enzymes. Arrowheads ( insert and the beginning of the genomic DNA sequence flanking the 5′ and 3′ end of the insert. The arrows (→) indicate t

87708 is shown. Identified on the map are gen

used in the Southern ana

Shown on 

ive restriction enzymes. Arrowheads ( 

) indicate t
Monsanto Company 10-SY-210U 

Table IV-1. Summary Chart of the Expected DNA Segments Based on Hybridizing Probes and RestrictiMON 87708 Analysis Southern Blot Figure IV-2 IV-3 IV-4 IV-5 IV-6 IV-7 IV-8 Probe Used 8

9 10

4

5

6 1, 2, 3

and 7

Probing Target Digestion Enzyme Expected Band Sizes (kb) on Each Southern Blot Plasmid Vector PV-GMHT4355 Aat II/Nde I ~7.4 ~7.4 ~4.0 ~7.4 ~4.0 ~7.4 ~4.0 ~7.4 ~7.4 ~4.0 ~7.4 Probe Templates1 N/A ~~2 ~~2 ~~2 ~~2 ~~2 ~~2

~0.2~0.9~1.5~1.8

Conventional Control A3525 

Bsp1286 I/Pvu II None None None None None None None Hpa I/Kpn I None None None None None None None 

MON 87708 

Bsp1286 I/Pvu II ~2.6 ~2.6 ~1.5 ~1.5 ~1.5 None None None Hpa I/Kpn I >2.7* >2.7* ~1.7 ~1.7 ~1.7 None None None 

1 Probe templates were spiked when multiple probes are used in Southern blot analysis.2 ‘~~’ indicates that only plasmid template was used since the Southern blot was hybridized with one probe. 3 ‘--’ indicates that the particular restriction enzyme or the combination of the enzymes was not used in the analysis. * Southern analysis indicatesthis segment to be ~5.6 kb. 
Monsanto Company 10-SY-210U 51 of 721 

Table IV-2. Summary of Genetic Elements in MON 87708 

Genetic Element

Location 

(bp) Function (Reference)

5′ Flanking Sequences 1-1048 DNA sequence adjacent to the 5′ end of the 

insertion site

B1-Right Border* 1049-1091

DNA region from Agrobacterium tumefaciens 

containing the Right Border sequence used for 

transfer of the T-DNA (Depicker et al., 1982; 

Zambryski et al., 1982)

Intervening sequence 1092-1136 Sequence used in DNA cloning

P2-PC1SV 1137-1569

Promoter for the Full-Length Transcript (FLt) of 

peanut chlorotic streak caulimovirus (Maiti and 

Shepherd, 1998) that directs transcription in plant 

cells

Intervening sequence 1570-1589 Sequence used in DNA cloning

L3-TEV 1590-1721

5' non-translated region from the Tobacco Etch 

virus genome (Niepel and Gallie, 1999) that is 

involved in regulating gene expression

Intervening sequence 1722-1722 Sequence used in DNA cloning

TS4-RbcS 1723-1965

Sequences encoding the transit peptide and the first 

24 amino acids of the mature protein of the RbcS

gene from Pisum sativum (pea) (Fluhr et al., 1986) 

that directs transport of the DMO precursor protein 

to the chloroplast 

Intervening Sequence 1966-1974 Sequence used in DNA cloning

CS5-dmo 1975-2997

Coding sequence for the dicamba mono-oxygenase 

from Stenotrophomonas maltophilia (Herman et al., 

2005; Wang et al., 1997)

Intervening Sequence 2998-3065 Sequence used in DNA cloning

T6-E9 3066-3708

3′ non-translated region from the RbcS2 gene of 

Pisum sativum (pea) encoding the Rubisco small 

subunit, which functions to direct polyadenylation 

of the mRNA (Coruzzi et al., 1984)

Intervening Sequence 3709-3797 Sequence used in DNA cloning

B-Left Border* 3798-4051

DNA region from Agrobacterium tumefaciens 

containing the Left Border sequence used for 

transfer of the T-DNA (Barker et al., 1983)

3′ Flanking Sequences 4052-5322 DNA sequence adjacent to the 3′ end of the 

insertion site

1

B-border. 

2

P-promoter. 

3

L-leader. 

4

TS- targeting sequence. 

5

S- coding sequence. 

6

T-3' non-translated transcriptional termination sequence and polyadenylation signal sequences. 

*These borders are truncated. 
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IV.A. Insert and Copy Number of T-DNA I in MON 87708 

The copy number and insertion site of T-DNA I was assessed by digesting MON 87708 

genomic DNA with the restriction enzyme combination Bsp1286 I/Pvu II or Hpa I/Kpn I 

and hybridizing Southern blots with probes that span T-DNA I (Figure III-1). Each 

restriction digest is expected to produce a specific banding pattern on the Southern blots 

(Table IV-1). Since each detected segment contains flanking genomic DNA, any 

additional integrated sites would produce a different banding pattern with additional 

bands. 

The restriction enzyme combination Bsp1286 I/Pvu II cuts once within T-DNA I and 

once within each of the known genomic DNA sequences flanking the 5′ and 3′ ends of 

T-DNA I (Figure IV-1). Therefore, if T-DNA I sequences are present at a single 

integration site in MON 87708, the digestion with Bsp1286 I/Pvu II was expected to 

generate two border segments with expected sizes of ~2.6 kb and ~1.5 kb (Figure IV-1, 

and Table IV-1). The ~2.6 kb restriction segment contained genomic DNA flanking the 

5′ end of T-DNA I, the Right Border, the PC1SV promoter, the TEV leader, the RbcS

targeting sequence, and a portion of the dmo coding sequence. The ~1.5 kb restriction 

segment contained a portion of the dmo coding sequence, the E9 3′ non-translated 

sequence, the Left Border, and genomic DNA flanking the 3′ end of T-DNA I. 

The restriction enzyme combination Hpa I/Kpn I cuts once within T-DNA I and once 

within the known genomic DNA flanking the 3′ end of T-DNA I (Figure IV-1). 

Therefore, if T-DNA I sequences are present at a single integration site in MON 87708, 

the digestion with Hpa I/Kpn I was expected to generate two border segments with 

expected sizes of ~1.7 kb and greater than 2.7 kb (Figure IV-1, and Table IV-1). Since 

the Hpa I/Kpn I restriction site in the genomic DNA flanking the 5′ end of the insert lies 

outside of the known sequence, it was not possible to predict a precise segment size. 

However, the segment size was determined by Southern blot analyses to be ~5.6 kb 

(Figures IV-2 and IV-3). The ~5.6 kb restriction segment contained genomic DNA 

flanking the 5′ end of T-DNA I, the Right Border, the PC1SV promoter, the TEV leader, 

the RbcS targeting sequence, and a portion of dmo coding sequence. The ~1.7 kb 

restriction segment contained a portion of the dmo coding sequence, the E9 3′

non-translated sequence, the Left Border, and genomic DNA flanking the 3′ end of 

T-DNA I. 

In the Southern blot analyses performed, each Southern blot contained a negative and a 

positive control. Conventional control genomic DNA digested with either the restriction 

enzyme combination Bsp1286 I/Pvu II or Hpa I/Kpn I was used as a negative control to 

determine if the probes hybridized to any endogenous soybean sequences. As a positive 

control on the Southern blots, PV-GMHT4355 digested with the restriction enzyme 

combination Aat II/Nde I was mixed with predigested conventional control DNA. The 

positive hybridization control was spiked at 0.1 and 1 genome equivalent to demonstrate 

sufficient sensitivity of the Southern blot. Individual Southern blots were hybridized 

with the following probes: probes 8, 9, and 10 (refer to Figure III-1 and Table IV-1). 

The results of this analysis are shown in Figures IV-2 through IV-4. 
Monsanto Company 10-SY-210U 53 of 721 

IV.A.1. Probe 8 

Conventional control DNA digested with the restriction enzyme combination 

Bsp1286 I/Pvu II (Figure IV-2, lanes 1 and 5) or Hpa I/Kpn I (Figure IV-2, lanes 3 and 7) 

and hybridized with probe 8 (Figure III-1) produced no detectable hybridization bands as 

expected for the negative control. PV-GMHT4355, digested with the restriction enzyme 

combination Aat II/Nde I and mixed with conventional control DNA predigested with the 

restriction enzyme combination Hpa I/Kpn I (Figure IV-2, lanes 10 and 11), produced the 

expected size band at ~7.4 kb (refer to Figure III-1 and Table IV-1). These results 

indicate that the probe is hybridizing to its target sequence. 

MON 87708 DNA digested with the restriction enzyme combination Bsp1286 I/Pvu II 

and hybridized with probe 8 (Figure III-1) produced one unique band at ~2.6 kb 

(Figure IV-2, lanes 2 and 6). The ~2.6 kb band is the expected size for the border 

segment containing the 5′ end of T-DNA I along with the adjacent genomic DNA 

flanking the 5′ end of T-DNA I (Figure IV-1). 

MON 87708 DNA digested with the restriction enzyme combination Hpa I/Kpn I and 

hybridized with probe 8 (Figure III-1) produced one unique band at ~5.6 kb (Figure IV-2, 

lanes 4 and 8). The ~5.6 kb band is consistent with the expected band being greater than 

2.7 kb for the border segment containing the 5′ end of T-DNA I along with the adjacent 

genomic DNA flanking the 5′ end of T-DNA I (Figure IV-1). 

No additional bands were detected using probe 8. Based on the results presented in 

Figure IV-2, it was concluded that T-DNA I sequences covered by probe 8 reside at a 

single integration locus as one copy in MON 87708. 

IV.A.2. Probe 9 

Conventional control DNA digested with the restriction enzyme combination 

Bsp1286 I/Pvu II (Figure IV-3, lanes 1 and 5) or Hpa I/Kpn I (Figure IV-3, lanes 3 and 7) 

and hybridized with probe 9 (Figure III-1) produced no detectable hybridization bands as 

expected for the negative control. PV-GMHT4355, digested with the restriction enzyme 

combination Aat II/Nde I and mixed with conventional control DNA predigested with the 

restriction enzyme combination Hpa I/Kpn I (Figure IV-3, lanes 10 and 11), produced the 

expected size band at ~7.4 kb (refer to Figure III-1 and Table IV-1). These results 

indicate that the probe is hybridizing to its target sequence. 

MON 87708 DNA digested with the restriction enzyme combination Bsp1286 I/Pvu II 

and hybridized with probe 9 (Figure III-1) produced two unique bands at ~1.5 kb and 

~2.6 kb (Figure IV-3, lanes 2 and 6). The ~1.5 kb band is the expected size for the border 

segment containing the 3′ end of T-DNA I along with the adjacent genomic DNA 

flanking the 3′ end of T-DNA I (Figure IV-1). The ~2.6 kb band is the expected size for 

the border segment containing the 5′ end of T-DNA I along with the adjacent genomic 

DNA flanking the 5′ end of T-DNA I (Figure IV-1). 

MON 87708 DNA digested with the restriction enzyme combination Hpa I/Kpn I and 

hybridized with probe 9 (Figure III-1) produced two unique bands at ~1.7 kb and ~5.6 kb 
Monsanto Company 10-SY-210U 54 of 721 

(Figure IV-3, lanes 4 and 8). The ~1.7 kb band is the expected size for the border 

segment containing the 3′ end of T-DNA I along with the adjacent genomic DNA 

flanking the 3′ end of T-DNA I (Figure IV-1). The ~5.6 kb band is consistent with the 

expected band being greater than 2.7 kb for the border segment containing the 5′ end of 

T-DNA I along with the adjacent genomic DNA flanking the 5′ end of T-DNA I 

(Figure IV-1). 

No additional bands were detected using probe 9. Based on the results presented in 

Figure IV-3, it was concluded that T-DNA I sequences covered by probe 9 reside at a 

single integration locus as one copy in MON 87708. 

IV.A.3. Probe 10 

Conventional control DNA digested with the restriction enzyme combination 

Bsp1286 I/Pvu II (Figure IV-4, lanes 1 and 5) or Hpa I/Kpn I (Figure IV-4, lanes 3 and 7) 

and hybridized with probe 10 (Figure III-1) produced no detectable hybridization bands 

as expected for the negative control. PV-GMHT4355, digested with the restriction 

enzyme combination Aat II/Nde I and mixed with conventional control DNA predigested 

with the restriction enzyme combination Hpa I/Kpn I (Figure IV-4, lanes 10 and 11), 

produced two bands at ~4.0 kb and ~7.4 kb. Both bands were expected because probe 10 

contains E9 and left border sequences that hybridized to both the ~4.0 kb and the ~7.4 kb 

fragments from the digested plasmid (refer to Figure III-1 and Table IV-1). These results 

indicate that the probe is hybridizing to its target sequence. 

MON 87708 DNA digested with the restriction enzyme combination Bsp1286 I/Pvu II 

and hybridized with probe 10 (Figure III-1) produced a unique band at ~1.5 kb 

(Figure IV-4, lanes 2 and 6). The ~1.5 kb band is the expected size for the border 

segment containing the 3′ end of T-DNA I along with the adjacent genomic DNA 

flanking the 3′ end of T-DNA I (Figure IV-1). 

MON 87708 DNA digested with the restriction enzyme combination Hpa I/Kpn I and 

hybridized with probe 10 (Figure III-1) produced a unique band at ~1.7 kb (Figure IV-4, 

lanes 4 and 8). The ~1.7 kb band is the expected size for the border segment containing 

the 3′ end of T-DNA I along with the adjacent genomic DNA flanking the 3′ end of 

T-DNA I (Figure IV-1). 

No additional bands were detected using probe 10. Based on the results presented in 

Figure IV-4, it was concluded that T-DNA sequences covered by probe 10 reside at a 

single integration locus as one copy in MON 87708. 
Monsanto Company 10-SY-210U 55 of 721 

 

Figure IV-2. Southern Blot Analysis to Determine Insert and Copy Number of 

T-DNA I in MON 87708: Probe 8 

The blot was hybridized with a 32P labeled T-DNA I probe that spans a portion of the T-DNA I 

sequence (Probe 8, Figure III-1). Each lane contains approximately 10 μg of digested genomic 

DNA isolated from leaf tissue. Lane designations are as follows: 

Lane Description 

 1. Conventional control (Bsp1286 I/Pvu II) 

2. MON 87708 (Bsp1286 I/Pvu II) 

3. Conventional control (Hpa I/Kpn I) 

4. MON 87708 (Hpa I/Kpn I) 

5. Conventional control (Bsp1286 I/Pvu II) 

6. MON 87708 (Bsp1286 I/Pvu II) 

7. Conventional control (Hpa I/Kpn I) 

8. MON 87708 (Hpa I/Kpn I) 

9. Blank 

10. Conventional control (Hpa I/Kpn I) spiked with PV-GMHT4355 (Aat II/Nde I) 

 (~1 genome equivalent) 

11. Conventional control (Hpa I/Kpn I) spiked with PV-GMHT4355 (Aat II/Nde I) 

 (~0.1 genome equivalent) 

Arrows denote sizes of DNA, in kilobase pairs, obtained from molecular weight markers on 

ethidium bromide stained gel. 

1 2 3 4 5 6 7 8 9 10 11

1.0

1.6

2.0

3.1

4.1

5.1

6.1

7.1

8.1

10

15 20

40

1.0

1.6

2.0

3.1

4.1

5.1 6.1 7.1 8.1 10

15 20 40

0.5

LONG RUN SHORT RUN
Monsanto Company 10-SY-210U 56 of 721 

Figure IV-3. Southern Blot Analysis to Determine Insert and Copy Number of 

T-DNA I in MON 87708: Probe 9 

The blot was hybridized with a 32P labeled T-DNA I probe that spans a portion of the T-DNA I 

sequence (Probe 9, Figure III-1). Each lane contains approximately 10 μg of digested genomic 

DNA isolated from leaf tissue. Lane designations are as follows: 

Lane Description 

1. Conventional control (Bsp1286 I/Pvu II) 

2. MON 87708 (Bsp1286 I/Pvu II) 

3. Conventional control (Hpa I/Kpn I) 

4. MON 87708 (Hpa I/Kpn I) 

5. Conventional control (Bsp1286 I/Pvu II) 

6. MON 87708 (Bsp1286 I/Pvu II) 

7. Conventional control (Hpa I/Kpn I) 

8. MON 87708 (Hpa I/Kpn I) 

9. Blank 

10. Conventional control (Hpa I/Kpn I) spiked with PV-GMHT4355 (Aat II/Nde I) 

 (~1 genome equivalent) 

11. Conventional control (Hpa I/Kpn I) spiked with PV-GMHT4355 (Aat II/Nde I) 

 (~0.1 genome equivalent) 

Arrows denote sizes of DNA, in kilobase pairs, obtained from molecular weight markers on 

ethidium bromide stained gel. 

1.0

1.6

2.0

3.1

4.1

5.1

6.1

7.1

8.1

10

15 20 40

1.0

1.6

2.0

3.1

4.1

5.1 6.1

7.1 8.1

10

15 20 40

0.5

1 2 3 4 5 6 7 8 9 10 11

LONG RUN SHORT RUN
Monsanto Company 10-SY-210U 57 of 721 

Figure IV-4. Southern Blot Analysis to Determine Insert and Copy Number of 

T-DNA I in MON 87708: Probe 10 

The blot was hybridized with a 32P labeled T-DNA I probe that spans a portion of the T-DNA I 

sequence (Probe 10, Figure III-1). Each lane contains approximately 10 μg of digested genomic 

DNA isolated from leaf tissue. Lane designations are as follows: 

Lane Description 

 1. Conventional control (Bsp1286 I/Pvu II) 

2. MON 87708 (Bsp1286 I/Pvu II) 

3. Conventional control (Hpa I/Kpn I) 

4. MON 87708 (Hpa I/Kpn I) 

5. Conventional control (Bsp1286 I/Pvu II) 

6. MON 87708 (Bsp1286 I/Pvu II) 

7. Conventional control (Hpa I/Kpn I) 

8. MON 87708 (Hpa I/Kpn I) 

9. Blank 

10. Conventional control (Hpa I/Kpn I) spiked with PV-GMHT4355 (Aat II/Nde I) 

 (~1 genome equivalent) 

11. Conventional control (Hpa I/Kpn I) spiked with PV-GMHT4355 (Aat II/Nde I) 

 (~0.1 genome equivalent) 

Arrows denote sizes of DNA, in kilobase pairs, obtained from molecular weight markers on 

ethidium bromide stained gel.

1.0

1.6

2.0

3.1

4.1

5.1

6.1

7.1

8.1

10

15 20 40

1.0

1.6

2.0

3.1

4.1

5.1

6.1 7.1

8.1 10

15

20 40

0.5

1 2 3 4 5 6 7 8 9 10 11

LONG RUN SHORT RUN
Monsanto Company 10-SY-210U 58 of 721 

IV.B. Southern Blot Analysis to Determine the Presence or Absence of T-DNA II 

Sequences in MON 87708 

To determine the presence or absence of T-DNA II sequences, MON 87708 and 

conventional control genomic DNA were digested with the restriction enzyme 

combination Bsp1286 I/Pvu II or Hpa I/Kpn I and Southern blots were hybridized with 

probes that span the T-DNA II sequence (Figure III-1). As a positive control on the 

Southern blots, PV-GMHT4355 digested with the restriction enzyme combination 

Aat II/Nde I was mixed with predigested conventional control DNA. The positive 

hybridization control was spiked at 0.1 and 1 genome equivalent to demonstrate 

sufficient sensitivity of the Southern blot. Each blot was hybridized with one of three 

overlapping probes spanning the T-DNA II sequence other than the two border regions 

that share the same sequences as present in T-DNA I (Probes 4, 5 and 6, Figure III-1). If 

T-DNA II sequences were present in MON 87708, then probing with the T-DNA II 

sequences should result in unique hybridizing bands. The results of this analysis are 

shown in Figures IV-5 through IV-7. 

IV.B.1. Probe 4 

Conventional control DNA digested with Bsp1286 I/Pvu II (Figure IV-5, lanes 1 and 5) 

or Hpa I/Kpn I (Figure IV-5, lanes 3 and 7) and hybridized with probe 4 showed no 

detectable hybridization bands, as expected for the negative control. PV-GMHT4355, 

previously digested with Aat II/Nde I and mixed with conventional control DNA 

predigested with Hpa I/Kpn I (Figure IV-5, lanes 10 and 11), produced two bands at 

~4.0 kb and ~7.4 kb. Both bands were expected because probe 4 contains E9 sequence 

that hybridized to both the ~4.0 kb and the ~7.4 kb fragments from the digested plasmid 

(refer to Figure III-1 and Table IV-1). These results indicate that the probe is hybridizing 

to its target sequence. 

MON 87708 DNA digested with the restriction enzyme combination Bsp1286 I/Pvu II 

and hybridized with probe 4 (Figure III-1) produced one unique band at ~1.5 kb 

(Figure IV-5, lanes 2 and 6). MON 87708 DNA digested with Hpa I/Kpn I and 

hybridized with probe 4 (Figure III-1) produced one unique band at ~1.7 kb (Figure IV-5, 

lanes 4 and 8). Probe 4 contains the E9 3′ non-translated region sequence that is also 

contained in T-DNA I (Figure III-1). Therefore, probe 4 was expected to hybridize to the 

~1.5 kb and ~1.7 kb fragments (Figure IV-1) derived from the T-DNA I insert. These 

bands were also detected by probe 10 (Figure IV-4, lanes 2 and 6, and lanes 4 and 8). 

Any T-DNA II sequences other than those associated with T-DNA I would be detected as 

novel bands. No unexpected bands were detected indicating that MON 87708 contains 

no detectable T-DNA II elements covered by probe 4. 

IV.B.2. Probe 5 

Conventional control DNA digested with the restriction enzyme combination 

Bsp1286 I/Pvu II (Figure IV-6, lanes 1 and 5) or Hpa I/Kpn I (Figure IV-6, lanes 3 and 7) 

and hybridized with probe 5 (Figure III-1) showed no detectable hybridization bands, as 

expected for the negative control. PV-GMHT4355, previously digested with Aat II/Nde I 
Monsanto Company 10-SY-210U 59 of 721 

and mixed with conventional control DNA predigested with Hpa I/Kpn I (Figure IV-6, 

lanes 10 and 11), produced two expected size bands at ~4.0 kb and ~7.4 kb (refer to 

Figure III-1 and Table IV-1). These results indicate that the probe is hybridizing to its 

target sequence. 

MON 87708 DNA digested with the restriction enzyme combination Bsp1286 I/Pvu II 

(Figure IV-6, lanes 2 and 6) or Hpa I/Kpn I (Figure IV-6, lanes 4 and 8) and hybridized 

with probe 5, produced no detectable hybridization bands. These results indicate that 

MON 87708 contains no detectable T-DNA II elements covered by probe 5. 

IV.B.3. Probe 6 

Conventional control DNA digested with Bsp1286 I/Pvu II (Figure IV-7, lanes 1 and 5) 

or Hpa I/Kpn I (Figure IV-7, lanes 3 and 7) and hybridized with probe 6 (Figure III-1) 

showed no detectable hybridization bands, as expected for the negative control. 

PV-GMHT4355 previously digested with Aat II/Nde I and mixed with conventional 

control DNA predigested with Hpa I/Kpn I (Figure IV-6, lanes 10 and 11) produced one 

expected size band at ~7.4 kb (refer to Figure III-1, and Table IV-1). These results 

indicate that the probe is hybridizing to its target sequence. 

MON 87708 DNA digested with the restriction enzyme combination Bsp1286 I/Pvu II 

(Figure IV-7, lanes 2 and 6) or Hpa I/Kpn I (Figure IV-7, lanes 4 and 8) and hybridized 

with probe 6 produced no detectable hybridization bands. These results indicated that 

MON 87708 contains no detectable T-DNA II elements covered by probe 6. 
Monsanto Company 10-SY-210U 60 of 721 

Figure IV-5. Southern Blot Analysis to Detect the Presence or Absence of T-DNA II 

Sequences in MON 87708: Probe 4 

The blot was hybridized with a 32P labeled T-DNA II probe that spans a portion of the T-DNA II 

sequence (Probe 4, Figure III-1). Each lane contains approximately 10 μg of digested genomic 

DNA isolated from leaf tissue. Lane designations are as follows: 

Lane Description 

 1. Conventional control (Bsp1286 I/Pvu II) 

2. MON 87708 (Bsp1286 I/Pvu II) 

3. Conventional control (Hpa I/Kpn I) 

4. MON 87708 (Hpa I/Kpn I) 

5. Conventional control (Bsp1286 I/Pvu II) 

6. MON 87708 (Bsp1286 I/Pvu II) 

7. Conventional control (Hpa I/Kpn I) 

8. MON 87708 (Hpa I/Kpn I) 

9. Blank 

10. Conventional control (Hpa I/Kpn I) spiked with PV-GMHT4355 (Aat II/Nde I) 

 (~1 genome equivalent) 

11. Conventional control (Hpa I/Kpn I) spiked with PV-GMHT4355 (Aat II/Nde I) 

 (~0.1 genome equivalent) 

Arrows denote sizes of DNA, in kilobase pairs, obtained from molecular weight markers on 

ethidium bromide stained gel. 

1.0

1.6

2.0

3.1

4.1

5.1

6.1

7.1

8.1

10

15 20

40

1.0

1.6

2.0

3.1

4.1

5.1

6.1

7.1 8.1 10

15

20 40

0.5

1 2 3 4 5 6 7 8 9 10 11

LONG RUN SHORT RUN
Monsanto Company 10-SY-210U 61 of 721 

Figure IV-6. Southern Blot Analysis to Detect the Presence or Absence of T-DNA II 

Sequences in MON 87708: Probe 5 

The blot was hybridized with a 32P labeled T-DNA II probe that spans the coding region of the 

T-DNA II sequence (Probe 5, Figure III-1). Each lane contains approximately 10 μg of digested 

genomic DNA isolated from leaf tissue. Lane designations are as follows: 

Lane Description 

 1. Conventional control (Bsp1286 I/Pvu II) 

2. MON 87708 (Bsp1286 I/Pvu II) 

3. Conventional control (Hpa I/Kpn I) 

4. MON 87708 (Hpa I/Kpn I) 

5. Conventional control (Bsp1286 I/Pvu II) 

6. MON 87708 (Bsp1286 I/Pvu II) 

7. Conventional control (Hpa I/Kpn I) 

8. MON 87708 (Hpa I/Kpn I) 

9. Blank 

10. Conventional control (Hpa I/Kpn I) spiked with PV-GMHT4355 (Aat II/Nde I) 

 (~1 genome equivalent) 

11. Conventional control (Hpa I/Kpn I) spiked with PV-GMHT4355 (Aat II/Nde I) 

 (~0.1 genome equivalent) 

Arrows denote sizes of DNA, in kilobase pairs, obtained from molecular weight markers on 

ethidium bromide stained gel. 

1.0

1.6

2.0

3.1

4.1

5.1

6.1

7.1

8.1

10

15

20

40

1.0

1.6

2.0

3.1

4.1

5.1 6.1 7.1 8.1 10

15 20 40

0.5

1 2 3 4 5 6 7 8 9 10 11

LONG RUN SHORT RUN
Monsanto Company 10-SY-210U 62 of 721 

Figure IV-7. Southern Blot Analysis to Detect the Presence or Absence of T-DNA II 

Sequences in MON 87708: Probe 6 

The blots were hybridized with a 32P labeled T-DNA II probe that spans a portion of the 

T-DNA II sequence (Probe 6, Figure III-1). Each lane contains approximately 10 μg of digested 

genomic DNA isolated from leaf tissue. Lane designations are as follows: 

Lane Description 

 1. Conventional control (Bsp1286 I/Pvu II) 

2. MON 87708 (Bsp1286 I/Pvu II) 

3. Conventional control (Hpa I/Kpn I) 

4. MON 87708 (Hpa I/Kpn I) 

5. Conventional control (Bsp1286 I/ Pvu II) 

6. MON 87708 (Bsp1286 I/Pvu II) 

7. Conventional control (Hpa I/Kpn I) 

8. MON 87708 (Hpa I/Kpn I) 

9. Blank 

10. Conventional control (Hpa I/Kpn I) spiked with PV-GMHT4355 (Aat II/Nde I) 

 (~1 genome equivalent) 

11. Conventional control (Hpa I/Kpn I) spiked with PV-GMHT4355 (Aat II/Nde I) 

(~0.1 genome equivalent) 

Arrows denote sizes of DNA, in kilobase pairs, obtained from molecular weight markers on 

ethidium bromide stained gel. 

1.0

1.6

2.0

3.1

4.1

5.1

6.1

7.1

8.1

10

15 20

40

1.0

1.6

2.0

3.1

4.1

5.1 6.1 7.1 8.1 10 15 20 40

0.5

1 2 3 4 5 6 7 8 9 10 11

LONG RUN SHORT RUN
Monsanto Company 10-SY-210U 63 of 721 

IV.C. Southern Blot Analysis to Determine the Presence or Absence of 

PV-GMHT4355 Backbone Sequences in MON 87708 

To determine the presence or absence of PV-GMHT4355 backbone sequences, 

MON 87708 and conventional control genomic DNA were digested with the restriction 

enzyme combination Bsp1286 I/Pvu II or Hpa I/Kpn I and Southern blots were 

hybridized with probes that span the plasmid vector backbone sequence (Figure III-1). 

As a positive control on the Southern blots, digested PV-GMHT4355 and probe 

templates generated from PV-GMHT4355 were used. Approximately 1 genome 

equivalent of PV-GMHT4355 digested with the restriction enzyme combination 

Aat II/Nde I was mixed with predigested conventional control DNA. As an additional 

positive control, approximately 0.1 and 1 genome equivalent of probe templates 

(Figure III-1, probes 1, 2, 3, and 7) generated from PV-GMHT4355 were mixed with 

predigested conventional control DNA. The blot was hybridized with probes 1, 2, 3, and 

7 (Figure III-1). If backbone sequences are present in MON 87708, then probing with 

backbone probes should result in hybridizing bands. The results of this analysis are 

shown in Figure IV-8. 

IV.C.1. Plasmid Vector Backbone Probes 1, 2, 3, and 7 

Conventional control DNA digested with the restriction enzyme combination 

Bsp1286 I/Pvu II (Figure IV-8, lanes 1 and 5) or Hpa I/Kpn I (Figure IV-8, lanes 3 and 7) 

and hybridized simultaneously with the probes 1, 2, 3, and 7 (Figure III-1) spanning the 

entire backbone sequence of PV-GMHT4355 showed no detectable hybridization bands, 

as expected for the negative control. PV-GMHT4355, previously digested with 

Aat II/Nde I and mixed with conventional control DNA predigested with Hpa I/Kpn I 

(Figure IV-8, lane 10), produced two expected size bands at ~4.0 kb and ~7.4 kb (refer to 

Figure III-1 and Table IV-1). In addition, there are two faint hybridization bands at 

~4.5 kb and ~11 kb (Figure IV-8, lane 10). The ~4.5 kb band was likely due to an artifact 

that occurred during the electrophoresis, and the ~11 kb band was likely due to 

undigested plasmid DNA or an artifact that occurred during the electrophoresis. Since 

these faint bands appeared only in the plasmid spike and the expected bands were 

observed, they have no negative impact on the conclusions made from this blot. Probe 

template spikes of probes 1, 2, 3, and 7 (Figure III-1) generated from PV-GMHT4355 

mixed with conventional control DNA predigested with Hpa I/Kpn I (Figure IV-8, 

lanes 11 and 12) produced the expected size bands at ~0.2 kb, ~0.9 kb, ~1.5 kb, and 

~1.8 kb, respectively. The 0.1 genome equivalent copy of the expected ~0.2 kb band was 

not observed on the exposure of the Southern blot that is reported in Figure IV-8, lane 12; 

however, the band was observed on the same blot with a longer exposure. These results 

indicate that the probes are hybridizing to their target sequences. 

MON 87708 DNA digested with the restriction enzyme combination Bsp1286 I/Pvu II 

(Figure IV-8, lanes 2 and 6) or Hpa I/Kpn I (Figure IV-8, lanes 4 and 8) and hybridized 

simultaneously with probes 1, 2, 3, and 7 produced no detectable bands. The data 

indicate MON 87708 contains no detectable backbone sequences from PV-GMHT4355. 
Monsanto Company 10-SY-210U 64 of 721 

Figure IV-8. Southern Blot Analysis to Determine the Presence or Absence of 

PV-GMHT4355 Backbone Sequences in MON 87708: Probes 1, 2, 3, and 7 

The blot was hybridized simultaneously with four 32P labeled backbone probes (Probes 1, 2, 3, 

and 7, Figure III-1). Each lane contains approximately 10 μg of digested genomic DNA isolated 

from leaf tissue. Lane designations are as follows: 

Lane Description 

 1. Conventional control (Bsp1286 I/Pvu II) 

2. MON 87708 (Bsp1286 I/Pvu II) 

3. Conventional control (Hpa I/Kpn I) 

4. MON 87708 (Hpa I/Kpn I) 

5. Conventional control (Bsp1286 I/Pvu II) 

6. MON 87708 (Bsp1286 I/Pvu II) 

7. Conventional control (Hpa I/Kpn I) 

8. MON 87708 (Hpa I/Kpn I) 

9. Blank 

10. Conventional control (Hpa I/Kpn I) spiked with PV-GMHT4355 (Aat II/Nde I) 

 (~1 genome equivalent) 

11. Conventional control (Hpa I/Kpn I) spiked with probe templates (~1 genome 

 equivalent) 

12. Conventional control (Hpa I/Kpn I) spiked with probe templates (~0.1 genome 

 equivalent) 

Arrows denote sizes of DNA, in kilobase pairs, obtained from molecular weight markers on 

ethidium bromide stained gel. 

23.1

9.4

6.6

4.4

2.3

2.0

1 2 3 4 5 6 7 8 9 10 11 12

23.1

9.4

6.6

4.4

2.3

0.6

2.0

LONG RUN SHORT RUN
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IV.D. Organization and Sequence of the Insert and Adjacent DNA in MON 87708 

The organization of the elements within the T-DNA I was confirmed by DNA sequence 

analyses. PCR primers were designed with the intent to amplify two overlapping regions 

of the DNA that span the entire length of T-DNA I (Figure B-1, Appendix B). The 

amplified DNA segments were subjected to DNA sequencing analyses. The T-DNA I in 

MON 87708 is 3003 bp and matches the sequence of plasmid vector PV-GMHT4355, as 

described in Tables III-1 and IV-2. 

IV.E. PCR and DNA Sequence Analyses to Examine the MON 87708 Insertion Site 

PCR and sequence analyses were performed on genomic DNA extracted from 

MON 87708 and conventional control to examine the insertion sites. The PCR was 

performed with one primer specific to the genomic DNA sequence flanking the 5′ end of 

T-DNA I paired with a second primer specific to the genomic DNA sequence flanking 

the 3′ end of T-DNA I (Figure B-2, Appendix B). A sequence comparison between the 

PCR product generated from the conventional control and the sequence generated from 

the 5′ and 3′ flanking sequences of T-DNA I in MON 87708 indicates there was an 899 

bp deletion and a 128 bp insertion just 5′ of T-DNA I, and a 35 bp insertion just 3′ of 

T-DNA I. These molecular rearrangements presumably resulted from double-stranded 

break repair mechanisms in the plant during the Agrobacterium-mediated transformation 

process (Salomon and Puchta, 1998). 

IV.F. Southern Blot Analysis to Examine Insert Stability in Multiple Generations of 

MON 87708 

In order to demonstrate the stability of the T-DNA I insert present in MON 87708 

through multiple generations, Southern blot analysis was performed using DNA obtained 

from five breeding generations of MON 87708. For reference, the breeding history of 

MON 87708 is presented in Figure IV-9. The specific generations tested are indicated in 

the legend of Figure IV-10. The R3 generation was used for the molecular 

characterization analyses shown in Figures IV-2 through IV-8. To analyze stability, four 

additional generations were evaluated by Southern blot analysis and compared to the 

fully characterized R3 generation. Genomic DNA, isolated from each of the selected 

generations of MON 87708 and the conventional control, was digested with the 

restriction enzyme combination Bsp1286 I/Pvu II (Figure IV-1) and hybridized with 

probe 9 (Figure III-1). Probe 9 will detect both border fragments generated by the 

Bsp1286 I/Pvu II digestion. Any instability associated with the T-DNA I insert would be 

detected as novel bands within the fingerprint on the Southern blot. The Southern blot 

has the same positive hybridization controls as described in Section IV.A. The results are 

shown in Figure IV-10. 
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IV.F.1. Probe 9 

Conventional control DNA digested with the restriction enzyme combination 

Bsp1286 I/Pvu II produced no hybridization signals (Figure IV-10, lane 1) as expected 

for the negative control. PV-GMHT4355, digested with the restriction enzyme 

combination Aat II/Nde I and mixed with conventional control DNA predigested with the 

restriction enzyme combination Bsp1286 I/Pvu II (Figure IV-10, lanes 8 and 9), produced 

the expected size band at ~7.4 kb (refer to Figure III-1 and Table IV-1). Additionally, 

there were two very faint hybridization bands in the ~1 genome equivalent plasmid 

vector PV-GMHT4355 spike at ~4.3 kb and ~6.5 kb observed in a longer exposure of the 

Southern blot (data not shown). These bands were likely due to an artifact that occurred 

during the electrophoresis. Since these faint bands appeared only in the plasmid vector 

spike and the expected ~7.4 kb band was observed, they do not have any negative impact 

on the conclusions from this Southern blot analysis. These results indicate that the probe 

is hybridizing to its target sequence. 

Digestion of MON 87708 genomic DNA from multiple generations with the restriction 

enzyme combination Bsp1286 I/Pvu II and hybridized with probe 9 (Figure III-1) 

produced two bands at ~1.5 kb and ~2.6 kb (Figure IV-10, lanes 2-6). The ~1.5 kb band 

is the expected size for the border segment containing the 3′ end of T-DNA I along with 

the adjacent genomic DNA flanking the 3′ end of T-DNA I (Figure IV-1). The ~2.6 kb 

band is the expected size for the border segment containing the 5′ end of T-DNA I along 

with the adjacent genomic DNA flanking the 5′ end of T-DNA I (Figure IV-1). The 

fingerprint of the Southern blot signals from multiple generations, R2, R4, R5, and R6

(Figure IV-10, lanes 2, 4, 5, and 6), of MON 87708 is consistent with the fully 

characterized generation R3 (Figure IV-3, lanes 2 and 6; Figure IV-10, lane 3). No 

unexpected bands were detected, indicating that MON 87708 contains one copy of 

T-DNA I that is stably maintained across multiple generations. 
Monsanto Company 10-SY-210U 67 of 721 

Figure IV-9. Breeding History of MON 87708 

The R3 generation was used for the molecular analyses reported in Figures IV-2 through IV-8 and 

is referred to as MON 87708 in all Southern blot figures. The R5 generation was used for 

development of all commercial products. MON 87708 from generations R2, R3, R4, R5, and R6

(bolded in the breeding tree) were used for analyzing the stability of T-DNA I in MON 87708 

across generations (Figure IV-10). 

 
Monsanto Company 10-SY-210U 68 of 721 

Figure IV-10. Southern Blot Analysis to Examine Insert Stability in Multiple 

Generations of MON 87708: Probe 9 

The blot was hybridized with a 32P labeled T-DNA I probe that spans the coding region of the 

T-DNA I (Probe 9, Figure III-1). Each lane contains approximately 10 μg of digested genomic 

DNA isolated from leaf tissue. Lane designations are as follows: 

Lane Description 

 1. Conventional control (Bsp1286 I/Pvu II) 

2. R2 generation of MON 87708 (Bsp1286 I/Pvu II) 

3. R3 generation of MON 87708 (Bsp1286 I/Pvu II) 

4. R4 generation of MON 87708 (Bsp1286 I/Pvu II) 

5. R5 generation of MON 87708 (Bsp1286 I/Pvu II) 

6. R6 generation of MON 87708 (Bsp1286 I/Pvu II) 

7. Blank 

8. Conventional control (Bsp1286 I and Pvu II) spiked with PV-GMHT4355 

 (Aat II/Nde I) (~1 genome equivalent) 

9. Conventional control (Bsp1286 I and Pvu II) spiked with PV-GMHT4355 

 (Aat II/Nde I) (~0.1 genome equivalent) 

Arrows denote sizes of DNA, in kilobase pairs, obtained from molecular weight markers on 

ethidium bromide stained gel. 

1 2 3 4 5 6 7 8 9 

1.0

1.6

2.0

3.1

4.1

5.1

6.1 7.1 8.1

10

15 20

40

0.5
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IV.G. Inheritance of the Genetic Insert in MON 87708 

During development of MON 87708, segregation data were generated to assess the 

heritability and stability of the T-DNA I present in MON 87708. Chi-square analysis was 

performed over several generations to confirm the segregation and stability of T-DNA I 

in MON 87708. The Chi-square analysis is based on testing the observed segregation 

ratio to the expected segregation ratio according to Mendelian principles. 

The MON 87708 breeding path, from which segregation data were generated, is 

described in Figure IV-11. The transformed R0 plant was self-pollinated to produce R1

seed. An individual plant (#2, designated as MON 87708), that was homozygous for a 

single copy of the dmo expression cassette, was identified from the R1 segregating 

population via Invader and Southern blot analysis. Invader is a non-PCR based assay 

that can be used to accurately quantify transgene copy number in plant genomes (Gupta 

et al., 2008). 

The selected R1 MON 87708 plant was self-pollinated to give rise to a population of R2

plants that were repeatedly self-pollinated through the R4 generation. At each generation, 

the fixed homozygous plants were tested for the expected segregation pattern of 1:0 

(positive:negative) for the dmo expression cassette using the Invader analysis, Southern 

blot analysis, and/or PCR. 

At the R4 generation, homozygous MON 87708 plants were bred via traditional breeding 

with a soybean variety that did not contain the dmo expression cassette to produce F1

hemizygous seed. The resulting F1 plants were then self-pollinated to produce F2 seed. 

The F2 plants were tested for the presence of the dmo expression cassette by Invader 

analysis, and hemizygous F2 plants were selected and self-pollinated to produce F3 seed. 

This process was repeated through the F4 generation. The heritability and stability of the 

dmo expression cassette in MON 87708 was assessed in the F2, F3, and F4 generations. A 

total of 2413 out of 3223 plants were positive for the presence of the dmo expression 

cassette in the F2 generation; however, the zygosity of 200 of those 2413 plants could not 

be determined from the assay. Exclusion of these dmo-positive plants from the analysis 

likely would have skewed the distribution of homozygous positive: 

hemizygous positive:homozygous negative plants. Therefore, the segregation assessment 

in the F2 generation was based on the presence or absence of the dmo expression cassette 

which was expected to segregate at a 3:1 (positive:negative) ratio according to Mendelian 

inheritance principles. Subsequently, assessment of segregation in the F3 and F4

generations was based on zygosity, and the dmo expression cassette was predicted to 

segregate at a 1:2:1 (homozygous positive:hemizygous positive :homozygous negative) 

ratio according to Mendelian inheritance principles. 

A Chi-square (χ

2

) analysis was used to compare the observed segregation ratios to the 

expected ratios according to Mendelian inheritance principles. The χ

2

 was calculated as: 

χ

2

 = ∑ [( | o - e | )2 / e] 
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where o = observed frequency of the phenotype and e = expected frequency of the 

phenotype. The level of statistical significance was predetermined to be 5%. 

The results of the χ

2

 analysis of the segregating progeny of MON 87708 are presented in 

Table IV-3. The χ

2 value for the F2, F3, and F4 generations indicated no significant 

difference between the observed and expected segregation ratios. These results support 

the conclusion that the dmo expression cassette in MON 87708 resides at a single locus 

within the soybean genome and is inherited according to expected Mendelian inheritance 

principles. These results are also consistent with the molecular characterization data that 

indicate MON 87708 contains a single, intact copy of the dmo expression cassette that 

was inserted into the soybean genome at a single locus. 
Monsanto Company 10-SY-210U 

Transformed and regenerated R0 plant 

 R1

 R2 (homozygous positive) 

 R3 (homozygous positive)  R4 (homozygous positive) F1 (hemizygous positive) 

 Breeding path continued F2 (expected segregation of 3:1)

 positive : negative  F3 (expected segregation of 1:2:1)

 homozygous positive : hemizygous positive : homozygous nega F4 (expected segregation of 1:2:1)

 homozygous positive : hemizygous positive : homozygSelf pollinated Figure IV-11. Breeding Path for Generating Segregation Data for MON 87708  Chi-square analysis conducted on segregation data from the F2, F3, and F4 generations. 

Note: Hemizygous positive plants in the F1, F2, F3, and F4 generations were selected and self-pollinated to produce seed of the subsequ



Plant #2 (MON 87708) selected and self pollinated 





cross with variety that did not contain the dmo expression 

cassette 




Monsanto Company 10-SY-210U 

Table IV-3. Segregation of the dmo Expression Cassette During the Development of MON 87708 

Generation1

Total 

Plants 

Tested2

Observed 

# Plants 

Positive 

Observed 

# Plants 

Negative 

3:1 Segregation3 

Expected 

# Plants 

Positive 

Expected 

# Plants 

Negative 

χ 2 Probability 

F2 3223 2413 810 2417.25 805.75 0.03 0.863 

Generation1

Total 

Plants 

Tested2

Observed 

# Plants 

Homozygous 

Positive

Observed 

# Plants 

Hemizygous 

Positive

Observed 

# Plants 

Homozygous 

Negative

1:2:1 Segregation 

Expected 

# Plants 

Homozygous 

Positive 

Expected 

# Plants 

Hemizygous 

Positive 

Expecte

# Plants 

Homozygo

Negativ

F3 118 29 52 37 29.5 59 29.5 F4 343 83 171 89 85.75 171.5 85.75 

1

F2, F3, and F4 progeny were from self-pollinated F1, F2, and F3 plants hemizygous positive for the dmo expression cassette, 

2Plants were tested for the presence of the dmo expression cassette by Invader analysis. 3Assessment of segregation in the F2 generation was based on the presence or absence of the dmo expression cassette 

number of dmo-positive plants for which zygosity could not be determined from the assay. 
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IV.H. Genetic Modification Characterization Conclusion 

Molecular characterization of MON 87708 by Southern blot analyses demonstrated that a 

single copy of the T-DNA I sequences from the plasmid vector PV-GMHT4355 was 

integrated into the soybean genome at a single locus. There were no additional genetic 

elements from the T-DNA II or backbone sequences of the plasmid vector 

PV-GMHT4355 detected, linked or unlinked to the intact T-DNA I present in 

MON 87708. 

The PCR and DNA sequence analyses performed on MON 87708, which confirmed the 

organization of the elements within T-DNA I, demonstrated the 5′ and 3′ insert-to-plant 

junctions and determined the complete DNA sequence of T-DNA I and adjacent DNA 

sequence flanking the insert in MON 87708. Analysis of the T-DNA I insertion site 

indicates that there was an 899 bp deletion of genomic DNA at the insert-to-plant DNA 

junction. Additionally, a 128 bp insertion was identified in the 5′ adjacent flanking 

sequence of MON 87708 and a 35 bp insertion was identified in the 3′ adjacent flanking 

sequence of MON 87708. 

Generational stability analysis by Southern blot demonstrated that MON 87708 has been 

maintained through five breeding generations, thereby confirming the stability of 

T-DNA I in MON 87708. Results from segregation analyses show heritability and 

stability of the insert occurred as expected across multiple generations, which 

corroborates the molecular insert stability analysis and establishes the genetic behavior of 

the T-DNA I in MON 87708 at a single chromosomal locus. 
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V. CHARACTERIZATION AND SAFETY ASSESSMENT OF THE 

MON 87708 DMO 

Characterization of the introduced protein in a biotechnology-derived crop product is 

important to establishing its food, feed, and environmental safety. As described in 

Section IV, MON 87708 contains a dmo expression cassette that upon translation results 

in two forms of the DMO protein; referred to as DMO and DMO+27 (Section V.A). The 

active form of these proteins, necessary to confer dicamba tolerance, is a trimer 

comprised of three DMO monomers (Chakraborty et al., 2005). In MON 87708, the 

trimer can be comprised of DMO, DMO+27, or a combination of both. Therefore, this 

document will refer to both forms of the protein and all forms of the trimer as 

MON 87708 DMO. 

This section summarizes: 1) the functionality of DMO; 2) the characterization of 

MON 87708 DMO; 3) the levels of MON 87708 DMO in plant tissues; 4) assessment of 

the potential allergenicity of MON 87708 DMO and 5) the food, feed, and environmental 

safety assessment of MON 87708 DMO. The data support a conclusion that MON 87708 

is safe for the environment and human or animal consumption based on several lines of 

evidence, all of which are summarized below. 

V.A. Function of DMO and MON 87708 DMO 

DMO was initially purified from Stenotrophomonas maltophilia (S. maltophilia)

strain DI-6, isolated from soil at a dicamba manufacturing plant (Krueger et al., 1989). 

DMO is an enzyme that catalyzes the demethylation of dicamba to the non-herbicidal 

compound DCSA and formaldehyde (Chakraborty et al., 2005). DMO is a Rieske-type 

non-heme iron oxygenase, that is part of a three component system comprised of a 

reductase, a ferredoxin, and a terminal oxygenase, in this case the DMO. These three 

enzymes work together in a redox system similar to many other oxygenases to transport 

electrons from nicotinamide adenine dinucleotide (NADH) to oxygen and catalyze the 

demethylation (Behrens et al., 2007) as presented in Figure V-1. 

Figure V-1. Three Components of the DMO Oxygenase System 

 

Reductase

(oxidized)

Flavin

Reductase

(reduced)

Ferredoxin

(reduced)

[2Fe-2S]

Ferredoxin

(oxidized)

NADH

+

H+

NAD+

Oxygenase

(oxidized)

[2Fe-2S]

Fe+2

Oxygenase

(reduced)

O2 O OH

OCH3

Cl

Cl

O OH

OH

Cl

Cl

3,6 DCSA

Dicamba

+ O

H H

Dicamba

Mono-Oxygenase

Dicamba

Mono-Oxygenase

Formaldehyde
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The crystal structure of a DMO has been solved (D'Ordine et al., 2009; Dumitru et al., 

2009) and shows that the DMO monomers contain a Rieske [2Fe-2S] cluster domain and 

a non-heme iron center domain typical of all Rieske-type mono-oxygenases (Ferraro et 

al., 2005). To catalyze the demethylation of dicamba, electrons transferred from NADH 

are shuttled through an endogenous reductase and ferredoxin to the terminal DMO. The 

electrons are received by the Rieske [2Fe-2S] cluster on one DMO monomer and 

transferred to the non-heme iron center at the catalytic site of an adjacent monomer 

(D'Ordine et al., 2009; Dumitru et al., 2009), where it reductively activates oxygen to 

catalyze the final demethylation of dicamba. As a result of the reaction, 3.6 

dichlorosalicylic acid (DCSA) and formaldehyde are formed. DCSA is a known soybean, 

soil, and livestock metabolite whose safety has been evaluated by the EPA. 

Formaldehyde is found naturally in many plants at levels up to several hundred ppm 

(Adrian-Romero et al, 1999). An assessment of the safety and potential effects of the 

DMO reaction products is provided in Section IX.B.3.6. 

V.A.1. Formation of MON 87708 DMO 

DMO is targeted to chloroplasts for co-localization with the endogenous reductase and 

ferredoxin enzymes that supply electrons for the DMO demethylation reaction as 

described by Behrens et al. (2007). The MON 87708 DMO precursor protein contains 84 

additional amino acids corresponding to a 57 amino acid Chloroplast Transit Peptide 

(CTP) from pea and 24 amino acids from the N-terminal coding region of the pea 

Rubisco small (RbcS) subunit to target the protein to the chloroplast (Comai et al., 1988), 

and three amino acids from an intervening sequence used for cloning purposes (Table IIII). It was anticipated that during translocation of the DMO precursor into chloroplasts 

the introduced 84 amino acids would be fully cleaved resulting in the predicted Nterminus of the DMO protein. However, analysis of mature seed extracts by western blot 

demonstrated the presence of two immunoreactive bands (Figure C-2, Appendix C). 

Analysis of these two bands determined that the lower molecular weight band 

corresponded to the full-length DMO protein (referred to as DMO). DMO has an 

apparent molecular weight of 39.8 kDa and is a single polypeptide chain of 339 amino 

acids. The higher molecular weight band of approximately 42 kDa corresponded to the 

full-length DMO protein plus 27 amino acids originating from the pea Rubisco small 

subunit and intervening sequence on its N-terminus (referred to as DMO+27; 367 amino 

acid polypeptide). Both forms of the DMO protein were characterized (Appendix C). 

As described previously the active form of DMO is a trimer (Chakraborty et al., 2005; 

Dumitru et al., 2009). For MON 87708 DMO to be functionally active and confer 

dicamba tolerance to MON 87708, a trimeric structure is required. This trimer contains 

either form of DMO or a mixture of both, and its activity was confirmed during 

characterization (Section V.B. and Appendix C). 

V.A.2. Specificity of MON 87708 DMO 

DMO has high specificity for its substrate dicamba (D'Ordine et al., 2009; Dumitru et al., 

2009). The specificity of DMO for dicamba is likely due to the specific interactions that 

occur in the catalytic site between the dicamba substrate and DMO. Dicamba interacts 

with amino acids in the active site of DMO through both the carboxylate moiety and the 
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chlorine atoms of dicamba, which are primarily involved in orienting the substrate in the 

catalytic pocket. These chlorine atoms are required for catalysis (D'Ordine et al., 2009; 

Dumitru et al., 2009). 

The possibility that MON 87708 DMO can metabolize plant endogenous substrates was 

tested in in vitro experiments using an Escherichia coli (E. coli)-produced DMO. The E. 

coli-produced DMO is similar in sequence and function to MON 87708 DMO, therefore 

it is appropriate to extend specificity data generated with the E. coli-produced DMO to 

MON 87708 DMO (Appendix C.2.). A set of potential substrates was selected based on 

structural similarity to dicamba and abundance in soybean (Janas et al., 2000), including 

o-anisic acid (2-methoxybenzoic acid), vanillic acid (4-hydroxy-3-methoxybenzoic acid), 

syringic acid (3,5-dimethoxy-4-hydroxybenzoic acid), ferulic acid [3-(4-hydroxy-3-

methoxy-phenyl)prop-2-enoic acid] and sinapic acid [3-(4-hydroxy-3,5-

dimethoxyphenyl)prop-2-enoic acid] (Figure V-2). The disappearance of potential 

substrates and the formation of potential oxidation products were monitored using LCUV and LC-MS (Appendix C). None of the tested substrates was metabolized by the 

E.coli-produced DMO in vitro. Therefore, DMO, though structurally similar to other 

Rieske mono-oxygenases, is specific for dicamba (see Section V.E.3 for additional 

details).

Figure V-2. Dicamba and Set of Potential Endogenous Substrates Tested in in vitro

Experiments with DMO 

The arrow indicates methyl group removed by DMO. 

The possibility that MON 87708 DMO can metabolize exogenous substrates was tested 

in in vivo experiments. In addition to dicamba, a total of 19 herbicides representing eight 

families with distinct modes-of-action, some of which are approved for use in soybean, 

were tested with MON 87708 and the near isogenic conventional soybean control A3525 

(Table V-1). Soybean naturally has varying levels of tolerance to different herbicides. 

For example, soybean is tolerant to in-crop postemergence applications of alachlor, but 

not atrazine. Each herbicide was applied at two spray rates, representative of potential 

commercial rates needed to control broadleaf weeds, at the V2-V3 soybean growth stage 

and then scored a visual rating based on the amount of injury observed on the plants. 

Across nearly all of the herbicides tested, MON 87708 and the conventional control were 

similar in their level of tolerance, indicating that these herbicides do not serve as a 

substrate for MON 87708 DMO. However, MON 87708 did show slightly more 

tolerance compared to the conventional control when treated with the three 

phenoxycarboxylic acid (phenoxy) synthetic auxin herbicides: 2,4-D 

(2,4-dichlorophenoxy acetic acid), MCPA (2-methyl-4-chlorophenoxy acetic acid) and 
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2,4-DB (2,4-dichlorophenoxy butanoic acid). Chloramben and TBA are no longer 

available in the U.S and were excluded from this testing. See Appendix C, Section C.2. 

As 2,4-D is the most structurally similar to dicamba of the three phenoxy auxin 

herbicides (2,4-D, 2,4-DB, and MCPA), it was selected as a representative for further in 

vitro experimentation. Subsequent experiments were performed to evaluate whether 

2,4-D can be metabolized by E. coli-produced DMO. The presumptive product of the 

oxidative reaction between 2,4-D and DMO is 2,4-dichlorophenol (2,4-DCP), formed 

from the dealkylation of 2,4-D. The potential disappearance of 2,4-D and formation of 

2,4-DCP were monitored using LC-UV and LC-MS (Appendix C, Section C.2). Neither 

the formation of 2,4-DCP nor any measurable decrease in 2,4-D were detected in the in 

vitro experiment. These results indicate that 2,4-D cannot be metabolized by E. coliproduced DMO, and demonstrate that DMO is specific to dicamba. The tolerance of 

MON 87708 to other herbicides at anticipated commercial application rates is no 

different than the conventional soybean, except for the phenoxy auxin herbicides where 

MON 87708 showed limited, but not commercially acceptable, tolerance when treated. 

Table V-1. Herbicides Applied to MON 87708 and Conventional Control 

Herbicide Active Ingredient Herbicide Chemical Family (MOA)1

Dicamba Benzoic acid (Synthetic Auxin) 

2,4-D Phenoxycarboxylic acid (Synthetic Auxin) 

2,4-DB Phenoxycarboxylic acid (Synthetic Auxin) 

MCPA Phenoxycarboxylic acid (Synthetic Auxin) 

Triclopyr Pyridinecarboxylic acid (Synthetic Auxin) 

Clopyralid Pyridinecarboxylic acid (Synthetic Auxin) 

Picloram Pyridinecarboxylic acid (Synthetic Auxin) 

Alachlor Chloroacetamide (Inhibition of VLCFAs) 

Acetochlor Chloroacetamide (Inhibition of VLCFAs) 

Atrazine Triazine (Inhibition of Photosynthesis at Photosytem II) 

Linuron Ureas (Inhibition of Photosynthesis at Photosytem II) 

Oxyfluorofen Diphenylether(Inhibition of PPO) 

Lactofen Diphenylether (Inhibition of PPO) 

Chlorimuron Sulfonylurea (Inhibition of ALS) 

Chlorsulfuron Sulfonylurea (Inhibition of ALS) 

Halosulfuron Sulfonylurea (Inhibition of ALS) 

Imazapyr Imidazolinone (Inhibition of ALS) 

Trifluralin Dinitroaniline (Microtubule Assembly Inhibition) 

Paraquat Bipyridilium (Photosystem I electron diversion) 

Glyphosate Glycine (Inhibition of EPSP synthase) 

1

HRAC (2009) 

V.B. Characterization of MON 87708 DMO 

The safety assessment of crops derived through biotechnology includes characterization 

of the functional and physicochemical properties, and confirmation of the safety of the 

introduced protein. As stated previously, both forms of the protein and all forms of the 
Monsanto Company 10-SY-210U 78 of 721 

trimer are referred to as the MON 87708 DMO. MON 87708 DMO was purified in 

sufficient quantities directly from the seed of MON 87708 and used in subsequent safety 

assessment studies. Typically protein safety studies are conducted on proteins produced 

in heterologous expression systems, such as E. coli. Since the MON 87708 DMO used in 

the subsequent safety studies was purified directly from MON 87708 DMO, equivalence 

evaluations between plant-produced and bacterial-produced MON 87708 DMO was not 

necessary. The physicochemical characteristics and functional activity of the 

MON 87708 DMO were determined by a panel of analytical techniques, including: 1) 

western blot analysis to establish identity and immunoreactivity of MON 87708 DMO 

using an anti-DMO antibody, 2) N-terminal sequence analysis, 3) matrix-assisted laser 

desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) to generate a 

tryptic peptide map of the MON 87708 DMO, 4) sodium dodecyl sulfate polyacrylamide 

gel electrophoresis (SDS-PAGE) to establish the apparent molecular weight of 

MON 87708 DMO, 5) glycosylation status of MON 87708 DMO, and 6) MON 87708 

DMO activity analysis to demonstrate functional activity. The details of the materials, 

methods, and results are described in Appendix C, while the conclusions of the 

MON 87708 DMO characterization are summarized below. 

The identities of both forms of the DMO protein produced in MON 87708 that constitute 

MON 87708 DMO were confirmed by western blot analysis by probing with an antiDMO antibody, N-terminal sequencing, and MALDI-TOF MS analysis of peptides 

produced after trypsin digestion. The antibody specifically detected DMO and DMO+27 

on a western blot. The N-terminal sequence of the first 15 amino acid residues of both 

DMO and DMO+27 was identical to the predicted amino acid sequence, with the 

exception of the N-terminal methionine residue. MALDI-TOF MS analyses of DMO and 

DMO+27 yielded peptide masses consistent with their expected sequence. The apparent 

molecular weights of DMO and DMO+27 were 39.8 and 42.0 kDa, respectively and 

neither were glycosylated. The MON 87708 DMO activity was determined by measuring 

the production of DCSA using dicamba as the substrate, resulting in a specific activity of 

62.21 nmoles DCSA/min/mg of MON 87708 DMO. Taken together, these data provide a 

detailed characterization of the MON 87708 DMO isolated from the seed of MON 87708. 

V.C. Expression Levels of MON 87708 DMO 

The levels of MON 87708 DMO in various tissues of MON 87708 that are relevant to the 

risk assessment were determined by a validated ELISA. Tissues of MON 87708 and the 

near isogenic conventional soybean control A3525 were collected during the 2008 

growing season from five field sites in the U.S.: Jefferson County, Iowa; Stark County, 

Illinois; Clinton County, Illinois; Parke County, Indiana; and Berks County, Pennsylvania. 

These field sites were representative of soybean producing regions suitable for 

commercial production. At each site, three replicated plots containing MON 87708, as 

well as the conventional control, were planted using a randomized complete block field 

design. Over-season leaf (OSL 1-4), root, forage, and seed tissues were collected from 

each replicated plot at all field sites (except for the conventional control from Berks 

County, Pennsylvania where only two replicates were collected). A description of tissues 

collected is provided in Table V-2. 
Monsanto Company 10-SY-210U 79 of 721 

Table V-2. Tissues Collected and Analyzed for MON 87708 DMO 

Tissue Soybean Development Stage1

 Days After Planting 

OSL-1 V3-V4 21-30 

OSL-2 V5-V8 31-42 

OSL-3 R2-V12 43-58 

OSL-4 R5-V16 55-78 

Root R6 70-91 

Forage R6 70-91 

Seed R8 109-147 

1

Soybean plant growth stages described in Soybean Growth and Development (ISU, 

2004). 

The levels of MON 87708 DMO were determined in all seven tissue types as described in 

Table V-3. The ELISA assay detected all forms of MON 87708 DMO and therefore the 

levels represent the total of MON 87708 DMO. The results obtained from the ELISA 

analysis are summarized in Table V-3 and the details of the materials and methods are 

described in Appendix D. In summary, expression analysis of the samples from the 2008 

U.S. field trial showed that MON 87708 DMO was detected in all tissue types across all 

five sites ranging from 3.9 – 180 μg/g dry weight (dwt). The mean levels of the 

MON 87708 DMO across the five sites were highest in leaf (ranging from OSL-1 at 17 

μg/g dwt, to OSL-4 at 69 μg/g dwt), followed by forage (53 μg/g dwt), seed 

(47 μg/g dwt), and root (6.1 μg/g dwt). As expected for the conventional control, the 

ELISA values for MON 87708 DMO were less than the limit of quantitation (LOQ) of 

the assay in all tissue types. 

 
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Table V-3. Summary of the Levels of MON 87708 DMO in Leaf, Root, Forage, and 

Seed from MON 87708 Grown in 2008 U.S. Field Trials 

Tissue 

Type

MON 87708 

DMO1

Mean (SD)2

(μg/g fwt) 3

Range4

(μg/g fwt)

MON 87708 

DMO 

Mean (SD) 

(μg/g dwt)5

Range

(μg/g dwt)

LOQ/LOD

(μg/g fwt)6,7

 

OSL-1 3.1 (1.9) 0.87 – 6.8 17 (7.7) 6.2 – 29 0.63/0.20 

OSL-2 5.2 (2.6) 1.4 – 9.8 31 (13) 12 – 54 0.63/0.20 

 

OSL-3 6.0 (2.2) 3.5 – 11 44 (14) 25 – 71 0.63/0.20 

 

OSL-4 16 (12) 4.6 – 43 69 (46) 23 – 180 0.63/0.20 

 

Root 1.9 (0.73) 1.2 – 3.6 6.1 (2.1) 3.9 – 11 0.031/0.015 

 

Forage 12 (2.5) 7.0 – 17 53 (18) 25 – 84 0.63/0.10 

Seed 43 (7.7) 31 – 55 47 (8.7) 34 – 59 1.3/0.21 

 1

Represents total for MON 87708 DMO. 2

The mean and standard deviation (SD) were calculated (n=15). The “n” values for the calculated 

mean and standard deviations represent the number of samples figured into the calculation. 

3

Protein levels are expressed as microgram (μg) of protein per gram (g) of tissue on a fresh 

weight (fwt) basis. 

4

Minimum and maximum values were determined for each tissue type. 

5

Protein levels are expressed as μg/g dwt. The dry weight values were calculated by dividing the 

μg/g fwt by the dry weight conversion factors obtained from moisture analysis data. 

6

The limit of quantitation (LOQ) was calculated based on the lowest DMO standard 

concentration. The “ng/ml” value was converted to “μg/g fwt” using the respective dilution 

factor and tissue-to-buffer ratio. 

7

The limit of detection (LOD) was calculated as the mean value plus three SD using the data 

generated with conventional control sample extracts for each tissue type. The LOD value in 

“ng/ml” was converted to “μg/g fwt” using the respective dilution factor and tissue-to-buffer 

ratio. 
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V.D. Assessment of Potential Allergenicity of MON 87708 DMO 

The allergenic potential of an introduced protein is assessed by comparing the 

biochemical characteristics of the introduced protein to biochemical characteristics of 

known allergens (Codex Alimentarius, 2003). A protein is not likely to be associated 

with allergenicity if: 1) the protein is from a non-allergenic source, 2) the protein 

represents a very small portion of the total plant protein, 3) the protein does not share 

structural similarities to known allergens based on the amino acid sequence, and 4) the 

protein is rapidly digested in mammalian gastrointestinal systems. MON 87708 DMO, as 

defined above, refers to all forms of the protein and the resulting trimer, has been 

assessed for its potential allergenicity according to these safety assessment guidelines. 

1) MON 87708 DMO originates from S. maltophilia, an organism that has not 

been reported to be a source of known allergens. 

2) MON 87708 DMO represents no more than 0.01% of the total protein in the 

seed of MON 87708. 

3) Bioinformatics analyses demonstrated that the DMO+27 form of MON 87708 

DMO, that also contains the DMO sequence, does not share amino acid sequence 

similarities with known allergens and, therefore, is highly unlikely for DMO or 

DMO+27 to contain immunologically cross-reactive allergenic epitopes. 

4) In vitro digestive fate experiments conducted with the MON 87708 DMO 

demonstrate that the proteins are rapidly digested in simulated gastric fluid (SGF) 

and in simulated intestinal fluid (SIF). 

Taken together, these data support the conclusion that MON 87708 DMO does not pose a 

significant allergenic risk to humans or animals. 

V.E. Safety Assessment Summary of MON 87708 DMO 

Numerous factors have been considered in the safety assessment of MON 87708 DMO. 

A comprehensive food, feed, and environmental safety assessment of the MON 87708 

DMO was conducted. The results are summarized below along with the conclusions 

reached from the assessment. 

V.E.1. The Donor Organism is Safe 

The dmo gene is derived from the bacterium Stenotrophomonas maltophilia (Palleroni 

and Bradbury, 1993). S. maltophilia is an aerobic, ubiquitous environmental gram 

negative bacterium commonly present in aquatic environments, soil, and plants. 

S. maltophilia is ubiquitously associated with plants and has been isolated from the 

rhizosphere of wheat, maize, grasses, beet, cucumber, chicory, potato, strawberry, 

sugarcane, and rapeseed (Berg et al., 1996; Berg et al., 1999; Berg et al., 2002; Denton et 

al., 1998; Echemendia, 2007; Juhnke and des Jardin, 1989; Juhnke et al., 1987; Lambert 

et al., 1987). S. maltophilia was isolated from cotton seed, bean pods, and coffee (Nunes 

and de Melo, 2006; Swings et al., 1983), thus, S. maltophilia can be found in a variety of 
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foods and feeds. It is also widespread in the home environment and can be found around 

dishwashers, sponges, toothbrushes, flowers, plants, fruits, vegetables, frozen fish, milk, 

and poultry (Ryan et al., 2009). Strains of S. maltophilia have been found in the transient 

flora of hospitalized patients as a commensal organism (Echemendia, 2007). Infections 

caused by S. maltophilia are extremely uncommon (Cunha, 2006) and S. maltophilia can 

be found in healthy individuals without causing any harm to human health (Denton et al., 

1998). Similar to the indigenous bacteria of the gastrointestinal tract, S. maltophilia can 

be an opportunistic pathogen (Berg, 1996). As such, S. maltophilia is of low virulence in 

immuno-compromised patients where a series of factors must occur for colonization by S. 

maltophilia on humans (Ryan et al., 2009). The ubiquitous presence of S. maltophilia in 

the environment, the presence in healthy individuals, and the incidental presence on foods 

without any adverse safety reports establishes the safety of the donor organism. 

V.E.2. MON 87708 DMO Belongs to a Common Class of Mono-Oxygenases 

MON 87708 DMO is classified as an oxygenase. Oxygenases are enzymes that 

incorporate one or two oxygen atoms into substrates and are widely distributed in many 

universal metabolic pathways (Harayama et al., 1992). Within this large enzymatic class 

are mono-oxygenases that incorporate a single oxygen atom as a hydroxyl group with the 

concomitant production of water and oxidation of NAD(P)H (Harayama et al., 1992). 

Non-heme iron oxygenases, where iron is involved in the catalytic site, are an important 

class of oxygenases. Within this class are Rieske oxygenases, which contain a Rieske 

iron-sulfur [2Fe-2S] cluster. All Rieske non-heme iron oxygenases contain two catalytic 

domains, a non-heme iron domain (nh-Fe) that is a site of oxygen activation, and a 

Rieske [2Fe-2S] domain (Ferraro et al., 2005). MON 87708 DMO belongs to this class 

of oxygenases which are found in diverse phyla ranging from bacteria to plants (Ferraro 

et al., 2005; Schmidt and Shaw, 2001). 

As discussed previously, the crystal structure of a DMO has been solved (D'Ordine et al., 

2009; Dumitru et al., 2009). The crystallography results demonstrated that, similar to all 

Rieske non-heme iron oxygenases, DMO contains two catalytically important and highly 

conserved domains; a mononuclear non-heme iron domain (nh-Fe) that is a site of 

oxygen activation, and a Rieske [2Fe-2S] domain (D'Ordine et al., 2009; Dumitru et al., 

2009; Ferraro et al., 2005). The amino acids binding the non-heme iron and those that 

constitute the Rieske [2Fe-2S] domain in the DMO protein are also highly conserved in 

these plant proteins, as is their spatial orientation (D'Ordine et al., 2009; Ferraro et al., 

2005). Rieske domains are ubiquitous in numerous bacterial and plant proteins like the 

iron-sulfur protein of the cytochrome bc1 complex, chloroplast cytochrome b6/f complex, 

and choline mono-oxygenases (Breyton, 2000; Darrouzet et al., 2004; Gray et al., 2004; 

Hibino et al., 2002; Rathinasabapathi et al., 1997; Russell et al., 1998). The presence of 

two conserved domains, a Rieske [2Fe-2S] domain and a mononuclear iron domain, 

suggests that all Rieske type non-heme iron oxygenases share the same reaction 

mechanism, by which the Rieske domain transfers electrons from the ferredoxin to the 

mononuclear iron to allow catalysis (Chakraborty et al., 2005; Dumitru et al., 2009; 

Ferraro et al., 2005). The structure and mechanistic homologies are further evidence of 

the evolutionary relatedness of all Rieske non-heme iron oxygenases to each other (Nam 

et al., 2001; Rosche et al., 1997; Werlen et al., 1996). Additionally, a FASTA alignment 
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search of publicly available databases using the DMO+27 sequence as a query yielded 

homologous sequences from many different species, predominantly bacteria, with amino 

acid sequence identity ranging up to approximately 42%. Homologous oxygenases are 

also present in plants, including such crops as rice (Orysa sativa), canola (Brassica 

napus), and corn (Zea mays), with sequence identity up to 24%. The highest homology 

was observed to pheophorbide A oxygenases from corn, canola and pea (Pisum sativum). 

Pheophorbide A oxygenase is also a Rieske-type oxygenase that plays a key role in the 

overall regulation of chlorophyll degradation in plants (Rodoni et al., 1997). The protein 

is constitutively present in all green tissues and, at slightly lower levels, in etiolated and 

non-photosynthetic tissues including seeds (Yang et al., 2004). 

Therefore, MON 87708 DMO shares sequence identity and many catalytic and domain 

structural similarities with a wide variety of oxygenases present in bacteria and plants 

currently widely prevalent in the environment and consumed, establishing that animals 

and humans are extensively exposed to these types of enzymes. 

V.E.3. DMO is a Dicamba-Specific Mono-Oxygenase

DMO converts dicamba to DCSA. This demethylation is very specific to dicamba, where 

both the carboxylate moiety and the chlorine atoms help position the substrate at the 

active site of the enzyme (D'Ordine et al., 2009; Dumitru et al., 2009). Crystallography 

studies of the substrate in the active site demonstrated that these chlorines function as 

steric “handles” that position the substrate in the proper orientation in the binding pocket 

(Dumitru et al., 2009). Potential substrates abundant in soybean (o-anisic acid, vanillic 

acid, syringic acid, ferulic acid and sinapic acid) that are structurally similar to dicamba, 

were not metabolized by an E. coli-produced DMO in laboratory tests indicating that the 

DMO enzyme is specific for dicamba (Section V.A.2). The E. coli-produced DMO is 

similar in sequence and function to MON 87708 DMO, therefore it is appropriate to 

extend specificity data generated with the E. coli-produced DMO to MON 87708 DMO.

Given the limited amount of chlorinated metabolites with structures similar to dicamba in 

plants and other eukaryotes (Wishart, 2010; Wishart et al., 2009) it is unlikely that 

MON 87708 DMO will catalyze the conversion of other endogenous substrates. 

Therefore, the activity of the enzyme is specific for dicamba while it maintains many 

structural properties common to oxygenases that are ubiquitous to all organisms. 

V.E.4. MON 87708 DMO is Not a Known Allergen or Toxin 

Bioinformatics analyses were performed to assess the allergenic potential, toxicity, or 

biological activity of MON 87708 DMO. The bioinformatics assessment was performed 

on DMO+27, which includes the amino acid sequence of DMO. The analysis 

demonstrated that MON 87708 DMO does not share amino acid sequence similarities 

with known allergens, gliadins, glutenins, or protein toxins which could have adverse 

effects to human or animal health (Section V.D). 
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V.E.5. MON 87708 DMO is Labile in in vitro Digestion Assays

MON 87708 DMO was readily digestible in SGF and SIF. Rapid degradation of the 

MON 87708 DMO in SGF and SIF makes it highly unlikely that the MON 87708 DMO 

would be absorbed in the small intestine and have any adverse effects on human or 

animal health. 

V.E.6. MON 87708 DMO is Not a Toxin 

An acute oral toxicology study was conducted with MON 87708 DMO. Results indicate 

that MON 87708 DMO did not cause any adverse effects in mice, with a No Observable 

Adverse Effect Level (NOAEL) of 140 mg/kg body weight (BW), the highest dose level 

tested. 

Potential human health risks from consumption of foods derived from MON 87708 were 

evaluated using a Margin of Exposure (MOE) approach. A MOE was calculated between 

the acute mouse NOAEL (140 mg/kg BW) for the MON 87708 DMO and 95th percentile 

“eater-only” estimates of acute dietary exposure determined using the Dietary Exposure 

Evaluation Model (DEEM-FCID version 2.03, Exponent Inc.). DEEM food consumption 

data are obtained from the 1994-1996 and 1998 USDA Continuing Survey of Food 

Intakes by Individuals (CSFII). The MOEs for acute dietary intake of MON 87708 DMO 

were estimated to be 24,800 and 600 for the general population and non-nursing infants, 

the sub-population with the highest estimated exposure, respectively. These very large 

MOEs, in addition to the above mentioned protein safety data for MON 87708 DMO, 

support the conclusion that there is no meaningful risk to human health from dietary 

exposure to MON 87708 DMO. 

Potential health risks to animals from the presence of MON 87708 DMO in feed were 

evaluated by calculating an estimate of daily dietary intake (DDI). In the worst case 

scenario, the percentage of MON 87708 DMO consumed from MON 87708 as a 

percentage of the daily protein intake for a dairy cow is 0.0396% and for both the broiler 

and pig is less than 0.0121%. These very small levels of exposure of animals to 

MON 87708 DMO in their feed, in addition to the above mentioned safety data for 

MON 87708 DMO, support the conclusion that there is no meaningful risk to animal 

health when MON 87708 is present in their diets. 

Using the guidance provided by the FDA in its 1992 Policy Statement regarding the 

evaluation of New Plant Varieties, a conclusion of “no concern” is reached for the donor 

organism and MON 87708 DMO. The food and feed products containing MON 87708 or 

derived from MON 87708 are as safe as soybean currently on the market for human and 

animal consumption. 

V.F. MON 87708 DMO Characterization and Safety Conclusion 

MON 87708 DMO is an oxygenase that catalyzes the O-demethylation of the herbicide 

dicamba. MON 87708 DMO was derived from S. maltophilia, which is an 

environmentally ubiquitous bacterium that does not pose a health risk to healthy 

individuals. MON 87708 DMO is a Rieske-type mono-oxygenase that has homologs in 
Monsanto Company 10-SY-210U 85 of 721 

bacteria and plants that share many of the typical structural and functional characteristics 

of these types of oxygenases, while maintaining specificity for its substrate. MON 87708 

DMO was fully characterized confirming both the N-terminal and internal amino acid 

sequence and the lack of glycosylation. MON 87708 DMO was isolated from 

MON 87708 and was used for the described safety studies; therefore an equivalence 

evaluation to the protein produced in a heterologous expression system was not required. 

Expression studies using ELISA demonstrated that MON 87708 DMO was expressed in 

all tissues assayed at levels ranging from 3.9 – 180 μg/g dwt, representing a low 

percentage of the total protein in soybean. Bioinformatics analysis determined that 

MON 87708 DMO does not share amino acid sequence similarities with known allergens, 

gliadins, glutenins, or protein toxins. MON 87708 DMO was rapidly digested in in vitro

assays using simulated gastric and intestinal fluids and did not show any adverse effects 

when administered to mice via oral gavage at levels that resulted in large MOEs. 

Together with the safety data, these data support a conclusion that there is no meaningful 

risk to human health from dietary exposure to MON 87708 DMO. Therefore, the food 

and feed products containing MON 87708 or derived from MON 87708 are as safe as 

soybean currently on the market for human and animal consumption. 
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VI. COMPOSITIONAL ASSESSMENT OF MON 87708 

Safety assessments of biotech crops typically include comparisons of the composition of 

forage and whole grain of the GM crop to that of conventional counterparts (Codex 

Alimentarius, 2003). Compositional assessments are performed using the principles and 

analytes outlined in the OECD consensus documents for soybean composition (OECD, 

2001). 

A recent review of compositional assessments conducted according to OECD guidelines 

that encompassed a total of seven GM crop varieties, nine countries and eleven growing 

seasons concluded that incorporation of biotechnology-derived agronomic traits has had 

little impact on natural variation in crop composition; most compositional variation is 

attributable to growing region, agronomic practices and genetic background (Harrigan et 

al., 2010). Numerous scientific publications have further documented the extensive 

variability in the concentrations of crop nutrients and anti-nutrients that reflect the 

influence of environmental and genetic factors as well as extensive conventional breeding 

efforts to improve nutrition, agronomics and yield. (Reynolds et al., 2005). 

Compositional equivalence between biotechnology-derived and conventional crops 

provides an “equal or increased assurance of the safety of foods derived from genetically 

modified plants” (OECD, 2001). The OECD consensus documents emphasize 

quantitative measurements of essential nutrients and known anti-nutrients. This is based 

on the premise that such comprehensive and detailed analyses will most effectively 

discern any compositional changes that imply potential safety and nutritional concerns. 

Levels of the components in seed and forage of the biotechnology-derived crop are 

compared to: 1) corresponding levels in a conventional comparator, the nonbiotechnology near isogenic line, grown concurrently, under identical field conditions, 

and 2) natural ranges generated from an evaluation of commercial reference varieties 

grown concurrently and from data published in the scientific literature. 

The latter comparison places any potential differences between the assessed crop and its 

comparator in the context of the well-documented variation in the concentrations of crop 

nutrients and anti-nutrients. 

VI.A. Compositional Equivalence of MON 87708 Seed and Forage to Conventional 

Soybean 

Seed and forage samples were collected from MON 87708 and the near isogenic 

conventional soybean control A3525 grown in a 2008 U.S. field production. Four 

different commercial reference varieties were included at each site of the field production 

to provide data on natural variability of each compositional component analyzed. The 

field production was conducted at five sites: Jefferson County, Iowa; Stark County, 

Illinois; Clinton County, Illinois; Parke County, Indiana; and Berks County, Pennsylvania. 

All soybean plants including MON 87708, the conventional control, and the commercial 

reference varieties were treated with maintenance pesticides as necessary throughout the 

growing season. In addition, MON 87708 plots were either treated at the V2-V3 growth 

stage with dicamba herbicide at the maximum in-crop label rate (0.5 lb acid equivalence 

(a.e.)/acre) or not treated with dicamba herbicide. 
Monsanto Company 10-SY-210U 87 of 721 

Compositional analyses were conducted to assess whether levels of key nutrients and 

anti-nutrients in MON 87708 were equivalent to levels in the conventional control and to 

the composition of the commercial reference varieties. A description of nutrients and 

anti-nutrients present in soybean is provided in the OECD consensus document on 

compositional considerations for soybean (OECD, 2001). Nutrients assessed included 

proximates (ash, carbohydrates by calculation, moisture, protein, and fat), fiber, amino 

acids (18 components), fatty acids (FA, C8-C22), and vitamin E (α-tochopherol) in seed, 

and proximates (ash, carbohydrates by calculation, moisture, protein, and fat) and fiber in 

forage. Anti-nutrients assessed in seed included raffinose, stachyose, lectin, phytic acid, 

trypsin inhibitors, and isoflavones (daidzein, genistein, and glycitein). 

In all, 64 different components were measured (seven in forage and 57 in seed). 

Components that had more than 50% of the observations below the assay limit of 

quantitation (LOQ) were excluded from statistical analysis. Therefore, 50 components 

for both dicamba-treated and untreated MON 87708 were statistically assessed using a 

mixed-model analysis of variance method. Values for all assessed components were 

reported on a dry weight basis with the exception of moisture, which was reported as % 

fresh weight (fwt) and fatty acids, which were reported as % of total FA. 

For MON 87708, six statistical comparisons to the conventional control were conducted. 

One comparison was based on compositional data combined across all five field sites 

(combined-site analysis) and five separate comparisons were conducted on data from 

each of the individual field sites. Statistically significant differences were identified at a 

5% level of significance. Data from the commercial reference varieties were combined 

across all sites and used to calculate a 99% tolerance interval for each compositional 

component to define the natural variability of each component in soybean varieties that 

have a history of safe consumption and that were grown concurrently with MON 87708 

and the conventional control in the same trial. 

For the combined-site analysis, statistically significant differences in nutrient and antinutrient components were further evaluated using considerations relevant to the safety 

and nutritional quality of MON 87708 when compared to the conventional control A3525, 

the conventional counterpart with a history of safe consumption: 1) the relative 

magnitude of the difference in the mean values of nutrient and anti-nutrient components 

of MON 87708 and the conventional control, 2) whether the MON 87708 component 

mean value is within the range of natural variability of that component as represented by 

the 99% tolerance interval of the commercial varieties grown concurrently in the same 

trial, 3) analyses of the reproducibility of the statistically significant combined-site 

component differences at individual sites, and 4) assessing the differences within the 

context of natural variability of commercial soybean composition published in the 

scientific literature and in the International Life Sciences Institute (ILSI) Crop 

Composition Database (ILSI, 2006; Ridley et al., 2004). 

This analysis provides a comprehensive comparative assessment of the levels of key 

nutrients and anti-nutrients in seed, and of key nutrients in forage of MON 87708 and the 

conventional control, discussed in the context of natural variability in commercial 
Monsanto Company 10-SY-210U 88 of 721 

soybean. Results of the comparison indicate that the composition of the seed and forage 

of MON 87708 is equivalent to that of the near isogenic conventional control A3525 and 

within the range of natural variability of the commercial reference varieties. 

VI.A.1. Composition of Soybean Seed and Forage (Treated) 

VI.A.1.1. Nutrient Levels in Soybean Seed (Treated) 

In the combined-site analysis of nutrient levels in seed, the following components 

showed no statistically significant differences in mean values between MON 87708 and 

the conventional control: moisture, total fat, six amino acids (alanine, lysine, methionine, 

serine, threonine, and tryptophan), and three fatty acids (18:0 stearic acid, 20:0 arachidic 

acid, and 20:1 eicosenoic acid) (Table VI-2). 

The components that showed statistically significant differences in mean values between 

MON 87708 and the conventional control in the combined-site analysis were: three 

proximates (ash, carbohydrates by calculation, and protein), 12 amino acids (arginine, 

aspartic acid, cystine, glutamic acid, glycine, histidine, isoleucine, leucine, phenylalanine, 

proline, tyrosine, and valine), three types of fiber (acid detergent fiber -ADF, neutral 

detergent fiber -NDF, and crude fiber), five fatty acids (16:0 palmitic acid, 18:1 oleic acid, 

18:2 linoleic acid, 18:3 linolenic acid, and 22:0 behenic acid), and vitamin E (Tables VI-1 

and VI-2). 

These statistically significant differences in nutrients were evaluated using considerations 

relevant to the safety and nutritional quality of MON 87708 when compared to the 

conventional control: 

1) All nutrient component differences observed in the combined-site analysis, 

whether reflecting increased or decreased MON 87708 mean values with respect 

to the conventional control were small. Relative magnitude of differences ranged 

from 2.65 to 7.91% for amino acids, 1.51 to 8.19% for fatty acids, 15.13% for 

vitamin E, and 2.41 to 12.37% for proximates and fibers. 

2) Mean values for all of these statistically different nutrient components from the 

combined-site analysis of MON 87708 were within the 99% tolerance interval 

established from the commercial reference varieties grown concurrently and were, 

therefore, within the range of natural variability of that component in commercial 

soybean varieties with a history of safe consumption (Tables VI-1 and VI-2). 

3) Assessment of the reproducibility of the combined-site differences at the five 

individual sites showed: statistically significant differences for carbohydrates by 

calculation, crude fiber, cystine, and glycine at one site; aspartic acid, 

phenylalanine, proline, tyrosine, valine, 16:0 palmitic acid, and 18:2 linoleic acid 

at two sites; protein, arginine, glutamic acid, histidine, isoleucine, leucine, and 

22:0 behenic acid at three sites; vitamin E at four sites; and 18:1 oleic acid and 

18:3 linolenic acid differed across all five sites. Although they were different in 

the combined-site analysis, no differences were observed for ash, ADF or NDF at 
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any of the individual sites. Individual site mean values of MON 87708 for all 

nutrient components with statistically significant differences fell within the 99% 

tolerance interval established from the commercial reference varieties grown 

concurrently and were, therefore, within the range of natural variability of that 

component in commercial soybean varieties with a history of safe consumption. 

4) All mean values of MON 87708 for all nutrient components were within the 

context of the natural variability of commercial soybean composition as published 

in the scientific literature and available in the ILSI Crop Composition Database 

(ILSI, 2006; Ridley et al., 2004). 

Thirteen of the 24 differences between MON 87708 and the conventional control 

observed in the combined-site data analysis were attributable to small differences in 

protein and 12 individual amino acids (all expressed as % dwt). The relative magnitude 

of the difference between the mean protein values for MON 87708 and the conventional 

control was small (a decrease of 3.65% in the combined-site analysis for MON 87708) 

and reached statistical significance at only three of the five individual sites. 

Correspondingly, differences in all amino acids were small and not observed consistently 

as statistically significant differences at all individual sites. Eleven of the 12 amino acids 

observed to be different in the combined-site analysis were decreased (2.65-7.91%) 

relative to the conventional control and, as with protein, statistically significant 

differences were not consistently observed at all individual sites. Cystine showed a 

relative increase of 3.01% but was statistically significantly different at only one site. 

Four of the six amino acids (alanine, lysine, serine, and threonine) not observed to be 

statistically different in the combined-site analysis also showed modest decreases ranging 

from ~ 1.5-2.3% (Table VI-2) consistent with the directionality of the changes observed 

in protein content. Overall, observed differences in protein and amino acid levels are not 

considered to be meaningful from a food and feed safety and nutritional perspective 

because they were small, and the mean MON 87708 values were within the 99% 

tolerance interval established by the commercial reference varieties grown concurrently 

in the same trial. 

Five of the combined-site differences between MON 87708 and the conventional control 

were attributable to fatty acid levels (all expressed as % total FA) in seed, whereas total 

fat content was not statistically significantly different. For 18:1 oleic acid and 18:3 

linolenic acid, the relative magnitude of differences between the mean values for 

MON 87708 and conventional control were small in the combined-site analysis (a 

decrease of 8.19% and an increase of 6.65% compared to the conventional control, 

respectively) and at the five individual sites (levels were <11% decreased for 18:1 oleic 

acid and <10% increased for 18:3 linolenic acid at all sites compared to conventional 

control) (Tables VI-2, E-4, E-7, E-10, E-13, and E-16). 

By comparison, the observed differences between MON 87708 and conventional control 

for 18:1 oleic and 18:3 linolenic acids are markedly less than differences in soybean 

varieties developed through conventional breeding (Fehr, 2007; Clemente and Cahoon, 

2009). The average relative levels of 18:3 linolenic acid in commercial soybean are 

approximately 10% total FA, while the average relative level of 18:1 oleic acid in 
Monsanto Company 10-SY-210U 90 of 721 

commercial soybean is approximately 18-25% total FA. In the compositional analysis 

presented here, the values of FA components in the conventional control, when assessed 

as individual replicates across all five individual sites, ranged from 19.6 to 22.4% total 

FA for 18:1 oleic acid and from 8.4 to 10.1% total FA for 18:3 linolenic acid 

(Table VI-2). The values from the commercial reference varieties ranged from 17.9 to 

25.3% total FA for 18:1 oleic acid and 7.4 to 11.4% total FA for 18:3 linolenic acid 

(Table VI-2). Additionally, literature data from Lundry et al. (2008) and Berman et al. 

(2009) and the ILSI Crop Composition Database (ILSI, 2006; Ridley et al., 2004) 

highlight the extensive natural variability in fatty acid levels in soybean, as presented in 

Table VI-9. The small relative magnitudes of the differences in 18:3 linolenic acid and 

18:1 oleic acid compared to the conventional control as well the broad range of these 

fatty acids present in commercial soybean varieties, suggest that the differences are not 

meaningful to food and feed safety and nutritional quality in MON 87708. 

The relative magnitudes of differences between the mean values for MON 87708 and the 

conventional control for the other three ftty acids observed in the combined-site analysis 

were small (2.29% increase for 16:0 palmitic acid, 1.51% increase for 18:2 linoleic acid 

and a 4.70% decrease for 22:0 behenic acid). The small magnitude of differences as well 

as the lack of statistical differences across all individual sites (Tables VI-2, E-4, E-7, 

E-10, E-13, and E-16) further confirmed that the differences observed in fatty acid 

composition are not meaningful to food and feed safety and nutritional quality. 

One of the combined-site differences observed between MON 87708 and the 

conventional control was attributable to vitamin E (expressed as mg/100g dwt). The 

relative magnitude of difference between the mean values of MON 87708 and 

conventional control for vitamin E in the combined-site analysis was an increase of 

15.1% with respect to the conventional control (Tables VI-1). 

Levels of vitamin E are known to be affected by environmental growing conditions (E) 

and germplasm (G) as demonstrated in results from recent assessments on soybean 

varieties grown at three locations in the U.S. over a period of four years (Britz et al., 

2008) and across six environments in Eastern Canada in a single year (Seguin et al., 

2009). Britz et al. (2008) showed more than a two-fold variation in levels across their 

study (units expressed as the ratio of α-tocopherol (vitamin E) to total tocopherol content). 

Vitamin E values in Seguin et al. (2009) ranged from 0.87 to 3.32 mg/100g dwt. Both 

assessments showed that G and E effects as well as G × E interaction effects influenced 

vitamin E content. In the compositional analysis presented here, values of vitamin E in 

the conventional control, when assessed as individual replicates across all sites, ranged by 

as much as 0.89 to 2.11 mg/100g dwt (Table VI-2). Ranges of vitamin E values from the 

concurrently grown commercial reference varieties were even greater and ranged from 

0.69 to 2.91 mg/100g dwt (Table VI-2). Literature data from other compositional 

assessments (Berman et al., 2009; Lundry et al., 2008; ILSI, 2006; Ridley et al., 2004) 

that further highlight the extensive natural variability in vitamin E levels in soybean are 

presented in Table VI-9. Therefore, given this established variability of vitamin E levels 

in conventional soybean and the fact that soybean is not an important nutritional source 

of vitamin E in human or animal diets, this increase in vitamin E levels in MON 87708 
Monsanto Company 10-SY-210U 91 of 721 

compared to the conventional control supports the conclusion that this observed 

difference is not meaningful to food and feed safety and nutritional quality. 

The remaining combined-site differences between MON 87708 and the conventional 

control were attributable to two proximates (ash and carbohydrates by calculation) and 

three fibers (ADF, NDF, and crude fiber). The relative magnitude of these increases 

were small (2.41% to 12.37%) and there was no consistency of these combined-site 

differences at the individual sites (carbohydrates by calculation and crude fiber were 

different at only one site, whereas ash, ADF and NDF were not different at any of the 

individual sites). The combined-site mean values for these nutrient components also 

were within the 99% tolerance interval established from the commercial reference 

varieties grown concurrently establishing that these differences are not meaningful to 

food and feed safety and nutrition. 

In summary, statistical analyses found no consistent differences across sites in the levels 

of nutrient components in seed from MON 87708 and the conventional control, except 

for differences in 18:1 oleic acid, 18:3 linoleic acid, and vitamin E levels that were of 

small magnitude and were within the natural variability of the concurrently grown 

commercial soybean varieties. These data support the conclusion that MON 87708 is 

compositionally equivalent to conventional soybean. 

VI.A.1.2. Anti-Nutrient Levels in Soybean Seed (Treated) 

In the combined-site analysis, no statistically significant differences were observed in 

four of the eight anti-nutrient component comparisons (lectin, trypsin inhibitors, genistein, 

and glycitein) between MON 87708 and the conventional control. Statistically 

significant differences were observed between MON 87708 and the conventional control 

in the other four anti-nutrient components that were measured (Tables VI-1 and VI-3). 

The differences included decreased mean values for phytic acid, raffinose, stachyose, and 

an increased mean level of daidzein, compared to the conventional control. 

The statistically significant differences in anti-nutrients were evaluated using 

considerations relevant to the safety and nutritional quality of MON 87708 when 

compared to the conventional control: 

1) All anti-nutrient component differences observed in the combined-site analysis, 

whether reflecting increased or decreased MON 87708 mean values with respect 

to the conventional control were small. Relative magnitude of differences in the 

combined-site analysis for the anti-nutrients that were decreased in MON 87708 

ranged from 6.1% (phytic acid) to 7.73% (raffinose). The relative magnitude of 

difference (increase) in daidzein was 11.5%. 

2) MON 87708 mean values for these anti-nutrient components from the combinedsite analysis were within the 99% tolerance interval established from the 

commercial reference varieties concurrently grown in the same trial and, therefore 

were within the range of natural variability of these components in commercial 

soybean varieties with a history of safe consumption (Tables V1-1 and V1-3). 
Monsanto Company 10-SY-210U 92 of 721 

3) Assessment of the reproducibility of the combined-site differences at the five 

individual sites showed no consistent pattern across sites. A statistically 

significant decrease was observed for stachyose at one site and phytic acid at two 

sites, whereas a significant increase was seen for daidzein at two sites. No 

differences for raffinose were observed at any of the individual sites. Mean 

values for all of the above anti-nutrient components in MON 87708 at the 

individual sites were within the 99% tolerance interval established from the 

concurrently grown commercial reference varieties. 

4) All mean values of MON 87708 for all anti-nutrients were within the context of 

the natural variability of commercial soybean composition as published in the 

scientific literature and available in the ILSI Crop Composition Database (ILSI, 

2006; Ridley et al., 2004). 

In summary, statistical analyses found no consistent differences across sites in the levels 

of anti-nutrient components in seed from MON 87708 and the conventional control. 

Thus, a comprehensive evaluation of anti-nutrient components in seed support the 

conclusion that MON 87708 is compositionally equivalent to conventional soybean. 

VI.A.1.3. Nutrient Levels in Soybean Forage (Treated) 

In the combined-site analysis of forage, six of the seven nutrient component comparisons 

did not have a statistically significant difference between MON 87708 and the 

conventional control (Tables VI-1 and VI-4). The only statistical difference was for the 

ADF mean value and it was evaluated using considerations relevant to the safety and 

nutritional quality of MON 87708 when compared to the conventional control. 

1) The relative magnitude of difference in ADF, with respect to the conventional 

control, was small with an increase of 10.45%. 

2) The mean value for ADF from the combined-site analysis of MON 87708 was 

within the 99% tolerance interval established from the commercial reference 

varieties grown concurrently in the same trial and, therefore within the range of 

natural variability of that component in commercial soybean varieties with a 

history of safe consumption (Tables V1-1 and V1-4). 

3) Assessment of the reproducibility of the combined-site difference of ADF across 

the individual sites showed no statistically significant differences at any of the 

five individual sites. 

4) The level of ADF was within the natural variability observed for commercial 

soybean varieties as published in the scientific literature and available in the ILSI 

Crop Composition Database (ILSI, 2006; Ridley et al., 2004). 

In summary, statistical analyses found no consistent differences across sites in the levels 

of nutrient components in forage from MON 87708 and the conventional control. Thus, a 

comprehensive evaluation of nutrient components in forage supports the conclusion that 

MON 87708 is compositionally equivalent to conventional soybean. 
Monsanto Company 10-SY-210U 93 of 721 

VI.A.2. Composition of Soybean Seed and Forage (Untreated) 

VI.A.2.1. Nutrient Levels in Soybean Seed (Untreated) 

In the combined-site analysis of nutrient levels in seed, the following components 

showed no statistically significant differences in mean values between MON 87708 

(untreated) and the conventional control: ash, carbohydrates by calculation, moisture, 

total fat, crude fiber, ten amino acids (alanine, aspartic acid, glycine, histidine, lysine, 

methionine, serine, threonine, tryptophan, tyrosine), and three fatty acids (18:0 stearic 

acid, 20:0 arachidic acid, and 20:1 eicosenoic acid) (Table VI-6). 

The components that showed statistically significant differences in mean values between 

MON 87708 (untreated) and the conventional control in the combined-site analysis were: 

protein, eight amino acids (arginine,cystine, glutamic acid, isoleucine, leucine, 

phenylalanine, proline, and valine), two types of fiber (acid detergent fiber (ADF), and 

neutral detergent fiber (NDF)), five fatty acids (16:0 palmitic acid, 18:1 oleic acid, 18:2 

linoleic acid, 18:3 linolenic acid, and 22:0 behenic acid), and vitamin E (Tables VI-5 and 

VI-6). 

These statistically significant differences in nutrients were evaluated using considerations 

relevant to the safety and nutritional quality of MON 87708 (untreated) when compared 

to the conventional control: 

1) All nutrient component differences observed in the combined-site analysis, 

whether reflecting increased or decreased MON 87708 (untreated) mean values with 

respect to the conventional control were small. Relative magnitude of differences ranged 

from 2.27 to 5.88% or protein and amino acids, 1.45 to 7.60% for fatty acids, 18.16% for 

vitamin E, and 3.99 to 6.33% for fibers. 

2) Mean values for all of these statistically different nutrient components from the 

combined-site analysis of MON 87708 (untreated) were within the 99% tolerance interval 

established from the commercial reference varieties grown concurrently and were, 

therefore, within the range of natural variability of that component in commercial 

soybean varieties with a history of safe consumption (Tables VI-5 and VI-6). 

3) Assessment of the reproducibility of the combined-site differences at the five 

individual sites showed: statistically significant differences for cystine, isoleucine, 

valine, and 16:0 palmitic acid at one site; protein, arginine, glutamic acid, and leucine at 

two sites; phenylalanine, proline, 18:2 linoleic acid, and 22:0 behenic acid at three sites; 

18:1 oleic acid, 18:3 linolenic acid, and vitamin E at four sites. No components were 

statistically significantly different at all five sites. Although they were different in the 

combined site analysis, no differences were observed for ADF or NDF at any of the 

individual sites. Individual site mean values of MON 87708 (untreated) for all nutrient 

components with statistically significant differences fell within the 99% tolerance interval 

established from the commercial reference varieties grown concurrently and were, 

therefore, within the range of natural variability of that component in commercial 

soybean varieties with a history of safe consumption. 
Monsanto Company 10-SY-210U 94 of 721 

4) All mean values of MON 87708 (untreated) for all nutrient components were 

within the context of the natural variability of commercial soybean composition as 

published in the scientific literature and available in the ILSI Crop Composition Database 

(ILSI, 2006; Ridley et al., 2004). 

Nine of the 17 differences between MON 87708 (untreated) and the conventional control 

observed in the combined-site data analysis were attributable to small differences in 

protein and eight individual amino acids (all expressed as % dwt). The relative 

magnitude of the difference between the mean protein values for MON 87708 (untreated) 

and the conventional control was small (a decrease of 2.94% in the combined-site 

analysis for MON 87708 (untreated) and reached statistical significance at only two of 

the five individual sites. Correspondingly, differences in all amino acids were small and 

not observed consistently as statistically significant differences at all individual sites. 

Seven of the 8 amino acids observed to be different in the combined-site analysis were 

decreased (2.27 to 5.88%) relative to the conventional control. Cystine showed a relative 

increase of 3.27% but was statistically significantly different at only one site. Overall, 

observed differences in protein and amino acid levels are not considered to be meaningful 

from a food and feed safety and nutritional perspective because they were small, and the 

mean MON 87708 (untreated) values were within the 99% tolerance interval established 

by the commercial reference varieties grown concurrently in the same trial. 

Five of the combined-site differences between MON 87708 (untreated) and the 

conventional control were attributable to fatty acid levels (all expressed as % total FA) in 

seed, whereas total fat content was not statistically significantly different. For 18:1 oleic 

acid and 18:3 linolenic acid, the relative magnitude of differences between the mean 

values for MON 87708 (untreated) and conventional control were small in the combinedsite analysis (a decrease of 7.60% and an increase of 5.78% compared to the conventional 

control, respectively) and at the individual sites (levels were <12% decreased for 18:1 

oleic acid and <8% increased for 18:3 linolenic acid at all sites compared to conventional 

control) (Tables VI-6, E-19, E-22, E-25, E-28, and E-31). 

By comparison, the observed differences between MON 87708 (untreated) and 

conventional control for 18:1 oleic and 18:3 linolenic acids are markedly less than 

differences in soybean varieties developed through conventional breeding (Clemente and 

Cahoon, 2009; Fehr, 2007). The average relative levels of 18:3 linolenic acid in 

commercial soybean are approximately 10% total FA, while the average relative level of 

18:1 oleic acid in commercial soybean is approximately 18-25% total FA. In the 

compositional analysis presented here, the values of FA components in the conventional 

control, when assessed as individual replicates across all five individual sites, ranged 

from 19.6 to 22.4% total FA for 18:1 oleic acid and from 8.4 to 10.1% total FA for 18:3 

linolenic acid (Table VI-6). The values from the commercial reference varieties ranged 

from 17.9 to 25.3% total FA for 18:1 oleic acid and 7.4 to 11.4% total FA for 18:3 

linolenic acid (Table VI-6). Additionally, literature data from Lundry et al. (2008) and 

Berman et al. (2009) and the ILSI Crop Composition Database highlight the extensive 

natural variability in fatty acid levels in soybean, as presented in Table VI-9. The small 

relative magnitudes of the differences in 18:3 linolenic acid and 18:1 oleic acid compared 

to the conventional control as well the broad range of these fatty acids present in 
Monsanto Company 10-SY-210U 95 of 721 

commercial soybean varieties, suggest that the differences are not meaningful to food and 

feed safety and nutritional quality in MON 87708 (untreated). 

The relative magnitudes of differences between the mean values for MON 87708 

(untreated) and the conventional control for the other three fatty acids observed in the 

combined-site analysis were small (2.37% increase for 16:0 palmitic acid, 1.45% increase 

for 18:2 linoleic acid and a 3.71% decrease for 22:0 behenic acid). The small magnitude 

of differences as well as the lack of statistical differences across all individual sites 

(Tables VI-6, E-19, E-22, E28 and E-28) further confirmed that the differences observed 

in fatty acid composition are not meaningful to food and feed safety and nutritional 

quality. 

One of the combined-site differences observed between MON 87708 (untreated) and the 

conventional control was attributable to vitamin E (expressed as mg/100g dwt). The 

relative magnitude of difference between the mean values of MON 87708 (untreated) and 

conventional control for vitamin E in the combined-site analysis was an increase of 

18.16% with respect to the conventional control (Table VI-6). 

Levels of vitamin E are known to be affected by environmental growing conditions and 

germplasm as demonstrated in results from recent assessments on soybean varieties 

grown at three locations in the U.S. over a period of four years (Britz et al., 2008) and 

across six environments in Eastern Canada in a single year (Seguin et al., 2009). Britz et 

al. (2008) showed more than a two-fold variation in levels across their study (units 

expressed as the ratio of α tocopherol (vitamin E) to total tocopherol content). Vitamin E 

values in Seguin et al. (2009) ranged from 0.87 to 3.32 mg/100g dwt. In the 

compositional analysis presented here, values of vitamin E in the conventional control, 

when assessed as individual replicates across all sites, ranged by as much as 0.89 to 2.11 

mg/100g dwt (Table VI-6). Ranges of vitamin E values from the concurrently grown 

commercial reference varieties were even greater and ranged from 0.69 to 2.91 mg/100g 

dwt (Table VI-6). Literature data from other compositional assessments (Berman et al., 

2009; ILSI, 2006; Lundry et al., 2008; Ridley et al., 2004) that further highlight the 

extensive natural variability in vitamin E levels in soybean are presented in Table VI-9. 

Therefore, given this established variability of vitamin E levels in conventional soybean 

and the fact that soybean is not an important nutritional source of vitamin E in human or 

animal diets, this increase in vitamin E levels in MON 87708 (untreated) compared to the 

conventional control supports the conclusion that this observed difference is not 

meaningful to food and feed safety and nutritional quality. 

The remaining combined-site differences between MON 87708 (untreated) and the 

conventional control were attributable to two fibers (ADF and NDF). The relative 

magnitude of these increases were small (3.99% to 6.33%) and ADF and NDF were not 

different at any of the individual sites. The combined-site mean values for these nutrient 

components also were within the 99% tolerance interval established from the commercial 

reference varieties grown concurrently establishing that these differences are not 

meaningful to food and feed safety and nutrition. 
Monsanto Company 10-SY-210U 96 of 721 

In summary, statistical analyses found no consistent differences across sites in the levels 

of nutrient components in seed from MON 87708 (untreated) and the conventional 

control, except for differences in 18:1 oleic acid, 18:3 linoleic acid, and vitamin E levels 

that were of small magnitude and were within the natural variability of the concurrently 

grown commercial soybean varieties. These data support the conclusion that 

MON 87708 (untreated) is compositionally equivalent to conventional soybean. 

VI.A.2.2. Anti-Nutrient Levels in Soybean Seed (Untreated) 

In the combined-site analysis, no statistically significant differences were observed in 

five of the eight anti-nutrient component comparisons (lectin, phytic acid, raffinose, 

stachyose, and glycitein) between MON 87708 (untreated) and the conventional control. 

Statistically significant differences were observed between MON 87708 (untreated) and 

the conventional control in the other three anti-nutrient components that were measured 

(Tables VI-5 and VI-7). The differences included increased mean values trypsin 

inhibitors, daidzein, and genistein compared to the conventional control. 

The statistically significant differences in anti-nutrients were evaluated using 

considerations relevant to the safety and nutritional quality of MON 87708 (untreated) 

when compared to the conventional control: 

1) All anti-nutrient component differences observed in the combined-site analysis 

reflected increased MON 87708 (untreated) mean values with respect to the conventional 

control, but were all less than 20%. Relative magnitude of differences in the combinedsite analysis for the anti-nutrients were 15.37% (trypsin inhibitor), 17.24% (daidzein) and 

11.59% (genistein). 

2) MON 87708 (untreated) mean values for these anti-nutrient components from the 

combined-site analysis were within the 99% tolerance interval established from the 

commercial reference varieties concurrently grown in the same trial and, therefore were 

within the range of natural variability of these components in commercial soybean 

varieties with a history of safe consumption (Tables VI-5 and VI-7). 

3) Assessment of the reproducibility of the combined-site differences at the five 

individual sites showed no consistent pattern across sites. None of the anti-nutrient 

components found to be different in the combined site analysis were observed to be 

statistically different at more than one of the five individual sites. Mean values for all of 

the above anti-nutrient components in MON 87708 (untreated) at the individual sites 

were within the 99% tolerance interval established from the concurrently grown 

commercial reference varieties. 

4) All mean values of MON 87708 (untreated) for all anti-nutrients were within the 

context of the natural variability of commercial soybean composition as published in the 

scientific literature and available in the ILSI Crop Composition Database (ILSI, 2006; 

Ridley et al., 2004). 

In summary, statistical analyses found no consistent differences across sites in the levels 

of anti-nutrient components in seed from MON 87708 (untreated) and the conventional 
Monsanto Company 10-SY-210U 97 of 721 

control. Thus, a comprehensive evaluation of anti-nutrient components in seed support 

the conclusion that MON 87708 is compositionally equivalent to conventional soybean. 

VI.A.2.3. Nutrient Levels in Soybean Forage (Untreated) 

In the combined-site analysis of forage, none of the seven nutrient component 

comparisons had a statistically significant difference between MON 87708 (untreated) 

and the conventional control (Tables VI-5 and VI-8). In summary, statistical analyses 

found no consistent differences across sites in the levels of nutrient components in forage 

from MON 87708 (untreated) and the conventional control. Thus, a comprehensive 

evaluation of nutrient components in forage supports the conclusion that MON 87708 

(untreated) compositionally equivalent to conventional soybean. 
Monsanto Company 10-SY-210U 

Table VI-1. Summary of Differences (

α=0.05) for the Comparison of Soybean Component Levels for M

vs. Conventional Control 

Mean Difference 

(MON 87708 minus Control)

Analytical Component (Units)1

MON 877082

Mean3

Control4

Mean

Mean Difference 

(% of Control)

Significance

(p-Value)

MON 8

Rang

Statistically Significant Differences Observed in Combined-Site AnalysisSeed Proximate (% dwt)Ash 5.24 5.12 2.41 0.031 4.94 - 5Carbohydrates 37.93 36.64 3.50 0.012 35.65 - 3Protein 40.86 42.41 -3.65 0.016 39.00 - 4Seed Fiber (% dwt)Acid Detergent Fiber 13.55 12.86 5.30 0.009 12.45 - Crude Fiber 8.29 7.37 12.37 <0.001 6.23 - 9Neutral Detergent Fiber 15.29 14.34 6.63 0.028 13.11 - Seed Amino Acid (% dwt)Arginine 3.30 3.58 -7.91 0.006 3.09 - 3
Monsanto Company 10-SY-210U 

Table VI-1 (continued). Summary of Differences (

α=0.05) for the Comparison of Soybean Component L

(Treated) vs. Conventional Control 

Mean Difference 

(MON 87708 minus Control)

Analytical Component (Units)1

MON 877082

Mean3

Control4

Mean

Mean Difference 

(% of Control)

Significance

(p-Value)

MON 877

Range

Statistically Significant Differences Observed in Combined-Site AnalysisSeed Amino Acid (% dwt)Aspartic Acid 4.63 4.78 -3.18 0.016 4.44 - 4.8Cystine 0.61 0.59 3.01 <0.001 0.58 - 0.6Glutamic Acid 7.38 7.69 -4.03 0.010 7.05 - 7.7Glycine 1.76 1.81 -2.65 0.020 1.67 - 1.8Histidine 1.06 1.09 -3.07 0.017 1.02 - 1.1Isoleucine 1.88 1.95 -3.58 0.006 1.75 - 1.9Leucine 3.06 3.17 -3.37 0.008 2.93 - 3.1Phenylalanine 2.06 2.13 -3.33 0.034 1.92 - 2.1
Monsanto Company 10-SY-210U 

Table VI-1 (continued). Summary of Differences (

α=0.05) for the Comparison of Soybean Component L

(Treated) vs. Conventional Control 

Mean Difference 

(MON 87708 minus Control)

Analytical Component (Units)1

MON 877082

Mean3

Control4

Mean

Mean Difference 

(% of Control)

Significance

(p-Value)

MON 877

Range

Statistically Significant Differences Observed in Combined-Site AnalysisSeed Amino Acid (% dwt)Proline 1.99 2.05 -3.24 0.017 1.90 - 2.0Tyrosine 1.37 1.42 -3.47 0.048 1.28 - 1.4Valine 1.98 2.06 -3.89 0.006 1.82 - 2.0Seed Fatty Acid (% Total FA)16:0 Palmitic 11.59 11.33 2.29 0.002 11.25 - 1218:1 Oleic 19.20 20.91 -8.19 <0.001 17.85 - 1918:2 Linoleic 54.40 53.59 1.51 0.010 53.42 - 5518:3 Linolenic 10.12 9.49 6.65 <0.001 8.99 - 10.22:0 Behenic 0.27 0.28 -4.70 0.001 0.25 - 0.2
Monsanto Company 10-SY-210U 

Table VI-1 (continued). Summary of Differences (

α=0.05) for the Comparison of Soybean Component 

(Treated) vs. Conventional Control 

Mean Difference 

(MON 87708 minus Control)

Analytical Component (Units)1

MON 877082

Mean3

Control4

Mean

Mean Difference 

(% of Control)

Significance

(p-Value)

MON 877

Range

Statistically Significant Differences Observed in Combined-Site AnalysisSeed Vitamin (mg/100g dwt)Vitamin E 1.41 1.23 15.13 0.001 1.08 - 2.1Seed Anti-nutrient (% dwt)Phytic Acid 1.30 1.39 -6.14 0.043 1.08 - 1.5Raffinose 0.43 0.47 -7.73 0.045 0.32 - 0.5Stachyose 3.36 3.62 -7.24 0.011 3.07 - 4.0Seed Isoflavone (μg/g dwt)Daidzein 1494.97 1340.71 11.51 0.046 899.83 - 230Forage Fiber (% dwt)Acid Detergent Fiber 30.58 27.69 10.45 0.021 23.30 - 45
Monsanto Company 10-SY-210U 

Table VI-1 (continued). Summary of Differences (

α=0.05) for the Comparison of Soybean Component 

(Treated) vs. Conventional Control 

Mean Difference 

(MON 87708 minus Control)

Analytical Component (Units)1

MON 877082

Mean3

Control4

Mean

Mean Difference 

(% of Control)

Significance

(p-Value)

MON 877

Range

Statistically Significant Differences Observed in Five Individual SitesSeed Fatty Acid (% Total FA)18:1 Oleic Site IARL 19.38 21.67 -10.58 0.001 19.07 - 1918:1 Oleic Site ILCY 19.74 21.57 -8.46 0.011 19.44 - 1918:1 Oleic Site ILWY 19.52 21.14 -7.66 0.010 19.34 - 1918:1 Oleic Site INRC 18.78 20.19 -6.96 <0.001 18.58 - 1818:1 Oleic Site PAHM 18.58 20.01 -7.13 0.015 17.85 - 1918:3 Linolenic Site IARL 10.64 10.04 5.94 0.033 10.58 - 1018:3 Linolenic Site ILCY 9.07 8.58 5.78 0.007 8.99 - 9.118:3 Linolenic Site ILWY 10.54 10.05 4.92 0.026 10.51 - 10
Monsanto Company 10-SY-210U 

Table VI-1 (continued). Summary of Differences (

α=0.05) for the Comparison of Soybean Component L

(Treated) vs. Conventional Control 

Mean Difference 

(MON 87708 minus Control)

Analytical Component (Units)1

MON 877082

Mean3

Control4

Mean

Mean Difference 

(% of Control)

Significance

(p-Value)

MON 877

Range

Statistically Significant Differences Observed in Five Individual SitesSeed Fatty Acid (% Total FA)18:3 Linolenic Site INRC 10.03 9.31 7.65 <0.001 9.89 - 10.18:3 Linolenic Site PAHM 10.33 9.47 9.02 0.006 9.91 - 10.Statistically Significant Differences Observed in Four Individual SitesSeed Vitamin (mg/100g dwt)Vitamin E Site IARL 1.15 0.94 22.25 0.033 1.10 - 1.2Vitamin E Site ILCY 2.13 1.86 14.43 0.038 2.10 - 2.1Vitamin E Site ILWY 1.18 0.94 24.64 0.011 1.08 - 1.2Vitamin E Site PAHM 1.32 1.23 7.90 0.010 1.21 - 1.5Statistically Significant Differences Observed in Three Individual SitesSeed Proximate (% dwt)Protein Site ILCY 40.17 41.72 -3.72 0.047 39.44 - 40
Monsanto Company 10-SY-210U 

Table VI-1 (continued). Summary of Differences (

α=0.05) for the Comparison of Soybean Component L

(Treated) vs. Conventional Control 

Mean Difference 

(MON 87708 minus Control)

Analytical Component (Units)1

MON 877082

Mean3

Control4

Mean

Mean Difference 

(% of Control)

Significance

(p-Value)

MON 877

Range

Statistically Significant Differences Observed in Three Individual SitesSeed Proximate (% dwt)Protein Site ILWY 40.88 41.99 -2.64 0.042 40.56 - 41Protein Site PAHM 40.25 43.69 -7.86 0.002 39.00 - 41Seed Amino Acid (% dwt)Arginine Site ILWY 3.30 3.57 -7.58 0.002 3.24 - 3.3Arginine Site INRC 3.44 3.72 -7.37 0.011 3.39 - 3.5Arginine Site PAHM 3.25 3.88 -16.13 0.001 3.09 - 3.3Glutamic Acid Site ILCY 7.43 7.61 -2.38 0.032 7.27 - 7.5Glutamic Acid Site ILWY 7.29 7.51 -2.86 0.002 7.20 - 7.3Glutamic Acid Site PAHM 7.28 8.00 -9.08 0.003 7.06 - 7.4
Monsanto Company 10-SY-210U 

Table VI-1 (continued). Summary of Differences (

α=0.05) for the Comparison of Soybean Component L

(Treated) vs. Conventional Control 

Mean Difference 

(MON 87708 minus Control)

Analytical Component (Units)1

MON 877082

Mean3

Control4

Mean

Mean Difference 

(% of Control)

Significance

(p-Value)

MON 877

Range

Statistically Significant Differences Observed in Three Individual SitesSeed Amino Acid (% dwt)Histidine Site ILCY 1.06 1.08 -1.84 0.022 1.04 - 1.0Histidine Site ILWY 1.05 1.07 -1.62 0.019 1.05 - 1.0Histidine Site PAHM 1.05 1.13 -7.52 0.002 1.02 - 1.0Isoleucine Site ILCY 1.89 1.97 -3.98 0.010 1.87 - 1.9Isoleucine Site ILWY 1.87 1.90 -1.22 0.004 1.85 - 1.8Isoleucine Site PAHM 1.85 2.00 -7.59 0.014 1.79 - 1.9Leucine Site ILCY 3.09 3.17 -2.42 0.002 3.04 - 3.1Leucine Site ILWY 3.02 3.10 -2.49 <0.001 3.00 - 3.0
Monsanto Company 10-SY-210U 

Table VI-1 (continued). Summary of Differences (

α=0.05) for the Comparison of Soybean Component L

(Treated) vs. Conventional Control 

Mean Difference 

(MON 87708 minus Control)

Analytical Component (Units)1

MON 877082

Mean3

Control4

Mean

Mean Difference 

(% of Control)

Significance

(p-Value)

MON 877

Range

Statistically Significant Differences Observed in Three Individual SitesSeed Amino Acid (% dwt)Leucine Site PAHM 3.03 3.28 -7.42 0.002 2.96 - 3.0Seed Fatty Acid (% Total FA)22:0 Behenic Site IARL 0.26 0.28 -5.49 0.022 0.25 - 0.222:0 Behenic Site ILWY 0.26 0.28 -6.67 0.008 0.26 - 0.222:0 Behenic Site INRC 0.28 0.29 -4.85 0.038 0.27 - 0.2Statistically Significant Differences Observed in Two Individual SitesSeed ProximateMoisture (% fwt) Site ILWY 6.96 6.16 12.99 0.022 6.80 - 7.1Moisture (% fwt) Site PAHM 7.84 10.50 -25.30 <0.001 7.38 - 8.4Seed Amino Acid (% dwt)Aspartic Acid Site ILWY 4.59 4.67 -1.90 0.011 4.55 - 4.6
Monsanto Company 10-SY-210U 

Table VI-1 (continued). Summary of Differences (

α=0.05) for the Comparison of Soybean Component L

(Treated) vs. Conventional Control 

Mean Difference 

(MON 87708 minus Control)

Analytical Component (Units)1

MON 877082

Mean3

Control4

Mean

Mean Difference 

(% of Control)

Significance

(p-Value)

MON 877

Range

Statistically Significant Differences Observed in Two Individual SitesSeed Amino Acid (% dwt)Aspartic Acid Site PAHM 4.56 4.94 -7.65 0.002 4.45 - 4.6Phenylalanine Site ILWY 2.01 2.07 -2.95 0.046 1.96 - 2.0Phenylalanine Site PAHM 2.04 2.21 -7.96 0.010 2.00 - 2.0Proline Site ILWY 1.94 2.05 -5.09 0.020 1.93 - 1.9Proline Site PAHM 1.98 2.10 -5.98 0.016 1.94 - 2.0Threonine Site ILWY 1.52 1.55 -1.69 0.005 1.51 - 1.5Threonine Site PAHM 1.55 1.62 -4.23 0.029 1.52 - 1.5Tyrosine Site INRC 1.38 1.44 -4.49 0.044 1.35 - 1.4
Monsanto Company 10-SY-210U 

Table VI-1 (continued). Summary of Differences (

α=0.05) for the Comparison of Soybean Component L

(Treated) vs. Conventional Control 

Mean Difference 

(MON 87708 minus Control)

Analytical Component (Units)1

MON 877082

Mean3

Control4

Mean

Mean Difference 

(% of Control)

Significance

(p-Value)

MON 877

Range

Statistically Significant Differences Observed in Two Individual SitesSeed Amino Acid (% dwt)Tyrosine Site PAHM 1.35 1.49 -9.43 0.011 1.28 - 1.4Valine Site ILCY 1.96 2.05 -4.37 0.013 1.94 - 2.0Valine Site PAHM 1.95 2.13 -8.17 0.012 1.89 - 2.0Seed Fatty Acid (% Total FA)16:0 Palmitic Site IARL 11.49 11.00 4.47 0.001 11.44 - 1116:0 Palmitic Site ILWY 11.26 11.04 2.02 0.017 11.25 - 1118:2 Linoleic Site ILCY 54.54 53.26 2.40 0.021 54.45 - 5418:2 Linoleic Site INRC 54.98 54.43 1.00 0.019 54.80 - 55
Monsanto Company 10-SY-210U 

Table VI-1 (continued). Summary of Differences (

α=0.05) for the Comparison of Soybean Component L

(Treated) vs. Conventional Control 

Mean Difference 

(MON 87708 minus Control)

Analytical Component (Units)1

MON 877082

Mean3

Control4

Mean

Mean Difference 

(% of Control)

Significance

(p-Value)

MON 877

Range

Statistically Significant Differences Observed in Two Individual SitesSeed Anti-nutrient (% dwt)Phytic Acid Site IARL 1.36 1.53 -11.28 0.018 1.33 - 1.3Phytic Acid Site ILWY 1.40 1.55 -9.34 0.030 1.33 - 1.4Seed Isoflavone (μg/g dwt)Daidzein Site ILWY 1458.08 1271.60 14.67 0.004 1416.31 

1535.98

Daidzein Site INRC 1683.50 1419.40 18.61 0.049 1593.24 

1777.49

Glycitein Site ILWY 111.77 79.70 40.23 <0.001 109.88 - 11Glycitein Site INRC 111.51 98.42 13.31 0.016 110.91 - 11Forage Proximate (% dwt)Protein Site IARL 25.21 23.00 9.63 0.043 24.71 - 25
Monsanto Company 10-SY-210U 

Table VI-1 (continued). Summary of Differences (

α=0.05) for the Comparison of Soybean Component L

(Treated) vs. Conventional Control 

Mean Difference 

(MON 87708 minus Control)

Analytical Component (Units)1

MON 877082

Mean3

Control4

Mean

Mean Difference 

(% of Control)

Significance

(p-Value)

MON 877

Range

Statistically Significant Differences Observed in Two Individual SitesForage Proximate (% dwt)Protein Site INRC 21.78 23.33 -6.63 0.019 20.99 - 22Statistically Significant Differences Observed in One Individual SiteSeed Proximate (% dwt)Carbohydrates Site PAHM 38.30 35.23 8.71 0.008 37.69 - 38Seed Fiber (% dwt)Crude Fiber Site INRC 8.06 6.89 17.03 0.009 7.76 - 8.4Seed Amino Acid (% dwt)Alanine Site PAHM 1.75 1.86 -5.81 0.010 1.74 - 1.7Cystine Site PAHM 0.62 0.59 4.79 0.024 0.60 - 0.6Glycine Site PAHM 1.73 1.86 -6.78 0.004 1.69 - 1.7
Monsanto Company 10-SY-210U 

Table VI-1 (continued). Summary of Differences (

α=0.05) for the Comparison of Soybean Component L

(Treated) vs. Conventional Control 

Mean Difference 

(MON 87708 minus Control)

Analytical Component (Units)1

MON 877082

Mean3

Control4

Mean

Mean Difference 

(% of Control)

Significance

(p-Value)

MON 877

Range

Statistically Significant Differences Observed in One Individual SiteSeed Amino Acid (% dwt)Lysine Site PAHM 2.60 2.75 -5.39 0.009 2.53 - 2.6Serine Site ILWY 1.98 2.06 -3.83 0.003 1.97 - 2.0Tryptophan Site ILCY 0.51 0.48 6.21 0.024 0.49 - 0.5Seed Anti-nutrient (% dwt)Lectin (H.U./mg dwt) Site ILWY 1.10 2.33 -52.88 0.045 0.59 - 1.5Stachyose Site INRC 3.14 3.46 -9.18 0.043 3.12 - 3.1Forage Proximate (% dwt)Carbohydrates Site PAHM 70.95 65.81 7.81 0.015 69.23 - 73Moisture (% fwt) Site PAHM 74.27 74.91 -0.86 0.021 73.40 - 751dwt = dry weight; fwt = fresh weight; FA = fatty acid; H.U. = Hemagglutinating Units. 2MON 87708 was treated with dicamba. 3Mean = least-square mean. 4Control refers to the near isogenic conventional soybean control A3525. 5With 95% confidence, interval contains 99% of the values expressed in the population of commercial reference varieties. Negative limits set to zero.
Monsanto Company 10-SY-210U 

Table VI-2. Statistical Summary of Combined-Site Soybean Seed Nutrients for MON 87708 (Treated) vs. C

Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 877082 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Proximate (% dwt)Ash 5.24 (0.067) 5.12 (0.067) 0.12 (0.055) 0.011, 0.24 0.031 

(4.94 - 5.69) (4.73 - 5.47) (-0.28 - 0.45) 

Carbohydrates 37.93 (0.50) 36.64 (0.50) 1.28 (0.40) 0.36, 2.20 0.012 

(35.65 - 39.21) (34.11 - 38.45) (-0.38 - 4.07) 

Moisture (% fwt) 6.88 (0.65) 7.14 (0.65) -0.26 (0.52) -1.46, 0.94 0.629 

(5.17 - 8.47) (5.79 - 10.60) (-3.12 - 1.43) 

Protein 40.86 (0.39) 42.41 (0.39) -1.55 (0.51) -2.73, -0.37 0.016 

(39.00 - 42.53) (40.69 - 43.85) (-4.84 - 0.088) 

Total Fat 15.97 (0.59) 15.84 (0.59) 0.13 (0.31) -0.58, 0.84 0.691 

(14.00 - 18.56) (14.40 - 18.39) (-1.90 - 2.37) 

Fiber (% dwt)Acid Detergent Fiber 13.55 (0.40) 12.86 (0.40) 0.68 (0.25) 0.18, 1.19 0.009 

(12.45 - 15.57) (11.62 - 14.57) (-0.71 - 2.13) 
Monsanto Company 10-SY-210U 

Table VI-2 (continued). Statistical Summary of Combined-Site Soybean Seed Nutrients for MON Conventional Control 

Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 877082 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Fiber (% dwt)Crude Fiber 8.29 (0.26) 7.37 (0.26) 0.91 (0.26) 0.40, 1.43 <0.00

(6.23 - 9.65) (6.05 - 8.64) (-0.34 - 2.67) 

Neutral Detergent Fiber 15.29 (0.59) 14.34 (0.59) 0.95 (0.41) 0.11, 1.79 0.028 

(13.11 - 17.83) (11.81 - 17.99) (-1.31 - 4.57) 

Amino Acid (% dwt)Alanine 1.76 (0.018) 1.80 (0.018) -0.037 (0.017) -0.075, 0.0018 0.059 

(1.66 - 1.83) (1.69 - 1.90) (-0.16 - 0.042) 

Arginine 3.30 (0.069) 3.58 (0.069) -0.28 (0.078) -0.46, -0.10 0.006 

(3.09 - 3.50) (3.19 - 3.93) (-0.83 - 0.0059) 

Aspartic Acid 4.63 (0.044) 4.78 (0.044) -0.15 (0.050) -0.27, -0.037 0.016 

(4.44 - 4.80) (4.46 - 5.01) (-0.56 - 0.12) 

Cystine 0.61 (0.0049) 0.59 (0.0049) 0.018 (0.0046) 0.0085, 0.027 <0.00

(0.58 - 0.63) (0.56 - 0.62) (-0.0071 - 0.053) 
Monsanto Company 10-SY-210U 

Table VI-2 (continued). Statistical Summary of Combined-Site Soybean Seed Nutrients for MON Conventional Control 

Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 877082 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Amino Acid (% dwt)Glutamic Acid 7.38 (0.085) 7.69 (0.085) -0.31 (0.093) -0.53, -0.095 0.010 

(7.05 - 7.73) (7.12 - 8.14) (-1.09 - 0.17) 

Glycine 1.76 (0.016) 1.81 (0.016) -0.048 (0.017) -0.086, -0.0096 0.020 

(1.67 - 1.83) (1.70 - 1.89) (-0.20 - 0.042) 

Histidine 1.06 (0.0095) 1.09 (0.0095) -0.033 (0.011) -0.059, -0.0076 0.017 

(1.02 - 1.10) (1.02 - 1.14) (-0.12 - 0.031) 

Isoleucine 1.88 (0.019) 1.95 (0.019) -0.070 (0.019) -0.11, -0.026 0.006 

(1.75 - 1.97) (1.79 - 2.04) (-0.24 - 0.11) 

Leucine 3.06 (0.029) 3.17 (0.029) -0.11 (0.031) -0.18, -0.035 0.008 

(2.93 - 3.19) (2.96 - 3.32) (-0.36 - 0.072) 

Lysine 2.64 (0.019) 2.68 (0.019) -0.041 (0.023) -0.094, 0.012 0.110 

(2.53 - 2.71) (2.54 - 2.77) (-0.23 - 0.090) 
Monsanto Company 10-SY-210U 

Table VI-2 (continued). Statistical Summary of Combined-Site Soybean Seed Nutrients for MON Conventional Control 

Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 877082 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Amino Acid (% dwt)Methionine 0.58 (0.0053) 0.58 (0.0053) 0.00012 (0.0062) -0.013, 0.013 0.985 

(0.53 - 0.60) (0.53 - 0.60) (-0.039 - 0.071) 

Phenylalanine 2.06 (0.028) 2.13 (0.028) -0.071 (0.028) -0.13, -0.0067 0.034 

(1.92 - 2.18) (1.95 - 2.27) (-0.27 - 0.048) 

Proline 1.99 (0.021) 2.05 (0.021) -0.067 (0.022) -0.12, -0.015 0.017 

(1.90 - 2.09) (1.89 - 2.13) (-0.17 - 0.065) 

Serine 2.04 (0.023) 2.09 (0.023) -0.048 (0.026) -0.11, 0.013 0.105 

(1.92 - 2.12) (1.95 - 2.21) (-0.19 - 0.054) 

Threonine 1.56 (0.015) 1.58 (0.015) -0.023 (0.015) -0.058, 0.012 0.169 

(1.48 - 1.62) (1.51 - 1.64) (-0.10 - 0.052) 

Tryptophan 0.47 (0.0085) 0.46 (0.0085) 0.0070 (0.0097) -0.015, 0.029 0.494 

(0.44 - 0.53) (0.43 - 0.50) (-0.035 - 0.064) 
Monsanto Company 10-SY-210U 

Table VI-2 (continued). Statistical Summary of Combined-Site Soybean Seed Nutrients for MON Conventional Control 

Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 877082 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Amino Acid (% dwt)Tyrosine 1.37 (0.018) 1.42 (0.018) -0.049 (0.021) -0.098, -0.00046 0.048 

(1.28 - 1.46) (1.34 - 1.52) (-0.20 - 0.078) 

Valine 1.98 (0.020) 2.06 (0.020) -0.080 (0.022) -0.13, -0.030 0.006 

(1.82 - 2.09) (1.90 - 2.17) (-0.27 - 0.13) 

Fatty Acid (% Total FA)16:0 Palmitic 11.59 (0.16) 11.33 (0.16) 0.26 (0.060) 0.12, 0.40 0.002 

(11.25 - 12.16) (10.92 - 12.08) (-0.15 - 0.62) 

18:0 Stearic 4.06 (0.10) 4.04 (0.10) 0.028 (0.049) -0.085, 0.14 0.584 

(3.60 - 4.40) (3.67 - 4.31) (-0.19 - 0.42) 

18:1 Oleic 19.20 (0.30) 20.91 (0.30) -1.71 (0.19) -2.15, -1.27 <0.00

(17.85 - 19.94) (19.60 - 22.44) (-2.71 - -0.90) 

18:2 Linoleic 54.40 (0.37) 53.59 (0.37) 0.81 (0.24) 0.25, 1.37 0.010 

(53.42 - 55.67) (52.33 - 54.99) (-0.59 - 1.68) 
Monsanto Company 10-SY-210U 

Table VI-2 (continued). Statistical Summary of Combined-Site Soybean Seed Nutrients for MON Conventional Control 

Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 877082 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Fatty Acid (% Total FA)18:3 Linolenic 10.12 (0.27) 9.49 (0.27) 0.63 (0.072) 0.46, 0.80 <0.00

(8.99 - 10.88) (8.42 - 10.14) (0.36 - 1.20) 

20:0 Arachidic 0.26 (0.0052) 0.26 (0.0052) -0.0012 (0.0031) -0.0082, 0.0059 0.707 

(0.23 - 0.27) (0.24 - 0.27) (-0.013 - 0.020) 

20:1 Eicosenoic 0.093 (0.017) 0.090 (0.017) 0.0029 (0.0042) -0.0056, 0.011 0.495 

(0.069 - 0.16) (0.068 - 0.17) (-0.010 - 0.050) 

22:0 Behenic 0.27 (0.0038) 0.28 (0.0038) -0.013 (0.0029) -0.020, -0.0066 0.001 

(0.25 - 0.29) (0.27 - 0.30) (-0.023 - 0.0024) 

Vitamin (mg/100g dwt)Vitamin E 1.41 (0.18) 1.23 (0.18) 0.19 (0.038) 0.098, 0.27 0.001 

(1.08 - 2.17) (0.89 - 2.11) (0.018 - 0.42) 

1dwt = dry weight; fwt = fresh weight; FA = fatty acid. 2MON 87708 was treated with dicamba. 3Mean (S.E.) = least-square mean (standard error). 4Control refers to the near isogenic conventional soybean control A3525. 5With 95% confidence, interval contains 99% of the values expressed in the population of commercial reference varieties. Negative limits set to zero.
Monsanto Company 10-SY-210U 

Table VI-3. Statistical Summary of Combined-Site Soybean Seed Anti-Nutrients for MON 87708 (TreaControl 

Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 877082 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Anti-nutrientLectin (H.U./mg dwt) 3.17 (0.76) 3.16 (0.76) 0.013 (0.67) -1.54, 1.57 0.984 

(0.59 - 10.27) (0.46 - 10.38) (-4.27 - 8.13) 

Phytic Acid (% dwt) 1.30 (0.071) 1.39 (0.071) -0.085 (0.035) -0.17, -0.0034 0.043 

(1.08 - 1.51) (1.09 - 1.62) (-0.29 - 0.15) 

Raffinose (% dwt) 0.43 (0.038) 0.47 (0.038) -0.036 (0.018) -0.072, -0.00077 0.045 

(0.32 - 0.59) (0.36 - 0.60) (-0.24 - 0.069) 

Stachyose (% dwt) 3.36 (0.078) 3.62 (0.078) -0.26 (0.099) -0.46, -0.062 0.011 

(3.07 - 4.02) (3.07 - 4.15) (-1.00 - 0.40) 

Trypsin Inhibitor (TIU/mg dwt) 32.27 (1.40) 30.37 (1.40) 1.90 (1.79) -2.23, 6.04 0.319 

(26.09 - 39.27) (25.22 - 34.22) (-4.76 - 8.72) 

Isoflavone (μg/g dwt)Daidzein 1494.97 (155.94) 1340.71 (155.94) 154.26 (65.62) 2.95, 305.57 0.046 

(899.83 - 2305.26) (762.49 - 1729.91) (-258.27 - 795.19) 
Monsanto Company 10-SY-210U 

Table VI-3 (continued). Statistical Summary of Combined-Site Soybean Seed Anti-Nutrients for MON Conventional Control 

Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 877082 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Isoflavone (μg/g dwt)Genistein 967.01 (90.36) 886.57 (90.36) 80.44 (41.86) -4.30, 165.19 0.062 

(594.13 - 1496.78) (588.17 - 1162.01) (-185.98 - 513.56) 

Glycitein 108.01 (5.24) 95.85 (5.24) 12.16 (6.91) -3.77, 28.09 0.116 

(77.67 - 119.09) (68.68 - 122.09) (-43.86 - 50.41) 

1dwt = dry weight; H.U. = Hemagglutinating Units; TIU = Trypsin Inhibitor Units. 2MON 87708 was treated with dicamba. 3Mean (S.E.) = least-square mean (standard error). 4Control refers to the near isogenic conventional soybean control A3525. 5With 95% confidence, interval contains 99% of the values expressed in the population of commercial reference varieties. Negative limits set to zero.
Monsanto Company 10-SY-210U 

Table VI-4. Statistical Summary of Combined-Site Soybean Forage Nutrients for MON 87708 (TreatControl 

Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 877082 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95%

Confidence Interval

Significa

(p-Valu

Proximate (% dwt)Ash 7.29 (0.54) 7.39 (0.54) -0.10 (0.27) -0.71, 0.51 0.712 

(5.94 - 9.65) (6.10 - 10.46) (-0.89 - 1.56) 

Carbohydrates 66.48 (1.03) 65.66 (1.04) 0.83 (0.96) -1.40, 3.05 0.414 

(62.21 - 73.31) (62.91 - 67.94) (-3.95 - 6.90) 

Moisture (% fwt) 75.63 (1.82) 75.55 (1.82) 0.081 (0.27) -0.55, 0.71 0.775 

(72.40 - 82.80) (71.60 - 82.70) (-1.40 - 1.30) 

Protein 21.52 (0.95) 22.32 (0.95) -0.80 (0.80) -2.67, 1.07 0.350 

(15.23 - 25.52) (20.88 - 24.11) (-6.26 - 2.75) 

Total Fat 4.67 (0.66) 4.64 (0.66) 0.032 (0.26) -0.57, 0.63 0.904 

(2.00 - 7.34) (2.01 - 6.72) (-0.68 - 1.96) 

Fiber (% dwt)Acid Detergent Fiber 30.58 (1.79) 27.69 (1.80) 2.89 (1.19) 0.45, 5.34 0.021 

(23.30 - 45.11) (21.79 - 38.15) (-4.78 - 16.24) 
Monsanto Company 10-SY-210U 

Table VI-4 (continued). Statistical Summary of Combined-Site Soybean Forage Nutrients for MON Conventional Control 

Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 877082 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95%

Confidence Interval

Significa

(p-Valu

Fiber (% dwt)Neutral Detergent Fiber 29.63 (1.68) 30.49 (1.70) -0.86 (1.22) -3.65, 1.94 0.503 

(24.21 - 38.51) (23.66 - 39.42) (-8.13 - 11.03) 

1dwt = dry weight; fwt = fresh weight. 2MON 87708 was treated with dicamba. 3Mean (S.E.) = least-square mean (standard error). 4Control refers to the near isogenic conventional soybean control A3525. 5With 95% confidence, interval contains 99% of the values expressed in the population of commercial reference varieties. Negative limits set to zero.
Monsanto Company 10-SY-210U 

Table VI-5. Summary of Differences (

α=0.05) for the Comparison of Soybean Component Levels for MON 

vs. Conventional Control 

 Mean Difference 

(MON 87708 minus Control)

Analytical Component (Units)1

MON 877082

Mean3

Control4

Mean

Mean Difference 

(% of Control)

Significance

(p-Value)

MON 877

Range

Statistical Differences Observed in Combined-Site AnalysisSeed Proximate (% dw)Protein 41.17 42.41 -2.94 0.040 39.96 - 43Seed Fiber (% dw)Acid Detergent Fiber 13.38 12.86 3.99 0.046 11.01 - 15Neutral Detergent Fiber 15.24 14.34 6.33 0.035 12.91 - 18Seed Amino Acid (% dw)Arginine 3.37 3.58 -5.88 0.026 3.12 - 3.6Cystine 0.61 0.59 3.27 <0.001 0.59 - 0.6Glutamic Acid 7.46 7.69 -3.03 0.037 7.25 - 7.8Isoleucine 1.90 1.95 -2.42 0.039 1.80 - 2.0
Monsanto Company 10-SY-210U 

Table VI-5 (continued). Summary of Differences (

α=0.05) for the Comparison of Soybean Component Levels fo

(Untreated) vs. Conventional Control 

 Mean Difference 

(MON 87708 minus Control)

Analytical Component (Units)1

MON 877082

Mean3

Control4

Mean

Mean Difference 

(% of Control)

Significance

(p-Value)

MON 877

Range

Statistical Differences Observed in Combined-Site AnalysisSeed Amino Acid (% dw)Leucine 3.10 3.17 -2.27 0.049 3.01 - 3.2Phenylalanine 2.06 2.13 -3.04 0.048 1.98 - 2.1Proline 1.98 2.05 -3.39 0.014 1.89 - 2.0Valine 2.00 2.06 -2.76 0.030 1.90 - 2.1Seed Fatty Acid (% Total FA)16:0 Palmitic 11.60 11.33 2.37 0.002 11.02 - 1218:1 Oleic 19.32 20.91 -7.60 <0.001 18.26 - 2018:2 Linoleic 54.37 53.59 1.45 0.012 52.18 - 5518:3 Linolenic 10.04 9.49 5.78 <0.001 8.94 - 10.
Monsanto Company 10-SY-210U 

Table VI-5 (continued). Summary of Differences (

α=0.05) for the Comparison of Soybean Component Levels f

(Untreated) vs. Conventional Control (continued) 

 Mean Difference 

(MON 87708 minus Control)

Analytical Component (Units)1

MON 877082

Mean3

Control4

Mean

Mean Difference 

(% of Control)

Significance

(p-Value)

MON 877

Range

Statistical Differences Observed in Combined-Site AnalysisSeed Fatty Acid (% Total FA)22:0 Behenic 0.27 0.28 -3.71 0.006 0.25 - 0.2Seed Vitamin (mg/100g dw)Vitamin E 1.45 1.23 18.16 <0.001 1.11 - 2.2Seed Anti-nutrientTrypsin Inhibitor (TIU/mg dw) 35.03 30.37 15.37 0.031 23.32 - 51Seed Isoflavone (μg/g dw)Daidzein 1571.79 1340.71 17.24 0.007 910.73 - 229Genistein 989.28 886.57 11.59 0.018 654.16 - 146Statistical Differences Observed in More than One Individual SiteSeed Fatty Acid (% Total FA)18:1 Oleic Site IARL 19.28 21.67 -11.02 0.001 18.77 - 19
Monsanto Company 10-SY-210U 

Table VI-5 (continued). Summary of Differences (

α=0.05) for the Comparison of Soybean Component Levels f

(Untreated) vs. Conventional Control 

 Mean Difference 

(MON 87708minus Control)

Analytical Component (Units)1

MON 877082

Mean3

Control4

Mean

Mean Difference 

(% of Control)

Significance

(p-Value)

MON 877

Range

Statistical Differences Observed in More than One Individual SiteSeed Fatty Acid (% Total FA)18:1 Oleic Site ILCY 19.85 21.57 -7.98 0.013 19.62 - 2018:1 Oleic Site INRC 18.80 20.19 -6.90 <0.001 18.63 - 1818:1 Oleic Site PAHM 18.45 20.01 -7.78 0.011 18.26 - 1818:3 Linolenic Site ILCY 9.09 8.58 5.96 0.006 8.94 - 9.218:3 Linolenic Site ILWY 10.65 10.05 5.95 0.014 10.37 - 1018:3 Linolenic Site INRC 10.05 9.31 7.94 <0.001 10.00 - 1018:3 Linolenic Site PAHM 10.03 9.47 5.87 0.026 9.74 - 10.Seed Vitamin (mg/100g dw)Vitamin E Site IARL 1.26 0.94 33.64 0.008 1.11 - 1.4
Monsanto Company 10-SY-210U 

Table VI-5 (continued). Summary of Differences (

α=0.05) for the Comparison of Soybean Component Levels f

(Untreated) vs. Conventional Control 

 Mean Difference 

(MON 87708 minus Control)

Analytical Component (Units)1

MON 877082

Mean3

Control4

Mean

Mean Difference 

(% of Control)

Significance

(p-Value)

MON 877

Range

Statistical Differences Observed in More than One Individual SiteSeed Vitamin (mg/100g dw)Vitamin E Site ILCY 2.21 1.86 18.85 0.016 2.11 - 2.2Vitamin E Site ILWY 1.15 0.94 21.98 0.017 1.11 - 1.2Vitamin E Site PAHM 1.37 1.23 12.00 0.002 1.26 - 1.5Seed Amino Acid (% dw)Phenylalanine Site ILCY 2.07 2.13 -3.03 0.024 2.05 - 2.0Phenylalanine Site INRC 2.08 2.20 -5.54 0.037 2.03 - 2.1Phenylalanine Site PAHM 2.08 2.21 -6.07 0.025 2.03 - 2.1Proline Site ILCY 1.95 2.06 -5.34 0.004 1.90 - 1.9Proline Site INRC 2.01 2.06 -2.63 0.020 1.99 - 2.0
Monsanto Company 10-SY-210U 

Table VI-5 (continued). Summary of Differences (

α=0.05) for the Comparison of Soybean Component Levels f

(Untreated) vs. Conventional Control 

 Mean Difference 

(MON 87708 minus Control)

Analytical Component (Units)1

MON 877082

Mean3

Control4

Mean

Mean Difference 

(% of Control)

Significance

(p-Value)

MON 877

Range

Statistical Differences Observed in More than One Individual SiteSeed Amino Acid (% dw)Proline Site PAHM 2.00 2.10 -5.09 0.028 1.93 - 2.0Seed Fatty Acid (% Total FA)18:2 Linoleic Site IARL 54.30 52.70 3.04 0.025 53.70 - 5518:2 Linoleic Site ILCY 54.31 53.26 1.97 0.040 53.67 - 5418:2 Linoleic Site INRC 54.94 54.43 0.93 0.024 54.79 - 5522:0 Behenic Site IARL 0.26 0.28 -4.96 0.030 0.26 - 0.222:0 Behenic Site INRC 0.28 0.29 -5.54 0.025 0.27 - 0.222:0 Behenic Site PAHM 0.26 0.27 -4.78 0.018 0.25 - 0.2
Monsanto Company 10-SY-210U 

Table VI-5 (continued). Summary of Differences (

α=0.05) for the Comparison of Soybean Component Levels f

(Untreated) vs. Conventional Control 

 Mean Difference 

(MON 87708 minus Control)

Analytical Component (Units)1

MON 877082

Mean3

Control4

Mean

Mean Difference 

(% of Control)

Significance

(p-Value)

MON 877

Range

Statistical Differences Observed in More than One Individual SiteSeed Proximate (% dw)Moisture (% fw) Site IARL 7.84 6.07 29.21 0.010 7.39 - 8.7Moisture (% fw) Site PAHM 9.53 10.50 -9.24 0.011 9.21 - 9.9Protein Site INRC 41.47 43.58 -4.85 0.049 40.22 - 43Protein Site PAHM 40.38 43.69 -7.57 0.002 39.96 - 40Seed Amino Acid (% dw)Arginine Site INRC 3.40 3.72 -8.50 0.007 3.35 - 3.5Arginine Site PAHM 3.39 3.88 -12.75 0.002 3.35 - 3.4Glutamic Acid Site ILCY 7.44 7.61 -2.24 0.039 7.33 - 7.5Glutamic Acid Site PAHM 7.42 8.00 -7.24 0.007 7.32 - 7.5
Monsanto Company 10-SY-210U 

Table VI-5 (continued). Summary of Differences (

α=0.05) for the Comparison of Soybean Component Levels f

(Untreated) vs. Conventional Control 

 Mean Difference 

(MON 87708 minus Control)

Analytical Component (Units)1

MON 877082

Mean3

Control4

Mean

Mean Difference 

(% of Control)

Significance

(p-Value)

MON 877

Range

Statistical Differences Observed in More than One Individual SiteSeed Amino Acid (% dw)Leucine Site ILCY 3.11 3.17 -1.82 0.008 3.08 - 3.1Leucine Site PAHM 3.10 3.28 -5.46 0.009 3.08 - 3.1Lysine Site ILWY 2.68 2.63 1.76 0.006 2.66 - 2.6Lysine Site PAHM 2.64 2.75 -4.02 0.025 2.62 - 2.6Seed Isoflavone (μg/g dw)Glycitein Site ILWY 96.96 79.70 21.65 0.001 92.19 - 103Glycitein Site INRC 119.27 98.42 21.19 0.003 113.04 - 12Statistical Differences Observed in One Individual SiteSeed Amino Acid (% dw)Alanine Site PAHM 1.79 1.86 -3.65 0.046 1.76 - 1.8
Monsanto Company 10-SY-210U 

Table VI-5 (continued). Summary of Differences (

α=0.05) for the Comparison of Soybean Component Levels f

(Untreated) vs. Conventional Control 

 Mean Difference 

(MON 87708 minus Control)

Analytical Component (Units)1

MON 877082

Mean3

Control4

Mean

Mean Difference 

(% of Control)

Significance

(p-Value)

MON 877

Range

Statistical Differences Observed in One Individual SiteSeed Amino Acid (% dw)Aspartic Acid Site PAHM 4.66 4.94 -5.62 0.008 4.64 - 4.6Cystine Site INRC 0.62 0.59 5.30 0.029 0.60 - 0.6Glycine Site PAHM 1.77 1.86 -4.84 0.014 1.76 - 1.7Histidine Site PAHM 1.07 1.13 -5.37 0.008 1.07 - 1.0Isoleucine Site PAHM 1.89 2.00 -5.67 0.035 1.88 - 1.9Serine Site ILWY 2.10 2.06 2.11 0.026 2.09 - 2.1Threonine Site ILWY 1.59 1.55 2.85 <0.001 1.58 - 1.6Valine Site PAHM 2.00 2.13 -5.93 0.035 2.00 - 2.0
Monsanto Company 10-SY-210U 

Table VI-5 (continued). Summary of Differences (

α=0.05) for the Comparison of Soybean Component Levels f

(Untreated) vs. Conventional Control 

 Mean Difference 

(MON 87708 minus Control)

Analytical Component (Units)1

MON 877082

Mean3

Control4

Mean

Mean Difference 

(% of Control)

Significance

(p-Value)

MON 877

Range

Statistical Differences Observed in One Individual SiteSeed Fatty Acid (% Total FA)16:0 Palmitic Site IARL 11.51 11.00 4.67 0.001 11.39 - 1120:1 Eicosenoic Site ILCY 0.15 0.16 -4.82 0.028 0.15 - 0.1Seed Anti-nutrientTrypsin Inhibitor (TIU/mg dw) Site ILWY 38.93 29.73 30.95 0.033 37.89 - 39Forage Proximate (% dw)Ash Site INRC 6.20 6.95 -10.70 0.047 5.87 - 6.7Moisture (% fw) Site IARL 83.47 81.97 1.83 0.027 82.70 - 84Protein Site IARL 25.76 23.00 12.02 0.022 24.63 - 271dw = dry weight; fw = fresh weight; FA = fatty acid; TIU = Trypsin Inhibitor Units. 2 MON 87708 plants were not sprayed with dicamba, but received another conventional treatment as was done for the conve3Mean = least-square mean. 4Control refers to the non-biotechnology derived, conventional control (A3525). 5With 95% confidence, interval contains 99% of the values expressed in the population of conventional substances. Negativ
Monsanto Company 10-SY-210U 

Table VI-6. Statistical Summary of Combined-Site Soybean Seed Nutrients for MON 87708 (Untreated) vControl 

 Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 87708 2 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Proximate (% dw)Ash 5.22 (0.067) 5.12 (0.067) 0.10 (0.055) -0.010, 0.21 0.073 

(4.89 - 5.51) (4.73 - 5.47) (-0.28 - 0.55) 

Carbohydrates 37.30 (0.50) 36.64 (0.50) 0.66 (0.40) -0.26, 1.58 0.138 

(35.27 - 39.79) (34.11 - 38.45) (-1.89 - 4.52) 

Moisture (% fw) 7.30 (0.65) 7.14 (0.65) 0.16 (0.52) -1.04, 1.36 0.771 

(5.68 - 9.91) (5.79 - 10.60) (-1.19 - 2.89) 

Protein 41.17 (0.39) 42.41 (0.39) -1.25 (0.51) -2.42, -0.069 0.040 

(39.96 - 43.06) (40.69 - 43.85) (-3.89 - 0.47) 

Total Fat 16.32 (0.59) 15.84 (0.59) 0.48 (0.31) -0.23, 1.19 0.155 

(13.95 - 18.66) (14.40 - 18.39) (-1.73 - 1.70) 

Fiber (% dw)Acid Detergent Fiber 13.38 (0.40) 12.86 (0.40) 0.51 (0.25) 0.0084, 1.02 0.046 

(11.01 - 15.72) (11.62 - 14.57) (-0.72 - 2.45) 
Monsanto Company 10-SY-210U 

Table VI-6 (continued). Statistical Summary of Combined-Site Soybean Seed Nutrients for MON 87708 (UntrConventional Control 

 Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 877082 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Fiber (% dw)Crude Fiber 7.50 (0.26) 7.37 (0.26) 0.13 (0.26) -0.39, 0.64 0.620 

(6.14 - 8.89) (6.05 - 8.64) (-1.33 - 1.19) 

Neutral Detergent Fiber 15.24 (0.59) 14.34 (0.59) 0.91 (0.41) 0.064, 1.75 0.035 

(12.91 - 18.38) (11.81 - 17.99) (-2.27 - 4.17) 

Amino Acid (% dw)Alanine 1.78 (0.018) 1.80 (0.018) -0.025 (0.017) -0.063, 0.014 0.174 

(1.72 - 1.84) (1.69 - 1.90) (-0.10 - 0.043) 

Arginine 3.37 (0.069) 3.58 (0.069) -0.21 (0.078) -0.39, -0.031 0.026 

(3.12 - 3.60) (3.19 - 3.93) (-0.55 - 0.051) 

Aspartic Acid 4.68 (0.044) 4.78 (0.044) -0.10 (0.050) -0.22, 0.014 0.078 

(4.57 - 4.90) (4.46 - 5.01) (-0.35 - 0.10) 

Cystine 0.61 (0.0049) 0.59 (0.0049) 0.019 (0.0046) 0.010, 0.029 <0.00

(0.59 - 0.64) (0.56 - 0.62) (-0.0068 - 0.047) 
Monsanto Company 10-SY-210U 

Table VI-6 (continued). Statistical Summary of Combined-Site Soybean Seed Nutrients for MON 87708 (UntrConventional Control 

 Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 87708 2 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Amino Acid (% dw)Glutamic Acid 7.46 (0.085) 7.69 (0.085) -0.23 (0.093) -0.45, -0.018 0.037 

(7.25 - 7.88) (7.12 - 8.14) (-0.70 - 0.12) 

Glycine 1.77 (0.016) 1.81 (0.016) -0.034 (0.017) -0.073, 0.0042 0.074 

(1.74 - 1.85) (1.70 - 1.89) (-0.13 - 0.041) 

Histidine 1.07 (0.0095) 1.09 (0.0095) -0.021 (0.011) -0.047, 0.0044 0.092 

(1.05 - 1.11) (1.02 - 1.14) (-0.073 - 0.029) 

Isoleucine 1.90 (0.019) 1.95 (0.019) -0.047 (0.019) -0.091, -0.0028 0.039 

(1.80 - 2.02) (1.79 - 2.04) (-0.22 - 0.085) 

Leucine 3.10 (0.029) 3.17 (0.029) -0.072 (0.031) -0.14, -0.00024 0.049 

(3.01 - 3.24) (2.96 - 3.32) (-0.23 - 0.055) 

Lysine 2.65 (0.019) 2.68 (0.019) -0.026 (0.023) -0.079, 0.027 0.295 

(2.60 - 2.76) (2.54 - 2.77) (-0.13 - 0.083) 
Monsanto Company 10-SY-210U 

Table VI-6 (continued). Statistical Summary of Combined-Site Soybean Seed Nutrients for MON 87708 (UntrConventional Control 

 Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 87708 2 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Amino Acid (% dw)Methionine 0.58 (0.0053) 0.58 (0.0053) 0.0013 (0.0062) -0.011, 0.014 0.834 

(0.55 - 0.61) (0.53 - 0.60) (-0.036 - 0.056) 

Phenylalanine 2.06 (0.028) 2.13 (0.028) -0.065 (0.028) -0.13, -0.00067 0.048 

(1.98 - 2.14) (1.95 - 2.27) (-0.18 - 0.023) 

Proline 1.98 (0.021) 2.05 (0.021) -0.070 (0.022) -0.12, -0.018 0.014 

(1.89 - 2.05) (1.89 - 2.13) (-0.17 - 0.043) 

Serine 2.07 (0.023) 2.09 (0.023) -0.021 (0.026) -0.081, 0.039 0.449 

(1.91 - 2.11) (1.95 - 2.21) (-0.19 - 0.073) 

Threonine 1.57 (0.015) 1.58 (0.015) -0.0093 (0.015) -0.044, 0.026 0.556 

(1.49 - 1.61) (1.51 - 1.64) (-0.088 - 0.052) 

Tryptophan 0.47 (0.0085) 0.46 (0.0085) 0.0095 (0.0097) -0.013, 0.032 0.359 

(0.41 - 0.50) (0.43 - 0.50) (-0.053 - 0.049) 
Monsanto Company 10-SY-210U 

Table VI-6 (continued). Statistical Summary of Combined-Site Soybean Seed Nutrients for MON 87708 (UntrConventional Control 

 Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 87708 2 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Amino Acid (% dw)Tyrosine 1.41 (0.018) 1.42 (0.018) -0.015 (0.021) -0.064, 0.034 0.506 

(1.35 - 1.48) (1.34 - 1.52) (-0.086 - 0.098) 

Valine 2.00 (0.020) 2.06 (0.020) -0.057 (0.022) -0.11, -0.0069 0.030 

(1.90 - 2.13) (1.90 - 2.17) (-0.25 - 0.097) 

Fatty Acid (% Total FA)16:0 Palmitic 11.60 (0.16) 11.33 (0.16) 0.27 (0.060) 0.13, 0.41 0.002 

(11.02 - 12.15) (10.92 - 12.08) (-0.094 - 0.59) 

18:0 Stearic 4.04 (0.10) 4.04 (0.10) 0.0057 (0.049) -0.11, 0.12 0.909 

(3.55 - 4.57) (3.67 - 4.31) (-0.23 - 0.38) 

18:1 Oleic 19.32 (0.30) 20.91 (0.30) -1.59 (0.19) -2.03, -1.15 <0.00

(18.26 - 20.73) (19.60 - 22.44) (-2.82 - -0.045) 

18:2 Linoleic 54.37 (0.37) 53.59 (0.37) 0.78 (0.24) 0.22, 1.34 0.012 

(52.18 - 55.62) (52.33 - 54.99) (-0.98 - 2.61) 
Monsanto Company 10-SY-210U 

Table VI-6 (continued). Statistical Summary of Combined-Site Soybean Seed Nutrients for MON 87708 (UntrConventional Control 

 Difference (MON 87708 minus Control)

Analytical Compoent (Units)1

MON 87708 2 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Fatty Acid (% Total FA)18:3 Linolenic 10.04 (0.27) 9.49 (0.27) 0.55 (0.072) 0.38, 0.72 <0.00

(8.94 - 10.90) (8.42 - 10.14) (-0.18 - 1.01) 

20:0 Arachidic 0.26 (0.0052) 0.26 (0.0052) -0.0013 (0.0031) -0.0083, 0.0058 0.684 

(0.23 - 0.29) (0.24 - 0.27) (-0.014 - 0.022) 

20:1 Eicosenoic 0.092 (0.017) 0.090 (0.017) 0.0011 (0.0042) -0.0074, 0.0096 0.795 

(0.065 - 0.16) (0.068 - 0.17) (-0.013 - 0.058) 

22:0 Behenic 0.27 (0.0038) 0.28 (0.0038) -0.010 (0.0029) -0.017, -0.0038 0.006 

(0.25 - 0.29) (0.27 - 0.30) (-0.023 - 0.0054) 

Vitamin (mg/100g dw)Vitamin E 1.45 (0.18) 1.23 (0.18) 0.22 (0.038) 0.14, 0.31 <0.00

(1.11 - 2.27) (0.89 - 2.11) (0.0086 - 0.49) 

1dw = dry weight; fw = fresh weight; FA = fatty acid. 2MON 87708 plants were not sprayed with dicamba, but received another conventional treatment as was done for the conve3Mean (S.E.) = least-square mean (standard error). 4Control refers to the non-biotechnology derived, conventional control (A3525). 5With 95% confidence, interval contains 99% of the values expressed in the population of conventional substances. Negativ
Monsanto Company 10-SY-210U 

Table VI-7. Statistical Summary of Combined-Site Soybean Seed Anti-Nutrients for MON 87708 (UntreaControl 

 Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 87708 2 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Anti-nutrientLectin (H.U./mg dw) 3.05 (0.76) 3.16 (0.76) -0.12 (0.67) -1.67, 1.44 0.867 

(1.18 - 6.35) (0.46 - 10.38) (-7.83 - 3.07) 

Phytic Acid (% dw) 1.33 (0.071) 1.39 (0.071) -0.060 (0.035) -0.14, 0.022 0.129 

(1.05 - 1.48) (1.09 - 1.62) (-0.21 - 0.26) 

Raffinose (% dw) 0.46 (0.038) 0.47 (0.038) -0.0069 (0.018) -0.042, 0.029 0.697 

(0.34 - 0.58) (0.36 - 0.60) (-0.065 - 0.056) 

Stachyose (% dw) 3.48 (0.078) 3.62 (0.078) -0.15 (0.099) -0.35, 0.054 0.147 

(2.94 - 3.85) (3.07 - 4.15) (-0.82 - 0.53) 

Trypsin Inhibitor (TIU/mg dw) 35.03 (1.40) 30.37 (1.40) 4.67 (1.79) 0.53, 8.80 0.031 

(23.32 - 51.50) (25.22 - 34.22) (-2.94 - 18.17) 

Isoflavone (μg/g dw)Daidzein 1571.79 (155.94) 1340.71 (155.94) 231.08 (65.62) 79.76, 382.39 0.007 

(910.73 - 2297.58) (762.49 - 1729.91) (-187.35 - 691.83) 
Monsanto Company 10-SY-210U 

Table VI-7 (continued). Statistical Summary of Combined-Site Soybean Seed Anti-Nutrients for MON 87708 (Conventional Control 

 Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 87708 2 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Isoflavone (μg/g dw)Genistein 989.28 (90.36) 886.57 (90.36) 102.71 (41.86) 17.96, 187.46 0.018 

(654.16 - 1469.13) (588.17 - 1162.01) (-116.63 - 400.27) 

Glycitein 110.40 (5.24) 95.85 (5.24) 14.55 (6.91) -1.38, 30.48 0.068 

(83.25 - 133.73) (68.68 - 122.09) (-2.04 - 37.48) 

1dw = dry weight; H.U. = Hemagglutinating Units; TIU = Trypsin Inhibitor Units. 2MON 87708 plants were not sprayed with dicamba, but received another conventional treatment as was done for the conve3Mean (S.E.) = least-square mean (standard error). 4Control refers to the non-biotechnology derived, conventional control (A3525). 5With 95% confidence, interval contains 99% of the values expressed in the population of conventional substances. Negativ
Monsanto Company 10-SY-210U 

Table VI-8. Statistical Summary of Combined-Site Soybean Forage Nutrients for MON 87708 (UntreatedControl 

 Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 877082 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Proximate (% dw)Ash 7.01 (0.54) 7.39 (0.54) -0.39 (0.27) -0.99, 0.22 0.183 

(4.92 - 9.45) (6.10 - 10.46) (-1.57 - 0.97) 

Carbohydrates 65.55 (1.03) 65.66 (1.04) -0.11 (0.96) -2.33, 2.12 0.913 

(61.64 - 71.05) (62.91 - 67.94) (-6.11 - 4.64) 

Moisture (% fw) 75.81 (1.82) 75.55 (1.82) 0.25 (0.27) -0.37, 0.88 0.379 

(72.30 - 84.10) (71.60 - 82.70) (-1.00 - 2.70) 

Protein 22.70 (0.95) 22.32 (0.95) 0.38 (0.80) -1.49, 2.25 0.648 

(16.28 - 27.04) (20.88 - 24.11) (-5.21 - 4.89) 

Total Fat 4.70 (0.66) 4.64 (0.66) 0.055 (0.26) -0.55, 0.66 0.838 

(2.61 - 6.52) (2.01 - 6.72) (-1.38 - 1.63) 

Fiber (% dw)Acid Detergent Fiber 27.72 (1.79) 27.69 (1.80) 0.036 (1.19) -2.41, 2.48 0.976 

(23.32 - 34.63) (21.79 - 38.15) (-7.68 - 4.38) 
Monsanto Company 10-SY-210U 

Table VI-8 (continued). Statistical Summary of Combined-Site Soybean Forage Nutrients for MON 87708 (UnConventional Control 

 Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 877082 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Fiber (% dw)Neutral Detergent Fiber 30.98 (1.68) 30.49 (1.70) 0.49 (1.22) -2.31, 3.29 0.698 

(25.38 - 37.80) (23.66 - 39.42) (-5.93 - 10.81) 

1dw = dry weight; fw = fresh weight. 2MON 87708 plants were not sprayed with dicamba, but received another conventional treatment as was done for the conve3Mean (S.E.) = least-square mean (standard error). 4Control refers to the non-biotechnology derived, conventional control (A3525). 5With 95% confidence, interval contains 99% of the values expressed in the population of conventional substances. Negativ
Monsanto Company 10-SY-210U 142 of 721 

Table VI-9. Literature and ILSI Database Ranges for Components in Soybean Seed 

and Forage 

Seed Tissue Components1 Literature Range2 ILSI Range3

Seed Nutrients 

Proximates (% dwt) 

Ash 4.61 – 6.32a

; 4.32 – 5.88b 3.89 – 6.99 

Carbohydrates by calculation 32.75 – 40.98a

; 29.88 – 43.48b 29.6 – 50.2 

Moisture (% fwt) 6.24 – 12.10a

; 5.44 – 11.70b 4.7 – 34.4 

Protein 34.78 – 43.35a; 32.29 – 42.66b 33.19 – 45.48 

Total Fat 14.40 – 20.91a

; 15.10 – 23.56b; 15.5c – 24.7c 8.10 – 23.56 

 

Fiber (% dwt) 

Acid Detergent Fiber 9.22 – 26.26a

; 11.81 – 19.45b 7.81 – 18.61 

Neutral Detergent Fiber 10.79 – 23.90a

; 13.32 – 23.57b 8.53 – 21.25 

 

Amino Acids (% dwt) 

Alanine 1.62 – 1.89a

; 1.43 – 1.93b 1.51 – 2.10 

Arginine 2.57 – 3.34a

; 2.15 – 3.05b 2.29 – 3.40 

Aspartic acid 4.16 – 5.02a

; 4.01 – 5.72b 3.81 – 5.12 

Cystine/Cysteine 0.52 – 0.69a

; 0.41 – 0.71b 0.37 – 0.81 

Glutamic acid 6.52 – 8.19a

; 5.49 – 8.72b 5.84 – 8.20 

Glycine 1.59 – 1.90a; 1.41 – 1.99b 1.46 – 2.00 

Histidine 0.96 – 1.13a

; 0.86 – 1.24b 0.88 – 1.18 

Isoleucine 1.59 – 2.00a

; 1.41 – 2.02b 1.54 – 2.08 

Leucine 2.79 – 3.42a; 2.39 – 3.32b 2.59 – 3.62 

Lysine 2.36 – 2.77a

; 2.19 – 3.15b 2.29 – 2.84 

Methionine 0.45 – 0.63a

; 0.39 – 0.65b 0.43 – 0.68 

Phenylalanine 1.82 – 2.29a; 1.62 – 2.44b 1.63 – 2.35 

Proline 1.83 – 2.23a

; 1.63 – 2.25b 1.69 – 2.28 

Serine 1.95 – 2.42a

; 1.51 – 2.30b 1.11 – 2.48 

Threonine 1.44 – 1.71a; 1.23 – 1.74b 1.14 – 1.86 

Tryptophan 0.30 – 0.48a

; 0.41 – 0.56b 0.36 – 0.50 

Tyrosine 1.27 – 1.53a; 0.74 – 1.31b 1.02 – 1.61 

Valine 1.68 – 2.11a; 1.50 – 2.13b 1.60 – 2.20 

 

Fatty Acids (% total FA) 

8:0 Caprylic not available 0.148 – 0.148 

10:0 Capric 0.15 – 0.27b

 not available 

12:0 Lauric not available 0.082 – 0.132 

14:0 Myristic 0.063 – 0.11b 0.071 – 0.238 

14:1 Myristoleic not available 0.121 – 0.125 

15:0 Pentadecanoic not available not available 

15:1 Pentadecenoic not available not available 

16:0 Palmitic 9.80 – 12.63b

 9.55 – 15.77 

16:1 Palmitoleic 0.055 – 0.14b

 0.086 – 0.194 

17:0 Heptadecanoic 0.076 – 0.13b

 0.085 – 0.146 

17:1 Heptadecenoic 0.019 – 0.064b

 0.073 – 0.087 

18:0 Stearic 3.21 – 5.63b

 2.70 – 5.88 

18:1 Oleic 16.69 – 35.16b

 14.3 – 32.2 

18:2 Linoleic 44.17 – 57.72b

 42.3 – 58.8 

18:3 Gamma Linolenic not available not available 

18:3 Linolenic 4.27 – 9.90b

 3.00 – 12.52 

 
Monsanto Company 10-SY-210U 143 of 721 

Table VI-9 (continued). Literature and ILSI Database Ranges for Components in 

Soybean Seed and Forage 

Seed Tissue Components1 Literature Range2 ILSI Range3

Seed Nutrients 

Fatty Acids (% total FA) 

20:0 Arachidic 0.35 – 0.57b

 0.163 – 0.482 

20:1 Eicosenoic 0.13 – 0.30b

 0.140 – 0.350 

20:2 Eicosadienoic 0.016 – 0.071b

 0.077 – 0.245 

20:3 Eicosatrienoic not available not available

20:4 Arachidonic not available not available

22:0 Behenic 0.35 – 0.59b

 0.277 – 0.595 

22:1 Erucic not available not available 

 

Vitamins (mg/100g dwt) 

Vitamin E 1.29 – 4.80a

; 1.12 – 8.08b 0.19 – 6.17 

Seed Anti-Nutrients 

Lectin (H.U./mg fwt) 0.45 – 10.87a

; 0.090 – 11.18b 0.09 – 8.46 

Trypsin Inhibitor 

 (TIU/mg dwt) 

20.79 – 59.03a

; 18.14 – 42.51b 19.59 – 118.68 

Phytic Acid (% dwt) 0.41 – 1.92a

; 0.81 – 2.66b 0.63 – 1.96 

Raffinose (% dwt) 0.26 – 0.84a

; 0.43 – 1.85b 0.21 – 0.66 

Stachyose (% dwt) 1.53 – 3.04a

; 1.97 – 6.65b 1.21 – 3.50 

Isoflavones (μg/g dwt) (mg/kg dwt)

Daidzein 224.03 – 1571.91a

; 198.95 – 1458.24b 60.0 – 2453.5 

Genistein 338.24 – 1488.89a; 148.06 – 1095.57b 144.3 – 2837.2 

Glycitein 52.72 – 298.57a

; 32.42 – 255.94b 15.3 – 310.4 

 

Forage Tissue Components1

 Literature Range2 ILSI Range3

Forage Nutrients 

Proximate (% dwt)

Ash 5.28 – 9.24a

; 4.77 – 8.54b 6.72 – 10.78 

Carbohydrates by calculation 62.25 – 72.30a

; 60.61 – 77.26b 59.8 – 74.7 

Moisture (% fwt) 68.50 – 78.40a

; 62.76 – 80.20b 73.5 – 81.6 

Protein 16.48 – 24.29a

; 12.68 – 23.29b 14.38 – 24.71 

Total Fat 2.654

 – 9.87a; 2.96 – 7.88b 1.302 – 5.132 

 

Fiber (% dwt) 

Acid Detergent Fiber 23.86 – 50.89a

; 25.49 – 47.33b not available 

Neutral Detergent Fiber 19.61 – 43.70a

; 30.96 – 54.55b not available 1

fwt = fresh weight; dwt = dry weight; H.U. = hemagglutinating unit; TIU = trypsin inhibitor unit. 

2

Literature range references: a(Lundry et al., 2008); b(Berman et al., 2009); c(OECD, 2001). 4 ILSI Crop Composition Database (2006). 
Monsanto Company 10-SY-210U 144 of 721 

VI.B. Compositional Assessment Conclusion 

Analyses of nutrient and anti-nutrient levels in both dicamba-treated and untreated 

MON 87708 and the near isogenic conventional control A3525 were conducted to assess 

compositional equivalence. The tissues analyzed included seed and forage harvested 

from plants grown at five field sites in the U.S. during the 2008 field season. The 

composition analysis, conducted in accordance with OECD guidelines, also included 

measurement of nutrients and anti-nutrients in the commercial reference varieties 

concurrently grown with MON 87708 to provide data on natural variability of each 

compositional component. All soybean plants including MON 87708, the conventional 

control, and the commercial reference varieties were treated with maintenance pesticides 

as necessary throughout the growing season. In addition, MON 87708 plots were either 

treated at the V2-V3 growth stage with dicamba herbicide at the maximum in-crop label 

rate (0.5 lb a.e./acre) or not treated with dicamba herbicide. 

For MON 87708 treated, the combined-site analysis of both seed and forage showed no 

statistically significant differences between MON 87708 and conventional control for 21 

(42.0%) of the 50 mean value comparisons. Of the statistically significant differences 

observed, one was from the forage analysis, and 28 were from the seed analysis. Nutrient 

component differences in seed included mean values for ash, carbohydrates by 

calculation, protein and 12 amino acids, five fatty acids, ADF, NDF, crude fiber, and 

vitamin E. In the combined-site analysis, all nutrient component differences in seed 

between MON 87708 and the conventional control were of small relative magnitude with 

respect to the conventional control and, whether increased or decreased, ranged from 

1.51% to 12.37% for the three proximates, amino acids, fatty acids, and fibers, and 

15.13% for vitamin E. Two of the nutrient components in the combined-site analysis 

(decreased levels of 18:1 oleic acid and increased levels of 18:3 linolenic acid) were also 

observed to be statistically different at all five individual sites, and one nutrient 

component (vitamin E) was observed to be increased at four of the five individual sites as 

in the combined-site analysis. The other combined-site differences occurred at fewer or 

none of the individual sites. Anti-nutrient component differences in seed were observed 

in mean values for phytic acid, raffinose, stachyose, and daidzein. In the combined-site 

analysis, all anti-nutrient component differences in seed between MON 87708 and the 

conventional control were of small relative magnitude, with respect to the conventional 

control, and ranged from a 6.14% decrease (phytic acid) to an 11.51% increase (daidzein). 

None of the anti-nutrient components were observed to be statistically different at more 

than two of the five individual sites. The only nutrient component difference in forage 

for the combined-site analysis was observed in ADF and its relative magnitude of 

difference, with respect to the conventional control, was 10.45%. No differences 

between MON 87708 and the conventional control ADF mean values were observed at 

any of the five individual sites. Mean values of MON 87708 components with 

statistically significant differences to the conventional control were all within the 99% 

tolerance interval established from the commercial reference varieties grown 

concurrently and at the same field sites, as well as ranges in the scientific literature and 

the ILSI Crop Composition Database. 
Monsanto Company 10-SY-210U 145 of 721 

For MON 87708 (untreated) the combined-site analysis of both seed and forage showed 

no statistically significant differences between MON 87708 (untreated) and conventional 

control for 30 (60.0%) of the 50 mean value comparisons. Of the statistically significant 

differences observed, none were from the forage analysis, and 20 were from the seed 

analysis. Nutrient component differences in seed included mean values for protein and 

eight amino acids, five fatty acids, ADF, NDF, and vitamin E. In the combined-site 

analysis, all nutrient component differences in seed between MON 87708 (untreated) and 

the conventional control were of small relative magnitude with respect to the 

conventional control and, whether increased or decreased, ranged from 1.45% to 7.60% 

for protein and amino acids, fatty acids, and fibers, and 18.16% for vitamin E. None of 

the nutrient components in the combined-site analysis were observed to be statistically 

different at all five individual sites. Anti-nutrient component differences in seed were 

observed in mean values for trypsin inhibitor, daidzein, and genistein. In the combinedsite analysis, all anti-nutrient component differences in seed between MON 87708 

(untreated) and the conventional control were of small relative magnitude, with respect to 

the conventional control, and ranged from an 11.59% increase (genistein), 15.37% 

increase (trypsin inhibitor), and a 17.24% increase (daidzein). None of the anti-nutrient 

components from the combined-site analysis were observed to be statistically different at 

more than one of the five individual sites. No nutrient component differences in forage 

for the combined-site analysis were observed. Mean values of MON 87708 components 

with statistically significant differences to the conventional control were all within the 

99% tolerance interval established from the commercial reference varieties grown 

concurrently and at the same field sites, as well as ranges in the scientific literature and 

the ILSI Crop Composition Database. 

In summary, a comprehensive evaluation of key nutrients and anti-nutrients in seed and 

key nutrients in forage for both dicamba-treated and untreated MON 87708 supports the 

conclusion that soybean seed and forage produced from MON 87708 are compositionally 

equivalent to that of conventional soybean and that neither the dicamba tolerance trait in 

MON 87708, nor the dicamba herbicide treatment, applied according to maximum incrop label rates (including the associated dicamba residue levels) have a meaningful 

impact on the composition and therefore on the food and feed safety or the nutritional 

quality of MON 87708 compared to conventional soybean. 
Monsanto Company 10-SY-210U 146 of 721 

VII. PHENOTYPIC, AGRONOMIC, AND ENVIRONMENTAL 

INTERACTIONS ASSESSMENT 

This section provides an assessment of the phenotypic, agronomic, and environmental 

interaction characteristics, including plant-symbiont associations of both dicamba treated 

and untreated MON 87708 compared to the near isogenic conventional soybean control 

A3525. Analysis of the untreated MON 87708 allows an assessment of the effect of the 

inserted gene only. However, for this product, since DCSA and formaldehyde are 

reaction products formed in the presence of dicamba herbicide, and since salicylic acid 

and formaldehyde are known to be involved in plant defense responses, dicamba treated 

MON 87708 data are supplied as well. The data support a determination that 

MON 87708 is similar to conventional soybean with the exception of the dicamba 

tolerance trait and, therefore, is no more likely to pose a plant pest risk than conventional 

soybean. These conclusions are based on the results of multiple evaluations. 

Phenotypic, agronomic, and environmental interaction characteristics of both 

dicamba-treated and untreated MON 87708 were evaluated in a comparative manner to 

assess plant pest potential. These assessments included evaluation of seed germination 

characteristics, plant growth and development characteristics, observations for plant 

responses to abiotic stress, plant-disease and plant-arthropod interactions, pollen 

characteristics, and plant-symbiont interaction characteristics. Results from the 

phenotypic, agronomic, and environmental interactions assessment demonstrate that 

MON 87708 does not possess weedy characteristics, increased susceptibility or tolerance 

to specific abiotic stress, diseases, or arthropods, or characteristics that would confer a 

plant pest risk compared to the conventional control. 

VII.A. Characteristics Measured for Assessment 

A detailed phenotypic description of the regulated article is requested as part of the 

petition for determination of nonregulated status in 7 CFR § 340.6 including differences 

from the unmodified recipient organism that would “substantiate that the regulated article 

is unlikely to pose a greater plant pest risk than the unmodified organism from which it 

was derived”. As part of the characterization of MON 87708, data were collected to 

provide a detailed phenotypic, agronomic, and environmental interaction description of 

both dicamba-treated and untreated MON 87708 and included an evaluation of specific 

characteristics related to altered weediness or plant pest potential. 

The plant characterization and assessment of MON 87708 encompassed six general data 

categories: 1) seed germination, dormancy, and emergence; 2) vegetative growth; 3) 

reproductive development (including pollen characteristics); 4) seed retention on the 

plant and lodging; 5) plant response to abiotic stress and interactions with diseases and 

arthropods; and 6) plant-symbiont interactions. An overview of the characteristics 

assessed is presented in Table VII-1. 
Monsanto Company 10-SY-210U 147 of 721 

The data were evaluated from a basis of familiarity (OECD, 1993) and were comprised of 

a combination of field, greenhouse, and laboratory assessments conducted by scientists 

who are familiar with the production and evaluation of soybean. In each of these 

assessments, the dicamba-treated and/or untreated MON 87708 was compared to the near 

isogenic conventional soybean control A3525 that has a genetic background similar to 

MON 87708 but does not possess the dicamba tolerance trait. In addition, multiple 

commercial reference varieties (see Appendices F-I and Tables F-1, G-1, and I-1) were 

included to provide a range of comparative values that are representative of existing 

commercial soybean varieties for each measured phenotypic, agronomic, and 

environmental interaction characteristic. Commercial reference soybean varieties are 

developed through a process of selecting and breeding for various desirable soybean 

characteristics and can provide a range of natural variability for characteristics and 

context for interpreting experimental results. 
Monsanto Company 10-SY-210U 148 of 721 

Table VII-1. Phenotypic, Agronomic, and Environmental Interaction 

Characteristics Evaluated in U.S. Field Trials, Laboratory, or Greenhouse Tests 

Data 

category 

Characteristics 

measured 

(associated section 

where discussed) 

Evaluation timing (setting of 

evaluation)1

Evaluation description 

(measurement endpoints) 

Seed 

germination, 

dormancy, 

and 

emergence 

Normal germinated 

(VII.C.1) 

Day 5 and 8 (20/30°C) 

(laboratory) 

Percentage of seed producing seedlings 

exhibiting normal developmental 

characteristics 

Abnormal 

germinated 

(VII.C.1) 

Day 8 (20/30°C) (laboratory) Percentage of seed producing seedlings 

that could not be classified as normal 

germinated 

Germinated 

(VII.C.1) 

Day 5, 8, and 13 (10, 20, 30, 

10/20 and 10/30°C) 

(laboratory) 

Percentage of seed that had germinated 

normally and abnormally 

Dead 

(VII.C.1) 

Day 5 and 8 (10, 20, 30, 10/20, 

10/30, and 20/30°C); Day 13 

(10, 20, 30, 10/20 and 10/30°C) 

(laboratory) 

Percentage of seed that had visibly 

deteriorated and become soft to the touch 

(also included non-viable hard and nonviable firm-swollen seed) 

Viable hard 

(VII.C.1) 

Day 8 (20/30°C); Day 13 (10, 

20, 30, 10/20 and 10/30°C) 

(laboratory) 

Percentage of seed that did not imbibe 

water and remained hard to the touch 

(viability determined by a tetrazolium 

test2

) 

Viable firm-swollen 

(VII.C.1) 

Day 8 (20/30°C); Day 13 (10, 

20, 30, 10/20 and 10/30°C) 

(laboratory) 

Percentage of seed that imbibed water and 

were firm to the touch but did not 

germinate (viability determined by a 

tetrazolium test2

) 

Early stand count 

(VII.C.2.3) 

V2 - V4 (Field) Number of emerged plants in two rows, 

standardized to 20 ft rows 

Final stand count 

(VII.C.2.3) 

Maturity, R8 (Field) Number of plants in two rows, 

standardized to 20 ft rows 

Vegetative 

growth 

Seedling vigor 

(VII.C.2.3) 

V2 - V4 (Field) Rated on a 1-9 scale, where 1 = excellent, 

5 = average, and 9 = poor vigor 

Growth stage 

assessment 

(VII.C.2.3) 

Every 2-3 weeks, V2-R8 

(Field) 

Average soybean plant growth stage per 

plot 

Flower color 

(VII.C.2.3) 

Flowering, R2 (Field) Color of flowers: purple, white, or mixed 

Plant pubescence 3

(VII.C.2.3) 

Maturity, R8 (Field) Pubescence on plants in each plot 

categorized as hairy or hairless 

Plant height 

(VII.C.2.3) 

Maturity, R8 (Field) Distance (in) from the soil surface to the 

uppermost node on the main stem of five 

representative plants per plot 

Reproductive 

development 

Days to 50% 

flowering 

(VII.C.2.3) 

Flowering, R1-R2 (Field) Calendar day number (days from January 

1) when approximately 50% of the plants 

in each plot were flowering 

Pollen viability 

(VII.C.3) 

Flowering, R1-R2 (laboratory) Percentage of viable pollen based on pollen 

grain staining characteristics 

Pollen morphology 

(VII.C.3) 

Flowering, R1-R2 (laboratory) Diameter (μm) of viable pollen grains 

Seed moisture 

(VII.C.2.3 

Harvest (Field) Percent moisture content of harvested seed 

100 seed weight 

(VII.C.2.3) 

Harvest (Field) Mass (g) of 100 harvested seed 

Test weight3

(VII.C.2.3) 

Harvest (Field) Mass (lb) of a bushel of harvested seed 

Yield 

(VII.C.2.3) 

Harvest (Field) Bushels of harvested seed per acre, 

adjusted to 13% moisture 
Monsanto Company 10-SY-210U 149 of 721 

Table VII-1 (continued). Phenotypic, Agronomic and Environmental Interaction 

Characteristics Evaluated in U.S. Field Trials, Laboratory or Greenhouse Tests 

Data 

category 

Characteristics 

measured 

(associated section 

where discussed) 

Evaluation timing (setting of 

evaluation)1

Evaluation description 

(measurement endpoints) 

Seed 

retention and 

lodging 

Lodging 

(VII.C.2.3) 

Maturity, R8 (Field) Rated on 0-9 scale, where 0 = completely 

erect and 9 = completely flat or lodged 

Pod shattering 

(VII.C.2.3) Maturity, R8 (Field) Rated on 0-9 scale, where 0 = no shattering 

and 9 = completely shattered

Plantenvironment 

interactions 

Plant response to 

abiotic stress 

(VII.C.2.4) 

Four times per growing season 

(Field) 

Qualitative assessment of each plot, with 

rating on a 0-9 scale, where 0 = no 

symptoms and 9 = severe symptoms 

Disease damage 

(VII.C.2.4) 

Four times per growing season 

(Field) 

Qualitative assessment of each plot, with 

rating on a 0-9 scale, where 0 = no 

symptoms and 9 = severe symptoms 

Arthropod-related 

damage (VII.C.2.4) 

Four times during growing 

season (Field) 

Damage assessed on upper four nodes of 

10 representative plants per plot using 

arthropod-specific 0-5 rating scales of 

increasing severity 

Arthropod 

abundance 

(VII.C.2.4) 3

Four times during growing 

season (Field) 

Quantitative assessment of pest and 

beneficial arthropods 

Plantsymbiont 

interactions3

Biomass 

(VII.C.4) 

6 weeks after emergence 

(Greenhouse) 

Nodule, root, and shoot dry weight 

(g/plant) 

Nodule number 

(VII.C.4) 

6 weeks after emergence 

(Greenhouse) 

Nodule number 

Total nitrogen 

(VII.C.4) 

6 weeks after emergence 

(Greenhouse) 

Shoot total nitrogen (% and g/plant) 

1

Soybean plant growth stages were determined using descriptions and guidelines outlined in Soybean 

Growth and Development (ISU, 2004). 

2

Viability of hard and firm-swollen seed were determined by a tetrazolium test (AOSA, 2000). 3

 Plant pubescence, test weight, arthropod abundance, and plant symbiont interactions were recorded only 

in the 2008 (untreated) field trials. 

 
Monsanto Company 10-SY-210U 150 of 721 

VII.B. Interpretation of Phenotypic and Environmental Interaction Data 

Plant pest risk assessments for biotechnology-derived crops are comparative assessments. 

Familiarity provides a basis from which the potential environmental impact of a 

biotechnology-derived plant can be evaluated. The concept of familiarity is based on the 

fact that the biotechnology-derived plant is developed from a well-characterized 

conventional plant variety. Familiarity considers the biology of the crop, the introduced 

trait, the receiving environment and the interaction of these factors, and provides a basis 

for comparative environmental risk assessment between a biotechnology-derived plant 

and its conventional counterpart. 

Expert knowledge and experience with conventionally bred soybean was the basis for 

selecting appropriate endpoints and estimating the range of responses that would be 

considered typical for soybean. As such, both dicamba-treated and untreated 

MON 87708 was compared to the conventional control in the assessment of phenotypic, 

agronomic, and environmental interaction characteristics. An overview of the 

characteristics assessed is presented in Table VII-1. A subset of the data relating to wellunderstood weedy characteristics (e.g., seed dormancy, pre-harvest seed loss 

characteristics, and lodging) was used to assess whether there is an increase in weediness 

of MON 87708, an element of APHIS’s plant pest determination. Evaluation of 

environmental interaction characteristics (e.g., plant-abiotic stress, plant-disease, and 

plant-arthropod interactions) was also considered in the plant pest assessment. Based on 

all of the data collected, an assessment was made to determine if MON 87708 is likely to 

pose an increased plant pest risk compared to conventional soybean. Prior to analysis, 

the overall dataset was evaluated for evidence of biologically relevant changes, and for 

possible evidence of an unexpected plant response. No unexpected observations or issues 

were identified. 

VII.B.1. Interpretation of Detected Differences Criteria 

Comparative plant characterization data from a biotechnology-derived crop and the 

conventional control are interpreted in the context of contributions to increased plant pest 

potential as assessed by APHIS. Under the framework of familiarity, characteristics for 

which no differences are detected support a conclusion of no increased plant pest 

potential of the biotechnology-derived crop compared to the conventional crop. 

Characteristics for which differences are detected are considered in a step-wise method 

(Figure VII-1). All detected differences for a characteristic are considered in the context 

of whether or not the difference would increase the plant pest potential of the 

biotechnology-derived crop. Ultimately, a weight of evidence approach considering all 

characteristics and data is used for the overall risk assessment of differences and their 

significance. Figure VII-1 illustrates the stepwise assessment process employed in detail: 
Monsanto Company 10-SY-210U 151 of 721 

Note: A “no” answer at any step indicates that the characteristic does not contribute to a biological or 

ecological change for the crop in terms of plant pest potential and subsequent steps are not considered. If 

the answer is “yes” or “uncertain”, the subsequent step is considered.

Figure VII-1. Schematic Diagram of Agronomic and Phenotypic Data 

Interpretation Methods 

Steps 1 and 2 - Evaluate Detected Statistically Significant Differences 

Data on each measured characteristic are statistically analyzed, where appropriate, within 

each individual site and in a combined-site analysis, in which the data are pooled among 

sites. All statistically significant differences are evaluated and considered in the context 

of a change in plant pest potential. Differences detected in individual-site analyses that 

are not consistently observed across multiple environments in the combined-site analysis 

are considered not biologically meaningful in terms of plant pest potential and, therefore, 

are not further considered in subsequent steps. Any difference detected in the combinedsite analysis is further assessed. 

Step 3 - Evaluate Differences Relative to Commercial Reference Varieties Range 

If a difference for a characteristic is detected in the combined-site analysis across 

multiple environments, then the mean value of the biotechnology-derived crop for the 

characteristic is assessed relative to the commercial reference varieties. 

Hazard identification and 

risk assessment on 

difference 

Outside variation for crop? No 

Yes 

No 

Yes 

Adverse in terms of plant pest 

potential? 

Yes 

Outside variation of study references 

No 

Not adverse; the direction 

or magnitude of the 

detected difference in the 

measured characteristic 

does not contribute to a 

biological or ecological 

change for the crop in 

terms of plant pest 

potential 

Yes 

Statistical differences detected 

in combined-site analysis? 

No 

Differences detected in the combined-site 

and individual site analyses are evaluated*

Step 2 

Step 4 

The measured

characteristic does not 

contribute to a biological 

or ecological change for 

the crop in terms of plant 

pest potential 

Step 5 

Step 6 

*See text for interpretation of differences 

 detected in the individual site analysis 

Step 3 

Step 1 
Monsanto Company 10-SY-210U 152 of 721 

Step 4 - Evaluate Differences in the Context of the Crop 

If the mean value of the biotechnology-derived crop is outside the variation of the 

commercial reference varieties (e.g., reference range), the mean value of the 

biotechnology-derived crop for the characteristic is assessed relative to known values 

common for the crop (e.g., published values). 

Step 5 - Plant Pest Potential 

If the mean value of the biotechnology-derived crop is outside the range of values 

common for the crop, the detected difference for the characteristic is then assessed for 

whether or not it is adverse in terms of plant pest potential. 

Step 6 - Conduct Risk Assessment on Identified Hazard 

If an adverse effect (hazard) is identified, risk assessment on the difference is conducted. 

The risk assessment considers contributions to enhanced plant pest potential of the crop 

itself, the impact of differences detected in other measured characteristics, and potential 

for and effects of trait transfer to feral populations of the crop or to a sexually-compatible 

species. 
Monsanto Company 10-SY-210U 153 of 721 

VII.C. Comparative Assessments of the Phenotypic, Agronomic, and Environmental 

Interaction Characteristics of MON 87708 

This section provides the results of comparative assessments conducted in replicated 

laboratory, greenhouse, and/or multi-site field experiments to provide a detailed 

phenotypic, agronomic, and environmental interaction description of MON 87708. The 

MON 87708 characteristics evaluated in these assessments included: seed dormancy and 

germination characteristics (Section VII.C.1), plant phenotypic and environmental 

interaction observations under field conditions (Section VII.C.2), pollen characteristics 

(Section VII.C.3), and symbiont interactions (Section VII.C.4). Additional details for 

each assessment are provided in Appendices F through I. 

VII.C.1. Seed Dormancy and Germination Characteristics 

USDA-APHIS considers the potential for weediness to constitute a plant pest factor 

(7 CFR § 340.6). Seed germination and dormancy mechanisms vary among species and 

their genetic basis tends to be complex. Seed dormancy (e.g., hard seed) is an important 

characteristic that is often associated with plants that are considered weeds (Anderson, 

1996; Lingenfelter and Hartwig, 2003); however, it is not uncommon to observe low 

levels of hard seed in conventional soybean (Mullin and Xu, 2001). Standardized 

germination assays are available and routinely used to measure the germination 

characteristics of soybean seed. The Association of Official Seed Analysts (AOSA), an 

internationally recognized seed testing organization, recommends a temperature range of 

20/30°C as optimal for testing the germination characteristics of soybean seed (AOSA, 

2007). 

Comparative assessments of seed dormancy and germination characteristics were 

conducted on MON 87708 and the conventional control. In addition, eight commercial 

reference varieties were included to provide a range of comparative values that are 

representative of existing commercial soybean varieties. The seed lots for MON 87708, 

conventional control, and the commercial reference varieties were produced in replicated 

field trials during 2008 in Iowa, Illinois, and Missouri, geographic areas which represent 

environmentally relevant conditions for soybean production for this product. In addition 

to the AOSA recommended temperature range of 20/30°C, seed was tested at five 

additional temperature regimes of 10, 20, 30, 10/20, and 10/30°C to assess seed 

germination properties. The details of the materials, experimental methods, and 

germination data from all individual production sites are presented in Appendix F. 

In a combined-site analysis, in which the data were pooled among the three seed 

production sites, no statistically significant differences (5% level of significance) were 

detected between MON 87708 and the conventional control for percent viable hard seed 

or percent viable firm-swollen seed in any temperature regime (Table VII-2). Within 

some temperature regimes, it was not possible to conduct an analysis of variance for 

percent viable firm-swollen seed due to no variance in the data. For these data, the values 

for MON 87708 and the conventional control were all zero, indicating no biological 

differences. No statistically significant differences were detected between MON 87708 
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and the conventional control for percent germinated or percent dead seed in the 20, 30, 

10/20, and 20/30°C temperature regimes. 

Four statistically significant differences were detected between MON 87708 and the 

conventional control in the combined-site analysis (Table VII-2). MON 87708 had lower 

percent germinated seed than the conventional control at 10°C (98.9% vs. 99.7%) and 

10/30°C (98.6% vs. 99.7%). Concurrently, MON 87708 had higher percent dead seed 

than the conventional control at 10°C (0.8% vs. 0.2%) and 10/30°C (1.4% vs. 0.3%). 

The differences in percent germinated and dead seed of MON 87708 were small in 

magnitude, and the mean values of MON 87708 were all within the range of the 

commercial reference varieties produced in the field trials along with MON 87708 and 

the conventional control. Furthermore, lower percent germinated seed and higher percent 

dead seed would not contribute to increased weediness. 

The biological characteristics evaluated were used to characterize MON 87708 in the 

context of plant pest risk assessment. Based on the seed dormancy and germination 

characteristics assessed, the results, particularly MON 87708’s lack of hard seed, 

demonstrate there were no changes indicative of increased weediness or plant pest 

potential of MON 87708 compared to conventional soybean. 
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Table VII-2. Combined-Site Comparison of MON 87708 to Conventional Control 

for Seed Dormancy and Germination Characteristics 

Temperature 

Regime 

Germination 

Characteristic1

Mean % (S.E.)2

 Reference Range3 

MON 87708 Control Min. Max. 

10 °C Germinated 98.9 (0.4)* 99.7 (0.2) 94.4 99.8 

Viable Hard 0.0 (0.0) 0.0 (0.0) 0.0 0.3 

Dead 0.8 (0.4)* 0.2 (0.1) 0.2 5.3 

Viable Firm-swollen 0.3 (0.1) 0.2 (0.1) 0.0 0.4 

20 °C Germinated 99.3 (0.4) 99.3 (0.3) 95.3 100.0 

Viable Hard 0.0 (0.0) 0.1 (0.1) 0.0 0.4 

Dead 0.8 (0.4) 0.6 (0.3) 0.0 4.8 

Viable Firm-swollen 0.0 (0.0) 0.0 (0.0) 0.0 0.3 

30 °C Germinated 98.7 (0.6) 99.3 (0.3) 93.3 100.0 

Viable Hard 0.0 (0.0) 0.0 (0.0) 0.0 0.1 

Dead 1.3 (0.6) 0.7 (0.3) 0.0 6.8 

Viable Firm-swollen 0.0 (0.0)† 0.0 (0.0) 0.0 0.0 

10/20 °C Germinated 99.3 (0.3) 99.3 (0.3) 95.9 100.0 

Viable Hard 0.1 (0.1) 0.1 (0.1) 0.0 0.4 

Dead 0.6 (0.3) 0.6 (0.3) 0.0 3.9 

Viable Firm-swollen 0.0 (0.0) 0.1 (0.1) 0.0 0.3 

10/30 °C Germinated 98.6 (0.5)* 99.7 (0.2) 95.9 99.8 

Viable Hard 0.0 (0.0) 0.1 (0.1) 0.0 0.0 

Dead 1.4 (0.5)* 0.3 (0.2) 0.3 4.1 

Viable Firm-swollen 0.0 (0.0) 0.0 (0.0) 0.0 0.1 

20/30 °C Normal Germinated 95.9 (1.2) 96.6 (1.0) 86.0 99.0 

 Abnormal Germinated 2.9 (0.7) 2.8 (0.8) 0.8 9.8 

Viable Hard 0.0 (0.0) 0.0 (0.0) 0.0 0.3 

Dead 1.2 (0.5) 0.7 (0.3) 0.3 4.1 

Viable Firm-swollen 0.0 (0.0)† 0.0 (0.0) 0.0 0.0 

Note: The data in this table are the combined-site results for the seeds from three 2008 field sites. The 

experimental design was a split-plot where the whole-plot treatment was seed production site and the subplot treatment was seed material (i.e., MON 87708, conventional control, or commercial reference 

varieties). 

*Indicates a statistically significant difference between MON 87708 and the conventional control (α=0.05). 

† No statistical comparisons were made due to lack of variability in the data. 

1

Germinated seed in the AOSA temperature regime (20/30°C) were categorized as either normal 

germinated or abnormal germinated seed. 2

Means based on twelve replicates (n = 12) of 100 seeds. The total percentage of all germination 

characteristics of MON 87708 or conventional control in some temperature regimes is greater than 100.0% 

due to numerical rounding of the means. S.E. = Standard Error. 

3

Minimum and maximum mean values from among eight commercial reference varieties.
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VII.C.2. Phenotypic, Agronomic, and Environmental Interaction Characteristics 

Evaluated under Field Conditions 

Plant growth, development, and yield characteristics were evaluated under field 

conditions as part of the plant characterization assessment of MON 87708. These data 

were developed to provide USDA-APHIS with a detailed description of MON 87708 

relative to the conventional --control and commercial reference varieties. According to 

7 CFR § 340.6, as part of the petition to seek deregulation, a petitioner must submit “a 

detailed description of the phenotype of the regulated article.” This information is being 

provided to assess whether there are phenotypic differences between MON 87708 and the 

conventional control that may impact its pest potential. Certain growth, reproduction, 

and pre-harvest seed loss characteristics (e.g., lodging, pod shattering) were used to 

assess whether there is an increase in weediness of MON 87708, an element of APHIS’s 

plant pest determination. Environmental interactions were also assessed as an indirect 

indicator of phenotypic changes to MON 87708 compared to the same comparators 

described above and are also considered in the plant pest assessment. 

VII.C.2.1. Phenotypic, Agronomic, and Environmental Interaction Characteristics 

for Untreated MON 87708 Evaluated under 2008 Field Conditions 

Data were collected from field trials located at 16 field sites in the U.S. and two field 

sites in Canada during 2008 to evaluate phenotypic, agronomic, and environmental 

interaction characteristics. These 18 field sites provided a diverse range of environmental 

and agronomic conditions representative of commercial soybean production areas in 

North America (Table VII-3). The experiments were arranged as randomized complete 

block designs with three replications at each field site. All plots of MON 87708, the 

conventional control, and the commercial reference varieties at each site were uniformly 

managed in order to assess whether the introduction of the dicamba tolerance trait altered 

the phenotypic and agronomic characteristics or the environmental interactions of 

MON 87708 compared to the conventional control. Therefore, dicamba herbicide was 

not applied to MON 87708 during the study. A description of the evaluated phenotypic 

and environmental interaction characteristics and the designated developmental stages 

when evaluations occurred are listed in Table VII-1. The methods and detailed results of 

the individual-site data comparisons are presented and discussed in Appendix G, while 

the combined-site analyses are summarized below. The results of this assessment 

demonstrated that the introduction of the dicamba tolerance trait did not alter 

MON 87708 compared to the conventional control in terms of weediness. The lack of 

differences in plant response to abiotic stress, disease damage, arthropod-related damage, 

and pest and beneficial arthropod abundance further support the conclusion that the 

introduction of the dicamba tolerance trait is not likely to result in increased plant pest 

potential or an altered environmental impact from untreated MON 87708 compared to 

conventional soybean. 
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Table VII-3. 2008 Field Phenotypic Evaluation Sites for Untreated MON 87708 

Location 

Location 

Code 

USDA-APHIS 

Notification Number 

Jackson County, Arkansas AR 08-072-110n 

Norfolk County, Ontario, Canada Can1 N/A 

Kent County, Ontario, Canada Can2 N/A 

Jefferson County, Iowa IA1 08-058-101n 

Benton County, Iowa IA2 08-058-101n 

Howard County, Iowa IA3 08-072-110n 

Clinton County, Illinois IL1 08-058-101n 

Stark County, Illinois IL2 08-058-101n 

Boone County, Indiana IN1 08-072-110n 

Clinton County, Indiana IN2 08-072-110n 

Parke County, Indiana IN3 08-058-101n 

Pawnee County, Kansas KS 08-072-110n 

Ottawa County, Michigan MI 08-072-110n 

Shelby County, Missouri MO1 08-072-110n 

Macon County, Missouri MO2 08-072-110n 

York County, Nebraska NE 08-072-110n 

Berks County, Pennsylvania PA 08-058-101n 

Walworth County, Wisconsin WI 08-058-101n 

N/A = Not applicable, trial was conducted under a Canadian confined research field 

testing permit. 

VII.C.2.2. Phenotypic, Agronomic, and Environmental Interaction Characteristics 

for Dicamba-Treated Evaluated under 2009 Field Conditions 

In addition to the data from 2008 field trials, data were collected from field trials 

conducted in 2009 at eight sites within U.S. soybean production regions (Table VII-4). 

MON 87708, the conventional control variety A3525, and three commercially-released 

reference soybean varieties were evaluated at each site. A total of 14 reference varieties 

were evaluated among the sites. The experimental design at each site was a randomized 

complete block with four replications. All plots of MON 87708, the conventional control, 

and the commercial reference varieties at each site were uniformly managed in order to 

assess whether the introduction of the dicamba tolerance trait in the presence of dicamba 

altered the phenotypic and agronomic characteristics or the environmental interactions of 

MON 87708 compared to the conventional control. Therefore, MON 87708, in each 

replication at all sites, received an application of a commercial formulation of dicamba 

(Clarity) at 0.5 pound a.e dicamba per acre. Treated MON 87708 was compared to the 

control within each site (i.e., individual-site analysis) and in a combined-site analysis, in 

which the data were pooled across the eight sites, for 12 plant characteristics. The 

minimum and maximum mean values (reference range) were determined from the 

references to provide phenotypic characteristic and environmental interaction values 

representative of commercial soybean varieties. 
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Table VII-4. 2009 Field Phenotypic Evaluation Sites for Dicamba Treated 

MON 87708 

County, State 

Location 

Code 

USDA-APHIS 

Notification Number 

Jackson County, Arkansas ARNE 09-061-108n 

Jefferson County, Iowa IARL 09-061-108n 

Clinton County, Illinois ILCY 09-061-108n 

Stark County, Illinois ILWY 09-061-108n 

Parke County, Indiana INRC 09-061-108n 

Boone County, Indiana INSH 09-061-108n 

Pawnee County, Kansas KSLA 09-061-108n 

York County, Nebraska NEYO 09-061-108n 

VII.C.2.3. Field Phenotypic and Agronomic Characteristics 

VII.C.2.3.1. 2008 Field Phenotypic and Agronomic Characteristics for Untreated 

MON 87708 

A total of 14 phenotypic and agronomic characteristics were evaluated (Table VII-5 and 

Table G-5 of Appendix G). In a combined-site analysis in which the data were pooled 

among the sites, no statistically significant differences were detected (5% level of 

significance) between untreated MON 87708 and the conventional control for early stand 

count, seedling vigor, days to 50% flowering, lodging, pod shattering, final stand count, 

seed moisture, test weight, or yield (Table VII-5). A statistically significant difference 

was detected between untreated MON 87708 and the conventional control for plant 

height and 100 seed weight in the combined-site analysis. Untreated MON 87708 was 

6% taller (33.5 vs. 31.6 inches) and had 3.3% lower 100 seed weight than the 

conventional control (15.0 vs. 15.5 grams). However, the differences in plant height and 

100 seed weight were small in magnitude, and the mean values of untreated MON 87708 

for both plant height and 100 seed weight were within the range of the commercial 

reference varieties grown in the trials along with untreated MON 87708 and the 

conventional control. The difference in 100 seed weight did not result in a statistically 

significant difference in final yield, and it is unlikely that a difference in seed weight or 

plant height would contribute to increased weediness of MON 87708 compared to the 

conventional control. 

Flower color, plant pubescence, and plant growth stage data were categorical and were 

not statistically analyzed; however, at each site, all plants of untreated MON 87708 and 

the conventional control had purple flowers and hairy pubescence as expected. 

Additionally, untreated MON 87708 and the conventional control were within the same 

range of plant growth stages for 131 out of the 132 growth stage gobservations among all 

sites (Appendix G; Table G-4). During the second observation at the WI site, untreated 

MON 87708 plants were at the V5 growth stage while the conventional control was at V4. 
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The growth stage of untreated MON 87708, however, was within the range of growth 

stages observed for the commercial reference varieties grown concurrently (V4 − V5). 

Thus, there were no biologically meaningful differences in plant development observed 

between untreated MON 87708 and the conventional control. 

Table VII-5. Combined-Site Comparison of Untreated MON 87708 to Conventional 

Control during 2008 for Phenotypic and Agronomic Characteristics 

Phenotypic 

Characteristic (units) 

Mean (S.E.) Reference Range1 

MON 87708 Control2 Minimum Maximum 

Early stand count (#/plot) 271.1 (9.0) 265.2 (9.8) 195.7 452.5 

Seedling vigor (1-9 scale) 3.5 (0.2) 3.4 (0.2) 1.8 4.5 

Days to 50% flowering3

206.4 (1.2) 206.2 (1.3) 199.6 216.7 

Flower color4 Purple Purple Purple Purple 

Plant pubescence4 Hairy Hairy Hairy Hairy 

Plant height (in.) 33.5 (0.9)* 31.6 (0.8) 25.4 42.4 

Lodging (0-9 scale) 1.2 (0.2) 0.9 (0.1) 0.1 2.9 

Pod shattering (0-9 scale) 0.1 (0.0) 0.1 (0.0) 0.0 0.3 

Final stand count (#/plot) 251.1 (8.4) 243.4 (8.7) 178.3 338.0 

Seed moisture (%) 11.8 (0.3) 11.7 (0.3) 10.4 13.9 

100 seed weight (g)5 15.0 (0.2)* 15.5 (0.2) 14.2 18.7 

Test weight (lb/bu) 56.2 (0.3) 55.9 (0.3) 53.6 57.6 

Yield (bu/A) 55.4 (2.0) 55.1 (2.3) 37.7 72.7 

Note: The experimental design was a randomized complete block. S.E. = Standard Error. Means 

based on n = 54 for MON 87708 and n = 53 for the conventional control with exception of 100 

seed weight for the control where n = 52. 

* Indicates a statistically significant difference between MON 87708 and the conventional control 

(α=0.05). 

1

Reference ranges were determined from the minimum and maximum mean values from among 

18 commercial reference varieties. 

2

Excessive water damage from heavy precipitation early in the season at the WI site resulted in a 

poor stand in one replicate of the conventional control. Therefore, the data for all characteristics 

from the single replicate at the WI site were excluded from the statistical analysis. 

3

Calendar day number (from 1 Jan 2008) when approximately 50% of the plants in each plot were 

flowering. 

4

Flower color and plant pubescence data were categorical and not statistically analyzed (see 

Appendix G, Table G-3). 

5

Data on 100 seed weight were inadvertently not collected from one replicate of the conventional 

control at the MO1 site. 

 
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VII.C.2.3.2. 2009 Field Phenotypic and Agronomic Characteristics for 

Dicamba-Treated MON 87708 

A total of 12 phenotypic and agronomic characteristics were evaluated (Table VII-6 and 

Table G-6 of Appendix G). In the combined-site analysis, no statistically significant 

differences were detected between treated MON 87708 and the conventional control for 

early stand count, seedling vigor, days to 50% flowering, plant height, lodging, pod 

shattering, final stand count, seed moisture, or yield (Table VII-6). One statistically 

significant difference was detected between treated MON 87708 and the control, where 

MON 87708 had a lower 100 seed weight than the control (14.6 vs. 15.6 g). The 

difference in 100 seed weight is relatively small in magnitude and the mean 100 seed 

weight of treated MON 87708 was slightly below the reference range. It is unlikely that 

a difference in 100 seed weight would contribute to increased weed potential of MON 

87708 when treated with dicamba compared to conventional soybean. Flower color and 

plant growth stage data were categorical and were not statistically analyzed; however, at 

each site, all plants of treated MON 87708 and the control had purple flowers as 

expected. Additionally, treated MON 87708 and the control were within the same range 

of plant growth stages for all growth stage observations among the sites. Thus, there 

were no biologically-meaningful differences in plant development observed between 

treated MON 87708 and conventional soybean. 

The results of this assessment demonstrated that the introduction of the dicamba tolerance 

trait and the associated dicamba application does not alter MON 87708 compared to the 

conventional control in terms of weediness. The lack of differences in plant response to 

abiotic stress, disease damage, and arthropod-related damage further support the 

conclusion that the introduction of the dicamba tolerance trait is not likely to result in 

increased plant pest potential or an altered environmental impact for dicamba-treated 

MON 87708 compared to conventional soybean. 
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Table VII-6. Combined-Site Comparison of Dicamba Treated MON 87708 to 

Conventional Control during 2009 for Phenotypic and Agronomic Characteristics 

Phenotypic 

Characteristic (units) 

MON 87708 (S.E.) Reference Range1

MON 87708 Control Minimum Maximum 

Early stand count (#/plot) 298.9 (5.20) 301.1 (3.58) 263.4 340.8 

Seedling vigor (1-9 scale) 3.0 (0.33) 3.0 (0.31) 1.0 4.5 

Days to 50% flowering2 214.7 (1.12) 214.6 (1.19) 205.0 226.0 

Plant height (in) 31.6 (0.78) 31.2 (0.92) 25.3 38.3 

Lodging (1-9 scale) 2.2 (0.27) 2.4 (0.31) 1.0 4.5 

Pod shattering (1-9 scale) 1.1 (0.05) 1.1 (0.07) 1.0 1.5 

Final stand count (#/plot) 264.8 (6.81) 266.8 (5.20) 219.5 305.5 

Seed moisture (%) 13.1 (0.38) 13.4 (0.37) 11.1 17.0 

100 seed weight (g) 14.6* (0.23) 15.6 (0.23) 15.0 17.7 

Yield (bu/ac) 46.7 (2.37) 46.8 (2.23) 29.4 59.9 

 

Note: The experimental design was a randomized complete block with four replications. Site 

codes are as follows: ARNE = Jackson County, AR; IARL = Jefferson County, IA; ILCY = 

Clinton County, IL; ILWY = Stark County, IL; INRC = Parke County, IN; INSH = Boone 

County, IN; KSLA = Pawnee County, KS; NEYO = York County, NE 

* Statistically significant differences (α=0.05) between MON 87708(T) and the conventional 

soybean control. 

1

 Reference range = Minimum and maximum mean values among the 14 commercially-released 

reference soybean varieties.

2

 Calendar day number (days after 1 Jan 2009) when approximately 50% of the plants in each plot 

were flowering. 
Monsanto Company 10-SY-210U 162 of 721 

VII.C.2.3.3. Field Phenotypic and Agronomic Characteristics for Both 

Dicamba-Treated and Untreated MON 87708 - Conclusion 

The phenotypic and agronomic characteristics were used to provide a detailed description 

of both dicamba-treated and untreated MON 87708 compared to the conventional control. 

A subset of these characteristics was useful to assess the weediness potential of 

MON 87708. Based on the assessed phenotypic and agronomic characteristics, the 

results support a determination that both dicamba-treated and untreated MON 87708 are 

similar to conventional soybean and are no more weedy or likely to pose a plant pest risk 

than conventional soybean. 

VII.C.2.4. Environmental Interaction Characteristics 

USDA-APHIS considers the environmental interaction of the biotechnology-derived crop 

compared to its conventional counterpart to determine the potential for increased plant 

pest characteristics. Evaluations of environmental interactions were conducted as part of 

the plant characterization for untreated MON 87708. In the 2008 North American field 

trials conducted for evaluation of phenotypic and agronomic characteristics of 

MON 87708, data were also collected on plant response to abiotic stress (drought, wind, 

nutrient deficiency, etc.), disease damage, arthropod-related damage, and arthropod 

abundance (Appendix G; Tables G-9, G-10, G-11, G-12, and G-13). Similarly these data, 

except for arthropod abundance, were also collected for the 2009 U.S. field trials 

(Appendix G; Tables G-14, G-15, and G-16). These data were used as part of the 

environmental consequences (Section IX) to assess plant pest potential and provide an 

indication of potential effects of untreated MON 87708 on non-target organisms (NTOs) 

and threatened and endangered species compared to the conventional control. In addition, 

multiple commercial reference varieties were included in the analysis to establish a range 

of natural variability for each assessed characteristic. The results of the field evaluations 

showed that the dicamba tolerance trait did not unexpectedly alter the assessed 

environmental interactions of untreated MON 87708 compared to the conventional 

control. Additionally, results of field evaluations showed that the dicamba tolerance trait 

in the presence of dicamba herbicide did not unexpectedly alter the assessed 

environmental interactions of treated MON 87708 compared to the conventional control. 

The lack of significant biologically meaningful differences in plant response to abiotic 

stress, disease damage, arthropod-related damage, and pest and beneficial arthropod 

abundance support the conclusion that the introduction of the dicamba tolerance trait is 

unlikely to result in increased plant pest potential or an altered environmental impact 

from MON 87708 compared to conventional soybean. 

In the 2008 field trials, the observations of plant response to abiotic stress, disease 

damage, and arthropod-related damage were performed four times during the growing 

season at all 18 sites, and arthropod abundance was assessed quantitatively from 

collections performed four times during the growing season at four of the 18 sites (i.e., 

IL2, IN1, MI, and MO1 sites). In the 2009 field trials, the observations of plant response 
Monsanto Company 10-SY-210U 163 of 721 

to abiotic stress, disease damage, and arthropod-related damage were performed four 

times during the growing season at all eight sites. 

The assessed stressors (abiotic, diseases, and arthropods) were at natural levels as no 

artificial infestation or imposed abiotic stress was used and, therefore, typically varied 

between observations at a site and among sites. Abiotic stress and disease damage data 

were collected from each plot using a non-specific 0 – 9 scale of increasing severity of 

observed damage. The 0 – 9 scale was not designed to rate specific damage symptoms. 

However, the non-specific scale was utilized to allow for the evaluation of the wide 

variety of potential abiotic stressor and disease damage symptoms potentially occurring 

across the season and across sites. Due to the non-specific nature of the scale used, the 

data were not statistically analyzed but rather assessed qualitatively and placed into one 

of the following categories: none, slight, moderate, or severe. The response of 

MON 87708 and the conventional control to an abiotic stress or disease were considered 

different on a particular observation date at a site if the range of injury severity to 

MON 87708 did not overlap with the range of injury severity to the control across all 

three replications (e.g., “none” vs. “slight-moderate” rating). For each observation at a 

site, the range of injury severity across the commercial reference varieties provided 

assessment data that are representative of commercial soybean varieties. Arthropodrelated damage was assessed from each plot on the upper four nodes of 10 representative 

plants using a 0 – 5 pest-specific rating scale of increasing severity of observed damage. 

These numerical data along with the quantitative arthropod abundance data were 

subjected to statistical analysis. 

VII.C.2.4.1. 2008 Environmental Interaction Characteristics for Untreated 

MON 87708 

In an assessment of abiotic stress response and disease damage, no differences were 

observed between untreated MON 87708 and the conventional control for 193 out of 194 

comparisons for the assessed abiotic stressors or for any of the 215 comparisons for the 

assessed diseases among all observations at the sites (Appendix G; Tables G-9 and G-10). 

One difference was observed in abiotic stress response during the fourth observation at 

the WI site where minor wind damage was observed in MON 87708 (“slight” rating) and 

no wind damage was observed in the conventional control or commercial reference 

varieties (“none” rating). The difference, however, was not observed during any of the 

other 29 wind damage observations among the sites. Thus, the small difference in wind 

damage rating during the single observation was not indicative of a consistent response 

associated with the trait and was considered not biologically meaningful in terms of 

increased weediness or plant pest potential or an altered environmental impact from 

untreated MON 87708 compared to conventional soybean. 

In an assessment of arthropod-related damage, no statistically significant differences 

were detected (5% level of significance) between untreated MON 87708 and the 

conventional control for 89 out of 95 comparisons for the assessed arthropods 

(Appendix G; Table G-11). Statistical comparisons could not be made between 

MON 87708 and the conventional control for 121 additional arthropod-related damage 

comparisons due to no variance in the data; however, the means for MON 87708 and the 
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conventional control were the same value for these comparisons, indicating no biological 

differences. A total of six statistically significant differences involving four taxa were 

detected between MON 87708 and the conventional control. MON 87708 had less 

damage than the conventional control from aphids in the third observation at the IA2 site 

(0.3 vs. 0.5 rating) and second observation at the IA3 site (0.8 vs. 0.9 rating). 

MON 87708 had less damage than the conventional control from blister beetles in the 

second observation at the MO1 site (0.1 vs. 0.4 rating) and more damage than the 

conventional control from potato leafhopper in the first observation at the PA site (1.1 vs. 

0.6 rating). MON 87708 had less damage than the conventional control from Japanese 

beetle in the second observation at the IN1 site (0.6 vs. 0.9 rating) and more damage in 

the fourth observation at the PA site (0.6 vs. 0.4 rating). The mean damage ratings for 

MON 87708 were within the range of the commercial reference varieties grown 

concurrently with MON 87708 and the conventional control for all differences detected 

in arthropod-related damage with the exception of the differences in Japanese beetle 

damage at the IN1 and PA sites. The mean Japanese beetle damage rating for 

MON 87708 at the IN1 site was slightly lower than the range of commercial reference 

varieties for the difference detected (MON 87708 mean = 0.6 rating; range of commercial 

reference varieties = 0.7 – 0.9 ratings) and slightly higher than the range of commercial 

reference varieties for the difference detected at the PA site (MON 87708 mean = 0.6 

rating; range of commercial reference varieties = 0.3 – 0.5 ratings). Furthermore, the 

differences detected in arthropod-related damage were all small in magnitude and were 

not consistent across observations or sites. These results support a conclusion that the 

detected differences in arthropod-related damage were not indicative of a consistent 

response associated with the trait and were not considered biologically meaningful in 

terms of increased weediness or plant pest potential or an altered environmental impact 

from untreated MON 87708 compared to conventional soybean. 

In an assessment of pest and beneficial arthropod abundance, no statistically significant 

differences were detected (5% level of significance) between untreated MON 87708 and 

the conventional control for 142 out of 151 comparisons, including 74 arthropod pest 

abundance comparisons and 77 beneficial arthropod abundance comparisons, among the 

collection intervals at the four sites (Appendix G; Tables G-12 and G-13). Statistical 

comparisons could not be made between MON 87708 and the conventional control for 

eight additional comparisons, including three arthropod pest abundance comparisons and 

five beneficial arthropod abundance comparisons, due to no variance in the data; however, 

the means for MON 87708 and the conventional control were the same value for these 

comparisons, indicating no biological differences. A total of nine statistically significant 

differences were detected between MON 87708 and the conventional control for 

arthropod abundance, including seven for pest arthropods and two for beneficial 

arthropods. 

The seven differences detected for pest arthropod abundance included observations for 

green cloverworm, Japanese beetles, and stink bugs (Table G-12). MON 87708 had 

lower green cloverworm abundance than the conventional control in the first collection 

from the IL2 site (0.0 vs. 2.0 per plot), third collection from the IN1 site (7.0 vs. 11.3 per 

plot), and fourth collection from the MI site (0.0 vs. 0.7 per plot); and higher green 

cloverworm abundance than the conventional control in the first collection from the MI 
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site (1.7 vs. 0.0 per plot). MON 87708 had lower Japanese beetle abundance than the 

conventional control in the first collection from the MI site (2.3 vs. 8.0 per plot). In 

addition, MON 87708 had lower stink bug abundance than the conventional control in the 

third collection from the IL2 site (0.0 vs. 1.3 per plot) and higher stink bug abundance in 

the third collection from the IN1 site (2.0 vs. 0.0 per plot). The mean arthropod 

abundance values for MON 87708 were within the range of the commercial reference 

varieties grown concurrently with MON 87708 and the conventional control for all 

differences detected with the exception of the difference detected for green cloverworm 

abundance at the IL2 site (MON 87708 mean = 0.0 per plot; range of commercial 

reference varieties = 0.3 – 2.7 per plot) and the IN1 site (MON 87708 = 7.0 per plot; 

range of commercial reference varieties = 8.0 – 13.0 per plot) and stink bug abundance at 

the IN1 site (MON 87708 = 2.0 per plot; range of commercial reference varieties = 0.0 – 

0.7 per plot). Furthermore, the differences detected in green cloverworm, Japanese beetle, 

and stink bug abundance were all small in magnitude and were not detected in other 

collections or sites where these pests were present. These results support a conclusion 

that the detected differences in pest arthropod abundance were not indicative of a 

consistent response associated with the trait and were not considered biologically 

meaningful in terms of increased weediness or plant pest potential or an altered 

environmental impact from MON 87708 compared to conventional soybean. 

The two differences detected for beneficial arthropod abundance included observations 

for Araneae (spiders) and Nabis spp. (Table G-13). MON 87708 had lower Araneae 

abundance (0.0 vs. 3.0 per plot) and higher Nabis spp. abundance (4.7 vs. 1.7 per plot) 

than the conventional control in the fourth collection from the MI site. The mean 

Araneae abundance value for MON 87708 was slightly lower than the range of 

commercial reference varieties (0.7 – 1.0 per plot), while the mean Nabis spp. abundance 

value for MON 87708 was within the range of commercial reference varieties (2.0 – 6.3 

per plot). Furthermore, the differences detected for both Araneae and Nabis spp. 

abundance were small in magnitude and were not consistent across collections or sites. 

The results support a conclusion that the detected differences in beneficial arthropod 

abundance were not indicative of a consistent response associated with the dicamba 

tolerance trait and were not considered biologically meaningful in terms of increased 

weediness or plant pest potential or an altered environmental impact from untreated 

MON 87708 compared to conventional soybean. 

VII.C.2.4.2. 2009 Environmental Interaction Characteristics for Treated 

MON 87708 

In an assessment of plant response to abiotic stressors and disease damage for the 2009 

field trials (Appendix G; Table G-14 and G-15), no differences were observed between 

treated MON 87708 and the conventional control for 181 of 182 comparisons (including 

89 abiotic stress response and 92 disease damage comparisons) among all observations at 

the sites. One difference was observed between treated MON 87708 and the control for 

white mold during a single observation (slight vs. none). The damage rating for treated 

MON 87708 was outside of the reference range (no damage was observed in the 

references), and this difference was not observed in any of the other two white mold 

evaluations across the sites. 
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In an assessment of arthropod-related damage (Appendix G; Table G-16) there were a 

total of 93 comparisons. No statistically significant differences were detected between 

treated MON 87708 and the control for 56 out of 59 comparisons. In addition, no 

numerical differences were observed for the 34 comparisons for which p-values could not 

be generated due to lack of variability in the data. Treated MON 87708 had lower bean 

leaf beetle damage than the control for Observation 3 at the KSLA site (0.00 vs. 0.08), 

and greater grasshopper damage for Observation 3 at the INRC site (0.45 vs. 0.10) and at 

the KSLA site (0.20 vs. 0.03). The mean damage ratings for bean leaf beetle damage and 

grasshopper damage at the KSLA site were within the respective reference ranges. For 

the remaining difference, the mean damage rating for grasshopper damage at the INRC 

site from treated MON 87708 was outside the reference range; however, this difference 

was not consistent across observations or sites. Thus, there was not a consistent response 

associated with the dicamba tolerance trait or the herbicide treatment, and the results are 

not considered biologically meaningful in terms of adverse environmental impacts of 

treated MON 87708 compared to the conventional soybean. 

VII.C.2.4.3. Environmental Interaction Characteristics for Both Dicamba Treated 

and Untreated MON 87708 - Conclusion 

The results of the 2008 field evaluations showed that the dicamba tolerance trait did not 

unexpectedly alter the assessed environmental interactions of untreated MON 87708 

compared to the conventional control. Additionally, the results of the 2009 field 

evaluations showed that the dicamba tolerance trait in the presence of dicamba herbicide 

did not unexpectedly alter the assessed environmental interactions of MON 87708 

compared to the conventional control. The lack of significant biological differences in 

plant responses to abiotic stress, disease damage, arthropod-related damage for both 

dicamba treated and untreated MON 87708, and pest and beneficial arthropod abundance 

for untreated MON 87708 support the conclusion that the introduction of the dicamba 

tolerance trait is unlikely to result in increased plant pest potential compared to 

conventional soybean. 

VII.C.3. Pollen Characteristics 

USDA-APHIS considers the potential for gene flow and introgression of the 

biotechnology-derived trait into other soybean varieties and wild relatives to determine 

the potential for increased weedy or invasive characteristics of the receiving species. 

Pollen morphology and viability information are pertinent to this assessment and, 

therefore, were assessed for MON 87708. In addition, characterization of pollen 

produced by MON 87708 and the conventional control is relevant to the plant pest risk 

assessment because it adds to the detailed description of the phenotype of MON 87708 

compared to the conventional control. 

The purpose of this evaluation was to assess the morphology and viability of pollen 

collected from MON 87708 compared to that of the conventional control. Pollen was 

collected from MON 87708, the conventional control, and four commercial reference 

varieties grown under similar agronomic conditions in a field trial in Illinois. The trial 

was arranged in a randomized complete block design with three replications. Twenty 
Monsanto Company 10-SY-210U 167 of 721 

flowers were collected from each plot. Pollen was extracted, combined among flowers 

collected from the same plot, and stained with Alexander’s stain (Alexander, 1980). 

Pollen viability was evaluated for each sample, and pollen grain diameter was measured 

for ten representative viable pollen grains per replication. General morphology of the 

pollen was observed for each of the three replications of MON 87708, the conventional 

control, and the commercial reference varieties (see Appendix H). 

No statistically significant differences were detected (5% level of significance) between 

MON 87708 and the conventional control for percent viable pollen or pollen grain 

diameter (Table VII-7). Furthermore, no visual differences in general pollen morphology 

were observed between MON 87708 and the conventional control. These results 

demonstrate that the introduction of the dicamba tolerance trait did not alter the overall 

morphology or viability of MON 87708 pollen compared to the conventional control. 

The pollen characterization data contribute to the detailed phenotypic description of 

MON 87708 compared to the conventional control. The result supports an overall 

conclusion that MON 87708 is similar to conventional soybean and is no more likely to 

pose a plant pest risk than conventional soybean. 

Table VII-7. Pollen Characteristics of MON 87708 Compared to Conventional 

Control 

Pollen 

Characteristic 

Mean (S.E.)1

 Reference Range2

MON 87708 Control Minimum Maximum 

 

Viability (%) 99.3 (0.3) 98.4 (0.9) 98.1 98.4 

Diameter (μm) 24.4 (0.5) 24.3 (0.7) 23.4 24.3 

Note: No statistically significant differences were detected between MON 87708 and the 

conventional control (α=0.05). 

1

Means based on n = 3. S.E. = Standard Error 

2

Reference ranges were determined from the minimum and maximum mean value from among 

the four commercial reference varieties.

VII.C.4. Symbiont Interactions 

As part of the plant pest risk assessment, USDA-APHIS considers the impact of the 

biotechnology-derived crop on plant pest potential and the environment as well as on 

agricultural or cultivation practices compared to its conventional counterpart. Changes in 

the symbiotic relationship with rhizosphere-inhabiting bacteria Rhizobiaceae and 

Bradyrhizobiaceae could directly impact pest potential, the environment, or cultivation 

practices (i.e., the need to add additional nitrogen to sustain soybean production). Thus, 

the purpose of this evaluation was to assess whether the introduction of the dicamba 

tolerance trait altered the symbiotic interaction of MON 87708 with Bradyrhizobium 

japonicum (B. japonicum) compared to that of the conventional control. 
Monsanto Company 10-SY-210U 168 of 721 

Members of the bacterial family Rhizobiaceae and Bradyrhizobiaceae form a highly 

complex and specific symbiotic relationship with leguminous plants, including soybean 

(Gage, 2004). The nitrogen-fixing plant-microbe symbiosis results in the formation of 

root nodules, providing an environment in which differentiated bacteria called bacteroids 

are capable of reducing or fixing atmospheric nitrogen. The product of nitrogen fixation, 

ammonia, then can be utilized by the plant. In soybean, atmospheric nitrogen is fixed 

into ammonia through a symbiotic association with the bacterium B. japonicum. As a 

result of this relationship, nitrogen inputs are typically not necessary for agricultural 

production of soybean. 

The relative effectiveness of the symbiotic association between a leguminous plant and 

its rhizobial symbiont can be assessed by various methods. Assessment of nodule 

number and mass along with plant growth and nitrogen status are commonly used to 

assess differences in the symbiotic association between a legume and its associated 

rhizobia (Israel et al., 1986). It should be noted, however, that nodule number relative to 

nodule dry weight may be variable in soybean experiments because nodules may be 

larger in diameter and less numerous, while others are not as developed (smaller) but 

more abundant (Appunu and Dhar, 2006; Israel et al., 1986). 

MON 87708, the conventional control, and six commercial reference varieties were 

produced from seeds planted in pots containing nitrogen-deficient potting medium grown 

in a greenhouse. Seeds were inoculated with a solution of B. japonicum. The pots were 

arranged in a randomized complete block design with eight replicates. At six weeks after 

emergence, plants were excised at the surface of the potting medium, and shoot and root 

plus nodule material were removed from the pots. Nodules were separated from roots 

prior to enumeration and determination of dry weight. MON 87708 was compared to the 

conventional control for key characteristics related to their association with the soybeanB. japonicum symbiosis. Detailed information on materials and methods used for the 

symbiont evaluation is presented in Appendix I. 

No statistically significant differences were detected (5% level of significance) between 

MON 87708 and the conventional control for any of the measured characteristics, 

including nodule number, nodule dry weights, root dry weights, shoot dry weights, and 

shoot total nitrogen (percent and mass) (Table VII-8). 

Based on the assessed characteristics, the results support the conclusion that the 

introduction of the dicamba tolerance trait does not alter the symbiotic relationship 

between B. japonicum and MON 87708 compared to the conventional control. Thus, 

these data further support a conclusion of no change in plant pest potential and no 

expected impact to cultivation practices relative to nitrogen inputs for MON 87708 

compared to conventional soybean. 
Monsanto Company 10-SY-210U 169 of 721 

Table VII-8. Symbiont Interaction Assessment of MON 87708 and Conventional 

Control 

Characteristic 

Mean (S.E.)1 Reference Range2 

MON 87708 Control Minimum Maximum 

Nodule Number

(per plant) 264 (25) 238 (13) 148 346 

Nodule Dry Weight

(g/plant) 0.58 (0.03) 0.56 (0.02) 0.43 0.66 

Root Dry Weight

(g/plant) 1.33 (0.07) 1.15 (0.04) 1.09 1.89 

Shoot Dry Weight

(g/plant) 6.95 (0.46) 6.03 (0.23) 5.93 8.68 

Shoot Total

Nitrogen (% dwt) 4.46 (0.05) 4.48 (0.06) 3.51 4.26 

Shoot Total

Nitrogen (g) 0.31 (0.02) 0.27 (0.01) 0.25 0.32 

Note: Pots were arranged in a greenhouse in a randomized complete block design. No 

statistically significant differences were detected between MON 87708 and the conventional 

control (α=0.05). 

1

Means based on n = 8. S.E. = Standard Error 

2

Reference ranges were determined from the minimum and maximum mean value from among 

the six commercial reference varieties. 

VII.D. Phenotypic, Agronomic, and Environmental Interactions Assessment 

Conclusion 

An extensive and robust set of information and data were used to assess whether the 

introduction of the dicamba tolerance trait or the introduction of the dicamba tolerance 

trait in the presence of dicamba herbicide altered the plant pest potential of MON 87708 

compared to the conventional control. Phenotypic, agronomic, and environmental 

interaction characteristics of both dicamba-treated and untreated MON 87708 were 

evaluated and compared to those of the conventional control and considered within the 

variation among commercial reference varieties. These assessments included plant 

growth and development characteristics; seed dormancy and germination characteristics; 

pollen characteristics; observations of abiotic stress response, disease damage, arthropodrelated damage and arthropod abundance; and plant-symbiont interaction characteristics. 

Results from the phenotypic, agronomic, and environmental interactions assessment 

demonstrate that MON 87708 does not possess weedy characteristics, increased 

susceptibility or tolerance to specific abiotic stress, diseases, or arthropods, or 

characteristics that would confer a plant pest risk or significant environmental impact 

compared to conventional soybean. 
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VIII. U.S. AGRONOMIC PRACTICES 

VIII.A. Introduction 

As part of the plant pest assessment required by 7 CFR § 340.6(c)(4), impacts to 

agricultural and cultivation practices must be considered. This section provides a 

summary of current agronomic practices in the U.S. for producing soybean and is 

included in this petition as a baseline to assess possible impacts to agricultural practices 

due to the cultivation of MON 87708. Discussions include soybean production, seed 

production, plant growth and development, general management practices during the 

season, management of weeds, insects and diseases, soybean rotational crops, and 

volunteer soybean management. Information presented in Section VII.C.2 demonstrated 

that MON 87708 is no more susceptible to diseases or pests than conventional soybean. 

Additionally data presented in Section VII.C show that, with the exception of tolerance to 

the herbicide dicamba, MON 87708 is phenotypically equivalent to conventional 

soybean. Thus, there are no changes to the inputs needed for MON 87708, and no 

specific impacts to most of the agronomic practices employed for production of soybean. 

In the areas where there is potential for impact on agronomic practices from the 

deregulation of MON 87708, the scope and magnitude of those impacts will be discussed. 

Soybean is planted in over 30 states, demonstrating its wide adaptation to varied soils and 

climate. The soil, moisture, and temperature requirements for producing soybean are 

generally similar to those for corn, and thus the two crops share a similar cultivation area. 

Proper seedbed preparation, appropriate variety selection, appropriate planting dates and 

plant population, and good integrated pest management practices are important for 

optimizing the yield potential and economic return for soybean. 

Annual and perennial weeds are perceived to be the greatest pest problem in soybean 

production (Aref and Pike, 1998). Weeds compete with soybean for water, nutrients, and 

light resulting in substantial yield losses when left uncontrolled. Weed species in 

soybean vary from region to region and state to state. Economic thresholds for 

controlling weeds in soybean require some form of weed management practice on all 

soybean acreage. Weed management practices include mechanical tillage, crop rotations, 

cultural practices, and herbicide application. Numerous selective herbicides are available 

for preplant, preemergence, and postemergence control of annual and perennial weeds in 

soybean. Approximately 98% of the soybean acreage in the U.S. receives an herbicide 

application (USDA-NASS, 2006). Soybean insects and diseases generally are considered 

less problematic, although infestations can reach economic thresholds requiring 

treatment. 

Volunteer soybean, i.e., soybean plants that have germinated and emerged unintentionally 

in a subsequent crop, are not considered a significant concern in rotational crops 

primarily because of climatic conditions and adequate control of volunteer soybean from 

tillage practices. Additionally, mechanical and chemical control methods are available to 

manage the occasional volunteer soybean plant. Due to its lack of weediness potential, 

introduction of MON 87708 in the soybean production system would have a negligible 

impact on managing soybean volunteer plants in rotational crops such as corn, cotton, 
Monsanto Company 10-SY-210U 171 of 721 

and wheat. The numerous control measures that are effective on conventional and 

Roundup Ready soybean volunteer plants will continue to be effective on volunteer 

MON 87708 plants when they arise. 

As shown in Sections VI and VII, with the exception of the dicamba tolerance trait, no 

phenotypic, compositional, or environmental differences between MON 87708 and 

conventional soybean have been observed. Moreover herbicide-tolerant soybean is 

currently grown on 91% of U.S. soybean acres (USDA-NASS, 2009c). MON 87708 will 

facilitate a wider window of application of dicamba (at planting and in-crop) and will 

replace or supplement the use of other soybean herbicides. Therefore, it is not anticipated 

that commercialization of MON 87708 in the U.S. would have a notable impact on 

current soybean cultivation practices, including the management of weeds, diseases, and 

insects other than the in-crop use of dicamba in soybean production. 

VIII.B. Overview of U.S. Soybean Production 

VIII.B.1. Soybean Production 

Soybean first entered North America in the 18th century (Hoeft et al., 2000). During the 

1930s, soybean started to be processed industrially in the U.S. for edible oil and protein 

meal. In 2008, soybean represented 56 percent of world oilseed production, and about a 

third of those soybeans were produced in the U.S. (ASA, 2009). In 2008, the U.S. 

exported 1.16 billion bushels (31.6 million metric tons) of soybean, which accounted for 

40 percent of the world's soybean exports (ASA, 2009). In total, the U.S. exported $20 

billion worth of soybean and soybean products globally in 2008 (ASA, 2009). China was 

the largest export market for U.S. soybean with purchases totalling $7.2 billion. Mexico 

was the second largest export market with purchases of $1.7 billion. Other significant 

markets include the European Union ($1.6 billion) and Japan ($1.3 billion). 

Approximately 94% of the world’s soybean seed supply was crushed to produce soybean 

meal and oil in 2008 (Soyatech, 2010), and the majority was used to supply the feed 

industry for livestock use or the food industry for edible vegetable oil and soybean 

protein isolates. 

The productivity of soybean is highly dependent upon soil and climatic conditions. In the 

U.S., the soil and climatic requirements for growing soybean are very similar to corn. 

The soils and climate in the Midwestern, Eastern, and portions of the Great Plains regions 

of the U.S. provide sufficient water under normal climatic conditions to produce a 

soybean crop. The general water requirement for a high-yielding soybean crop is 

approximately 20 inches of water during the growing season (Hoeft et al., 2000). Soil 

texture and structure are key components determining water availability in soils, where 

medium-textured soils hold more water, allowing soybean roots to penetrate deeper in 

medium-textured soils than in clay soils. Irrigation is used on approximately 9% of the 

soybean acreage to supplement the water supply during dry periods in the Western and 

Southern soybean growing regions (USDA-ERS, 2008). 
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Most of the soybean acreage is grown as a full-season crop. Approximately 8% of the 

soybean acres are planted in a double-crop system following winter wheat south of 35o 

North latitude (Boerma and Specht, 2004). However, this percentage can vary 

significantly from year to year. The decision to plant double-crop soybean is influenced 

by both agronomic and economic factors. Agronomic factors include harvest date of the 

wheat crop, which determines the double-crop soybean planting date, and available soil 

moisture. Economic factors include expected soybean price and anticipated economic 

return (Boerma and Specht, 2004). 

The U.S. soybean acreage in the past 10 years has varied from approximately 64.7 to 75.7 

million acres, with the lowest acreage recorded in 2007 and the highest in 2008 (Table 

VIII-1). Average soybean yields have varied from 33.9 to 43.3 bushels per acre over this 

same time period. Annual soybean production ranged from 2.45 to 3.19 billion bushels 

over the past ten years. According to data from USDA-NASS (2009a), soybean was 

planted on approximately 75.7 million acres in the U.S. in 2008, producing 2.96 billion 

bushels of soybean (Table VIII-1). Soybean acreage and production in 2008 was up from 

2007, mainly due to a decrease in corn acreage. The value of soybean reached $27.4 

billion in the U.S. in 2008 (USDA-NASS, 2009b). In comparison, corn and wheat values 

in 2008 were $47.37 and $16.57 billion, respectively (USDA-NASS, 2009a, b). 

For purposes of this agronomic practices discussion, soybean production is divided into 

three major soybean growing regions accounting for 99.1% of the 2008 U.S. soybean 

acreage: Midwest/Great Plains region (IL, IN, IA, KS, KY, MI, MN, MO, NE, ND, OH, 

SD, and WI), Southeast region (AL, AR, GA, LA, MS, NC, SC, and TN) and the Eastern 

Coastal region (DE, MD, NJ, NY, PA, and VA) (Table VIII-2). The vast majority of 

soybean was grown in the Midwest region, representing 82.1% of the total U.S. acreage. 

The Southeast and Eastern Coastal regions represented 14.3% and 2.7% of the acreage, 

respectively. Among the three regions, the Midwest region produced the highest average 

yield at 38.6 bushels per acre in 2008, and average state yields in this region ranged from 

28.0 to 47.0 bushels per acre. The average yield in the Southeast region was 34.4 bushels 

per acre, with states within this region averaging from 30.0 to 40.0 bushels per acre. The 

average yield in the Eastern Coastal region was 34.1 bushels per acre, with individual 

state averages ranging from 27.5 to 46.0 bushels per acre. 

Managing input costs is a major component to the economics of producing a soybean 

crop (Helsel and Minor, 1993). Key decisions on input costs include choosing what 

soybean varieties to plant, amounts of fertilizer to apply, and what herbicide program to 

use. The average operating cost for producing soybean in the U.S. in 2008 was $127.79 

per acre, according to statistics compiled by the USDA-Economic Research Service 

(USDA-ERS, 2008). The value of the production less operating cost was reported to be 

$318.66 per acre. A summary of potential production costs and returns are presented in 

Table VIII-3. 
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Table VIII-1. Soybean Production in the U.S., 1999 – 20081

Year 

Acres 

Planted 

(×1000) 

Acres 

Harvested 

(×1000) 

Average 

Yield 

(bushels/acre) 

Total 

Production 

(×1000 bushels) 

Value 

(billions $) 

2008 75,718 74,641 39.6 2,959,174 27.40 

2007 64,741 64,146 41.7 2,677,117 26.97 

2006 75,522 74,602 42.7 3,188,247 20.42 

2005 72,142 71,361 43.3 3,086,432 16.93 

2004 75,208 73,958 42.2 3,123,686 17.89 

2003 73,404 72,476 33.9 2,453,665 18.01 

2002 73,963 72,497 38.0 2,756,147 15.25 

2001 74,075 72,975 39.6 2,890,682 12.61 

2000 74,266 72,408 38.1 2,757,810 12.47 

1999 73,730 72,446 36.6 2,653,758 12.21 

Ave. 73,277 72,151 39.6 2,854,672 18.02 

1

Source is USDA-NASS (2009a,b). 

 
Monsanto Company 10-SY-210U 174 of 721 

Table VIII-2. U.S. Soybean Production by Region and State in 20081

Region/State

Acres Planted

(thousands)

Acres 

Harvested

(thousands)

Average Yield

(bushels/acre)

Total Production

(×1000 bushels)

Value

(billions $)

Midwest Region

Illinois 9,200 9,100 47.0 427,700 4.00

Indiana 5,450 5,430 45.0 244,350 2.27

Iowa 9,750 9,670 46.0 444,820 4.29

Kansas 3,300 3,250 37.0 120,250 1.03

Kentucky 1,390 1,380 34.0 46,920 0.42

Michigan 1,900 1,890 37.0 69,930 0.64

Minnesota 7,050 6,950 38.0 264,100 2.54

Missouri 5,200 5,030 38.0 191,140 1.72

Nebraska 4,900 4,860 46.5 225,990 2.12

North Dakota 3,800 3,760 28.0 105,280 0.96

Ohio 4,500 4,480 36.0 161,280 1.55

South Dakota 4,100 4,060 34.0 138,040 1.25

Wisconsin 1,610 1,590 35.0 55,650 0.51

Region Totals 62,150 61,450 38.6 2,495,450 23.30

Southeast Region

Alabama 360 350 35.0 12,250 0.12

Arkansas 3,300 3,250 38.0 123,500 1.09

Georgia 430 415 30.0 12,450 0.11

Louisiana 1,050 950 33.0 31,350 0.29

Mississippi 2,000 1,960 40.0 78,400 0.69

North Carolina 1,690 1,670 33.0 55,110 0.47

South Carolina 540 530 32.0 16,960 0.15

Tennessee 1,490 1,460 34.0 49,640 0.43

Region Totals 10,860 10,585 34.4 379,660 3.35

Eastern Coastal Region

Delaware 195 193 27.5 5,308 0.05

Maryland 495 485 30.0 14,550 0.13

New Jersey 92 90 29.0 2,610 0.02

New York 230 226 46.0 10,396 0.09

Pennsylvania 435 430 40.0 17,200 0.15

Virginia 580 570 32.0 18,240 0.16

Region Totals 2027 1994 34.1 68,304 0.60
Monsanto Company 10-SY-210U 175 of 721 

Table VIII-3. U.S. Soybean Production Costs and Returns in 20081 

Production Cost or Return Category Itemized Costs

Return per 

Planted Acre

($ USD)

 

Total Gross Value of Production 446.45

Operating Costs: Seed 44.35

Fertilizer 25.12

Chemicals 15.73

Custom operations 6.56

Fuel, lube and electricity 20.20

Repairs 12.91

Purchased irrigation water 0.12

Interest on operating capital 2.80

Total, operating costs 127.79

 

Allocated overhead: Hired labor 2.07

Opportunity cost of unpaid 

grower’s labor

16.77

Capital recovery of machinery 

and equipment

70.98

Opportunity cost of land (rental 

rate)

94.58

Taxes and insurance 9.64

General farm overhead 14.29

Total, allocated overhead 208.35

 

Total cost listed 336.13

Value of production less total cost 

listed

110.32

 

Value of production less operating 

costs

318.66

Supporting Information: Yield = 43 bushels/acre, Price = $10.48/bushel, Enterprise size = 303 

planted acres, Irrigated = 9%, Dry land = 91%. 

1

Source is USDA-ERS (2008). 
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VIII.B.2. Soybean Seed Production 

Standardized seed production practices are responsible for maintaining high-quality seed 

stocks, an essential basis for U.S. agriculture. By the early 20th century, agronomists 

learned how to develop specific plant varieties with desirable traits. In the U.S., state 

agricultural experiment stations developed many seed varieties that were distributed to 

growers for use. Seed was saved by growers and later sold to neighbors; however, the 

desirable traits of the varieties often were lost through random genetic changes and 

contamination with other crop and weed seed (Sundstrom et al., 2002). The value of seed 

quality (including genetic purity, vigor, and presence of weed seed, seed-borne diseases, 

and inert materials, such as dirt) was quickly identified as a major factor impacting crop 

yields. States developed seed laws and certification agencies to ensure that purchasers 

who received certified seed could be assured that the seed met established seed quality 

standards (Bradford, 2006). The federal government passed the U.S. Federal Seed Act of 

1939 to recognize seed certification and the establishment of official certifying agencies. 

Regulations first adopted in 1969 under the Federal Seed Act recognize land history, field 

isolation, and varietal purity standards for foundation, registered, and certified seed. 

Under international agreements such as the Organization for Economic Co-Operation and 

Development (OECD) scheme, the U.S. and other countries mutually recognize 

minimum seed quality standards (Bradford, 2006). The Association of Official Seed 

Certifying Agencies (AOSCA) represents state and private seed certification 

organizations in the U.S., and includes international member countries in North and 

South America, Australia, and New Zealand. 

Soybean seed is separated into four seed classes: 1) breeder, 2) foundation, 3) registered, 

and 4) certified (AOSCA, 2009). Breeder seed is seed directly controlled by the 

originating or sponsoring plant breeding organization or firm. Foundation seed is firstgeneration seed increased from breeder seed and is handled in a manner to maintain 

specific levels of varietal purity and identity. Registered seed is the progeny of 

foundation seed that is handled to maintain satisfactory varietal purity and identity. 

Certified seed is the progeny of breeder, foundation or registered seed, and is typically 

two generations removed from foundation seed. While not all soybean seed sold to 

growers is officially certified, commercial soybean seed sold and planted for typical 

soybean production is produced predominately to meet or exceed certified seed standards. 

This section of the petition will provide a broad overview of the practices used in 

producing certified seed. 

Soybean seed breeders and producers have put in place practical measures to assure the 

quality and genetic purity of soybean varietal seed for commercial planting. The need for 

such systems arose from the recognition that the quality of improved soybean varieties 

quickly deteriorated in the absence of monitoring for quality and genetic purity (CAST, 

2007). Seed certification programs were initiated in the early 1900s in the U.S. to 

preserve the genetic identity and variety purity of seed. There are special land 

requirements, seed stock eligibility requirements, field inspections and seed labeling 

standards for seed certification. Seed certification services are available through various 

state agencies affiliated with AOSCA. Large seed producers implement their own seed 

quality assurance programs. However, large seed producers often will utilize the services 
Monsanto Company 10-SY-210U 177 of 721 

of state certifying agencies as a third party source to perform certain field inspections and 

audits. 

U.S soybean production for all purposes has varied from approximately 64.7 to 75.7 

million acres in the past ten years (USDA-NASS, 2009a; Table VIII-1). To plant this 

area of soybean acreage requires 105 to 125 million units (50 lbs/unit) of soybean seed. 

This seed volume includes allowances for seed losses due to weather, poor yields, and 

quality issues. Additional allowances are included for distribution excess, seed returns, 

replants, and potential increases in soybean acreage. Assuming an average soybean yield 

of 45 bushels, or 54 units (50 lbs/unit) per acre, 1.9 to 2.3 million acres would be required 

to produce this volume of commercial certified soybean seed each year. 

Certified soybean seed is produced throughout most of the U.S. soybean-growing 

regions. Soybean varieties are developed and adapted to certain geographical zones and 

are separated into ten maturity groups – Group 00 to Group VIII (see Section VIII.C). 

Seed production for these maturity groups is grown in the respective geographical zone 

for each maturity group. However, the production areas generally are on the northern 

edge of the respective zone to minimize incidences of disease. 

Soybean seed is produced by a number of companies that produce and sell seed, such as 

Monsanto Company, Pioneer Hi-Bred International, Syngenta Seeds, Kruger Seed Co., 

and Becks Hybrids. In addition, certified seed is produced by toll seed producers, or 

tollers, which are companies that produce but do not directly sell certified seed, such as 

Remington Seeds LLC and Precision Soya. Seed companies and tollers in turn contract 

acreage with growers to produce the needed amount of soybean seed. Seed production or 

processing plants at these seed companies identify local soybean growers to produce the 

seed and also monitor and inspect seed fields throughout the growing season. The seed 

production plants also clean, condition, and bag the harvested soybean seed as well as 

monitor and inspect all the processes at the plant. Production plants typically produce 

between 100,000 units to 2,000,000 units of soybean seed. Production plants will 

produce the various soybean varieties in different climates or environments to spread 

production risks. 

The entire seed production process at the majority of the seed companies and tollers 

operate using International Organization for Standardization (ISO) certification standards 

and, therefore, include internal and external audits (ISO, 2009). ISO standards ensure 

desirable characteristics of seeds and services, such as quality, safety, reliability, and 

efficiency. The ISO standards represent an international consensus on good management 

practices with the aim of ensuring that the organization can consistently deliver excellent 

product or services. The standards not only must meet the customer’s requirements and 

applicable seed regulatory requirements, but must aim to enhance customer satisfaction 

and achieve continual improvement of its performance in pursuit of these objectives 

(ISO, 2009). 

The field operations and management practices for producing soybean seed are similar to 

normal soybean production. However, special attention is needed in certain areas to 

produce seed with high quality, high germination rates, and high genetic purity (Helsel 
Monsanto Company 10-SY-210U 178 of 721 

and Minor, 1993). General guidelines specific for seed production are discussed below. 

Importantly, the seed production field should not have been planted with soybean in the 

previous crop season in order to avoid potential volunteer soybean plants (even though 

the risk of soybean volunteer plants is negligible) and to ensure genetic purity. 

Very early planting is typically avoided because the seed produced from early planting 

often results in poorer quality seed (Helsel and Minor, 1993). Every effort must be made 

to eliminate weeds in a seed field through the use of herbicides and cultivation practices 

to prevent weed seed in the harvested soybean seed. Fields are scouted frequently for 

insect pests and insecticides are applied when insect pest infestations reach economical 

threshold levels. Foliar-applied fungicides should be considered when disease 

infestations are predicted in the area. Harvest should occur as soon as the mature 

soybean seed reaches 13% moisture content. Harvesting soybean seed with less than 

13% moisture can cause damage to the seed coat and result in split soybean seed that can 

affect germination and viability. Harvesting equipment must be adjusted to minimize or 

avoid seed damage. Harvesting equipment must be cleaned before entering the seed 

fields to assure genetic purity. Certain seed handling equipment, such as auger elevators, 

should be avoided because they can increase seed damage. 

Field inspections are vital to ensure the soybean seed meets seed certification 

requirements, ISO certification standards, regulatory standards, and trait licensing 

agreement standards. Field inspections are conducted on seed production fields 

throughout the soybean growing season to visually evaluate variety purity, ensure 

soybean plants are developing properly, and fields are maintained free of weeds, insects, 

and diseases. The fields are also mapped to ensure the seed field has the minimum 

federal isolation requirement of five feet as a physical barrier (AOSCA, 2009). Some 

states and seed producers have a stricter isolation requirement of 10 feet. 

Production plant personnel make every effort to avoid mechanical damage to the 

harvested seed during the screening, cleaning, and bagging process. Specific methods are 

used to assure the genetic purity and the identity of the seed is maintained throughout the 

handling and storage operation. Bin inspections and sample collections are conducted at 

storage locations at the seed production plant to examine the physical characteristics of 

the soybean seed and to ensure proper bin cleanout. Seed is inspected for appearance, 

disease, discoloration, seed coat, mechanical damage, inert matter, and weed seed. Warm 

and cold germination tests are conducted on all seed lots to verify acceptable germination 

rates. Many seed companies will also conduct tetrazolium staining tests to assess seed 

viability. 

Commercially certified soybean seed must meet state and federal seed standards and 

labeling requirements. AOSCA standards for certified soybean seed are as follows: 98% 

pure seed (minimum), 2% inert matter (maximum), 0.05% weed seed (maximum, not to 

exceed 10 per lb.), 0.60% total of other crop seeds (maximum), 0.5% other varieties 

(maximum, includes off-colored beans and off-type seeds), 0.10% other crop seeds 

(maximum, not to exceed three per lb.), and 80% germination and hard seed (minimum) 

(AOSCA, 2009). State seed certification standards vary slightly from state to state and 

can be more restrictive than the seed standards of AOSCA. 
Monsanto Company 10-SY-210U 179 of 721 

When deregulated, MON 87708 seed will be produced in the same manner as 

commercially certified soybean seed, such that it will meet all state and federal seed 

standards and labeling requirements. 

VIII.C. Production Management Considerations 

VIII.C.1. Pre-Season 

Well in advance of planting a soybean crop, decisions are made regarding the planned 

crop rotation, the tillage system and row spacing that will be implemented, the planting 

equipment that will be used, the seed or variety that will be planted, and soil fertility 

management requirements. Many of the decisions in this area are made prior to or 

immediately after harvest of the previous crop. There are many benefits to crop rotation, 

with the majority of the soybean acreage planted in a two-year corn-soybean rotation (see 

Section VIII.I). Crop rotation is generally a long-term decision, but the rotation sequence 

can be modified to take advantage of a particular economic or market opportunity. The 

decision to plant soybean in a conservation tillage or no-till system may require special 

equipment and will be made long before planting. In addition, this decision on tillage 

system usually will be a long-term commitment, provided the system is successful. A 

decision to change row spacing is a similar long-term commitment that generally requires 

new equipment. 

The benefits of conservation tillage or no-till systems are well documented and include 

reduced soil erosion, reduced fuel and labor costs, and conservation of soil moisture 

(CTIC, 2011). In 2004, approximately 27.5 million acres (39.6%) of soybean were 

planted in a no-till system (CTIC, 2007). Slow soybean emergence and growth leading to 

lower yields have been some of the concerns associated with adoption of conservation 

tillage systems in soybean, especially no-till. Research in Wisconsin and Minnesota 

shows that soil temperatures can be four to five degrees colder in no-till than 

conventional tillage systems, which can slow seedling emergence, but have little effect on 

soybean yield (Pedersen, 2008a). Improved planters for establishment of good soybean 

populations and planting Roundup Ready soybean allowing the use of glyphosate to 

effectively control weeds in no-till fields have made no-till a viable production system for 

soybean (Pedersen, 2008a). Extension specialists still recommend some spring tillage on 

fine-textured and poorly drained soils for proper seedbed preparation (Pedersen, 2008a). 

Most field crops, including soybean, respond well to fertilizer when planted in soils with 

low fertility levels. Soybean requires 16 essential elements for growth and development. 

Deficiencies in any of these elements can reduce yields (Hoeft et al., 2000). The primary 

or major essential nutrients are nitrogen, phosphorus and potassium. The soybean plant 

is a member of the legume family, like alfalfa and clover, and fixes a significant portion 

of its own nitrogen through the symbiotic relationship with the nitrogen-fixing 

Bradyrhizobia bacteria (Bradyrhizobium japonicum) that live in the nodules on its roots. 

Bradyrhizobia are unicellar, microscopic bacteria that invade the soybean plant through 

its root hairs (Hoeft et al., 2000). The plant responds to this invasion by forming nodules 

which contain colonies of bacteria. Once established on the soybean root, bacteria in the 

nodule take gaseous nitrogen from the atmosphere and fix it in forms easily used by the 
Monsanto Company 10-SY-210U 180 of 721 

soybean plant. Since these bacteria are not native to U.S. soils and would not normally 

be found in these soils, inoculation of the soybean seed with these bacteria is 

recommended when soybean has not been grown in a field for three to five years. 

Nitrogen fertilizer applications at planting generally do not improve yield and decrease 

nodulation while increasing the plant’s dependency on the soil for nitrogen (Pedersen, 

2008a). Therefore, nitrogen fertilizer is seldom applied prior to planting a soybean crop. 

Soil tests are the only reliable way to determine the pH, phosphorus, and potassium levels 

in the soil. Liming and fertilizer requirements subsequently are determined based on soil 

test results. Ideal soil test results for corn are also ideal for soybean (Scott and Aldrich, 

1970). In corn-soybean rotations in the Midwest, phosphorus and potassium fertilizers 

are applied prior to a corn crop in accordance with soil test recommendations, but are 

seldom applied prior to a soybean crop. However, in some of the southern growing areas, 

differences in crop rotations and soil types may require a fertilizer application prior to 

planting soybean. 

Although not common, deficiencies in soil can occur in secondary nutrients (calcium, 

magnesium, and sulfur) or micronutrients (boron, chloride, copper, iron, manganese, 

molybdenum, and zinc). The availability of soil nutrients is dependent on soil acidity or 

pH level. Because soybean is adversely affected when the pH is below approximately 5.8 

(Hoeft et al., 2000), soil pH should be maintained at about 6.0 to 6.5 through the addition 

of limestone. 

Soybean varieties are developed and adapted to certain geographical zones and are 

separated into ten maturity groups – Group 00 to Group VIII (Pimentel, 1991; Zhang et 

al., 2004). Groups 00 and 0 are the earliest maturity groups and are adapted best to the 

area north of latitude 46o North. Succeeding groups are adapted further south with 

Groups I and II within latitudes 41o and 46o North, and Group III within latitudes 38o and 

41o North. Group 00 through Group IV soybean varieties are planted in the Midwest and 

Eastern Coastal regions. Groups II, III and IV account for approximately 76% (24%, 

36%, and 16%, respectively) of the soybean planted in the U.S. (T. Schlueter, personal 

communication, August 2008). Groups IV through VIII are planted in the southern states 

with Groups V, VI and VII representing 7%, 2%, and 2% of the planted soybean, 

respectively (T. Schlueter, personal communication, August 2008). 

Soybean variety selection is crucial for high yield and quality, and is the foundation of an 

effective management plan (Pedersen, 2008a). Characteristics to consider in selecting a 

variety include maturity, yield potential, disease and pest resistance, iron deficiency 

tolerance (chlorosis), lodging score, height, and specific soybean quality traits, such as 

protein and oil content. If a field has a history of a particular disease or pest, planting 

soybean varieties that have resistance or tolerance to these pests and diseases can be an 

effective and economical method of control. 

VIII.C.2. Planting and Early Season 

An understanding of the growth stages of soybean is also important for the proper timing 

of certain management practices, such as herbicide and insecticide applications. In 
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addition, the impact of certain weather conditions, insect pests, and diseases on soybean 

yield is dependent on growth stage. The system of soybean growth stages divides plant 

development into vegetative (V) and reproductive (R) stages (Pedersen, 2008a). The 

vegetative stages begin with VE, which designates emergence. V stages continue and are 

numbered according to how many fully developed trifoliate leaves are present (i.e., V1, 

V2, etc.). The reproductive (R) stages begin at flowering (R1) and include pod 

development and plant maturation. Full maturity is designated as R8. 

Adequate soil moisture and warm temperatures facilitate rapid seed germination and 

emergence. The ideal soil temperature for soybean germination and emergence is 77oF 

(Pedersen, 2008a). However, waiting for soils to reach this soil temperature will delay 

planting beyond the optimum planting date that will maximize yield. Soybean can 

germinate at a soil temperature of 50oF when planted at a depth of two inches. However, 

emergence is slow and can take up to three weeks in northern climates. Because of 

fluctuations in soil temperature in early spring, soil temperature should not be the only 

criteria for optimum planting time. Planting into a good seedbed is the most important 

consideration. Planting into soil that is too wet will reduce emergence and plant 

population, and can lead to reduced yield. 

Planting date has the greatest impact on yield, according to research conducted in the 

Northern states (Hoeft et al., 2000). Highest yields are generally obtained when planting 

in early to mid May. Yields begin to decline quite rapidly when planting is delayed until 

late May. For example, the optimum planting dates for soybean in Iowa are the last week 

of April in the southern two-thirds of the state and the first week of May in the northern 

one-third of the state (Pedersen, 2008a). In the Southern U.S., planting adapted varieties 

before late April results in shorter plants and, in many cases, lower yields than when the 

same varieties are planted in May or early June. Planting after early June generally 

decreases plant height and yield due to water shortages in July and August. 

Variations in plant spacing through row spacing and plant population have a significant 

effect on canopy development and soybean yield. Row spacing is important to maximize 

soybean yield. Research in the Midwest over the past 20 years consistently shows that 

row spacing of less than 20 inches is preferred for soybean regardless of tillage system, 

rotation sequence or planting date (Pedersen, 2008a). In the Southern states, the 

advantage from narrow rows is less consistent and less beneficial. In 2000, 

approximately 40% of soybean was planted in row spacing of 10 inches or less, 27% in 

10.1 to 28.5 inches, and 33% in rows wider than 28.5 inches (Hoeft et al., 2000). 

Soybean has the ability to produce good yield over a wide range of plant populations. 

Most soybean varieties have the ability to branch and adjust the number of pods on 

branches to compensate for large differences in seeding rate. Maximum yields generally 

require planting rates that result in about 2.5 to 5 plants per square foot (Hoeft et al., 

2000). Therefore, a full stand of soybean is approximately eight to ten plants per foot of 

row at harvest for 40-inch rows, six to eight plants per foot of row in 30-inch rows, four 

to six plants in 20-inch rows, and two to three plants in 10-inch rows. This translates to 

109,000 to 218,000 plants per acre at harvest. Higher populations are recommended in 

narrow rows for maximum yields because plants are more uniformly spaced in narrow 
Monsanto Company 10-SY-210U 182 of 721 

rows. Seeding rates are generally 10 to 25% higher than the desired harvest population, 

especially in no-till fields, to account for the losses in germination, emergence, and 

seedling diseases. The accuracy of the planting equipment also can impact the decision 

on seeding rate. Soybean seed traditionally has been sold by weight. Therefore, the 

grower must know the number of seeds per pound for the particular soybean varieties 

being planted for accurate seeding rates. 

Treating soybean seed with a fungicide (e.g., metalaxyl or mefenoxam) to prevent 

damping-off diseases may be beneficial when planting in cold, wet soils, using reduced 

till and no-till planting systems, and when planting seed with a low germination rate 

(<80%) or low seed vigor (Pedersen, 2008a). 

Annual and perennial weeds are considered to be the greatest pest problem in soybean 

production (Aref and Pike, 1998). In order to maximize yields, weeds must be controlled 

during the early growth stages of soybean because weeds compete with soybean for 

water, nutrients, and light. There have been many studies examining the loss in soybean 

yield due to weed competition. The amount of loss is dependent upon the species, time 

when the weed is growing with the crop, and crop cultural practices, particularly row 

spacing. A sampling of these studies show that soybean yield reductions are generally in 

the range from 10% to 50% (Norsworthy et al., 2002; Shurtleff and Coble, 1985; Vail et 

al., 1993, Hock et al., 2006). In one study, yield loss due to several annual grasses ranged 

from 13% to 16% while yield losses due to various annual broadleaf weeds ranged from 

23% to 52% (Hock et al. 2006). A combination of tillage and herbicides are used to 

control weeds throughout the growing season (Section VIII.F). 

VIII.C.3. Mid to Late Season 

Ideal daytime temperatures for soybean growth are between 75oF and 85oF (Hoeft et al., 

2000). Warmer temperatures result in larger plants and earlier flowering. Sustained 

temperatures below 75oF will delay the beginning of flowering significantly. Seed set 

also is affected by temperature. Seed set is generally good when pollination follows 

night temperatures around 70oF. Soybean varieties differ in their response and tolerance 

to temperatures. 

Soybean is photoperiod sensitive, which means that it transitions from vegetative to 

flowering stage in direct response to length of daylight (Scott and Aldrich, 1970). Most 

soybean varieties begin flowering soon after the day length begins to shorten. Flowering 

of southern varieties is initiated by a shorter day than that of varieties adapted to the 

north. The extent of vegetative growth occurring after the initiation of flowering depends 

not only on environmental factors but also the growth habit. Soybean varieties are 

described as either indeterminate or determinate in their growth habit (Scott and Aldrich, 

1970). Indeterminate varieties increase their height by two to four times after flowering 

begins. Indeterminate varieties are typically grown in the northern and central U.S. 

Determinate varieties increase their height very little after flowering and generally are 

grown in the southern U.S. Indeterminate and determinate varieties also differ in 

flowering characteristics. Indeterminate plants generally bloom first at the fourth or fifth 
Monsanto Company 10-SY-210U 183 of 721 

node and progress upward. Flowering on determinate plants begins at the eight or tenth 

node and progresses both downward and upward. 

The first appearance of flowers signals the beginning of the reproductive stage, namely 

the R1 stage (Hoeft et al., 2000). The reproductive period consists of flowering, pod set, 

and seed formation. Climatic conditions such as temperature and moisture supply during 

the flowering period will affect the number of flowers. The soybean plant does not form 

a pod from each flower. It is common for the soybean plant to have 75% of the flowers 

fail to develop a pod (Scott and Aldrich, 1970). This characteristic makes soybean less 

susceptible than corn to short periods of adverse weather during flowering. Under 

normal conditions, pod set occurs over about a three week period. Good soil moisture is 

most critical during the pod-filling stages to prevent pod abortion and to ensure high 

yields (Hoeft et al., 2000). Another critical requirement during the seed-filling stages is a 

high rate of photosynthesis to maximize yield. High humidity and temperatures during 

seed development and maturity can result in poor seed quality because these conditions 

promote the development of reproductive-stage diseases. 

VIII.C.4. Harvest Season 

When dry matter accumulation ends, the plant is considered to be physiologically mature. 

The seed moisture content is approximately 55 to 60% at this stage (Hoeft et al., 2000). 

At this stage, namely R7, at least one normal pod on the plant reaches the mature pod 

color. Under warm and dry weather conditions, seed moisture content will drop to 13 to 

14% in 10 to 14 days from physiological maturity (Hoeft et al., 2000). Soybean can be 

harvested when the moisture content drops below 15%. However, soybean should be at 

13% moisture to be stored without artificial drying (Scott and Aldrich, 1970). Moisture 

content below 12% may increase seed cracking and seed coat damage. 

Pre-harvest losses are influenced by soybean variety, weather, and timeliness of harvest 

(Scott and Aldrich, 1970). Timely harvest when the moisture content is 13 to 14% also 

will minimize losses. Proper operation and adjustment of the combine is essential to 

minimizing harvest losses in the field. 

VIII.D. Management of Insects 

Although insects are rated as less problematic than weeds in U.S. soybean production, 

management of insect pests during the growth and development of soybean is important 

for protecting the yield of soybean (Aref and Pike, 1998). Understanding the impact of 

insects on soybean growth is essential for proper management (Higley and Boethel, 

1994). It is important to understand the way that insects injure soybean as well as how 

the soybean plant responds to insect injury. Insect injury can impact yield, plant 

maturity, and seed quality. Insect injury in soybean seldom reaches levels to cause an 

economic loss, as indicated by the low percentage (16%) of soybean acreage that receives 

an insecticide treatment (USDA-NASS, 2007b). 

Characterizing soybean responses to insect injury is essential in establishing economic 

injury levels (Higley and Boethel, 1994). Most often, soybean insects are categorized or 
Monsanto Company 10-SY-210U 184 of 721 

defined by the plant parts they injure, namely root-feeding, stem-feeding, leaf-feeding, or 

pod-feeding insects. The root- and stem-feeding insect groups are often the hardest to 

scout and typically are not detected until after they have caused their damage. The leaffeeding insects comprise the biggest group of soybean insect pests, but not necessarily the 

most economically damaging insects. Research on defoliation has determined that a 

major effect of leaf injury is to reduce light interception by the soybean canopy which in 

turn can have a significant effect on yield (Higley and Boethel, 1994). Soybean has an 

extraordinary capacity to withstand considerable defoliation early in the season without 

significant yield loss. By contrast, defoliation during the flowering and pod filling stages 

poses a greater threat to yield because the soybean plant has less time to compensate for 

injury compared to other growth stages. Research indicates that the soybean plant can 

sustain a 35% leaf loss prior to the pre-bloom period without lowering yield (NDSU, 

2002). However, from pod-set to maturity, the plant can tolerate only a 20% defoliation 

level before yield is impacted. 

VIII.E. Management of Diseases and Other Pests 

More than 100 pathogens are known to affect soybean, of which 35 are considered to be 

of economic importance (Heatherly and Hodges, 1999). The estimated yield losses to 

soybean diseases in the U.S. were 12.5, 13.2, and 13.0 million metric tons in 2008, 2009, 

and 2010, respectively ( Wrather and Koenning, 2011), which equated to 15.5%, 14.4%, 

and 14.4% of total soybean production, respectively (ASA, 2011). Pathogens can affect 

all parts of the soybean plant, resulting in reduced quality and yield. The extent of losses 

depends upon the pathogen, the state of plant development and health when infection 

occurs, the severity of the disease on individual plants, and the number of plants affected 

(Heatherly and Hodges, 1999). 

One or more diseases can generally be found in fields wherever soybean is grown 

(Heatherly and Hodges, 1999). However, a pathogen may be very destructive one season 

and difficult or impossible to find the next season. The extent and severity of soybean 

diseases depend on the degree of compatibility between the host and the pathogen and the 

influence of the environment. 

According to field surveys conducted in soybean-producing states during 1996 to 2010, 

soybean cyst nematode (SCN), Heterodera glycines, caused the greatest soybean yield 

losses (Wrather and Koenning, 2011). Phytophthora root and stem rot (Phytophthora 

sojae), brown spot (Septoria glycines), charcoal rot (Macrophomina phaseolina), 

Sclerotinia stem rot (Sclerotinia sclerotiorum), seedling diseases, and sudden death 

syndrome (Fusarium solani f.sp. glycines) followed in economical importance. As 

expected, yield losses vary by region. Sclerotinia stem rot caused yield losses in several 

Northern states, but not in other states. Rhizoctonia foliar blight losses were greatest in 

Arkansas, Louisiana, and Texas where humidity and temperature conditions are suitable 

for disease development (Wrather et al., 2001). 

Selecting resistant varieties is the primary tool growers have for disease control 

(Heatherly and Hodges, 1999). Resistant varieties may have morphological or 

physiological characteristics that provide immunity, resistance, tolerance or avoidance to 
Monsanto Company 10-SY-210U 185 of 721 

certain pathogens. Cultural practices can also play an important role in disease 

management by reducing initial inoculums or reducing the rate of disease development 

(Heatherly and Hodges, 1999). Preplant tillage can bury crop residue, which encourages 

the decomposition of fungal-resting structures. Crop rotation is routinely recommended 

as a disease-management strategy. Rotating crops interrupts the disease cycle and allows 

time for the decomposition of inoculums. One exception is Rhizontonia spp., a soilinhabitant pathogen that grows on a wide variety of crops and can survive sufficiently in 

the soil to make crop rotation as a means of controlling this pest impractical. Row 

spacing, plant population, and planting date also can be changed to manage soybean 

diseases. 

Soybean cyst nematode is one of the most damaging pathogens of soybean throughout 

the soybean growing regions of the U.S. (Pedersen, 2008b). Losses have been estimated 

to be at about $1.5 billion in the U.S. (Pedersen, 2008a). SCN can cause yield losses up 

to 50%, where this pest in 2004 alone caused an estimated loss of 50 million bushels of 

soybean in Iowa (Pedersen, 2008c). Soybean cyst nematodes feed on the roots, causing 

severely stunted and yellow plants. The simplest, least expensive method to reduce 

populations of this pest is to rotate soybean with a non-host crop such as corn, small 

grains, or sorghum. Planting resistant varieties is regarded as the best and most effective 

management practice to prevent losses from this pest. Several public and private soybean 

varieties offer sources of resistance to certain races of nematode. Alternating varieties 

with different sources of resistance also is beneficial. 

High-quality seed is essential for controlling seedling diseases. The most important 

seedling diseases in soybean are Phytophthora spp., Pythium spp., Rhizoctonia spp., and 

Fusarium spp. (Pedersen, 2008a). Many soybean varieties demonstrate resistance to 

specific taxonomic races of Phytophthora. Treating soybean seed with a fungicide (e.g., 

metalaxyl or mefenoxam) is effective against damping-off disease (seedling blight) 

caused by common soil fungi, such as Phytophthora spp. and Pythium spp. Fungicide 

seed treatments are recommended where there is a history of these seedling diseases. 

Asian soybean rust is a foliar fungal disease that typically infests soybean during 

reproductive stages of development and can cause defoliation and reduce yields 

significantly in geographies such as Brazil (Dorrance et al., 2007). Soybean rust is 

caused by the fungus Phakopsora pachyrhizi. This disease in the U.S. was first detected 

in Louisiana in 2004 (LSU, 2009). At this time, foliar application of fungicides is the 

standard disease-management practice to limit yield losses due to soybean rust. 

Foliar fungicide applications can effectively reduce the incidence of many fungal 

diseases (Heatherly and Hodges, 1999). However, the economic return from a fungicide 

application may be limited to select soybean production systems; for example, high-yield 

environments or when producing soybean seed. According to USDA-NASS (2007b) 

statistics, fungicides were applied on approximately 4% of the soybean acreage in 2006. 
Monsanto Company 10-SY-210U 186 of 721 

VIII.F. Weed Management 

Annual weeds are perceived to be the greatest pest problem in soybean production, 

followed by perennial weeds (Aref and Pike, 1998). Soybean insects and diseases are 

rated less problematic but may reach economic thresholds requiring treatment. Weed 

control in soybean is essential to optimizing yields. Weeds compete with soybean for 

light, nutrients, and soil moisture. Weeds can harbor insects and diseases, and also can 

interfere with harvest, causing extra wear on harvest equipment (Pedersen, 2008a). The 

primary factors affecting soybean yield loss from weed competition are the weed species, 

weed density, and the duration of the competition. When weeds are left to compete with 

soybean for the entire growing season, yield losses can exceed 75% (Dalley et al., 2001). 

Generally, the competition between crops and weeds increases with higher levels of weed 

density. The time period that weeds compete with the soybean crop influences the level 

of yield loss. In general, early season weed competition will have the greatest negative 

impact on yield (Dalley et al., 2001). Soybean plants withstand early-season weed 

competition longer than corn without affecting yield, and the canopy closes earlier in 

soybean than corn. In addition, canopy closure is much sooner when soybean is drilled 

or planted in narrow rows. The most common weeds in soybean for each of the three 

major U.S. growing regions are presented in Tables VIII-4, VIII-5 and VIII-6. 

Crop rotations and environment have a significant impact on the adaptation and 

occurrence of weeds in soybean. Foxtail spp. (Setaria spp.), pigweed (Amaranthus spp.), 

velvetleaf (Abutilon theophrasti), lambsquarters (Chenopodium album), and cocklebur 

(Xanthium strumarium) are common weeds in Midwest corn and soybean fields. 

However, growers consider giant ragweed (Ambrosia artemisiifolia), lambsquarters, 

Canada thistle (Cirsium arvense), cocklebur, and velvetleaf to be the top five most 

problematic weeds in corn and soybean because of difficulty controlling these weeds 

(Nice and Johnson, 2005). In a recent survey of growers utilizing glyphosate-tolerant 

crops, pigweed, morningglory (Ipomoea spp.), Johnsongrass (Sorghum halepense), 

ragweed spp. (Ambrosia spp.), foxtail, and velvetleaf were mentioned as the most 

problematic weeds, depending on the state and cropping system (Kruger et al., 2009). 

With the exception of morningglory and pigweed, these problematic weed species were 

present before the introduction of glyphosate-tolerant crops, and some improvement in 

weed control was realized after the implementation of glyphosate-tolerant cropping 

systems (Kruger et al., 2009). Common waterhemp (Amaranthus rudis) and ragweed 

were the most frequently mentioned problematic weeds in glyphosate-tolerant crops in 

Illinois, Indiana and Iowa. 

The most frequently reported common weeds in the Southeast region were morningglory 

(Ipomoea spp.), prickly sida (Sida spinosa), johnsongrass (Sorghum halepense), 

sicklepod (Cassia obtusifolia), and broadleaf signalgrass (Brachiaria platyphylla) 

(Webster et al., 2005, 2009). Morningglory, sicklepod, and pigweed are the most 

frequently mentioned problematic weeds in glyphosate-tolerant crops in Mississippi and 

North Carolina (Kruger et al., 2009). 

Cultural and mechanical weed control practices can be important components of an 

effective weed management program (Loux et al., 2009). Crop rotation, narrow row 
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spacing and planting date are a few of the crop management practices that are 

implemented to provide the crop with a competitive edge over weeds. Although the 

primary purpose of tillage is for seedbed preparation, tillage is still used to supplement 

weed control with selective herbicides in soybean production. Approximately 98% of the 

soybean acreage received an herbicide application in 2006, indicating the importance of 

excellent weed control in maximizing soybean yield (USDA-NASS, 2007b). 

Herbicide-tolerant soybean was introduced to provide growers with additional options to 

improve crop safety and/or improve weed control. The Roundup Ready soybean system 

(planting Roundup Ready soybean and applying glyphosate in crop to provide primary 

weed control) was introduced in 1996 and has become the standard weed control program 

in U.S. soybean production and is utilized on 91% of U.S. soybean acreage (USDANASS, 2009c). 

Herbicides provide effective and economical control of weeds in soybean. The risk of 

weeds developing resistance to herbicides and the potential impact of resistance on the 

usefulness of an herbicide vary greatly across different mechanisms of action and are 

dependent on a combination of factors, such as selection pressure, herbicide soil residual 

activity, herbicide chemistry, prolific seed production and high genetic variation in plants 

(see Appendix K for a more detailed discussion of herbicide resistance in weeds). Weedresistance management programs that integrate the use of herbicides with different 

mechanisms of action and short residual activity times in soil reduce selection pressure 

exerted on weed species (Prather et al., 2000). Crop rotation can also be beneficial in 

managing resistance because it may allow the grower to manipulate planting times to 

avoid early-season weed germination and to use mechanical as well as chemical weed 

control methods (Jordan et al., 1995). As described in Appendix K, when utilized in an 

integrated manner, these management practices can be used to impede the development 

of herbicide resistance in weeds. 
Monsanto Company 10-SY-210U 188 of 721 

Table VIII-4. Common Weeds in Soybean Production: Midwest Region 

Foxtail spp. (12)1

 Ragweed, giant (3) Dandelion (1) 

Pigweed spp. (11) Shattercane (3) Johnson grass (1) 

Velvetleaf (11) Quackgrass (3) Milkweed, honeyvine (1) 

Lambsquarters (10) Buckwheat, wild (2) Nightshade, hairy (1) 

Cocklebur (9) Crabgrass spp. (2) Oats, wild (1) 

Ragweed, common (7) Kochia (2) Pokeweed, common (1) 

Smartweed spp. (6) Mustard, wild (2) Prickly sida (1) 

Morningglory spp. (5) Nightshade, Eastern black (2) Proso millet, wild (1) 

Sunflower, spp. (5) Palmer pigweed (2) Sandbur, field (1) 

Waterhemp spp. (5) Canada thistle (1) Venice mallow (1) 

Horseweed (marestail) (3) Chickweed (1) Volunteer cereal (1) 

Panicum, fall (3) Cupgrass, woolly (1) Volunteer corn (1) 

1

Number provided in parenthesis is the number of states out of the thirteen total states in the 

Midwest region reporting each weed as a common weed. 

Sources: 

IL: University of Illinois (2002) and Aaron Hager, Extension Weed Specialist, University of 

Illinois - Personal Communication (2006). 

IN: 2003-2005 Statewide Purdue Horseweed Weed Survey, Special database query and personal 

communication (2006), Bill Johnson, Extension Weed Specialist, Purdue University. 

IA, MN, OH, WI: WSSA, 1992. 

KS: Dallas Petersen, Extension Weed Specialist, Kansas State - Personal communication (2006). 

KY, MO: Webster et al., 2005. 

MI: Davis et al., 2005. 

NE: Alex Martin, Extension Weed Specialist, University of Nebraska – Personal communication 

(2006).

ND: Zollinger, 2000. 

SD: Michael Moechnig, Extension Weed Specialist, South Dakota State University – Personal 

communication (2006). 
Monsanto Company 10-SY-210U 189 of 721 

Table VIII-5. Common Weeds in Soybean Production: Southeast Region 

Morningglory spp. (8)1

 Goosegrass (3) Cutleaf evening-primrose (1) 

Crabgrass spp. (6) Johnsongrass (3) Groundcherry (1) 

Prickly sida (6) Ragweed, common (3) Henbit (1) 

Nutsedge spp. (6) Cocklebur (2) Lambsquarters (1) 

Sicklepod (5) Florida beggarweed (2) Ragweed, giant (1) 

Signalgrass, broadleaf (5) Hemp sesbania (2) Smartweed (1) 

Palmer pigweed (4) Horseweed (marestail) (2) Spurge, nodding/hyssop (1) 

Pigweed spp. (4) Texas millet (2) Spurge, Prostrate (1) 

Barnyard grass (3) Browntop millet (1) Tropic croton (1) 

Florida pusely (3) Copperleaf, hophorn (1) 

1 Number provided in parenthesis is the number of states out of the eight total states in the 

Southeast region reporting each weed as a common weed. 

Sources: 

AL, AR, GA, LA, NC, SC: Webster et al., 2009. 

MS, TN: Webster et al., 2005. 

Table VIII-6. Common Weeds in Soybean Production: Eastern Coastal Region 

Foxtail spp. (6)1 Morningglory spp. (4) Dandelion (1) 

Ragweed, common (6) Panicum, fall (4) Goosegrass (1) 

Velvetleaf (6) Crabgrass spp. (3) Johnson grass (1) 

Lambsquarters (5) Nutsedge spp. (3) Nightshade, Eastern black (1) 

Pigweed spp. (5) Quackgrass (2) Prickly sida (1) 

Cocklebur (4) Canada thistle (1) Shattercane (1) 

Jimson weed (4) Burcucumber (1) Smartweed spp. (1) 

1

Number provided in parenthesis is the number of states out of the six total states in the Eastern 

Coastal region reporting each weed as a common weed. Data were not available for DE in 

soybean. 

Sources: 

DE, MD, NJ, PA: WSSA, 1992. 

NY: Russell Hahn, Extension Weed Specialist, Cornell University – Personal Communication 

(2006). 

VA: Webster et al., 2009. 

VIII.F.1. Methods of Weed Control in Soybean 

Mechanical methods of weed control including tillage have been used for centuries to 

control weeds in crop production. Spring or fall preplant tillage and in-crop shallow 

cultivation can effectively reduce the competitive ability of weeds by burying the plants, 

disturbing or weakening their root systems, or causing sufficient physical injury to kill 

the weeds. Research in the early 1900s centered on determining the economic benefits of 

removing weeds with the use of cultivation (Klingman et al., 1975). A consequence of 

in-crop cultivation for weed control can be injury to crop roots and moisture loss. 

Selective herbicides have proved more efficacious and reduced the need for in-crop 

tillage or cultivation to control weeds in soybean production. The development of 

selective herbicides has progressed rapidly since the introduction of the first synthetic 
Monsanto Company 10-SY-210U 190 of 721 

herbicide (2,4-D) for weed control in corn in the early 1940s. Although the primary 

purpose of tillage is for seedbed preparation, tillage still is used to supplement weed 

control with selective herbicides in soybean production. 

Alanap (1949), amiben (1958), trifluralin (1959), linuron (1960), and alachlor (1966) led 

the way for numerous selective herbicides in soybean (Agranova, 2010). Bentazone 

(1968) was one of the early selective postemergence herbicides used in soybean 

production. By the early 1990s, there were over 70 registered herbicides or premix 

herbicides for weed control in soybean (Gianessi et al., 2002). Table VIII-7 provides a 

summary of herbicide use in soybean production in the U.S. from 1995 through 2001. 

Weed control programs in soybean production during this time period consisted of 

preemergence herbicides used alone or in a tank mixture with other preemergence 

herbicides. Applications were made as preplant incorporated or preemergence surface 

applications prior to or at planting. Tank mixtures of two preemergence herbicides were 

used to broaden the spectrum of control to both grasses and broadleaf weed species. 

Preemergence herbicides are followed by postemergence applications to control weed 

escapes that emerge later in the crop. Total postemergence programs were seldom used 

in soybean production prior to 1995. For soybean planted in a no-till system, an 

additional preplant burndown herbicide application for broad-spectrum control of 

existing weeds at time of planting was also applied. Therefore, multiple herbicides 

and/or multiple applications were generally made in soybean production. The average 

number of herbicide applications per acre in soybean rose from 1.5 in 1990 to 1.7 

applications in 1995 reflecting the use of at-plant and postemergence applications or two 

postemergence applications (Gianessi et al., 2002). 

It is important to understand herbicide use in 1995, as this was prior to the introduction of 

Roundup Ready soybean system. The most widely used herbicides in 1995 were the 

sulfonylurea (chlorimuron, thifensulfuron) and imidazolinone (imazethapyr, imazaquin) 

herbicide classes that are applied preemergence and postemergence in a soybean crop. 

These two classes of herbicides, both acetolactate synthase (ALS) inhibitors, were 

applied on approximately 87% of the soybean treated acres in 1995 (Table VIII-7). The 

dinitroanaline herbicides (trifluralin and pendimethalin) were the second most widely 

used preemergence herbicides. Selective postemergence herbicides were used on 52% of 

the treated acres and were generally either effective on grass species or broadleaf species. 

Sethoxydim, clethodim, quizalofop, and fluazifop were among the postemergence grass 

herbicides. Acifluorfen and bentazon were the main postemergence broadleaf herbicides. 

Glyphosate was used on 20% of the treated acres, mainly as a preplant burndown 

treatment, but it also was used in spot treatments or ropewick applications to control 

weed escapes or volunteer corn in soybean. 

Herbicide programs used in conventional soybean have not changed significantly since 

1995, with many of the traditional herbicides still in use. Although, new active 

ingredients have been introduced, including carfentrazone, sulfentrazone, flufenacet, 

flumetsulam, flumiclorac, flumioxazin, cloransulam, and imazamox. These new active 

ingredients improve the level or spectrum of weed control. Numerous products have 

been introduced that are a pre-mixture product of two active ingredients for broad 

spectrum weed control. Some of the new active ingredients and pre-mixtures are more 
Monsanto Company 10-SY-210U 191 of 721 

effective in controlling waterhemp, ALS-resistant weeds, and other hard-to-control weeds. 

Hard-to-control weeds generally require a higher rate and/or application at a smaller 

growth stage in order to consistently achieve commercially acceptable control. Refer to 

the Roundup WeatherMax label (U.S. EPA Reg. No. 524-537) for a listing of these 

weeds. Herbicide resistant weeds are those listed on the International Survey of Resistant 

Weeds website (www.weedscience.org). 

Table VIII-8 provides a summary of the herbicide use in soybean in the U.S. in 2006. In 

2006, herbicide-tolerant soybean (glyphosate-tolerant) was planted on 89% of the 75.5 

million acres of soybean (USDA-NASS, 2007a). With the high percentage of 

glyphosate-tolerant soybean and the additional use of glyphosate for preplant burndown 

applications on both glyphosate-tolerant and conventional soybean, it is not surprising 

that glyphosate was used on 97% of the total soybean acres in 2006. The percentage of 

herbicide-tolerant soybean has subsequently increased to 91% in 2009 (USDA-NASS, 

2009c). The remaining preemergence and postemergence herbicides are utilized in 

conventional soybean as well as glyphosate-tolerant soybean. A grower survey 

conducted in 2006 showed that 15 to 21% of growers applied non-glyphosate herbicides 

as another mode-of-action in addition to glyphosate for weed control in glyphosatetolerant soybean (Givens et al., 2009). These non-glyphosate herbicides were applied 

prior to planting, at planting and postemergence in soybean. The non-glyphosate 

herbicides mainly included applications of chlorimuron, flumiclorac, pendimethalin, 

imazethapyr, and 2,4-D, which were commonly used herbicides in weed management 

programs prior to the introduction of glyphosate-tolerant soybean. Although these nonglyphosate herbicides were applied to supplement the weed control provided by 

glyphosate, researchers report that approximately 40 to 55% of the growers utilizing 

glyphosate-tolerant crops indicate that rotating herbicides or tank mixing glyphosate with 

other herbicides is an effective management practice to minimize glyphosate resistance 

development (Johnson et al., 2009). It should be noted that in 2006 approximately 

16,000 lbs of dicamba was used in soybean production which would be a sufficient 

amount of dicamba to treat 64,000 acres assuming the average application rate of 0.25 lb 

dicamba acid equivalent (a.e.) per acre (see TableVIII-7). Dicamba is currently labeled 

only for preplant and preharvest applications in soybean, where restrictions on days after 

preplant treatment are required due to insufficient ability of soybean to tolerate 

applications of the herbicide, referred to as “crop tolerance.” Similarly, dicamba 

currently cannot be used in-crop postemergence applications on soybean due to a lack of 

crop tolerance. 

Tables VIII-9 and VIII-10 provide a summary of the crop tolerance to herbicides applied 

in soybean production and the efficacy of these herbicides on 25 of the common weed 

species identified in Section VIII.F. These tables list only the most commonly used 

herbicides in soybean production. Glyphosate applied postemergence (as part of the 

Roundup Ready soybean system) and four other herbicides applied either preemergence 

or postemergence have the highest crop tolerance rating of excellent. The other 

herbicides are rated only good to poor. Seldom would one field or farm have all 25 weed 

species, but they generally have a mixture of grass and broadleaf weed species. These 

ratings can be used by growers to facilitate the selection of an herbicide program for a 

soybean crop, which offers the best overall control of the weed species. Based on Tables 
Monsanto Company 10-SY-210U 192 of 721 

VIII-9 and VIII-10, glyphosate is considered to have better control (>80%) on more grass 

and broadleaf weed species than any other herbicide. Glyphosate/imazethapyr has the 

next highest overall rating, but it is rated only good on crop tolerance. S-Metolachlor and 

pendimethalin are rated high on many grass species, but are rated low on most of the 

broadleaf weed species. Chlorimuron/tribenuron, fomesafen, and 

flumioxazin/cloransulam are rated high on the broadleaf species, but are rated low on 

grass species. 

Table VIII-7. Herbicide Use in Soybean in the U.S. from 1995 through 20011

 Percent-Treated Acres 

Active Ingredient 1995 1996 1997 1998 1999 2000 2001 

2,4-D 10 13 8 7 5 5 4 

2,4-DB 1 <1 1 <1 <1 NA NA 

Acifluorfen 12 11 12 7 3 3 3 

Alachlor 4 5 3 2 2 1 <1 

Bentazon 12 11 11 7 4 2 1 

Chlorimuron 16 14 13 12 12 10 5 

Clethodim 5 7 4 4 5 4 4 

Clomazone 4 3 5 4 1 <1 <1 

Cloransulam NA NA NA 1 5 4 5 

Dimethenamid 1 1 1 1 <1 <1 NA 

Ethalfluralin 1 1 <1 NA <1 <1 NA 

Fenoxaprop 6 4 6 4 4 4 3 

Fluazifop 10 7 7 5 4 5 3 

Flumetsulam 2 2 4 2 2 2 <1 

Flumiclorac NA 2 1 <1 <1 <1 <1 

Fomesafen 4 5 6 6 4 7 7 

Glyphosate 20 25 29 47 62 66 76 

Imazamox NA NA NA 7 3 6 5 

Imazaquin 15 15 13 8 5 4 2 

Imazethapyr 44 43 38 17 16 12 9 

Lactofen 5 8 4 2 2 2 1 

Linuron 2 1 1 <1 <1 <1 NA 

Metolachlor 7 5 7 4 4 2 NA 

Metribuzin 11 9 10 6 5 4 2 

Paraquat 2 1 2 1 1 <1 NA 

Pendimethalin 26 27 25 18 14 11 10 

Quizalofop 6 7 4 3 1 <1 <1 

S-Metolachlor NA NA NA NA NA NA <1 

Sethoxydim 7 9 7 5 3 2 1 

Sulfentrazone NA NA NA 3 4 4 5 

Thifensulfuron 12 10 9 5 5 6 2 

Trifluralin 20 22 21 16 14 14 7 

1

Source is Gianessi et al. (2002).
Monsanto Company 10-SY-210U 

Table VIII-8. Agricultural Chemical Applications Registered for Soybean Use in AR, IA, IL, IN, KS, KNE, NC, ND, OH, SD, TN, VA, and WI in 20061Herbicide Chemical Family

Mode-of-Action (MOA)

PercentTreated Acres

Total Area 

Applied

(Percent/MOA)

Quantity 

Applied

(1000 lbs)

Glyphosate glycine

EPSPS inhibitor

4

97

2,841

Glyphosate, amm. Salt glycine * 142 Glyphosate, iso. salt glycine 92 88,903

Sulfosate glycine 1 970Pendimethalin dinitroanaline Tubulin inhibitor 3 5 1,894 

Trifluralin dinitroanaline

2 1,454

Bentazon benzothiadiazinone

PSII inhibitor

*

3

70

Metribuzin triazinone

2 437 

Sulfentrazone triazolinone 1 70

Chlorimuron-ethyl sulfonylurea

ALS inhibitor

4

11

52

Cloransulam-methyl triazolopyrimidine 1 17

Flumetsulam triazolopyrimidine * 8

Imazamox imidazolinone * 9Imazaquin imidazolinone 1 66

Imazethapyr imidazolinone 3 100

Imazethapyr, ammon. Imidazolinone * 5Thifensulfuron sulfonylurea 1 3Tribenuron-methyl sulfonylurea 1 5Alachlor chloroacetamide Cell division

inhibitor 

*

1

485

S-Metolachlor chloroacetamide 1 837 Flufenacet oxyacetamide * 80Paraquat bipyridilium PSI disruption 1 1 335 
Monsanto Company 10-SY-210U 

Table VIII-8 (continued). Agricultural Chemical Applications Registered for Soybean Use in AR, IA, ILMS, MO, NE, NC, ND, OH, SD, TN, VA, and WI in 20061Herbicide Chemical Family

Mode-of-Action 

(MOA)

PercentTreated Acres

Total Area 

Applied

(Percent/MOA)

Quantity 

Applied

(1000 lbs)

Clethodim cyclohexenone

ACCase inhibitor

3

4

190

Fenoxaprop aryloxyphenoxy propionate * 9Fluazifop-P-butyl aryloxyphenoxy propionate 1 43Quizalofop-P-ethyl aryloxyphenoxy propionate * 14Sethoxydim cyclohexenone * 10Acifluorfen diphenyl ether

PPO inhibitor

*

6

47

Carfentrazone-ethyl triazolinones * 10 Flumiclorac-pentyl N-phenylphthalimide 1 17Flumioxazin N-phenylphthalimide 3 138Fomesafen diphenyl ether

2 330

Lactofen diphenyl ether * 232,4-D, 2-EHE phenoxy

Synthetic auxin

7

10

2,505

2,4-D, dimeth. salt phenoxy 3 953 2,4-D (butoxy ester) phenoxy * 68Dicamba, digly salt benzoic acid * 16

Total 

* Area receiving application is less than 0.5 percent. 1Data derived from USDA-NASS (2007b). Planted acreage for the nineteen primary soybean production states was 

represented 96.5% of total planted acres. 
Monsanto Company 10-SY-210U 

Table VIII-9. Crop Tolerance and Common Grass Weed Responses to Herbicides Applied in Soybean Pro

 Common Grass Weeds1,2

Herbicide/Application CT3 BY BS CG FP FT GG SC JGs JGr 

Preplant Incorporated Trifluralin 1 9 9 9 9 9 9 7 7 3 Preplant or Preemergence Chlorimuron/tribenuron 2 - 8 - - - 7 - - 2 Cloransulam 0 - NA - - - NA - - NA Flumioxazin 2 - 5 - - - 5 - - 0 Flumioxazin/cloransulam 2 - 5 - - - 5 - - 0 Imazaquin 1 - 7 - - - 5 - - 2 Imazethapyr 1 6 NA 7 7 7 NA 6 6 NA Metribuzin 2 2 6 6 5 6 7 6 - 0 Pendimethalin 2 8 9 9 9 8 9 7 7 3 s-Metolachlor 1 8 8 9 8+ 8+ 9 - - 0 Postemergence Bentazon 1 - 0 - - - 0 - - 0 Chlorimuron 2 - 0 - - - 0 - - 0 Clethodim 0 9 9 8+ 9 9 9 9 9 9 Cloransulam 1 - 0 - - - 0 - - 0 Clorimuron/thifensulfuron 0** - NA - - - NA - - NA Fluazifop/fenoxaprop 0 9 8 8+ 9 9 9 9 9 9 Flumiclorac 2 - NA - - - NA - - NA Fomesafen 2 - 3 - - - 3 - - 3 Glyphosate 0* 8+ 9 8+ 8+ 9 8 8 9 9 Glyphosate/imazethapyr 1* 9 NA 8+ 9 9 NA 9 9 NA Imazamox 2 6 NA 7 7 7-8+ NA - - NA Imazethapyr 1 6 7 7 7 7-8 5 8 8 6 Lactofen 3 - 4 - - - 4 - - 2 Thifensulfuron 2 - NA - - - NA - - NA 

1All weed control ratings except for BS, GG, JGr and RR are from the 2009 Weed Control Guide for Ohio and Indiana, Ohio State University and Purdue University (Lo

Ratings for BS, GG, JGr and RR are from the 2009 Weed Control Guidelines for Mississippi, Mississippi State University (MSU, 2010). Weed control rating for weedsRR, are: 9 = 90% to 100%, 8 = 80% to 90%, 7 = 70% to 80%, 6 = 60% to 70%, - = less than 60% control, not recommended. Weed control ratings for BS, GG, and RR 

7-8 = good, 4-6 = fair, 0-3 = none to slight. Ratings assume the herbicides are applied in the manner suggested in the guidelines and according to the label under optimu

2Weed species: BY = barnyardgrass, BS = broadleaf signalgrass, CG = crabgrass, FP = fall panicum, FT = giant and yellow foxtail, GG = goosegrass, SC = shattercane, 

Johnsongrass, JGr = rhizome Johnsongrass, RR = red rice, and NSy = yellow nutsedge. 3All crop tolerance ratings are from the 2009 Weed Control Guide for Ohio and Indiana, Ohio State University and Purdue University (Loux et al., 2009). Crop toleranc

good, 2 = fair, 3 = poor. 

NA denotes not available. *Rating based on application to Roundup Ready soybean. **Ratings based on application to STS soybean.
Monsanto Company 10-SY-210U 

Table VIII-10. Common Broadleaf Weed Responses to Herbicides Applied in Soybean Production 

Common Broadleaf Weeds1,2

Herbicide/Application BN CB CR GR HS LQ MG PA PW PS SPreplant Incorporated OnlyTrifluralin - - - - 0 8+

2

7

9

0

4 

Preplant or Preemergence

Chlorimuron/tribenuron - 8 9

7

9

9 8 8 9

7 N

Cloransulam - 8 9

7 NA

9 NA NA

9 NA N

Flumioxazin

9 - 7 - 9

9 6-8

9

9 8 7

Flumioxazin/cloransulam

9 8 9

7

9

9 7-8

9

9 8 7

Imazaquin

9 8 8 7

0

9 6-8

9

9

9 5 

Imazethapyr

9

7

6 - NA

9 NA NA

9 NA N

Metribuzin - - - 7

9

7 2-8

9 8 9 8 

Pendimethalin - - - - 0 8+

2

7

9

4

2 

s-Metolachlor 8 - - - 0

6

0 8 8 4

2 

Postemergence

Bentazon - 9

7

6

4

7 2-9

4 - 8 0

Chlorimuron - 9 8 7+ 8 - 8-9

6

9

2

7 

Cloransulam - 9

9

9 3 - 8-9

2 - 2

7 

Clorimuron/thifensulfuron - 9 8 7+ NA 8 NA NA

9 NA N

Flumiclorac - 7

7 - NA

7 NA NA

7 NA N

Fomesafen 8 7 8+ 8 9 - 8-9 8 9

2 3 

Glyphosate 8 9 8+ 8 7 8 7-9

9

9

7 8 

Glyphosate/imazethapyr

9

9 8+ 8+ NA 8+ NA NA

9 NA N

Imazamox

9 8 7 8 NA 8 NA NA

9 NA N

Imazethapyr

9

9

6

7

0

6 7-9

6

9

6

0

Lactofen 8+ 8 9 8 9 - 8-9 8 9 8 5 Thifensulfuron - 6 - - NA 8 NA NA

9 NA N

1All weed control ratings except for HS, MG, PA, PS, and SP are from the 2009 Weed Control Guide for Ohio and Indiana, Ohio State 

University (Loux et al., 2009). Ratings for HS, MG, PA, PS, and SP are from the 2009 Weed Control Guidelines for Mississippi, Miss

(MSU, 2010). Weed control ratings for weeds, except HS, MG, PA, PS, and SP, are: 9 = 90% to 100%, 8 = 80% to 90%, 7 = 70% to 8less than 60% control, not recommended. Weed control ratings for HS, MG, PA, PS, and SP are: 9-10 = excellent, 7-8 = good, 4-6 = fRatings assume the herbicides are applied in the manner suggested in the guidelines and according to the label under optimum growing 

2Weed species: BN = black nightshade, CB = cocklebur, CR = common ragweed, GR = giant ragweed, LQ = lambsquarters, MG = mo

hemp sesbania, PA = palmer and spiny pigweed, PW = pigweed, PS= prickly sida, SP = sicklepod, SW = smartweed, VL = velvetleaf, NA denotes not available. 
Monsanto Company 10-SY-210U 197 of 721 

VIII.G. Dicamba Herbicide Use in the U.S. 

Dicamba was approved by the U.S. EPA for agricultural uses in 1967 (U.S. EPA, 2009). 

Dicamba is formulated as a stand-alone herbicide product and marketed by several 

companies under various trade names such as Banvel®, Clarity®, Diablo®, Rifle®, and 

Sterling® that are various salt formulations of dicamba. These dicamba products can be 

tank mixed with one or more active ingredients depending on the treated crop. For 

example, Clarity can be tank mixed with over 75 herbicide products in labeled crops. 

Additionally, dicamba is formulated as a registered premix product with one or more 

other herbicide active ingredients such as glyphosate, 2,4-D, diflufenzopyr, atrazine, 

nicosulfuron, metsulfuron, primsulfuron, triazulfuron, rimsulfuron and halosulfuron. 

Dicamba herbicide (e.g., Clarity – diglycolamine (DGA) salt of dicamba) is currently 

labeled for weed control in soybean, corn, cotton, sorghum, wheat, barley, oats, millet, 

pasture, rangeland, asparagus, sugarcane, turf, grass grown for seed, conservation reserve 

programs, and fallow croplands. Table VIII-11 provides a summary of dicamba-treated 

acres (crop acreage that has dicamba applied to it) and the amount of dicamba active 

ingredient applied for all labeled crops each year from 1990 through 2008. Dicambatreated acreage has ranged from 17.4 to 36.3 million acres during this period. Usage of 

dicamba peaked during the period of 1994 through 1997, where 1994 was the peak year 

when 36.3 million acres were treated with 9.4 million pounds of dicamba. Since 1994, 

the use of dicamba has steadily declined to 20.2 million treated-acres with 2.7 million 

pounds in 2008 due to the competitive market introductions of sulfonylurea herbicides 

(chlorsulfuron, metsulfuron-methyl, and thifensulfuron-methyl) in wheat, new broadleaf 

herbicide active ingredients in corn, and Roundup Ready corn. Usage in cotton is one 

exception, where dicamba-treated acres (preplant applications) have increased from 

140,000 to 590,000 acres from 2004 to 2008 (AgroTrak, 2009). 

Table VIII-12 provides a summary of the dicamba-treated acres by crop in 2008. 

Approximately 20.2 million acres were treated with dicamba in 2008. Over 8 million 

acres of corn were treated, which is 40.1% of the total dicamba-treated acres for all crops. 

The next highest levels of treated acres are in wheat (25.2%) and fallow land (14.9%). 

The crops with the highest percentage of dicamba-treated acres are sugarcane (21.9%), 

fallow land (19.2%), sorghum (15.8%), and wheat (8.4%). Although corn represents the 

crop with the highest dicamba-treated acres, only 9.4% of the total corn acreage was 

treated with dicamba in 2008. For comparison, the treated percentage in corn was at 

approximately 29% as recently as 2000 (USDA-NASS, 2001). 

Approximately 2.67 million pounds of dicamba active ingredient were applied for all 

agricultural uses in 2008 (Table VIII-12). The distribution of dicamba active ingredient 

across the various labeled uses is similar to the distribution of treated acres. Based on 

USDA-NASS (2004, 2006, 2007b, 2008) statistics, dicamba application rates ranged 

from 0.03 to 0.25 pounds per acre with the number of applications ranging from 1 to 1.2 

applications per cropping season (Table VIII-13). Dicamba rates are the lowest in barley, 

wheat, and oats, where typically more than one application is made in these crops per 

cropping season. The average application rate in corn is 0.19 pounds of dicamba per acre 

with slightly over one application per season. 
Monsanto Company 10-SY-210U 198 of 721 

Dicamba is currently labeled for use in conventional or Roundup Ready soybean, 

although dicamba use is extremely limited because applications are restricted to very 

early preplant and/or preharvest applications due to soybean tolerance concerns. The 

dicamba-treated acreage in 2008 soybean production was approximately 530,000 acres 

that represented 0.7% of the total soybean acreage. 

Table VIII-11. Dicamba Use in All Labeled Crops from 1990 to 20081

Year 

Treated Acres 

(000,000 acres) 

Dicamba (a.e.) 

(000,000 lbs) 

1990 26.8 6.7 

1991 24.5 6.3 

1992 30.3 7.4 

1993 27.7 7.0 

1994 36.3 9.4 

1995 34.3 8.7 

1996 33.3 8.2 

1997 33.1 8.6 

1998 32.2 8.0 

1999 29.8 6.3 

2000 29.4 5.4 

2001 30.6 5.4 

2002 29.4 5.0 

2003 27.1 4.3 

2004 22.3 3.9 

2005 21.3 3.4 

2006 17.4 2.7 

2007 18.6 2.7 

2008 20.2 2.7 

1

Source is AgroTrak (2009). 

Shaded bar indicates the year with maximum dicamba-treated acres. 
Monsanto Company 10-SY-210U 

Table VIII-12. Dicamba-Treated Acres and Amounts Applied to Labeled Crops and Uses in 20081

Crop 

Total Crop 

Acres (000) 

DicambaTreated Acres 

(000) 

DicambaTreated Acres 

(% of Total)2

Dicamba 

Treated Crop 

(%) 

Dicamba Pounds 

(000 a.e.) 

Asparagus 34 1 <0.1 1.8 0.1 Barley 3,868 211 1.0 5.5 23 Corn 87,245 8,115 40.1 9.4 961 Cotton 9,309 590 2.9 6.3 139 Fallow 15,751 3,018 14.9 19.2 420 Pastureland 96,151 1,218 6.0 1.3 254 Sorghum 7,035 1,114 5.5 15.8 137 Soybean 74,405 530 2.6 0.7 118 Sugarcane 810 177 0.9 21.9 40 Wheat, all 60,835 5,094 25.2 8.4 549 All other uses NA 139 NA NA 30 Total 20,207 2,670 NA denotes not applicable. 1Source is AgroTrak (2009). 2The percentage of the total dicamba-treated acres for all labeled crops and uses. Table VIII-13. Dicamba Applications – Average Number and Rates to Labeled Crops1

Crop 

# of Dicamba 

Applications 

Rate of Dicamba per 

Application 

Rate of Dicamba per 

Crop Year 

Corn 1.02 0.188 0.192 Cotton 1.00 0.191 0.192 Sorghum 1.05 0.205 0.215 Soybean 1.00 0.250 0.250 Barley 1.20 0.060 0.080 Wheat, spring 1.13 0.032 0.085 Wheat, winter 1.20 0.122 0.149 Oats 1.00 0.088 0.088 1USDA-NASS, 2004 (sorghum), 2006 (corn and oats), 2007b (soybean, barley and wheat), and 2008 (cotton) 
Monsanto Company 10-SY-210U 200 of 721 

VIII.G.1. Dicamba Application Timing for Labeled Crops 

Label recommendations on the application timing of dicamba are highly dependent on the 

crop being treated to ensure adequate crop safety. Many of the field crops currently labeled 

for dicamba, such as soybean and cotton, include preplant applications but certain timing 

intervals are required between application and planting to avoid crop injury. However, 

dicamba can be applied in other field crops such as corn, sorghum, barley, and wheat either 

as preplant or postemergence applications with restrictions regarding the crop stage of 

growth. Broadcast applications in corn can be made up to the 5-leaf stage or 8 inches tall, 

whichever occurs first. Sorghum can be treated with broadcast applications from after the 

spike stage up to the 5-leaf stage or 8 inches tall3. These applications are considered earlypostemergence since they occur relatively early in the growing season, typically ending 

sometime in June. Post-directed applications can be made in corn (up to 36 inches tall) and 

sorghum (15 inches tall), which would be much later in the season than an earlypostemergence application. However, growers seldom make these applications in corn and 

sorghum since post-directed applications require special spraying equipment and equipment 

setup. Applications in wheat and barley must be made prior to the jointing stage except in 

spring seeded wheat where application can be made up to the 6-leaf stage of wheat. These 

applications are relatively early in the spring or late in the fall. 

Preharvest applications of dicamba are permitted in several crops including soybean, 

sorghum, barley, and wheat. Preharvest applications are only allowed after the crops reach a 

certain maturity stage and then harvesting must be delayed for a given time period 

depending on the crop4

. This type of application is infrequently used since it is considered a 

rescue or harvest-aid treatment intended to remove weeds which interfere with the 

harvesting equipment or operation. 

Table VIII-14 provides a summary of the application timings of dicamba in labeled crops. 

Approximately 24% of the dicamba treated acres are treated either in the fall or spring as 

preplant applications to the crop. Over 50% of the treated acres are treated postemergence. 

Postemergence applications represent the primary timing in corn, sorghum, and wheat. 

 

3

 Clarity product label can be found at: http://www.cdms.net/LDat/ld797002.pdf. 

4

 Clarity label specifies that preharvest application requires that soybean pods must have reached mature 

brown color and at least 75% leaf drop has occurred. Harvest of soybean is allowed 14 or more days after 

preharvest application. 
Monsanto Company 10-SY-210U 

Table VIII-14. Dicamba-Treated Acres (000) by Application Timing and Crop in 20081

 Application Timing2

Crop 

Fall 

Preplant 

Spring 

Preplant At Planting 

Pre-Crop 

Emergence 

In-Crop 

Postemergence 

Other 

Timing

Asparagus - - - - - 1 Barley 8 41 - - 162 - Corn 136 851 71 285 6,771 - Cotton 41 549 - - - - Fallow 647 - - - - 2,371 Sorghum 439 380 12 81 202 - Soybean 43 486 - - - - Sugarcane 13 - - 15 149 - Wheat, spring 341 193 - 26 791 - Wheat, winter 608 - 13 71 3,050 - Pastures 84 - - - - 1,134 Totals 2,360 2,500 96 478 11,125 3,506 % of Applications 11.8 12.5 0.5 2.4 55.4 17.5 - denotes no application at the timing for the listed crop. 1Source is AgroTrak (2009). 2 Dicamba-treated acres are expressed as dicamba a.e. 3Application timing could be throughout the season since applications are made between harvests or are not de

stage of growth. 
Monsanto Company 10-SY-210U 202 of 721 

VIII.G.2. Distribution of Dicamba Use in the U.S. 

Table VIII-15 provides a summary of the dicamba-treated acres by crop for each of the 

states in the U.S. soybean growing regions. As expected, based on the dicamba use data 

previously presented, over one-half of the dicamba-treated acres (58%) are in the Midwest 

region representing approximately 11.8 million acres. The primary dicamba-treated 

crops/uses in this region are corn, wheat, barley, and fallow. Although the Plains and 

Western States regions are not considered a soybean producing region, it represents a large 

portion (32%) of the dicamba-treated acres (6.4 million). Only 1.4 and 0.6 million acres are 

treated with dicamba in the Southeast and Eastern Coastal regions, respectively. Soybean 

producing states with over one million dicamba-treated acres include Illinois, Kansas, and 

North Dakota. The state of Kansas, which grows many of the primary dicamba-labeled 

crops, has the largest amount of dicamba-treated acres at approximately 4.6 million acres 

(22.6% of total dicamba-treated acres), which is 2.5 times more than the next largest use 

states of North Dakota, Colorado, Montana, and Texas. Dicamba is used on less than 1% of 

U.S. soybean acres, but in Arkansas dicamba is used on 12.6% of the soybean acres, and 

accounts for the majority of dicamba-treated soybean acres in the U.S. (~78%). The 

primary reason for the relatively higher use of dicamba in Arkansas, compared to other 

soybean producing states, is that dicamba is being recommended by academics and 

extension agents in the Delta to control glyphosate resistant marestail. 

The maps presented in Figures VIII-1 and VIII-2 provide a visual illustration of the 

historical distribution and intensity of dicamba-treated acres for all labeled crops and the 

current U.S. soybean production acres at the county level, respectively. By overlaying the 

geographical representations from Figures VIII-1 and VIII-2, it can be concluded that the 

historical use and intensity of total dicamba-treated acres align with soybean acreage. 
Monsanto Company 10-SY-210U 

Table VIII-15. Dicamba-Treated Acres by State and Labeled Crop in 20081

 Cro

p Acres Treated With Dicamba (000

Re

gion/State 

Soybean 

Acres 

(000)

Total 

Dicamba

Acres 

(000)

Percent 

U.S. 

Dicamba 

Acres2 Corn

Small 

Grains3 Fallow Pasture Cotton Sor

ghum 

Midwest Re

gion

Illinois 9,200 1,213 6.0 1,204 - - - - 3 Indiana 5,450 494 2.4 489 - - - - - Iowa 9,750 580 2.9 576 - - 4 - - Kansas 3,300 4,558 22.6 658 2,198 901 30 2 675 Kentucky 1,390 36 0.2 36 - - - - - Michigan 1,900 294 1.5 285 8 - - - - Minnesota 7,050 628 3.1 606 21 - <1 - - Missouri 5,200 341 1.7 205 <1 - 36 99 - Nebraska 4,900 946 4.7 558 222 134 6 - <1 North Dakota 3,800 1,670 8.3 740 846 64 - - - Ohio 4,500 270 1.3 247 23 - - - - South Dakota 4,100 502 2.5 183 88 198 - - 16 Wisconsin 1,610 261 1.3 250 - - 11 - - Region Totals 62,150 11,793 58.4 6,037 3,407 1,297 88 101 695 
Monsanto Company 10-SY-210U 

Table VIII-15 (continued). Dicamba-Treated Acres by State and Labeled Crop in 20081

 Cro

p Acres Treated With Dicamba (000)

Re

gion/State 

Soybean 

Acres 

(000)

Total 

Dicamba

Acres 

(000)

Percent 

U.S. 

Dicamba 

Acres2 Corn

Small 

Grains3 Fallow Pasture Cotton Sor

ghum 

Southeast RegionAlabama 360 119 0.6 54 - - 14 51 - Arkansas 3,300 791 3.9 - - - 92 262 - Georgia 430 32 0.2 17 - - 15 - - Louisiana 1,050 215 1.1 17 - 23 - - - Mississippi 2,000 102 0.5 - - - 50 52 - North Carolina 1,690 53 0.3 39 - - 14 - - South Carolina 540 6 0.0 - - - - - - Tennessee 1,490 102 0.5 5 - - <1 96 - Region Totals 10,860 1,420 7.0 132 - 23 186 461 - Eastern Coastal RegionDelaware 195 0 0 - - - - - - Maryland 495 268 1.3 268 - - - - - New Jersey 92 0 0 - - - - - - New York 230 62 0.3 62 - - - - - Pennsylvania 435 170 0.8 168 - - 2 - - Virginia 580 118 0.6 104 5 - 9 - - Region Totals 2,027 618 3.1 602 5 - 11 - - 
Monsanto Company 10-SY-210U 

Table VIII-15 (continued). Dicamba-Treated Acres by State and Labeled Crop in 20081

 Cro

p Acres Treated With Dicamba (000) ex

press

Re

gion/State 

Soybean 

Acres 

(000)

Total 

Dicamba

Acres 

(000)

Percent 

U.S. 

Dicamba 

Acres2 Corn

Small 

Grains3 Fallow Pasture Cotton Sor

ghum 

Plains and Western States RegionCalifornia 0 95 0.5 32 63 - - - - Colorado 0 1,606 8.0 248 253 958 3 - 122 Florida 327 142 0.7 - - - 140 - - Idaho 0 170 0.8 114 51 <1 <1 - - Montana 0 1,657 8.2 - 955 686 - - - New Mexico 0 46 0.2 37 - - - - 9 Oklahoma 400 622 3.1 8 84 5 409 - 113 Oregon 0 222 1.1 - 218 - 4 - - Texas 230 1,641 8.1 850 212 14 360 27 175 

Utah 0 22 0.1 3 <1 12 - - - 

Washington 0 96 0.5 - 55 31 5 - - Wyoming 0 57 0.3 54 - 3 - - - Region Totals 957 6,376 31.6 1,346 1,892 1,710 934 27 419 

U.S. Totals 75,994 20,207 100 8,117 5,304 3,030 1,207 589 1,114 

- denotes no dicamba-treated acres for listed crop/use. 1Source is AgroTrak (2009). 2Percent of total U.S. Dicamba Acres = Total Dicamba-Treated Acres (per state)/U.S. Total Dicamba-Treated 

100 

3Small grains include barley, winter wheat and spring wheat. 4Other labeled crops include asparagus and sugarcane.
Monsanto Company 10-SY-210U 206 of 721 

Figure VIII-1. Mean Annual Dicamba Use for Agricultural Uses in the U.S. During 

1999-20041

1

Based on 2002 Census of Agriculture county crop acreage (USGS, 2004). 

Figure VIII-2. Planted Soybean Acreage by County in the U.S. in 20091

1

Source is USDA-NASS (2010).
Monsanto Company 10-SY-210U 207 of 721 

VIII.G.3. Potential Impacts to Adjacent Crops 

U.S. EPA considers possible effects from offsite movement as part of the pesticide 

registration process. In order to approve the use of a pesticide (herbicide) under FIFRA, 

U.S. EPA must conclude that no unreasonable adverse effects on non-target vegetation will 

result from offsite movement when the herbicide is used according to the product label. 

Thus, when herbicides are applied in accordance with the pertinent label restrictions, offsite 

impacts can be avoided. EPA reassessed the potential risks to non-target plants in its 

analysis in the dicamba RED, and it concluded that no specific additional drift mitigations 

were needed to support the continued registration of all dicamba uses (U.S. EPA, 2009). 

Since the proposed use pattern for dicamba on MON 87708 is consistent with use patterns 

evaluated and deemed eligible for reregistration in the dicamba RED, it is reasonable to 

conclude that dicamba use on MON 87708 meets the FIFRA standards related to offsite 

movement and does not pose any greater risk to non-target vegetation over existing dicamba 

agricultural uses approved by EPA when used according to the product label. 

Growers and commercial herbicide applicators have been applying dicamba to agricultural 

row crops for over 40 years. This practice has provided valuable experience and knowledge 

on the proper application of dicamba for effective weed control and also for minimizing 

offsite movement to sensitive crops. Dicamba herbicide spray drift can be reduced during 

application by using industry standard procedures for minimizing spray drift. These 

procedures include making applications with coarse droplet size through appropriate nozzle 

selection, using lower spray pressure, applying at the lowest nozzle height that provides 

uniform coverage, and making applications when wind speeds are low and consistent in 

direction (SDTF, 1997). In addition, growers and commercial applicators are legally 

required to follow label requirements and are educated by university specialists and industry 

representatives on the proper application equipment, equipment setup, and climatic 

conditions to maximize herbicide performance and minimize offsite movement of 

herbicides. For example, with the introduction of Roundup Ready crops and the subsequent 

increase in glyphosate use, university specialists conducted extensive education programs 

on proper application procedures and precautions (Dr. W. Johnson – Purdue University, 

2010 personal communication). Equipment manufacturers have developed spray nozzles 

that provide uniform coverage for effective weed control while applying larger spray 

droplets to reduce the potential for particle drift. Similarly, offsite movement of dicamba 

has been managed with the knowledge of the proper spray equipment and equipment setup, 

climatic conditions for accurate, on-target applications, and based on the requirements for 

applying dicamba at an appropriate distance from adjacent crops and plants (Jordan et al., 

2009). 

Monsanto plans to further address the use of dicamba on MON 87708 with US EPA to 

evaluate whether any additional measures may be appropriate to further address potential 

drift and offsite movement. 
Monsanto Company 10-SY-210U 208 of 721 

VIII.H. Dicamba-Tolerant Soybean MON 87708 

Monsanto has developed a new herbicide-tolerant soybean, MON 87708, to offer growers 

an expanded use of the herbicide dicamba in soybean production. MON 87708 will 

facilitate a wider window of dicamba application in soybean, allowing preemergence 

application up to the day of crop emergence (cracking) and in-crop postemergence 

applications through the R1/R2 growth stage. MON 87708 will be combined with 

MON 89788 (Roundup Ready 2 Yield soybean) utilizing traditional breeding techniques. 

The combination of herbicide-tolerance traits will allow the pre- and postemergence use of 

both dicamba and glyphosate herbicides in an integrated weed management program to 

control a broad spectrum of grass and broadleaf weed species (Johnson et al., 2010). 

Increasing postemergence herbicide options is important, especially in conservation tillage 

situations, where the performance consistency of postemergence herbicides has generally 

been greater than that of soil active residual products. Dicamba will improve the control of 

glyphosate’s hard-to-control broadleaf weeds (e.g., common lambsquarters, hemp sesbania, 

morningglory species, nightshade, Pennsylvania smartweed, prickly sida ̧ and wild 

buckwheat) and also offer an effective control option for glyphosate-resistant broadleaf 

weed species, namely marestail, common ragweed, giant ragweed, palmer pigweed, and 

waterhemp (Johnson et al., 2010). Dicamba will also offer an effective control option for 

broadleaf species resistant to ALS and PPO chemistries. In the case of PPO resistance, a 

primary dicamba benefit will be to provide options for delaying the further spread of PPO 

resistant amaranthus species (University of Tennessee, 2010).

Upon integration of MON 87708 into the Roundup Ready soybean system, growers will 

have the ability to continue to use established soybean production practices including crop 

rotation, tillage systems, labeled herbicides, and row spacing, thereby using the same 

planting and harvesting machinery currently being utilized. Growers will also continue to 

have the flexibility and simplicity in weed control provided by glyphosate that will allow 

growers to continue to reap the environmental benefits associated with the use of 

conservation-tillage that is facilitated by the use of glyphosate for postemergence weed 

control in the Roundup Ready soybean system (CTIC, 2011; CTIC, 2004). 

Current labeled uses of dicamba in soybean are limited to early preplant and late 

postemergence (preharvest) applications. Significant planting restrictions exist in soybean 

for preplant applications of dicamba, including a maximum application rate of 0.5 lbs a.e. 

per acre, a 28-day interval between application and planting soybean, and a minimum of one 

inch of rainfall must occur before planting soybean to avoid soybean injury. Monsanto has 

submitted an application to U.S. EPA to amend Registration Number 524-582, a DGA salt 

formulation, to remove all preemergence planting restrictions (intervals and rainfall) and to 

allow in-crop postemergence dicamba applications to MON 87708 through the R1/R2 

growth stage of soybean. Once approved, growers would be authorized to apply dicamba 

alone or in mixtures with glyphosate or other herbicides for preplant or in-crop 

postemergence applications on MON 87708. Dicamba would be authorized to be applied 

preemergence up to crop emergence as a single application or split applications up to a total 

of 1.0 lb a.e. per acre, and up to two postemergence applications up to 0.5 lb a.e. per acre 

each through the R1/R2 growth stage of soybean. The maximum annual application rate of 

dicamba on MON 87708 is 2.0 lb dicamba a.e. per acre. Furthermore, Monsanto’s proposed 
Monsanto Company 10-SY-210U 209 of 721 

dicamba label does not allow aerial applications of dicamba on MON 87708, a stewardship 

measure intended to address potential offsite impacts. As indicated, Monsanto also plans to 

further address the use of dicamba on MON 87708 with US EPA to evaluate whether any 

additional measures may be appropriate to further address potential drift and offsite 

movement. 

The potential increase in dicamba use in U.S. soybean production upon deregulation of 

MON 87708 was assessed by estimating the total dicamba use across soybean acres. 

Assuming 100% adoption of MON 87708 across all U.S. soybean acreage 6 and an 

application of dicamba at the maximum labeled use rate on all soybean acres, dicamba use 

on MON 87708 could potentially total 150 million pounds. In practice, however, a single 

early season in-crop application per year of dicamba at 0.38 lb a.e. per acre is expected on 

the majority of MON 87708 planted acres. However, in no-till or conservation tillage 

soybean systems, an additional preplant application at 0.50 lb a.e. per acre could also be 

common practice, and in areas where glyphosate resistant weeds, especially Ambrosia and 

Amaranthus species, are present two in-crop applications at 0.5 lb a.e. each may be needed 

in some situations. These anticipated use patterns represent a high-end estimate for 

prediciting dicamba use associated with MON 87708 integrated with the Roundup Ready 

soybean system. 

Furthermore, consistent with recommendations by academics and weed scientists, Monsanto 

will recommend the use of a third herbicide mode-of-action with soil residual activity as 

part of a comprehensive weed resistance management program to assure that at least two 

effective modes-of-action are always used in the cultivated soybean field. A summary of 

the anticipated weed control recommendations for the combined MON 87708 and Roundup 

Ready soybean system is provided in Table VIII-16. 

In 2010, Monsanto conducted an informal survey of weed scientists across the country to 

estimate the number of crop acres with glyphosate resistant weed populations. Based upon 

this survey it was estimated that approximately 14-16 million acres of planted row-crops 

(i.e. corn, soybeans, cotton) had populations of glyphosate resistant weeds. Of these acres, 

the majority of acres (~ 10 million) are infested with glyphosate resistant marestail 

populations where a preplant application of dicamba and glyphosate described above will be 

effective for control. The remainder of resistant acres (~5 million) have resistant Ambrosia

(common and giant ragweed) and Amaranthus (palmer pigweed and water hemp,) species 

present. A conservative estimate of 5 million resistant acres is assumed for this assessment, 

which overestimates current resistant acres in soybean producing areas and also accounts for 

potential increases in resistant acres because not all resistant crop acres would be planted to 

soybean in any given year. 

 

 

6

 Based on approximately 75 million acres planted to soybean in 2008, see Table VIII-1. 
Monsanto Company 10-SY-210U 210 of 721 

Table VIII-16. Anticipated Weed Management Recommendations for MON 87708 

Combined with the Roundup Ready 2 Yield Soybean System1

Application 

Timing 

Conventional Tillage2 Conservation Tillage 2

(No-till or reduced till) 

No GR 

Weeds 

GR Weeds or Suspected 

GR Weeds No GR 

Weeds 

GR Weeds or Suspected 

GR Weeds 

Option 13 Option 24 Option 13 Option 24

Preemergence 

(burndown, at 

planting) 5

Residual Residual Residual 

Residual 

+ 

Glyphosate 

+ 

Dicamba 

Residual 

+ 

Glyphosate 

+ 

Dicamba 

Residual 

+ 

Glyphosate 

+ 

Dicamba 

Postemergence 1 

(V1-V3) 

Glyphosate 

+ 

Dicamba 

Glyphosate 

+ 

Dicamba 

Glyphosate 

+ 

Dicamba 

Glyphosate 

+ 

Dicamba 

Glyphosate 

+ 

Dicamba 

Glyphosate 

+ 

Dicamba 

Postemergence 2 

(V4-R2) --- 

Glyphosate 

+ 

Dicamba 

--- --- 

Glyphosate 

+ 

Dicamba 

--- 

1 The anticipated use patterns represent a high-end estimate for prediciting dicamba use associated with 

MON 87708 integrated with the Roundup Ready soybean system. Actual weed control practices by growers 

will vary depending on the specific weed spectrum and agronomic situation of the individual soybean field, 

specifically dicamba use could be lower especially for the preemergence and second postemergence 

applications. 2 Average rate for dicamba is 0.38 pound a.e. per acre except for fields with glyphosate resistant (GR) species 

where a 0.5 pound a.e. per acre postemergence application rate will be recommended. In some situations, 

the second postemergence application may not be needed. 3 Option 1 would be used for more aggressive glyphosate resistant weed species, such as Ambrosia or 

Amaranthus species. 

4 Option 2 would be used for less aggressive glyphosate resistant weed species, such as marestail. 

5 Monsanto and academics recommend the use of soil residuals as part of a comprehensive weed resistance 

management program to ensure that two effective herbicide modes-of-action are used in soybean and to 

provide protections against additional resistance development to existing soybean herbicides. 

Assuming the anticipated use rate of 0.5 lb a.e. per acre dicamba for preemergence 

applications and 0.38 lb a.e. per acre dicamba for postemergence applications, and using a 

conservative assumption that MON 87708 has 100% adoption across all U.S. geographies 

and conservation tillage systems are used on approximately 40% of the U.S. soybean acres 

(CTIC, 2007), dicamba use on MON 87708 would total approximately 44 million pounds. 

When considering a more realistic adoption rate for MON 87708 of 40% (refer to Section 

VIII.H.2), dicamba use on MON 87708 would total approximately 17 million pounds. In 

areas where resistant Ambrosia and Amaranthus species are present requiring two in-crop 

applications, the use of an additional 5 million pounds of dicamba per year is estimated. 

It is anticipated that dicamba applications will continue for currently labeled crops at the 

dicamba-treated acreage levels and amounts presented in Table VIII-12, such that the 

dicamba treatment to MON 87708 will thereby result in a total U.S. dicamba use of 

approximately 25 million pounds annually. This level of dicamba use would be 
Monsanto Company 10-SY-210U 211 of 721 

approximately double the historical peak level (Table VIII-11) experienced since dicamba’s 

introduction in 1967. 

Upon integration of MON 87708 into the Roundup Ready soybean system, dicamba will 

provide excellent control of numerous annual and perennial broadleaf weed species, 

including populations of broadleaf weeds that are resistant to ALS, atrazine, or glyphosate 

herbicides. Table VIII-17 shows weed control ratings for dicamba, glyphosate and several 

glyphosate tank mixtures when applied as a preplant burndown application to common 

broadleaf weed species found in soybean fields of the Midwest and Southeast regions. An 

application of dicamba alone provides effective control of a broad spectrum of winter and 

summer annual and perennial broadleaf weed species. In comparison, glyphosate alone 

provides excellent control of many grass species in addition to many of the annual and 

perennial broadleaf species listed. However, dicamba provides a higher level of control of 

certain broadleaf weeds including common lambsquarters, Pennsylvania smartweed, red 

clover, alfalfa, marestail, hairy vetch, and prickly lettuce. Dicamba will be very 

complementary in mixtures with glyphosate for weed control in a preplant application 

(Johnson et al, 2010) and will offer growers equal or superior weed control to other 

glyphosate mixtures because it offers reduced potential herbicide antagonism, improved 

efficacy and broader weed spectrum. 

The dicamba tolerance trait in MON 87708 will permit in-crop applications of dicamba to 

soybean with excellent crop safety (crop tolerance). Dicamba will also complement the 

weed control of in-crop applications of glyphosate when applied as a mixture or in 

sequence. Table VIII-18 shows common broadleaf weed responses to dicamba compared to 

glyphosate and several glyphosate labeled tank mixtures in soybean. Since dicamba is not 

currently labeled for in-crop applications in soybean, weed control ratings for dicamba were 

taken from labeled in-crop applications of dicamba in corn for comparison purposes. 

Glyphosate will continue to provide broad spectrum control of annual grasses and broadleaf 

weeds, while dicamba will provide improved control of common ragweed, giant ragweed, 

hemp sesbania, morningglory species, and prickly sida. As presented in Table VIII-17, 

dicamba is more effective in controlling marestail then glyphosate. Likewise, in comparison 

to glyphosate, dicamba is expected to also improve the control of lambsquarters, eastern 

black nightshade, kochia, palmer pigweed, and wild buckwheat. In addition to 

complementing the weed control of glyphosate, dicamba will provide another mode of 

action in the Roundup Ready soybean system to lower the potential risk of weeds 

developing resistance to glyphosate (see Section L.5.3.3 of Appendix L). Furthermore, 

dicamba will provide an alternative mode of action for control of broadleaf weeds with 

populations known to be resistant to ALS and PPO classes of herbicides (see Table VIII-8 

for herbicide listings). 

Application of both dicamba and glyphosate to MON 87708 integrated with the Roundup 

Ready soybean system will provide effective control of both dicamba- and glyphosateresistant broadleaf weeds (Johnson et al, 2010). In the U.S., kochia (Kochia scoparia) and 

prickly lettuce (Lactula serriola) are the only species with biotypes confirmed to be resistant 

to dicamba after 40+ years of use (Heap, 2009). Additionally, a population of lambsquarters 

(Chenopodium album) has been confirmed as resistant in New Zealand, and in Canada 

common hempnettle (Galeopsis tetrahit) and wild mustard (Sinapis arvensis) have been 
Monsanto Company 10-SY-210U 212 of 721 

confirmed as resistant, for a total of five species worldwide with confirmed resistance to 

dicamba. Glyphosate has been shown to provide good to excellent control of all five of 

these broadleaf weeds. In addition, there are 3 species (spreading dayflower (Commelina 

diffusa), field bindweed (Convolvulus arvensis) and wild carrot (Daucus carota)) in the U.S. 

with confirmed resistance to 2,4-D. Of the dicamba and 2,4-D species with known 

resistance in the U.S. and Canada, cross resistance between dicamba and 2,4-D has only 

been documented in wild mustard. However, cross resistance within the other species can 

not be totally ruled out nor assumed to be present. Currently in the U.S., six grass species 

and seven broadleaf species have been confirmed to have resistance to glyphosate. Dicamba 

provides good to excellent control of all seven of these broadleaf species. None of these 

broadleaf weed biotypes have been shown to have populations that are resistant to both 

glyphosate and dicamba. However, there are known resistant populations of kochia that are 

either resistant to glyphosate or to dicamba, but no population with known resistance to both 

glyphosate and dicamba. Since there is no cross resistance between dicamba and glyphosate 

either product can be effective on kochia populations resistant to the other. A more detailed 

discussion regarding the potential development of dicamba resistance in weeds can be found 

in Appendix K. 

The introduction of MON 87708 into the Roundup Ready soybean system will allow 

dicamba to effectively compete with and provide an additional mode-of-action to the 

alternative herbicides that are currently used in combination with glyphosate in preplant and 

in-crop postemergence applications in soybean (for a comparison of dicamba to alternative 

non-glyphosate herbicides, see Appendix L). This will provide an additional mode-of action 

into the soybean weed management system that was previously not available. A comparison 

of dicamba to alternative herbicides, in terms of human health effects (acute toxicity, cancer 

risk, chronic risk, and risk to infants and children), ecological effects (aquatic animal risk 

and aquatic plant risk), weed management efficacy, herbicide-resistant weed frequency, 

rotational crop restrictions, and the potential for injury to the soybean crop, is presented in 

Table VIII-19. In this analysis, dicamba offers an improved risk profile over each 

alternative herbicide in at least one and up to five of the comparative categories; however all 

alternative herbicides are safe when used according to label directions. Additionally, 

dicamba is expected to offer greater benefits than some alternative herbicides as a 

supplement to glyphosate for in-crop applications on MON 87708 since planting interval 

restrictions following preplant applications of dicamba in soybean will be removed and 

allowing dicamba to be applied through planting and up to crop emergence (cracking). 

Dicamba will have greater flexibility for preplant applications than current preplant 

applications of 2,4-D and potentially will replace some 2,4-D applications in soybean. The 

superior broadleaf weed control provided by dicamba and excellent crop tolerance when 

applied to MON 87708 will provide an additional mode-of-action and an alternative to the 

herbicides used for broadleaf weed control in soybean, particularly acifluorfen, lactofen, 

chlorimuron, and flumiclorac. The human health and environmental safety of dicamba 

relative to other non-glyphosate alternative herbicides is discussed in greater detail in 

Appendix L. Considering the characteristics of dicamba from a weed control, compatibility 

with glyphosate, and human and environmental safety perspective, it is concluded that 

MON 87708 will complement the established safety and efficacy of glyphosate use in the 

Roundup Ready soybean system. 
Monsanto Company 10-SY-210U 

Table VIII-17. Common Broadleaf Weed Responses to Preplant Burndown Herbicides 

 Common Broadleaf Weeds1,2Herbicide/Application LQ CR GR SW CC 

M, 

SP CT RC AL HV MT PL 

Spring Preplant Application2,4-D (0.5 lb/1.0 lb) - - - - - 9 -/6 6/8 -/7 6/8 8/9 8/9 Dicamba 9 9 9 9 6 7 - 9 8 8 7 9 

Dicamba + 2,4-D 9 9 9 9 6 9 6 9 8 9 9 9 

Glyphosate 8 9 8 7 7 8 6 7 6 6 6 8 Glyphosate + 2,4-D 9 9 9 8 7 9 6 8 8 8 8+ 9 Glyphosate + Canopy 8 9 9 9 7 8 6 7 6 6 8 8+ Glyphosate + Canopy + 2,4-D 9 9 9 9 7 9 6 8 8 8 9 9 Glyphosate + Gangster + 2,4-D 9 9 9 9 7 9 6 8 8 8 9 9 Glyphosate + Python + 2,4-D 9 9 9 8 7 9 6 8 8 8 9 9 Glyphosate + Scepter + 2,4-D 9 9 9 8 7 9 6 8 8 8 8+ 9 Gly + Sonic/Authority First + 2,4-D 9 9 9 9 7 9 6 8 8 8 9 9 

Glyphosate + Valor + 2,4-D 9 9 9 8 7 9 6 8 8 8 8+ 9 

1All weed control ratings are from the 2009 Weed Control Guide for Ohio and Indiana – Weed Responses to Bur

Ohio State University and Purdue University (Loux et al., 2009). Weed control ratings for weeds are: 9 = 90% t90%, 7 = 70% to 80%, 6 = 60% to 70%, and - = less than 60% control, not recommended. Ratings assume the hin the manner suggested in the guidelines and according to the label under optimum growing conditions. 2Weed species: LQ = lambsquarters, CR = common ragweed, GR = giant ragweed, SW = annual smartweed, CC 

chickweed, M & SP = mustard and shephard’s purse, CT = Canada thistle, RC = red clover, AL = alfalfa, HV = h

marestail, PL = prickly sida, DN & HB = deadnettle & henbit, DL = dandelion, and CG = crested groundsel 
Monsanto Company 10-SY-210U 

Table VIII-18. Common Broadleaf Weed Responses to Dicamba Compared to Labeled Postemergence H

Production 

 Common Broadleaf Weeds1,2

Herbicide/Application BN CB CR GR HS LQ MG PA PW PS SPostemergence Bentazon - 9 7 6 4 7 2-9 4 - 8 Chlorimuron - 9 8 7+ 8 - 8-9 6 9 2 Cloransulam - 9 9 9 3 - 8-9 2 - 2 Chlorimuron/thifensulfuron - 9 8 7+ NA 8 NA NA 9 NA NDicamba3 8 9 9 9 9 8 9 9 8 8 

Flumiclorac - 7 7 - NA 7 NA NA 7 NA NFomesafen 8 7 8+ 8 9 - 8-9 8 9 2 

Glyphosate 8 9 8+ 8 7 8 7-9 9 9 7 Glyphosate/imazethapyr 9 9 8+ 8+ NA 8+ NA NA 9 NA N

Imazamox 9 8 7 8 NA 8 NA NA 9 NA NImazethapyr 9 9 6 7 0 6 7-9 6 9 6 Lactofen 8+ 8 9 8 9 - 8-9 8 9 8 Thifensulfuron - 6 - - NA 8 NA NA 9 NA N

1All weed control ratings except for HS, MG, PA, PS, and SP are from the 2009 Weed Control Guide for Oh

State University and Purdue University (Loux et al., 2009). Ratings for HS, MG, PA, PS, and SP are from th

Guidelines for Mississippi, Mississippi State University (MSU, 2010), except for dicamba ratings for PA are 

Control Manual for Tennessee (University of Tennessee, 2010). Weed control ratings for weeds, except HS, are: 9 = 90% to 100%, 8 = 80% to 90%, 7 = 70% to 80%, 6 = 60% to 70%, and - = less than 60% control, not 

control ratings for HS, MG, PA, PS, and SP are: 9-10 = excellent, 7-8 = good, 4-6 = fair, 0-3 = none to sligh

herbicides are applied in the manner suggested in the guidelines and according to the label under optimum gr

denotes not available. 

2Weed species: BN = black nightshade, CB = cocklebur, CR = common ragweed, GR = giant ragweed, LQ = 

morningglory spp., HS = hemp sesbania, PA = palmer and spiny pigweed, PW = pigweed, PS= prickly sida, S

smartweed, VL = velvetleaf, and WH = waterhemp 

3Weed control ratings for dicamba are from postemergence applications in corn. 
Monsanto Company 10-SY-210U 

Table VIII-19. Summary of Comparative Analysis of Dicamba and Alternative Herbicides 

Herbicide Active 

Ingredient 

Mode-ofAction1

Human Health Risk Measures 

Aquatic NonTarget Species 

Risk Measures Known 

Resistant 

Weed 

Species2 

Herbicidal 

Efficacy 

(< 50% of 

dicamba) 

Lon

Rotatio

Cro

Restric

Acute 

Toxicity 

Risk 

Cancer 

Risk 

Chronic 

Risk 

Infants & 

Children 

Risk 

Aquatic 

Animal 

Risk 

Aquatic 

Plant 

Risk 

2,4-D acid / esters Aux (4) Yes Yes Yes Neutral Yes Yes 28 Neutral Neutr

2,4-DB Aux (4) Neutral Neutral Neutral Neutral Neutral Neutral Yes Neutr

imazethapyr ALS (2) No Neutral Neutral Neutral Neutral Neutral 

 Yes (107) 

Yes Yes 

cloransulam-methyl ALS (2) No Neutral Neutral Neutral Neutral Neutral Yes Yes 

chlorimuron-ethyl ALS (2) No Neutral Neutral Neutral Neutral NA Yes Yes 

thifensulfuron ALS (2) No Neutral Neutral Neutral Neutral Yes Yes Yes 

imazaquin ALS (2) No No Neutral Neutral Neutral NA Yes Yes 

imazamox-ammonium ALS (2) No Neutral No Neutral Neutral Neutral Yes Yes 

flumioxazin PPO (14) Neutral Neutral Yes Neutral Yes Yes 

5 

Yes Yes 

fomesafen PPO (14) Yes Neutral Yes Neutral Neutral Neutral Neutral Yes 

flumiclorac-pentyl PPO (14) No No Neutral Neutral Neutral NA Yes Neutr

sulfentrazone PPO (14) Yes Neutral Neutral Neutral Neutral Yes Yes Neutr

lactofen PPO (14) No Yes Neutral Yes Yes Yes 5 Neutral Neutr

fluthiacet-methyl PPO (14) No Yes Neutral Neutral Neutral Neutral 5 Yes Neutr

acifluorfen sodium PPO (14) Yes Yes Neutral Yes Neutral Neutral 5 Yes Neutr

glufosinate-ammonium Glu (10) Yes No Yes Neutral Neutral Neutral No reports Neutral Neutr

paraquat dichloride BiPyr (22) Yes No Yes Neutral Neutral Yes 24 Yes Neutr

mesotrione HPPD (28) No Neutral Neutral Yes Neutral Yes No reports Yes Yes 

Refer to Appendix L for additional details on the comparison of alternative herbicides to dicamba. “Yes”, indicates that dicamba has an improved risk profile based on presented categories. Entries not indicated with a “Ye

either comparable or less favorable than the alternative herbicide. ”Neutral” entries indicate similar risks exist for dica

herbicide. “No” means the alternative herbicide offers a risk benefit compared to dicamba 1Mode-of-Action based upon WSSA (2010) classifications. 

2A listing of the worldwide numbers of known resistant weeds for each herbicide based on its mode-of-action group. Dicam

resistant weed biotypes worldwide (www.weedscience.org/summary/MOASummary.asp Accessed May 28, 2010). A “Yes
Monsanto Company 10-SY-210U 

number of resistant weeds in this herbicide class is many more than the known five dicamba resistant species biotypes. A c

individual herbicide in the class is not provided. See Section L.5.3.3 in Appendix L. 3Number of “Yes” entries in each row is a summation for all the categories assessed, indicating a total score for improved r

4Number of “No” entries in each row is a summation for all the categories assessed, indicating a total score for worse risk p

NA – not available 
Monsanto Company 10-SY-210U 217 of 721 

VIII.H.1. Potential Impact of Dicamba Application Timing to MON 87708 

The proposed label directions for dicamba applications in MON 87708 will permit preplant 

applications up to the crop emergence and in-crop postemergence applications up to the 

R1/R2 soybean growth stage. Most in-crop applications are expected to be made up to the 

V4 growth stage, when germinated weeds are more easily and economically controlled and 

broadcast applications can be uniformly applied to the weeds (reduced crop canopy 

compared to later vegetative growth stages); with limited applications up to R1/R2, where 

this application is primarily anticipated to control missed weeds, treat a flush of broadleaf 

weeds germinating after the V4 application, or for control of more aggressive glyphosateresistant weed species such as Amaranthus or Ambrosia species. Currently, in-crop 

postemergence applications of dicamba, except for preharvest applications, are not 

permitted or practical on commercially available soybean due to the lack of crop tolerance. 

An application of dicamba at or following the V4 soybean growth stage is expected to be 

later in the growing season than the current latest in-crop application timings to corn, 

sorghum, and wheat. 

To assess the potential impact related to an anticipated difference in application timing of 

dicamba to MON 87708 compared to current labeled uses of dicamba, the latest timing of 

in-crop postemergence dicamba applications in corn was assessed. The comparison to corn 

application timing is pertinent since corn is grown in the same general areas where soybean 

is grown and a high percentage of the total dicamba-treated acres occur in corn (see Table 

VIII-15). Table VIII-20 provides the typical date and days from planting for corn and 

soybean to reach various growth stages. According to USDA-NASS (2009e) planting 

progress reports, the dates at which 50% of the corn and soybean are planted in Illinois are 

April 25 and May 17, respectively. The current dicamba label7

 specifies an allowable use 

pattern relative to corn growth stages as defined by the number of leaves and plant height. 

Dicamba can be broadcast applied to corn up to a height of 8 inches, which is roughly 

equivalent to the 5-leaf stage. To relate these application timings to soybean, it is necessary 

to convert the timing of the 5-leaf stage of corn to a comparable soybean vegetative growth 

stage (V stage). The 5-leaf stage of corn is equivalent to a V4 growth stage (Personal 

communication, Dr. R. Nielsen – Purdue University, 2010). Corn will generally reach this 

growth stage in 37 days from planting based on central Illinois temperatures. Presuming 

corn planting occurs on April 25, corn would reach the comparable V4 stage on June 1. 

Soybean will typically reach the V4 growth stage in 35 days. Presuming soybean planting 

on average occurs on May 17, soybean will reach the V4 stage on approximately June 21. 

Therefore, based on planting and environmental data from central Illinois, the most likely 

application timing for dicamba to MON 87708 is approximately 20 days later than the 

current latest application timing for corn. The timing difference is expected to be applicable 

to much of the Midwest region where the majority of the soybean is grown. 

 

7

 Clarity product label can be found at:http://www.cdms.net/Images/acroiconwblue.gif. Label specifies when 

applying to corn not to apply Clarity when soybean are growing nearby if any of the following conditions 

exist: 1) corn is more than 24” tall, 2) soybean is more than 10” tall, and 3) soybean has begun to bloom. 
Monsanto Company 10-SY-210U 218 of 721 

Table VIII-20. Dates and Days to Reach Various Growth Stages in Corn and Soybean 

in Central Illinois 

 Corn1 Soybean2

Growth 

Stage 

Date & Calendar 

Day 

Days From 

Planting 

Date & Calendar 

Day 

Days 

From Planting 

 

Planting April 25 (115)3 May 17 (137)3 

VE May 10 (130) 15 May 27 (147) 10 

VC June 1 (152) 15 

V1 May 27 (137) 22 June 6 (157) 20 

V2 May 22 (142) 27 June 11(162) 25 

V3 May 27 (147) 32 June 16 (167) 30 

V4 June 1 (152) 37 June 21 (172) 35 

V5 June 5 (156) 41 June 26 (177) 40 

V6 June 9 (160) 45 June 29 (180) 43 

V7 June 13 (164) 49 July 2 (183) 46 

V8 June 17 (168) 53 July 5 (186) 49 

1

Corn growth based on Nielsen (2008) and average growing degree days at Champaign, IL 

(1971-2000). 

2

Soybean growth is based on Naeve (2005). 

3

Average date when 50% of the crop acreage was planted in Illinois during the 5-year period 

2004-2008 (USDA-NASS, 2009d). 

Corn and soybean planting dates and climates are considerably different in the Southeast 

region states than the Midwest region states. Thus, a second analysis was conducted to 

assess the potential difference in timing between current dicamba applications in corn and 

the likely dicamba application to MON 87708 in a southern location. Table VIII-21 

provides the typical date and days for plantings of corn and soybean to reach various growth 

stages in western Tennessee. The dates at which 50% of the corn and soybean are planted in 

Tennessee are April 15 and May 25, respectively. Corn will generally reach the 5-leaf 

growth stage (comparable to the V4 soybean growth stage) in approximately 30 days from 

planting based on western Tennessee’s climate. Typical corn plantings occur on April 15, 

such that corn will reach the V4 stage on May 15. Soybean will reach the V4 growth stage 

in 27 days. Typical soybean planting occurs on May 25, such that soybean will typically 

reach the V4 stage on June 21. Therefore, based on planting and environmental data from 

western Tennessee, the expected postemergence dicamba application timing (V4 growth 

stage) to MON 87708 is approximately 37 days later than the current latest application 

timing for corn. Although the difference between application timings is greater in this 

southern location compared to a northern location, the estimated dates for the V4 stage in 

soybean for the two locations are the same. 

As presented in Appendix M, Monsanto has submitted an application to EPA to amend 

Registration Number 524-582 to add the new use on MON 87708 to the product label. 

Registration Number 524-582 is a DGA salt formulation of dicamba, which is a formulation 

with low volatility. Monsanto also plans to further address the use of dicamba on 

MON 87708 with EPA to evaluate whether any additional measures may be appropriate. In 

addition, Monsanto in cooperation with public sector scientists and organizations will 
Monsanto Company 10-SY-210U 219 of 721 

implement robust stewardship programs to assist in the education of applicators on the best 

application practices to minimize the potential for offsite movement, including the use of 

low volatility formulations. 
Monsanto Company 10-SY-210U 220 of 721 

Table VIII-21. Dates and Days to Reach Various Growth Stages in Corn and Soybean 

in Western Tennessee 

 Corn1 Soybean2

Growth 

Stage 

Date & Calendar 

Day 

Days From 

Planting 

Date & Calendar 

Day 

Days From 

Planting 

 

Planting April 15 (108)3 May 25(148)3 

VE April 25 (118) 10 June 1 (155) 7 

VC June 5 (159) 11 

V1 May 1 (124) 16 June 9 (163) 15 

V2 May 6 (129) 21 June 13 (167) 19 

V3 May 10 (133) 25 June 17 (171) 23 

V4 May 15 (138) 30 June 21 (175) 27 

V5 May 19 (142) 34 July 25 (179) 31 

V6 May 23 (146) 38 July 28 (182) 34 

V7 May 26 (149) 41 July 1 (185) 37 

V8 May 30 (153) 45 July 4 (188) 40 

1

Corn growth based on Nielsen (2008) and 30-year average growing degree days at West 

Memphis, TN. 

2

Soybean growth is based on Naeve (2005). 

3

Average date when 50% of the crop acreage was planted in Tennessee during the 5-year 

period 2004-2008 (USDA-NASS, 2009d). 

Table VIII-22. Average Daily Temperatures in Major Soybean Producing States1

Average Daily Temperature (°F) 

April May June July 

IN 51 62 71 74 

IA 49 60 70 74 

IL 52 62 72 75 

MN 42 56 65 69 

MO 55 64 73 78 

OH 50 60 69 73 

AR 60 69 76 81 

NE 48 59 69 74 

ND 42 55 64 69 

SD 45 57 66 72 

1

 Earth System Research Laboratory of National Oceanic and Atmospheric Administration 

(NOAA-ESRL 2011). 
Monsanto Company 10-SY-210U 221 of 721 

Table VIII-23. Average Relative Humidity (1971-2000) in Major Soybean Producing 

States1

 Morning Afternoon 

 April May June July April May June July 

IN 78 80 81 84 56 57 57 59 

IA 77 78 81 85 61 62 64 66 

IL 73 75 76 79 60 60 61 63 

MN 75 76 81 84 59 58 62 64 

MO 76 82 84 84 61 66 66 65 

OH 76 78 80 83 54 54 55 55 

AR 80 86 86 85 61 61 64 63 

NE 77 79 81 81 56 58 58 58 

ND 77 76 81 81 59 56 60 58 

SD 78 79 81 82 58 58 60 57 

1

 National Climatic Data Center of National Oceanic and Atmospheric Administration 

(NOAA-NCDC 2011). 

VIII.H.2. Impact on Dicamba Use in U.S. Soybean Production Following Integration 

of MON 87708 into the Roundup Ready Soybean System 

Upon integration of MON 87708 into the Roundup Ready soybean system, dicamba 

herbicide use will be expanded to in-crop postemergence applications for those hard-tocontrol and herbicide-resistant broadleaf weeds found in U.S. soybean production. The 

impact that MON 87708 will have on overall dicamba use will be dependent upon the level 

of MON 87708 adoption by growers. Thus, the extent of dicamba-treated soybean acreage 

following the deregulation of MON 87708 is difficult to forecast. Monsanto estimates 

dicamba-treated acres to ultimately be in the range of 30 to 50% of the total U.S. soybean 

acres. This estimate is based on a number of factors: 1) the percentage of non-glyphosate 

herbicides currently used in Roundup Ready soybean, 2) current and historical use of 

dicamba in corn, 3) the development of glyphosate-resistant weeds in soybean cultivation 

areas, 4) the effectiveness of other non-glyphosate herbicides used in the Roundup Ready 

soybean system, 5) compliance with EPA mandatory label instructions may have the 

practical affect of limiting the use in some soybean growing areas; and 6) the foreseeable 

future introduction of new competitive biotechnology-derived traits in soybean. 

Approximately 15 to 21% of growers used a non-glyphosate herbicide in addition to 

glyphosate in the Roundup Ready soybean system in 2005 (Givens et al., 2009). The 

reasons growers stated for their use of a non-glyphosate herbicide in the Roundup Ready 

soybean system were generally for early weed control, residual weed control, improvement 

in the control of specific hard-to-control weeds, and/or glyphosate-resistant weed 

management. Frisvold et al. (2009) reported that in 2007 approximately 30% of the growers 

often or always use herbicides with different modes-of-action in the Roundup Ready 

soybean system. The future use of non-glyphosate herbicides would be expected to increase 
Monsanto Company 10-SY-210U 222 of 721 

in order to support the management of glyphosate-resistant weeds. Additionally, grower 

educational programs on weed resistance management conducted by industry and 

universities encourage the use of non-glyphosate herbicides with alternative modes-ofaction in Roundup Ready cropping systems as a proactive measure to minimize the potential 

for development of glyphosate-resistant weeds (Beckie, 2006; Powles, 2008). These 

programs, along with Monsanto’s support for the use of another herbicide mode-of-action to 

control glyphosate-resistant weeds, will likely drive a further increase in non-glyphosate 

herbicides applied in soybean production. 

A second factor that can be used as an indicator of potential future dicamba-treated soybean 

acreage is the current and historical use of dicamba in corn. Dicamba has been applied for 

many years in conventional and Roundup Ready corn for supplemental broadleaf weed 

control very similar to the proposed uses of dicamba on MON 87708. Therefore, 

information on dicamba use in corn will provide an indication of the potential demand for 

dicamba applied to MON 87708 when planted as a combined trait with Roundup Ready 2 

Yield soybean. The use of non-glyphosate herbicides in Roundup Ready corn was reported 

to be 43 to 44% in 2005, which is significantly higher than presently used in Roundup 

Ready soybean (Givens et al., 2009). More recent market research data indicate a higher 

adoption rate of non-glyphosate herbicides in the corn, where market estimates were around 

70% in 2010 (Agro Trak, 2010). Although, the reasons for using a non-glyphosate 

herbicide in the Roundup Ready corn system are the same as in Roundup Ready soybean, 

the benefits of using residual herbicides are greater in Roundup Ready corn because of the 

need for early season weed control and greater application flexibility with postemergence 

application of glyphosate (Dalley et al., 2001). Arguably for these reasons, the use of nonglyphosate herbicides applied to MON 87708 as a combined trait with Roundup Ready 2 

Yield soybean will not likely exceed the current levels in Roundup Ready corn. 

Prior to the introduction of Roundup Ready corn, the use of dicamba in corn production was 

greater since it was used mainly to complement the control of residual grass herbicides by 

providing postemergence control of annual broadleaf weed escapes. Dicamba-treated acres 

in conventional corn peaked at 29% of the corn acres in 1994. Dicamba use in corn dropped 

to 9.4% of the corn acres in 2008 following the commercial introduction of Roundup Ready 

corn (Table VIII-12). Therefore, considering the current use of dicamba in Roundup Ready 

corn (conservatively assumed to be 43 to 70%), the historical use of dicamba in 

conventional corn (peaking at 29% of the corn acres in 1994), and the current use of nonglyphosate herbicides in soybean (15 to 21% of growers applied a non-glyphosate herbicide 

in 2005), it can reasonably be concluded that dicamba-treated acres of MON 87708 will not 

likely exceed 50% of the total soybean acres following introduction of MON 87708. 

A third factor impacting dicamba-treated soybean acreage is the current and future need for 

control of glyphosate-resistant weeds. Glyphosate-resistant weeds have been identified in 

multiple states (Heap, 2009). When a glyphosate-resistant weed biotype has been confirmed 

to be present in a geographical area, growers in that area are recommended by Monsanto, 

glyphosate distributors, and university specialists to proactively implement glyphosateresistant weed management programs to ensure effective control of the resistant weed 

biotype regardless of whether the weed species has been confirmed to be resistant on a 

grower’s farm. Therefore, the acreage in an area where responsive weed resistance 
Monsanto Company 10-SY-210U 223 of 721 

management practices are implemented is potentially greater than the actual acres known to 

be impacted by glyphosate-resistant weeds. University weed scientists are recommending 

growers proactively implement best management practices, including a non-glyphosate 

herbicide with a second mode-of-action, in their cropping systems to minimize the 

development and potential spread of glyphosate-resistant weeds in the future (Owen et al., 

2009; Frisvold et al., 2009). Monsanto supports this recommendation where glyphosateresistant biotypes are present, such that MON 87708 integrated into the Roundup Ready 

soybean system should be treated with glyphosate and dicamba plus another herbicide 

mode-of-action to ensure at least two effective modes-of-action cover the spectrum of weeds 

present in a grower’s soybean field. In this case, the other herbicide mode-of-action that 

will be recommended in the MON 87708 plus Roundup Ready soybean system is likely to 

be a soil active residual preemergence herbicide. 

It is anticipated that even in locations where glyphosate-resistant weeds are present, 

glyphosate will continue to be the base herbicide applied to MON 87708 as a combined trait 

product with MON 89788 (Roundup Ready 2 Yield soybean), thereby providing broad 

spectrum control of grass and broadleaf weeds. MON 87708 allows the use of dicamba in 

the Roundup Ready soybean system for control of glyphosate-resistant weeds and to 

improve the control of dicotyledonous weeds that are hard to control with glyphosate alone. 

As described in Section VIII.F.1 and Table VIII-8, numerous residual and postemergence 

herbicides are utilized currently to complement the control of glyphosate for hard-to-control 

weeds and to mitigate or control glyphosate-resistant weeds. These alternative herbicides 

will compete with dicamba and undoubtedly will reduce the potential dicamba use on 

MON 87708 integrated into the Roundup Ready soybean system and in future combined 

trait products containing dicamba tolerance. 

An additional factor influencing the number of dicamba-treated soybean acres in the future 

will be the introduction of competing herbicide-tolerant traits in soybean. Currently, there 

are numerous herbicide-tolerant soybean products under regulatory review or recently 

authorized (see Appendix J, Table J-4). This includes several products that have tolerance 

to multiple herbicides with different modes-of-action. These new biotechnology-derived 

herbicide-tolerant soybean products are anticipated to be introduced in future years and will 

compete with Monsanto’s MON 87708 × MON 89788 combined-trait soybean product, 

further reducing the ultimate potential of dicamba applications in soybean. A grower may 

chose to not cultivate MON 87708 or to use an alternative soybean herbicide for their weed 

control. Furthermore, compliance with EPA mandatory label instructions may have the 

practical affect of limiting the use in some soybean growing areas. 

Taking into consideration the above assessment, the potential dicamba-treated MON 87708 

acreage is estimated to be 40% of the U.S. soybean acres, and would represent 

approximately 30 million dicamba-treated soybean acres. Considering the acreage currently 

treated with dicamba (20.2 million acres of which 0.53 million acres area soybean), this 

would potentially result in a total of 50.2 million acres treated with dicamba. As presented 

previously, dicamba was used on approximately 36 million acres at its peak use in 1994. 

Furthermore, considering the anticipated weed control recommendations for dicamba use in 

MON 87708, as outlined in Section VIII.H, combined with current dicamba usage in other 

crops (2.7 million pounds), a total of 25 million pounds a.e. of dicamba is estimated (high-
Monsanto Company 10-SY-210U 224 of 721 

end) to be applied annually, compared to 9.4 million pounds a.e. of dicamba applied at its 

peak use in 1994. 

VIII.H.3. Conclusions on Dicamba-Tolerant Soybean MON 87708 

Upon integration of MON 87708 into the Roundup Ready soybean system, growers will 

have the ability to continue use of established soybean production practices including tillage 

systems; the same planting and harvesting machinery; traditional management of insects, 

diseases, and other pests; and many of the current herbicides used for weed control, 

including glyphosate with its established environmental benefits. Similarly, certified seed 

production will continue to use well-established industry practices to deliver high quality 

seed to growers. Due to the excellent crop safety of MON 87708 to dicamba, growers will 

have a new herbicide mode-of-action for in-crop control of hard-to-control and herbicide 

resistant broadleaf weeds that are present in U.S. soybean production. As expected with a 

new use of herbicide in U.S. soybean production, the number of dicamba-treated soybean 

acres and the total dicamba use will increase. The total dicamba use is expected to be about 

double the historical peak levels experienced since dicamba’s introduction in 1967. 

Additionally, due to the expanded timing of in-crop applications to soybean, dicamba 

treatments may be later in the growing season than currently occurs for most labeled 

dicamba uses, including applications to corn. Monsanto has requested EPA approval for 

the use of a low volatility dicamba (DGA salt) formulation, together with allowing only 

ground application on MON 87708, subject to certain limitations specified on the label. 

With these additional limitations, the potential for offsite movement (spray drift and 

volatility) onto adjacent crops due to applications on MON 87708 later in the season is not 

expected to significantly impact adjacent crops. In addition, Monsanto will consult with 

U.S. EPA to identify what additional measures, if any, are appropriate to address any 

potential impact of dicamba offsite movement. In addition, to reinforce anticipated EPA 

label requirements, Monsanto will implement a robust stewardship program that will include 

a strong emphasis on grower and applicator training. 

VIII.I. Crop Rotation Practices in Soybean 

The well-established farming practice of crop rotation is still a key management tool for 

growers. The purpose of growing soybean in rotation is to improve yield and profitability of 

one or both crops over time, decrease the need for nitrogen fertilizer on the crop following 

soybean, increase residue cover, mitigate or break disease, insect, and weed cycles, reduce 

soil erosion, increase soil organic matter, improve soil tilth and soil physical properties, and 

reduce runoff of nutrients, herbicides, and insecticides (Boerma and Specht, 2004; Al-Kaisi 

et al., 2003). According to USDA Economic Research Service, 95% of the soybean-planted 

acreage has been in some form of a crop rotation system since 1991 (USDA-ERS, 2001). 

Corn- and wheat-planted acreage has been rotated at a slightly lower level of 75% and 70%, 

respectively. Although the benefits of crop rotations can be substantial, the grower must 

make cropping decisions by evaluating both the agronomic and economic returns of various 

cropping systems. Crop rotations also afford growers the opportunity to diversify farm 

production in order to minimize market risks. 
Monsanto Company 10-SY-210U 225 of 721 

Continuous soybean production is not a common practice in the Midwest and is discouraged 

by most extension soybean specialists to reduce the risk of damage from diseases and 

nematodes(Hoeft et al., 2000; Al-Kaisi et al., 2003). Corn and soybean occupy more than 

80% of the farmland in many of the Midwestern states, and the two-year cropping sequence 

of soybean-corn is used most extensively in this region. However, a soybean crop 

sometimes is grown after soybean and then rotated to corn in a 3-year rotation sequence 

(soybean-soybean-corn) in the Midwest. Compared to corn, soybean shows a greater yield 

response to being grown after a number of years without soybean. The yield of both corn 

and soybean is approximately 10% higher when grown in rotation than when either crop is 

grown continuously (Hoeft et al., 2000). 

A combination of conservation tillage practices and crop rotation has been shown to be very 

effective in improving soil physical properties. Long-term studies in the Midwest indicate 

that the corn-soybean rotation improves yield potential of no-till systems compared to 

continuous corn production (Al-Kaisi, 2001). The reduction in yield of continuous corn 

production in no-till systems is attributed to low soil temperature during seed germination, 

which is evident on poorly drained soils under no-till practices. 

Unique to the southern portion of the Midwest and the Southeast regions, soybean is grown 

in a double-cropping system. Double-cropping refers to the practice of growing two crops 

in one year. This practice can improve income and reduce soil and water losses by having 

the soil covered with a plant canopy most of the year (Hoeft et al., 2000). In the Midwest, 

winter wheat is harvested in late June or July, and then soybean is planted into the wheat 

residue in a no-till system to conserve moisture. Due to the uncertainty of double-cropping 

yields, growers sometimes do not plant if soils are too dry at the time of wheat harvest. 

Soybean typically is grown in a corn-wheat-soybean rotation sequence when soybean is 

grown in a double-cropping system. 

Rotation practices for soybean vary from state to state. However, there are similarities 

among states within certain growing regions. This section provides a detailed description 

and quantitative assessment of the rotational cropping practices immediately following 

soybean production, by region and state. This assessment accounts for about 99% of total 

U.S. soybean acreage. This assessment is based on current agronomic practices following 

soybean production and current dicamba herbicide usage in labeled crops. USDA-NASS 

(2004, 2006, 2007b, 2008) and AgroTrak (2009) data on dicamba herbicide usage for corn, 

sorghum, cotton, wheat, barley, and oats were utilized for this assessment. For the purpose 

of this assessment, a 50% adoption rate in U.S. soybean production was assumed for 

MON 87708. These data on rotational patterns are presented in Tables VIII-24 through 

VIII-27. 

The majority of the U.S. soybean acreage (68.6%) is rotated to corn (Table VIII-24). The 

second largest rotational crop following soybean is soybean. Approximately 14.5% of the 

soybean acreage is rotated back to soybean the following year. Wheat follows soybean on 

approximately 11.2% of the U.S. soybean acreage, with cotton, rice, and sorghum the next 

largest rotational crops. However, these three crops were planted on only 4.6% of the 

soybean acreage. Other minor rotational crops that follow soybean production are listed in 

Table VIII-24. 
Monsanto Company 10-SY-210U 226 of 721 

Column J of each table provides the percentage of soybean acreage as a function of the total 

rotational crop acreage to indicate the level that soybean is the primary crop preceding the 

rotational crops. For the entire country (Table VIII-24), this percentage is 35.3% indicating 

that soybean is a major crop preceding these rotational crops. The percentage of soybean as 

a preceding crop varies widely in different states, which ranges from 16.8% (KS, Table 

VIII-25) to 95.2% (NJ, Table VIII-27). In the Midwest region where 82% of the soybean is 

grown, 34.6% of the rotational crop area was planted with soybean during the previous 

growing season. 

Dicamba-tolerant soybean can be followed by dicamba-tolerant soybean or another 

rotational crop with a labeled application of dicamba. To determine the likelihood that the 

rotational crops planted after MON 87708 will be a rotational crop where dicamba 

application is possible, an assessment also has been provided in Column K of Tables VIII24 through VIII-27. This assessment showed that the percentage of the total rotational crop 

acreage that may be rotated from MON 87708 to another crop with a potential for dicamba 

application (Table VIII-24 - Column K) is estimated to be 4.9% in the U.S., with ranges 

from32.3% in Mississippi to Georgia and New Jersey having essentially no rotational crop 

acres with dicamba usage since these states have essentially no continuous soybean acres 

and USDA-NASS and AgroTrak statistics show no use of dicamba currently in labeled 

crops in these states. The percentage is 4.1% in the Midwest region, which is the largest 

soybean production region. 
Monsanto Company 10-SY-210U 

Table VIII-24. Rotational Practices in the U.S. Following Soybean Production 

A B C D E F G H I J State 

Total Soybean Acres1

Major Crops Following Soybean in Rotation 

Total Acreage of Rotational Crop in the U.S.1

Rotational Crop Acres Following Soybean2

% Rotational Crop Following Soybean3

% Rotational Crop of Total Soybean4

Acreage of Dicamba in Rotational Crop Option5

% Dicamba Usage in Rotational Crop Option6

% AcPrMRo

United States 

75037 Corn 

Soybean Sorghum Cotton Wheat Barley Oats Rice Alfalfa Sugar Beets Potatoes Dry Beans Dry Peas Millet Flax Other9

80130 75037 4020 3767 37414 2159 1995 2301 1864 830 334 1183 520 250 345 452 

Total: 212601

51500 10866 841 1570 8396 41 98 1042 162 144 32 35 38 41 76 155 

Total: 75037

64.3 14.5 20.9 41.7 22.4 1.9 4.9 45.3 8.7 17.3 9.6 3.0 7.3 16.4 22.0 34.3 

68.6 14.5 1.1 2.1 11.2 0.05 0.1 1.4 0.2 0.19 0.04 0.05 0.05 0.05 0.10 0.2 

4431 5438 153 448 Total: 10470

8.6% 40.0% 9.7% 5.3% 

35

This U.S. summary was developed by compiling the data from all three regional summaries. All acreage is expressed as 1000s of acres1Acreage planted of the specific crops is based on 2008 planting data (USDA-NASS, 2009a); “other” crop and newly seeded alfalfa a

planting data from the Individual States data which was obtained from Quick Stat searches on http://www.nass.usda.gov/Data_and_Statistics/Quick_Stats/index.asp. 2Column E is obtained by compiling the data from all three regional summaries. 

3Column F is obtained by dividing Column E by Column D. 4Column G is obtained by dividing Column E by Column B. 5Column H is obtained by compiling the data from all three regional summaries. 6Column I is obtained by dividing Column H by Column E.7Column J is obtained by dividing Column B by Column D Total. 8Column K is obtained by dividing Column H Total by Column D Total. 9Various vegetables.
Monsanto Company 10-SY-210U 

Table VIII-25. Rotational Practices Following Soybean Production in the Midwest Region A B C D E F G H I J State 

Total Soybean Acres1

Major Crops Following Soybean in Rotation 

Total Acreage of Rotational Crop in States1

Rotational Crop Acres Following Soybean2

% Rotational Crop Following Soybean3

% Rotational Crop of Total Soybean4

Acreage of Dicamba in Rotational Crop Option5

% Dicamba Usage in Rotational Crop Option6

% SoyAcres PrecedMajor Rotatio

Region 62150 Corn 

Soybean Sorghum Cotton Wheat Barley Oats Rice Alfalfa9

Sugar Beets Potatoes Dry Beans Dry Peas Millet Flax Other10

72260 62150 3553 341 32039 1929 1590 200 1617 830 278 1166 520 250 345 342 Total: 179410

47480 4885 670 78 8102 41 98 182 162 144 32 3538417687Total: 62150

65.7 7.9 18.8 22.9 25.3 2.1 6.2 91.0 10.0 17.3 11.6 3.0 7.3 16.4 22.0 25.3 

76.47.91.10.113.00.070.20.30.30.20.050.060.060.070.10.1

4591 1954 313 14 448 6 Total: 7326

9.7 40 46.8 18 5.5 15 * NA NA NA NA NA NA * NA NA 

34.6 

IL 9200 Corn 

Soybean Sorghum Wheat 

12100 9200 80 1200 Total: 22580

8556 230 74 340Total: 9200

71 3 92 28 

93.0 2.5 0.8 3.7 

1711 92 265 Total: 2068

20 40 46

* 

40.7 

IN 5450 Corn 

Soybean Wheat 

5700 5450 580 Total: 11730 

4905 273 273 Total: 5450

86 5 47 

90 5 5 

343 109 Total: 453

7 40 * 

46.5 
Monsanto Company 10-SY-210U 

Table VIII-25 (continued). Rotational Practices Following Soybean Production in the Midwest Region A B C D E F G H I J State 

Total Soybean Acres1

Major Crops Following Soybean in Rotation 

Total Acreage of Rotational Crop in States1

Rotational Crop Acres Following Soybean2

% Rotational Crop Following Soybean3

% Rotational Crop of Total Soybean4

Acreage of Dicamba in Rotational Crop Option5

% Dicamba Usage in Rotational Crop Option6

% SoyAcres PrecedMajor Rotati

IA 9750 Corn 

Soybean Alfalfa9

13300 9750 125 Total: 23175

9458 195 98 

Total: 9750

71 2 78 

97 2 1 

378 78 Total: 456

4 40 NA 

42.1 

KS 3300 Corn 

Soybean Sorghum Wheat 

3850 3300 2900 9600 Total: 19650

1650 330 165 1155 

Total: 3300

43 10 6 12 

50 10 5 35 

297 132 13 58 Total: 500

18 40 8 5 

16.8 

KY 1390 Corn 

Soybean Wheat 

1210 1390 580 Total: 3180

1182 139 70 

Total: 1390

98 10 12 

85 10 5 

106 56 Total: 162

9 40 * 

43.7 

MI 1900 Corn 

Soybean Wheat 

2400 1900 730 Total: 5030

1330 95 475 

Total: 1900

55 5 65 

70 5 25 

160 38 5 Total: 202

12 40 16

37.8 

MN 7050 Corn 

Soybean Wheat Sugar beets Dry Beans Other11

7700 7050 1925 440 150 203 Total: 17468

5358 212 1269 106 35 71 

Total: 7050

70 3 66 24 24 35 

76 3 18 1.5 0.5 1 

697 85 14 Total: 796

13 40 16

NA NA NA 

40.4 
Monsanto Company 10-SY-210U 

Table VIII-25 (continued). Rotational Practices Following Soybean Production in the Midwest Region A B C D E F G H I J State 

Total Soybean Acres1

Major Crops Following Soybean in Rotation 

Total Acreage of Rotational Crop in States1

Rotational Crop Acres Following Soybean2

% Rotational Crop Following Soybean3

% Rotational Crop of Total Soybean4

Acreage of Dicamba in Rotational Crop Option5

% Dicamba Usage in Rotational Crop Option6

% SoybeAcres PrecedinMajor Rotation

MO 5200 Corn 

Soybean Sorghum Cotton Wheat Rice 

2800 5200 90 306 1250 200 Total: 9846

2756 1560 104 78 520 182 Total: 5200

98 30 116 25 42 91 

53 30 2 1.5 10 3.5 

201 624 14 Total: 839

76

40 * 18 * NA 

52.8 

NE 4900 Corn 

Soybean Sorghum Wheat 

8800 4900 300 1750 Total: 15750

3675 490 245 490 Total: 4900

42 10 82 28 

75 10 5 10 

147 196 22 15 Total: 380

4 40 9 3 

31.1 

ND 3800 Corn 

Soybean Wheat Sugar Beets Dry Peas Flax 

2550 3800 9230 208 520 335 Total: 16643

1140 798 1710 38 38 76 Total: 3800

45 21 19 18 7 23 

30 21 45 1 1 2 

331 319 274 Total: 923

29 50 16 NA NA NA 

22.8 

OH 4500 Corn 

Soybean Wheat 

3300 4500 1120 Total: 8920

3150 450 900 Total: 4500

95 10 80 

70 10 20 

32 180 18 Total: 230

1 40 25

50.4 
Monsanto Company 10-SY-210U 

Table VIII-25 (continued). Rotational Practices Following Soybean Production in the Midwest Region A B C D E F G H I J State 

Total Soybean Acres1

Major Crops Following Soybean in Rotation 

Total Acreage of Rotational Crop in States1

Rotational Crop Acres Following Soybean2

% Rotational Crop Following Soybean3

% Rotational Crop of Total Soybean4

Acreage of Dicamba in Rotational Crop Option5

% Dicamba Usage in Rotational Crop Option6

% SoybeaAcres Preceding Major Rotations7

SD 4100 Corn 

Soybean Sorghum Wheat Barley Oats Millet 

4750 4100 170 3661 63 220 110 Total: 13074

2952 82 82 820 41 82 41 Total: 4100

62 2 48 22 65 37 37 

72 2 2 20 1 2 1 

148 33 13 66 6 Total: 266

5 40 16 8 15 * * 

31.4 

WI 1610 Corn 

Soybean Wheat Oats Alfalfa9

Potatoes Other12

3800 1610 373 270 420 63.5 139.2 Total: 6676

1369 32 81 16 64 32 16 Total: 1610

36 2 22 6 15 51 12 

85 2 5 1 4 2 1 

41 13 Total: 54

3 40 * * NA NA NA

24.1 

This Midwest region summary table was developed by compiling the data from all the states within the region except where noted in fApplicable. An asterisk (*) indicates no usage reported. All acreage are expressed as 1000s of acres. 1Acreage planted of the specific crops is based on 2008 planting data (USDA-NASS, 2009a); “other” crop and newly seeded alfalfa a

planting data from the Individual States data which were obtained from Quick http://www.nass.usda.gov/Data_and_Statistics/Quick_Stats/index.asp. 2Column E for the regional summary is obtained by compilin

within the region. Column E for the individual states is obtained by multiplying Column B by Column G. 

3Column F is obtained by dividing Column E by Column D. 4Column G for the regional summary is obtained by dividing Column E by Column B. The rotational crop percentages in Column G f

based on estimates from personal communications (2006) with individual state Extension Crop Production Specialist; Extension Agrand Cotton; Extension Weed Control Specialist on Soybean and Corn; and/or Monsanto Technology Development Representatives. 

5Column H for the regional summary is obtained by compiling the data from all the states within the region. Column H for the indiv

multiplying Column E by Column I. 
Monsanto Company 10-SY-210U 

6Column I for the regional summary is obtained by dividing Column H by Column E. Dicamba usage except for individual states for 

recent statistics from USDA-NASS (2004 -sorghum, 2006 - corn and oats, 2007b - wheat and barley, 2008 -cotton). Percentages 

usage statistics from AgroTrak (2009), due to USDA-NASS data not being reported for the particular crop and state. Dicamba usfuture market adoption estimate.7Column J is obtained by dividing Column B by Column D Total. 8Column K is obtained by dividing Column H Total by Column D Total. 

9Newly seeded alfalfa. 10Various vegetables. 11Sweet corn and green peas. 12Sweet corn, green peas and onions.
Monsanto Company 10-SY-210U 

Table VIII-26. Rotational Practices Following Soybean Production in the Southeast Region A B C D F E G H I J State 

Total Soybean Acres1

Major Crops Following Soybean in Rotation 

Total Acreage of Rotational Crop in States1

Rotational Crop Acres Following Soybean2

% Rotational Crop of Total Soybean3

% Rotational Crop Following Soybean4

Acreage of Dicamba in Rotational Crop Option5

% Dicamba Usage in Rotational Crop Option6

% Soybean Acres Preceding Major Rotations7

Region 10860 Corn 

Soybean Sorghum Cotton Wheat Rice Other* 

3535 10430 245 2380 1530 1631 65 Total: 19816 

2284 5748 171 1469 294 860 34 Total:10860

21.0 52.9 1.6 13.5 2.7 7.9 0.3 

64.6 55.1 69.8 61.7 19.2 52.7 52.0

72 2299139 Total: 2510 

3.2 40 * 9.4 * NA

54.8 

AL 360 Corn 

Soybean Cotton Wheat 

260 360 290 240 Total: 1150

126 18 180 36 Total: 360

35 5 50 10 

48 5 62 15 

26 7 13 Total: 46 

216

40 7 * 

31.3 

AR 3300 Corn 

Soybean Sorghum Wheat Rice 

440 3300 125 1070 1401 Total: 6336

231 2112 66 231 660 Total: 3300

7 64 2 7 20 

53 64 53 22 47 

845 Total: 845 

* 40 * * NA 

52.1 

GA 430 Corn 

Cotton 

370 940 Total: 1310

43 387 Total: 430

10 90 

12 41 

2 Total: 2 

56

* 

32.8 

LA 1050 Corn 

Soybean Sorghum Cotton 

520 1050 120 300 Total: 1990

105 683 105 158 Total: 1050

10 65 10 15 

20 65 88 53 

3 273 Total: 276 

36

40 * * 

52.8 
Monsanto Company 10-SY-210U 

Table VIII-26 (continued). Rotational Practices Following Soybean Production in the Southeast Region A B C D E F G H I J State 

Total Soybean Acres1

Major Crops Following Soybean in Rotation 

Total Acreage of Rotational Crop in States1

Rotational Crop Acres Following Soybean2

% Rotational Crop of Total Soybean3

% Rotational Crop Following Soybean4

Acreage of Dicamba in Rotational Crop Option5

% Dicamba Usage in Rotational Crop Option6

% SoAcres PreceMajoRotat

MS 2000 Soybean 

Rice

2000 230 Total: 2230

1800 200 Total: 2000

90 10 

90 87 

720 Total: 720

40 NA 

89.7 

NC 1690 Corn 

Soybean Cotton Other9

900 1690 430 65 Total: 3085

811 423 423 34 Total: 1690

48 25 25 2 

90 25 98 52 

36 169 Total: 205

46

40 * NA 

54.8 

TN 1490 Corn 

Soybean Cotton 

690 1490 285 Total: 2465

671 551 268 Total: 1490

45 37 18 

97 37 94 

5 220 126 Total: 351

16

40 47 

60.4 

SC 540 Corn 

Soybean Cotton Wheat

355 540 135 220 Total: 1250

297 162 54 27 Total: 540

55 30 10 5 

84 30 40 12 

65 Total: 65

* 40 * * 

43.2 

This Mid-South region summary table was developed by compiling the data from all the states within the region except where noted othdenotes not applicable. An asterisk (*) indicates no usage reported. All acreage are expressed as 1000s of acres. 1Acreage planted of the specific crops is based on 2008 planting data (USDA-NASS, 2009a); “other” crop and newly seeded alfalfa acr

planting data from the Individual States data which were obtained from Quick Stat searches on http://www.nass.usda.gov/Data_and_Statistics/Quick_Stats/index.asp. 2 Column E for the regional summary is obtained by compiling d

within the region. Column E for individual states is obtained by multiplying Column B by Column G. 

3Column F is obtained by dividing Column E by Column D. 4Column G for the regional summary is obtained by dividing Column E by Column B. The rotational crop percentages in Column G fo

based on estimates from personal communications (2006) with individual state Extension Crop Production Specialist; Extension Agronand Cotton; Extension Weed Control Specialist on Soybean and Corn ;and/or Monsanto Technology Development Representatives. 5Column H for the regional summary is obtained by compiling the data from all the states within the region. Column H for the individu

multiplying Column E by Column I. 
Monsanto Company 10-SY-210U 

6Column I for the regional summary is obtained by dividing Column H by Column E. Dicamba usage except for individual sites for soy

recent statistics from USDA-NASS (2004 - sorghum, 2006 - corn and oats, 2007b - wheat and barley, 2008 - cotton). Percentages w

usage statistics from AgroTrak (2009) because USDA-NASS data was not reported for the particular crop and state. Dicamba usage 

future market adoption estimate. 7Column J is obtained by dividing Column B by Column D Total. 8Column K is obtained by dividing Column H Total by Column D Total. 

9Cucumbers and sweet potatoes. 
Monsanto Company 10-SY-210U 

Table VIII-27. Rotational Practices Following Soybean Production in the East Coastal Region A B C D E F G H I J State 

Total Soybean Acres1

Major Crops Following Soybean In Rotation 

Total Acreage of Rotational Crop in States1

Rotational Crop Acres Following Soybean2

% Rotational Crop Following Soybean3

% Rotational Crop of Total Soybean4

Acreage of Dicamba in Rotational Crop Option5

% Dicamba Usage in Rotational Crop Option6

% SoyAcres PrecedMajor Rotatio

Region 4687 Corn 

Soybean Cotton Other9

3615 1705 61 45 Total: 5426 

1741 242 23 21 

Total: 2027

48.2 14.2 38.0 45.9 

85.9 11.9 1.1 1.0 

390 97 Total: 487

22.4 40 * NA 

37.4 

DE 195 Corn 

Soybean 

160 195 Total: 355

156 39 

Total: 195

98 20 

80 20 16 

Total: 16

* 40 

54.9 

MD 495 Corn 

Soybean 

460 495 Total: 955

446 50 

Total: 495

97 10 

90 10 

258 20Total: 278

586

40 

51.8 

NJ 92 Corn 

Other9

85 11.6 Total: 97

83 9 

Total: 92

97 79 

90 10 

Total: 0

* NA 

95.2 

NY 230 Corn 

Other10

1090 33.5 Total: 1124

219 12 

Total: 230

20 34 

95 5 

11 Total: 11

5 * 

20.5 

PA 435 Corn 

Soybean 

1350 435 Total: 1785

426 9 

Total: 435

32 2 

98 2 

30 4 Total: 34

7 40 

24.4 

VA 580 Corn 

Soybean Cotton 

470 580 61 Total: 1111

412 145 23 

Total: 580

88 25 38 

71 25 4 

91 58 Total: 149

22640 * 

52.2 

This Eastern Coastal region summary table was developed by compiling the data from all the states within the region except where notedenotes not applicable. An asterisk (*) indicates no usage reported. All acreage are expressed as 1000s of acres. 
Monsanto Company 10-SY-210U 

1Acreage planted of the specific crops is based on 2008 planting data (USDA-NASS, 2009a); “other” crop and newly seeded alfalfa acr

planting data from the Individual States data which were obtained from Quick Stat searches on http://www.nass.usda.gov/Data_and_Statistics/Quick_Stats/index.asp. 2Column E for the regional summary obtained by compiling data from all of the states within the region. Column E for individual state

multiplying Column B by Column G. 

3Column F is obtained by dividing Column E by Column D. 4Column G for the regional summary is obtained by dividing Column E by Column B. The rotational crop percentages in Column G fo

based on estimates from personal communications (2006) with individual state Extension Crop Production Specialist; Extension Agronand Cotton; Extension Weed Control Specialist on Soybean and Corn; and/or Monsanto Technology Development Representatives.5Co

summary is obtained by compiling the data from all the states within the region. Column H for the individual states is obtained by mulColumn I. 

6Column I for the regional summary is obtained by dividing Column H by Column E. Dicamba usage except for individual states for s

recent statistics from USDA-NASS (2004 -sorghum, 2006 -corn and oats, 2007b -wheat and barley, 2008 -cotton). Percentages with th

statistics from AgroTrak (2009) because USDA-NASS data was not reported for the particular crop and state. Dicamba usage in soybemarket adoption estimate. 

7Column J is obtained by dividing Column B by Column D Total. 8Column K is obtained by dividing Column H Total by Column D Total. 9Sweet corn and other vegetables. 10Sweet corn and onions. 
Monsanto Company 10-SY-210U 238 of 721 

VIII.J. Soybean Volunteer Management 

Volunteer soybean is defined as a plant that has germinated and emerged unintentionally in 

a subsequent crop. Soybean seeds can remain in a field after soybean harvest as a result of 

pods splitting before or during harvest. Soybean seeds also can remain in a field when pod 

placement on the plants is too close to the ground for the combine head to collect all the 

pods or when the combine is improperly adjusted for efficient harvesting. Volunteer 

soybean in rotational crops is not a concern in the Midwest region because the soybean seed 

is typically not viable after the winter period (Carpenter et al., 2002; OECD, 2000). In 

southern soybean growing areas of the U.S. where the winter temperatures are milder, it is 

possible for soybean seed to remain viable over the winter and germinate the following 

spring. 

Volunteer soybean normally is not a concern in rotational crops, such as corn, cotton, rice, 

and small grains (e.g., wheat, barley, sorghum, and oats), that are the significant rotational 

crops following soybean due to control measures that are available for volunteer soybean 

when they arise (Carpenter et al., 2002; OECD, 2000). Because of these control measures 

and field testing which confirmed that MON 87708 has equivalent volunteer potential as 

other soybean, the introduction of MON 87708 plus Roundup Ready soybean will not 

elevate concerns about managing volunteer soybeans nor will it result in more dependence 

on preplant or in-crop tillage and cultivation because there are adequate alternative herbicide 

options. Preplant tillage and/or herbicides are the first management tool for control of 

emerging volunteer soybean in the spring, where this may be an issue, such as in the south. 

If volunteer soybean should emerge after planting, shallow cultivation and/or use of another 

herbicide will control volunteers and effectively reduce competition with the crop. Several 

postemergence herbicides also are available to control volunteer soybean (conventional or 

glyphosate-tolerant soybean, and by extension dicamba-tolerant soybean) in each of the 

major soybean rotational crops. Table VIII-28 provides control ratings on volunteer 

soybean for several herbicides used in the major rotational crops. Additionally, 19 

commonly used herbicides in agriculture, representing eight modes-of-action (i.e., ALS 

inhibitor, chloroacetamide, EPSPS, PPO inhibitor, PSI disruption, PSII inhibitor, synthetic 

auxin, and tubulin inhibitor classes) were tested as potential substrates for the MON 87708 

DMO present in MON 87708 (Section V.A). Across nearly all of the herbicides tested, 

MON 87708 and the conventional control were similar in their level of tolerance, except for 

the three phenoxy synthetic auxin herbicides (2,4-D, 2,4-DB, and MCPA). These results 

indicate that herbicides which are effective for the control of volunteer soybean can still be 

used to control MON 87708. 

To provide control of volunteer soybean in corn, postemergence applications of AAtrex®

(atrazine), Hornet® (flumetsulam + clopyralid) and Widematch® (clopyralid + fluroxypyr) 

provide excellent control (Zollinger, 2009). AAtrex and Permit® (halosufuron) provide 

excellent volunteer control in sorghum. In small grains (wheat, barley, oats), Bronate®

Advanced (bromoxynil/MCPA), and Widematch® applied postemergence provide excellent 

control of volunteer soybean (Zollinger, 2009). 

Volunteer soybean in cotton is normally not a concern. However, hurricanes or other 

extreme weather conditions can damage a soybean crop preceding cotton production in the 
Monsanto Company 10-SY-210U 239 of 721 

Mid-South region states, where the unharvested soybean seed can produce volunteer plants. 

Preplant applications of paraquat or herbicide mixtures containing paraquat will effectively 

control volunteer glyphosate-tolerant soybean (Montgomery et al., 2002; Murdock et al., 

2002). Recent research in North Carolina indicates Envoke (trifloxysulfuron) will provide 

excellent postemergence control of (volunteer) soybean containing traits for glyphosate and 

sulfonylurea herbicide tolerance in Roundup Ready cotton (York et al., 2005). 

Volunteer soybean in rice is rarely a concern due to the combination of preplant tillage, 

flooding practices, and herbicides used in producing rice. If volunteer plants should emerge 

in rice, postemergence applications of Grandstand (triclopyr), Regiment (bispyribac), Grasp 

(penoxsulam), and Permit (halosulfuron) typically used for weed control in rice will 

effectively alleviate competition from volunteer soybean (Dillon et al., 2006; Bond and 

Walker, 2009). 

Given the low potential for soybean to volunteer in subsequent crops, the availability of 

multiple herbicidal and cultivation methods for controlling volunteers, as well as the 

demonstrated lack of difference in germination of MON 87708 compared to conventional 

soybean (see Section VII.C.1), the introduction of MON 87708 into the Roundup Ready 

soybean system is not expected to impact the management of soybean volunteer plants. 
Monsanto Company 10-SY-210U 240 of 721 

Table VIII-28. Ratings for Postemergence Control of Volunteer Soybean in Labeled 

Rotational Crops1 

Product 

Rate 

(Product/Acre) 

Soybean 

V2 – V3 

Soybean 

V4- V6 

 

Corn2 

AAtrex 4L (atrazine) 0.38 qts E P 

0.50 qts E F 

Hornet WDG (flumetsulam/clopyralid) 1 – 2 oz E F-G 

Widematch (clopyralid/fluroxypyr) 0.25 pt E G 

Sorghum2,4 

AAtrex 4L (atrazine) 0.38 qts E P 

0.50 qts E F 

Permit (halosulfuron) 2/3 oz E E 

Buctril® (bromoxynil) 1 pt 

Wheat, Barley & Oats2 

Buctril (bromoxynil) 1 pt E E 

Widematch (clopyralid/fluroxypyr) 0.25 pt E G 

Cotton3 

Envoke® (trifloxysulfuron) 0.1 oz E E 

Rice4 

Grandstand® CA (triclopyr) 0.5 pint E E 

Regiment® (bispyribac) 0.4 oz E E 

Grasp® SC (penoxsulam) 2 oz E E 

Permit (halosulfuron) 2/3 oz E E 

 

NA denotes “not applicable.” 

1

Weed control ratings: E = Excellent (90 to 99% control), G = Good (80 to 90% control), F = Fair 

(65 to 80 control), and P = Poor (40 to 65% control). 

2

Source is Zollinger (2009). 

3

Source is York et al. (2005). 

4

Sources are Dillon et al. (2006); Bond and Walker (2009). 
Monsanto Company 10-SY-210U 241 of 721 

VIII.K. Weed Resistance to Dicamba Herbicide 

The risk of weeds developing resistance and the potential impact of resistance on the 

usefulness of an herbicide vary greatly across different herbicide modes-of-action and is 

dependent on a combination of different factors. Monsanto considers product 

stewardship to be a fundamental component of customer service and business practices, 

and invests considerably in research to understand the proper uses and stewardship of our 

herbicide-tolerant soybean systems. This research includes an evaluation of the factors 

that can contribute to the development of weed resistance. Detailed information 

regarding dicamba stewardship is presented in Appendix K.

VIII.L. Stewardship of MON 87708 

Monsanto Company develops effective products and technologies and is committed to 

assuring that its products and technologies are safe and environmentally 

responsible. Monsanto demonstrates this commitment by implementing product 

stewardship processes throughout the lifecycle of a product and by participation in the 

Excellence Through StewardshipSM (ETS) Program 8 . These policies and practices 

include rigorous field compliance and quality management systems and verification 

through auditing. Monsanto’s Stewardship Principles are also articulated in Technology 

Use Guides9 that are distributed annually to growers who utilize Monsanto branded traits. 

Growers who purchase seeds containing Monsanto’s proprietary biotechnology traits are 

contractually required to comply with stewardship measures outlined in the Technology 

Use Guide. 

As an integral action of fulfilling our stewardship commitment, Monsanto will seek 

biotechnology regulatory approvals for MON 87708 in all key soybean import countries 

with a functioning regulatory system to assure global compliance and support the flow of 

international trade. These actions will be consistent with the Biotechnology Industry 

Organization (BIO) Policy on Product Launch.10 Monsanto continues to monitor other 

countries that are key importers of soybean from the U.S. for the development of formal 

biotechnology approval processes. If new functioning regulatory processes are 

developed, Monsanto will make appropriate and timely regulatory submissions. 

Monsanto also commits to best industry practices on seed quality assurance and control 

to ensure the purity and integrity of MON 87708 seed. As with all of Monsanto’s 

products, before commercializing MON 87708 in any country, a MON 87708 detection 

method will be made available to soybean producers, processors, and buyers. 

Dicamba is a selective herbicide registered with the U.S. EPA for the preemergent and 

postemergent control of certain broadleaf weeds in agriculture. Dicamba has a long

 

8

 Excellence Through Stewardship Program can be found at: 

http://www.excellencethroughstewardship.org/. 9

 Monsanto Technology Use Guides can be found 

at:http://www.monsanto.com/SiteCollectionDocuments/Technology-Use-Guide.pdf. 10 BIO’s Product Launch guidelines can be found at: 

http://www.excellencethroughstewardship.org/LinkClick.aspx?fileticket=ppgyTABguQs%3d&tabid=84. 
Monsanto Company 10-SY-210U 242 of 721 

 history of effective use in U.S. crop production, including corn, soybean, cotton, 

sorghum, wheat, barley, oats, millet, pasture, rangeland, asparagus, sugarcane, turf, grass 

grown for seed, conservation reserve programs, and fallow croplands (as described in 

Section VIII.G). Although herbicide resistance may eventually occur in a weed species 

when an herbicide is widely used, research, education and best management practices can 

be utilized to potentially delay, contain and manage resistance. The addition of dicamba 

tolerance to the Roundup Ready soybean system will provide an efficient method for 

incorporation of an additional herbicide mode-of-action in the system, and reduce the 

potential for further resistance development to glyphosate and dicamba as well as other 

critical soybean herbicides. Current research conducted by Monsanto to define the 

optimum weed management systems indicate the following: 1) in the absence of 

glyphosate resistant populations, the recommendation will be to apply a soil active 

preemergence residual herbicide followed by an in-crop postemergence application of 

glyphosate plus dicamba to control weed escapes, and 2) in the presence of glyphosate 

resistant populations, the same system will be recommended with a potential second 

application of glyphosate plus dicamba if needed. In this latter case, the preemergence 

herbicide to be recommended will be one with activity against the targeted glyphosate 

resistant species. This will ensure more than one mode of action against the targeted 

species. These management systems will reduce the potential for further resistance 

development to glyphosate, dicamba and other critical soybean herbicides. In 

conservation tillage systems, a preplant application of glyphosate plus dicamba may be 

recommended in some situations in addition to the in-crop applications described above. 

This is not expected to increase selection pressure on either product since the preplant 

weed spectrum is generally different from the in-crop spectrum. 

Stewardship of dicamba to preserve its usefulness for growers is an important aspect of 

Monsanto’s stewardship commitment, as is discussed in Appendix K. Specifically, 

Monsanto will develop weed resistance management practices, and utilize multiple 

methods to distribute technical and stewardship information to growers, academics and 

grower advisors through a variety of communication tools. Monsanto’s Technology Use 

Guide (TUG) will set forth the requirements and best practices for the cultivation of 

MON 87708 including recommendations on weed resistance management practices. 

Growers purchasing products containing dicamba tolerance will be obligated to follow all 

practices outlined in the TUG as required in the Monsanto Technology Stewardship 

Agreement (MTSA). Furthermore, Monsanto is committed to actively evaluate herbicide 

performance and weed efficacy on a continuing basis, and develop additional mitigation 

plans as necessary to manage resistance development for glyphosate and dicamba. 

Dicamba tolerance integrated into the Roundup Ready soybean system will enable 

expanded use of dicamba herbicide in soybean production when and in the manner 

appropriate. Monsanto is seeking regulatory approvals with the U.S. EPA for the 

expanded application of dicamba herbicide, as a weed control tool, as well as establishing 

appropriate dicamba Maximum Residue Levels (MRLs) for key soybean import 

countries, including the EU, Canada, and Japan, and CODEX, to support importing 

countries that do not have an established regulatory system to set MRLs including China.

As with all U.S. EPA registered herbicides for agricultural use, it is possible that offsite 

movement during and/or following application can occur such that non-target plants may 
Monsanto Company 10-SY-210U 243 of 721 

be exposed to direct spray or to spray drift. Research has demonstrated that herbicide 

formulation, application equipment and application procedures can be optimized to 

significantly reduce spray drift potential in most circumstances (Jordan et al, 2009; 

STDF, 1997). 

Monsanto is addressing such issues in its application to U.S. EPA to amend Registration 

Number 524-582. Specifically, Monsanto seeks approval of a low volatility dicamba 

(DGA salt) formulation that will only be applied to MON 87708 using ground application 

equipment. In addition, Monsanto will consult with U.S. EPA to identify what additional 

measures, if any, are necessary to address any potential impact of dicamba offsite 

movement. To reinforce EPA’s label requirements, Monsanto will implement a robust 

stewardship program that will include a strong emphasis on grower and applicator 

training. Such training will, for example, teach growers and applicators that dicamba 

herbicide spray drift can be further reduced during ground application by using industry 

standard procedures for minimizing spray drift, e.g. making applications with coarse 

droplet size through appropriate nozzle selection, using lower spray pressure, applying at 

the lowest nozzle height that provides uniform coverage, and making applications when 

wind speeds are low and consistent in direction (SDTF, 1997). 

VIII.M. Impact of the Introduction of MON 87708 on Agricultural Practices 

Introduction of MON 87708 is expected to have no impact on current cultivation and 

management practices for soybean, with the exception of expanded dicamba application 

timings. Monsanto recommends the use of preemergent soil residual herbicides as part of 

our current weed control programs for soybean. Dicamba has been used in corn, soybean 

and small grain cropping systems since 1967. MON 87708 with its excellent crop 

tolerance to dicamba allows preemergence applications through crop emergence 

(cracking) and in-crop postemergence applications through the R1/R2 growth stage. 

MON 87708 will be combined with MON 89788 (Roundup Ready 2 Yield) utilizing 

traditional breeding techniques. Soybean containing both MON 87708 and MON 89788 

will allow the use of glyphosate and dicamba herbicides in an integrated weed 

management program to control a broad spectrum of grasses and broadleaf weed species, 

and sustain and compliment the benefits and value of the glyphosate use in the Roundup 

Ready soybean system. The addition of another herbicide with a different mode-ofaction for preemergence and in-crop postemergence control of broadleaf weeds in 

soybean that is compatible with glyphosate will help to preserve the growth in 

conservation tillage acres associated with Roundup Ready cropping systems. For an 

overview of the cumulative impacts on agricultural practices (weed control, tillage and 

crop rotation) from deregulation of MON 87708, see Appendix J. 

MON 87708 has been shown to be no different from conventional soybean in its 

agronomic, phenotypic, and compositional characteristics (refer to Sections VI, VII, and 

VIII), and has the same levels of susceptibility to insects and diseases as commercial 

soybean. Like other herbicide-tolerant soybean, such as Roundup Ready and Roundup 

Ready 2 Yield that have been cultivated and consumed in the U.S. since 1996, dicambatolerant soybean MON 87708 will improve the current agricultural practices for U.S. 

soybean growers by providing another preemergence and in-crop postemergence 
Monsanto Company 10-SY-210U 244 of 721 

herbicide mode-of-action for the control of hard-to-control and herbicide resistant 

broadleaf weeds, thereby improving the efficiency in the U.S. soybean production system 

to maximize or maintain soybean yield potential. 
Monsanto Company 10-SY-210U 245 of 721 

IX. ENVIRONMENTAL CONSEQUENCES 

IX.A. Introduction 

This section provides a brief review and assessment of the plant pest potential of 

MON 87708 and its impact on current agronomic practices. USDA-APHIS has 

responsibility, under the Plant Protection Act (7 U.S.C. § 7701-7772), to prevent the 

introduction and dissemination of plant pests into the U.S. APHIS regulation 

7 CFR § 340.6 provides that an applicant may petition APHIS to evaluate submitted data 

to determine that a particular regulated article does not present a plant pest risk and no 

longer should be regulated. If APHIS determines that the regulated article does not 

present a plant pest risk, the petition must be granted, thereby allowing unrestricted 

introduction of the article. 

The definition of “plant pest” in the Plant Protection Act (PPA) includes living organisms 

that could directly or indirectly injure, damage, or cause disease in any plant or plant 

product (7 U.S.C. § 7702[14]). 

The regulatory endpoint under the PPA for biotechnology-derived crop products is that 

deregulation of the regulated article is not likely to pose a plant pest risk. The approach 

used to assess the plant pest potential of MON 87708 is a weight of the evidence 

approach based primarily on eight lines of evidence: 1) insertion of a single functional 

copy of the dmo expression cassette, 2) characterization of MON 87708 DMO expressed 

in MON 87708, 3) safety of MON 87708 DMO, 4) compositional equivalence of 

harvested MON 87708 seed and forage to conventional soybean, 5) phenotypic, 

agronomic, and environmental interaction characteristics demonstrating no increased 

plant pest potential compared to conventional soybean, 6) negligible risk to NTOs and 

threatened or endangered species, 7) familiarity with soybean as a cultivated crop and the 

inherently low plant pest potential of soybean, and (8) no greater likelihood to impact 

agronomic practices, including land use, cultivation practices, or the management of 

weeds (other than the intended benefit of dicamba weed control), diseases, and insects 

than conventional soybean. 

Using the assessment above, the data and analysis presented in this petition lead to a 

conclusion that MON 87708 is unlikely to be a plant pest and, therefore, should no longer 

be subject to regulation under 7 CFR § 340. 

Under current regulations, APHIS’s noxious weed authorities are not at issue unless a 

petition is filed under7 C.F.R. Part 360. Although no such issues are posed by a Part 340 

petition for non-regulated status, the data presented in this petition nevertheless 

demonstrate that MON 87708 is neither noxious nor a weed and could not be designated 

as such even in the appropriate procedural context. 
Monsanto Company 10-SY-210U 246 of 721 

IX.B. Plant Pest Assessment of the MON 87708 Insert and Expressed Protein 

IX.B.1. Characteristics of the Genetic Insert and the Expressed Protein 

This section summarizes the details of the genetic insert, characteristics of the genetic 

modification, and safety and expression of the MON 87708 DMO used to evaluate the 

food, feed, and environmental safety of MON 87708. 

IX.B.1.1. Genetic Insert 

As described in more detail in Section III, MON 87708 was produced by Agrobacterium 

tumefaciens-mediated transformation of soybean with PV-GMHT4355, which is a binary 

vector containing two T-DNAs (Figure III-1). T-DNA I contains the dmo expression 

cassette and T-DNA II contains the cp4 epsps expression cassette. During plant 

transformation, both T-DNAs were inserted into the soybean genome, with the cp4 epsps

expression cassette functioning as a selectable marker. Subsequently, conventional selfpollinated breeding methods and segregation were used to isolate those plants that 

contain the dmo expression cassette (T-DNA I) and do not contain the cp4 epsps

expression cassette (T-DNA II), resulting in the production of marker-free MON 87708. 

T-DNA I contains the dmo coding sequence under the regulation of the PC1SV promoter, 

TEV leader, the RbcS targeting sequence, and the E9 3′ non-translated region. In addition, 

T-DNA I contains Left and Right Border sequences. The promoter, leader, targeting, and 

border sequences of T-DNA I are not known to cause plant disease. Furthermore, these 

sequences are well characterized, are noncoding regions, and will not cause MON 87708 

to promote plant disease. 

Molecular analyses demonstrated that MON 87708 contains one copy of the inserted 

T-DNA I at a single integration locus. No T-DNA II or backbone sequences from 

PV-GMHT4355 were detected in the genome of MON 87708. Additionally, the data 

confirmed the organization and sequence of the insert and demonstrated the stability of 

the insert over several generations. These data demonstrated that there are no unintended 

changes in the MON 87708 genome as a result of the insertion of the dmo expression 

cassette, and support the overall conclusion that MON 87708 is unlikely to be a plant pest. 

IX.B.1.2. Protein Safety 

MON 87708 exhibits tolerance to the herbicide dicamba through the insertion of a 

demethylase gene from Stenotrophomonas maltophilia that encodes for DMO. S. 

maltophilia is an environmentally ubiquitous bacterium. Infections caused by S. 

maltophilia are extremely uncommon and it can be found in healthy individuals without 

any harmful effects. DMO is a Rieske type non-heme iron oxygenase that catalyzes the 

demethylation of dicamba to the non-herbicidal compound DCSA (Section V.A). DMO 

is specific for dicamba (Dumitru et al., 2009). 

DMO is a trimer comprised of three DMO monomers (Chakraborty et al., 2005). 

MON 87708 DMO is comprised of two forms of the DMO protein; denoted as DMO and 

DMO+27. Since DMO can be formed by DMO, DMO+27, or a combination of both, all 

forms of the trimer are referred to as MON 87708 DMO (Section V). MON 87708 DMO 
Monsanto Company 10-SY-210U 247 of 721 

was fully characterized confirming the N-terminal and internal sequence of both protein 

forms, neither of which were glycosylated. MON 87708 DMO is structurally and 

functionally similar to oxygenase homologs present in bacteria and plants, where a 

history of safe use is established (Section V.E.2). 

The MON 87708 DMO levels in MON 87708 tissue samples ranged from 6.1 to 69 μg/g 

dwt in root, forage, harvested seed, and overseason leaf, and represent a low percentage 

of the total protein in soybean (Section V.C). MON 87708 DMO was rapidly digested in 

simulated gastric fluids, lacked homology with known toxins and allergens, and lacked 

acute toxicity in a mouse oral gavage study, which taken together, support the conclusion 

that there is no meaningful risk to human or animal health from dietary exposure to 

MON 87708 DMO. 

The low level of MON 87708 DMO expressed in MON 87708 tissues taken together with 

the safety of MON 87708 DMO support the conclusion that food and feed products 

containing or derived from MON 87708 are as safe for human and animal consumption 

as soybean currently on the market. Therefore, unintended environmental effects are not 

anticipated from dietary exposure to MON 87708 DMO, and support the overall 

conclusion that MON 87708 is unlikely to be a plant pest. 

IX.B.2. Compositional Characteristics 

Detailed compositional analyses in accordance with OECD guidelines were conducted to 

assess whether levels of key nutrients and anti-nutrients in MON 87708, both 

dicamba-treated and untreated, were comparable to levels present in the near isogenic 

conventional soybean control A3525 and several commercial reference varieties. Seed 

and forage were harvested from five individual sites in which MON 87708, the 

conventional control, and a range of commercial reference varieties were grown 

concurrently in the same field trial. The commercial reference varieties were used to 

establish a range of natural variability for the key nutrients and anti-nutrients in 

commercial soybean varieties that have a history of safe consumption. 

The combined-site analysis was conducted to determine statistically significant 

differences (5% level of significance) between MON 87708, both dicamba-treated and 

untreated, and the conventional control A3525. The results from the combined-site data 

were reviewed using considerations relevant to food and feed safety and nutritional 

quality. These considerations included the relative magnitudes of the difference in the 

mean values of nutrient and anti-nutrient components of MON 87708 and the 

conventional control, whether the MON 87708 component mean value was within the 

range of natural variability of that component as represented by the 99% tolerance 

interval of the commercial reference varieties grown concurrently in the same field trial, 

and analyses of the reproducibility of the statistically significant combined-site 

component differences at individual sites. 

Assessment of the analytical results confirmed that the differences observed in the 

combined-site analysis were not meaningful to food and feed safety or the nutritional 

quality of MON 87708 soybeans. In addition, the levels of assessed components in
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 MON 87708 were compositionally equivalent to the conventional control and within the 

range of variability of commercial soybeans that were grown concurrently in the same 

field trial. These results support the overall conclusion that MON 87708 is unlikely to be 

a plant pest. 

IX.B 3. Phenotypic and Agronomic and Environmental Interaction Characteristics 

An extensive set of comparative plant characterization data were used to assess whether 

the introduction of the dicamba tolerance trait and the associated treatment of dicamba 

herbicide altered the plant pest potential of MON 87708 compared to the conventional 

control (Section VII). Phenotypic, agronomic, and environmental interaction 

characteristics of MON 87708 were evaluated and compared to those of the conventional 

control (Section VII.B). As described below, these assessments included: seed dormancy 

and germination characteristics; plant growth and development characteristics; 

observations for abiotic stress response, disease damage, arthropod-related damage; 

pollen characteristics; and arthropod abundance; and plant-symbiont interaction 

characteristics. Results from the phenotypic, agronomic, and environmental interaction 

assessments demonstrated that MON 87708 does not possess weedy characteristics, or 

increased susceptibility or tolerance to specific diseases, insects, or abiotic stressors, or 

altered symbiont interactions compared to conventional soybean, and the dicamba 

treatment does not alter the plant pest potential of MON 87708. Taken together, the 

results of the analysis support a determination that MON 87708 is no more likely to pose 

a plant pest risk or have a biologically meaningful change in environmental impact than 

conventional soybean. 

IX.B.3.1. Seed Dormancy and Germination 

Seed dormancy and germination characterization demonstrated that MON 87708 seed 

had germination characteristics similar to those of the conventional control (Section 

VII.C.1). In particular, the lack of hard seed, a well-accepted characteristic often 

associated with plants that are weeds, supports a conclusion of no increased weediness or 

plant pest potential of MON 87708 compared to conventional soybean. 

IX.B.3.2. Plant Growth and Development 

Evaluations of plant growth and development characteristics in the field are useful for 

assessing potential weediness characteristics such as lodging and pod shattering (Section 

VII.C.2.3). Of the growth and development characteristics assessed between 

MON 87708 and the conventional control, no statistically significant differences were 

detected (5% level of significance) with the exception of plant height and 100 seed 

weight in a combined-site analysis of the data. The differences in these parameters were 

relatively small in magnitude, and the mean values of MON 87708 were within the range 

of the commercial reference varieties for these characteristics. Thus, the differences in 

these parameters are not considered to be biologically meaningful in terms of increased 

weediness or plant pest potential of MON 87708 compared to conventional soybean. 
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IX.B.3.3. Response to Abiotic Stressors 

No biologically meaningful differences were observed during comparative field 

observations between MON 87708 and the conventional control and their response to 

abiotic stressors, such as drought, heat stress, high winds, nutrient deficiency, etc. 

(Section VII.C.2.4). The lack of significant biologically meaningful differences in the 

MON 87708 response to abiotic stress support the conclusion that the introduction of the 

dicamba tolerance trait is unlikely to result in increased weediness or plant pest potential 

compared to conventional soybean. 

IX.B.3.4. Pollen Morphology and Viability 

Evaluations of pollen morphology and viability from field-grown plants provide 

information useful in a plant pest assessment as it relates to the potential for gene flow to, 

and introgression of, the biotechnology-derived trait into other soybean varieties and wild 

relatives (Section VII.C.3). Pollen morphology and viability evaluations demonstrated 

no statistically significant differences between MON 87708 and the conventional control. 

Taken together, these comparative assessments indicate that MON 87708 is not likely to 

have increased weediness or plant pest potential compared to conventional soybean. 

IX.B.3.5. Interactions with Non-Target Organisms Including Threatened and 

Endangered Species 

Evaluation of MON 87708 for potential adverse impacts on NTOs is a component of the 

plant pest risk assessment. Since MON 87708 is a product with no pesticidal activity, all 

organisms that interact with MON 87708 are considered to be NTOs. In a 2008 U.S. 

phenotypic and agronomic assessment, observational data on environmental interactions 

were collected for MON 87708 and the conventional control. In addition, multiple 

commercial reference varieties were included in the analysis to establish a range of 

natural variability for each characteristic. The environmental interactions assessment 

(Section VII.C.2.4) included data collected on plant-arthropod and plant-disease 

interactions. The results of this assessment indicated that the presence of the dicamba 

tolerance trait did not alter plant-arthropod interactions, including beneficial arthropods 

and arthropod pests, nor did it alter disease susceptibility of MON 87708 compared to 

conventional soybean. The lack of differences in disease damage, arthropod-related 

damage, and pest and beneficial arthropod abundance demonstrate that the introduction 

of the dicamba tolerance trait is unlikely to be biologically meaningful in terms of 

increased plant pest potential. 

In the field, soybean forms a complex symbiotic relationship with members of the 

bacterial family Rhizobiaceae and Bradyrhizobiaceae. This symbiosis results in the 

formation of root nodules in which the bacteria reduce or fix atmospheric nitrogenproducing ammonia that can be used by the plant. MON 87708 was assessed for changes 

in the symbiotic relationship with B. japonicum relative to the conventional control by 

evaluating shoot total nitrogen, nodule number, and nodule, root, and shoot dry weights 

(Section VII.C.4). No statistically significant differences were detected between 

MON 87708 and the conventional control for the parameters measured, indicating no 
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impact on either the symbiotic relationship or the symbiotic nitrogen-fixing bacteria. 

These data support a conclusion of no change in plant pest potential and no expected 

impact to cultivation practices relative to nitrogen inputs for MON 87708 compared to 

conventional soybean. 

The potential for MON 87708 to harm NTOs was evaluated using a combination of 

biochemical information and experimental data. The biochemical information and 

experimental data included molecular characterization, the MON 87708 DMO safety 

assessments, the history of environmental exposure to mono-oxygenases (the class of 

enzymes to which DMO belongs), results from the environmental assessment described 

above, and the demonstration of compositional, agronomic and phenotypic equivalence to 

conventional soybean. Taken together, these data support the conclusion that 

MON 87708 is unlikely to adversely affect NTOs, or pose an additional risk to threatened 

and endangered species above those posed by the cultivation of conventional soybean. 

Furthermore, according to APHIS, the only listed threatened or endangered animal that 

occupies a habitat where it is likely to include soybean fields, and that might feed on 

soybean, is the federally endangered Delmarva Peninsula Fox Squirrel, (Sciurus niger 

cinereus), found in areas of the mid-Atlantic Eastern seaboard (USDA-APHIS, 2007). It 

is known to utilize certain agricultural lands readily, but its diet includes acorns; 

nuts/seeds of hickory, beech, walnut, and loblolly pine; buds and flowers of trees; fungi; 

insects; fruit; and an occasional bird egg (NatureServe, 2007). The safety of the 

MON 87708 DMO, the compositional, agronomic and phenotypic equivalence of 

MON 87708 to conventional soybean, and the diversity of the Fox Squirrel diet, support a 

conclusion that no biologically significant changes to the habitat or diet of the Delmarva 

Peninsula Fox Squirrel are expected. Consequently, the planting of MON 87708 is not 

expected to affect the Delmarva Peninsula Fox Squirrel. 

IX.B.3.6. Effects of dicamba reaction products 

MON 87708 DMO rapidly demethylates dicamba rendering it inactive, thereby 

conferring tolerance to dicamba in MON 87708. In dicamba-treated MON 87708 the 

demethylation of dicamba produces 3,6-dichlorosalicylic acid (DCSA), a known soybean, 

soil, and livestock metabolite whose safety has been evaluated by the Environmental 

Protection Agency (U.S. EPA), and formaldehyde. In the absence of a dicamba 

treatment on MON 87708, DCSA and formaldehyde would not be produced. DCSA is 

structurally similar to salicylic acid (SA). Numerous studies have reported on the stress 

defense activities of SA, although most studies have looked at the protective effects of 

exogenously applied SA (Janda et al, 2007). Formaldehyde has a potential linkage to 

apoptosis in plants (Szende and Tyihak, 2010), and formaldehyde concentrations in 

plants have been found to increase under certain stress conditions (Szabo et al, 2003). 

The National Toxicology Program (NTP) 12th Report on Carcinogens has reclassified 

formaldehyde as a known human carcinogen (USHHS-NTP, 2011). The relevant route of 

exposure for this health risk is from repeated inhalation at levels associated with indoor 

or occupational environments, which are generally higher than outdoor environments 

(USHHS-NTP, 2011). Formaldehyde is present in food and in the human body naturally, 
Monsanto Company 10-SY-210U 251 of 721 

and there is no evidence to suggest that dietary intake of formaldehyde is important 

(USHHS-NTP, 2011). 

A full discussion on DCSA, and the safety of this metabolite, is provided in Appendix M. 

DCSA as well as other dicamba metabolite products were measured in the residue study 

provided to the EPA to demonstrate that dicamba and dicamba metabolite residues are 

well below the current MRL set for dicamba in soybeans in the U.S. 

Formaldehyde is ubiquitous in the environment and present in plants and animals. 

Formaldehyde was not considered a relevant metabolite in the demethylation of dicamba 

by the EPA. According to the guidelines published by Office of Prevention, Pesticides 

and Toxic Substances, United States Environmental Protection Agency (US EPA OPPTS 

860.1300), the methoxy sidechain that is cleaved from dicamba to form formaldehyde 

would specifically not be chosen to be labeled in a metabolism study (U.S. EPA, 1996). 

This is because it is not metabolically stable and would not be considered a significant 

moiety as it would be readily metabolized and incorporated into the 1-carbon pool of the 

plant through known pathways. Therefore, formaldehyde was not measured in the 

residue study when dicamba was applied to MON 87708. 

Data from both dicamba-treated and non-treated MON 87708 compared to a conventional 

control are available from multiple sites across the U.S. where agronomic, phenotypic 

and observational environmental interaction data were collected. The results of this 

assessment demonstrate no biologically meaningful difference between MON 87708 

treated with dicamba, or MON 87708 not treated with dicamba compared to the 

conventional control, and support a conclusion that the formation of DCSA and 

formaldehyde does not alter the weedy characteristics, or increased susceptibility and 

tolerance to diseases, insects or abiotic stresses. Therefore, MON 87708, as cultivated, is 

no more likely to be a plant pest risk or have a biologically meaningful change in 

environmental impact than conventional soybean. 

Further, the metabolism of formaldehyde in plants is well understood, and there are a 

number of natural occurring sources of formaldehyde in plants. For example, it is 

produced during the process of photorespiration (Oliver, 1994) and during oxidative 

demethylation of DNA (Zhu, 2009), which supports why it is not considered a byproduct 

of interest as dicamba is demethylated. It is well known and understood that 

formaldehyde is rapidly metabolized in plants through two basic routes: 1) it can be 

incorporated into the one-carbon folate pool via spontaneous or enzyme-mediated formation of methylene tetrahydrofolate (Hanson and Roje, 2001); or 2) it can be oxidized to 

formate by a detoxification pathway that begins with spontaneous formation of the 

glutathione adduct S-hydroxymethylglutathione (Hanson and Roje, 2001). In each case, 

formaldehyde is further metabolized to carbon dioxide or entered into the 1C folate pool 

(Hanson and Roje, 2001; Giese et al., 1994). The maximum theoretical production of 

formaldehyde produced from dicamba-treated MON 87708 is estimated to be 16.7 and 

37.5 mg/kg11. This is well within the range of formaldehyde concentrations measured for 

 

11 Calculation based on an assumption that the entire 0.56 kg/ha (0.5 lb/acre a.e.) application of dicamba to 

MON 87708 soybean at the V3 growth stage is intercepted by the soybean plants, is instantaneously and 
Monsanto Company 10-SY-210U 252 of 721 

a variety of agricultural commodities – up to 60 mg/kg in fruits and vegetables (WHOIPCS, 1989). Plants have a large capacity to metabolize formaldehyde naturally 

produced from internal processes (A. Hanson (2011), C.V. Griffin, Sr. Eminent Scholar, 

Horticulture Department, University of Florida, Personal Communication), and any 

additional amount of formaldehyde that could be theoretically produced in the plant by 

dicamba treatment in MON 87708 would be metabolized very quickly. Additionally, as 

dicamba would not be instantaneously absorbed and metabolized, the incremental 

increase in formaldehyde over and above the levels already presumed to be present in the 

soybean plant would be small and transient. Further, since current literature supports that 

formaldehyde is only emitted from foliage under certain conditions (Nemecek-Marshall 

et al., 1995; Cojocariu et al., 2004; Cojocariu et al., 2005) and that emission rates are low 

(Nemecek-Marshall et al., 1995), little opportunity exists for formaldehyde to be released 

from MON 87708 after dicamba treatment. 

In addition to formaldehyde production in plants, plants and animals are constantly 

exposed to low levels of formaldehyde. Formaldehyde is already present in the 

environment and the atmosphere from a variety of biogenic (e.g. plant and animal) and 

anthropogenic (e.g. automotive or industrial emissions) sources. Additionally, 

formaldehyde degrades rapidly in environmental compartments (air, soil, and water). In 

water, formaldehyde dissipates through biodegradation to low levels in a few days 

(USHHS-ATSDR, 1999). Aerobic biodegradation half-lives are estimated to be 1-7 days 

for surface water and 2-14 days for ground water (US EPA, 2008). The half-life of 

formaldehyde in air is dependent on a number of factors (light intensity, temperature and 

location). Through reaction with hydroxyl radical, the half-life of formaldehyde in air 

varies from 7 to 70 hours (US EPA, 2008). The photolytic half-life of formaldehyde in 

air (e.g., in the presence of sunlight) is estimated to be 1.6-6 hours (US EPA, 2008; 

USHHS-ATSDR, 1999). The rapid degradation of formaldehyde in the environment 

combined with the understanding that formaldehyde is widely used by living organisms 

as a 1C source, support a conclusion that any environmental effects of formaldehyde, 

including effects on other plants and NTO’s, resulting from dicamba-treated MON 87708 

would be negligible. 

Humans are also constantly exposed to low levels of formaldehyde. Human exposure to 

formaldehyde is primarily due to indoor air exposures (USHHS-ATSDR, 1999). 

Formaldehyde is also found in a variety of consumer products such as cosmetics and 

paints, often as an antimicrobial agent, and is used extensively in urea-formaldehyde 

“slow-release” fertilizer formulations and adhesives (USHHS-ATSDR, 1999). Indoor 

formaldehyde air concentrations are generally significantly higher than outdoor air 

concentrations (USHHS-ATSDR, 1999) as a result of combustion (cooking, heating, 

tobacco use) and the emission of formaldehyde from a variety of construction materials 

(e.g., particle board, plywood or foam insulation) as well as permanent press fabrics 

(clothing or draperies) (US CPSC, 1997). Formaldehyde present in outdoor air results 

 completely absorbed, and then instantaneously metabolized by the DMO enzyme (Complete demethylation 

of 560 g (2.5 mol)/ ha dicamba would yield 2.5 mol/ha formaldehyde). Above ground biomass of V3 plants 

is estimated at 2 metric tons/ha, and results in 37.5 mg/kg formaldehyde in planta. For dicamba 

applications at R1 growth stage, the crop biomass is estimated to be 4.5 metric tons/ha, and level of 

formaldehyde produced in planta is 16.7 mg/kg. 
Monsanto Company 10-SY-210U 253 of 721 

from a number of sources, and levels of formaldehyde are generally higher in urban areas 

than in rural areas (WHO-IPCS, 1989). Direct contributions of formaldehyde to the 

atmosphere (i.e., those in the form of formaldehyde itself) from man-made sources are 

present, but are generally considered to be small relative to natural sources or indirect 

production of formaldehyde in the atmosphere (WHO, 2002). Formaldehyde is rapidly 

consumed in the atmosphere through direct photolysis or by oxidation with hydroxyl or 

nitrate radicals (USHHS-ATSDR, 1999). 

The National Toxicology Program (NTP) 12th Report on Carcinogens has reclassified 

formaldehyde as a known human carcinogen (USHHS-NTP, 2011). However, the 

relevant route of exposure for this health risk is from repeated inhalation of concentrated 

levels associated with indoor or occupational environments. As previously discussed, 

formaldehyde may only be released by plants in very small quantities and under certain 

conditions. Any incremental exposure to formaldehyde associated with the application of 

dicamba to MON 87708 would occur outdoors, would be minimal, and also transient in 

nature. Therefore human safety concerns of formaldehyde released from dicamba treated 

MON 87708 are considered to be negligible. USHHS-NTP (2011) has already stated that 

there is no evidence to suggest that dietary intake of formaldehyde is important. In 

addition, commodity soybean seed is not directly consumed by humans, and would be 

processed into food products, limiting potential exposure to humans to any formaldehyde 

in dicamba-treated MON 87708 seed. 

IX.C. Weediness Potential of MON 87708 

The commercial Glycine species in the U.S. (Glycine max L.) does not exhibit weedy 

characteristics and is not effective in invading established ecosystems. Soybean is not 

listed as a weed in the major weed references (Crockett, 1977; Holm et al., 1979; Holm et 

al., 1997), nor is it present on the lists of noxious weed species distributed by the federal 

government (7 CFR Part 360). Soybean does not possess any of the attributes commonly 

associated with weeds (Baker, 1965), such as long persistence of seed in the soil, the 

ability to disperse, invade, and become a dominant species in new or diverse landscapes 

or the ability to compete well with native vegetation. Due to the lack of dormancy, which 

is a trait that has been removed from soybean through commercial breeding, soybean 

seed can germinate quickly under adequate temperature and moisture conditions, and 

potentially grow as volunteer plants. However, volunteer plants likely would be killed by 

frost during autumn or winter of the year they were produced. If they did become 

established, volunteer plants would not compete well with the succeeding crop, and could 

be controlled readily by either mechanical or chemical means (Carpenter et al., 2002; 

Dillon et al., 2006; OECD, 2000; York et al., 2005; Zollinger, 2009). In addition, since 

wild populations of Glycine species are not known to exist in the U.S., the potential does 

not exist for MON 87708 to outcross to wild or weedy relatives and to alter their 

weediness potential. 

In comparative studies between MON 87708 and the conventional control, phenotypic, 

agronomic, and environmental interaction data were evaluated (Section VII) for changes 

that would impact the plant pest potential and, in particular, plant weediness potential. 
Monsanto Company 10-SY-210U 254 of 721 

Results of these evaluations show that there is no biologically meaningful difference 

between MON 87708 and the conventional control for characteristics potentially 

associated with weediness. Furthermore, comparative field observations between 

MON 87708 and its conventional control in their response to abiotic stressors, such as 

drought, heat stress, and high winds, indicated no biologically meaningful differences and, 

therefore, no increased weediness potential. Data on environmental interactions also 

indicate that MON 87708 does not confer any biologically meaningful increased 

susceptibility or tolerance to specific diseases or insect pests. Collectively, these findings 

support the conclusion that MON 87708 has no increased weediness potential compared 

to conventional soybean. 

Because MON 87708 has the same limited weediness potential as conventional soybean, 

it is similarly unlikely to survive as a volunteer plant. Under 7 C.F. R. Part 340, no 

“noxious weed” issues are reviewed by APHIS. Nevertheless, even if APHIS’s Part 360 

noxious weed authorities were at issue here, MON 87708 could pose no “noxious weed” 

risks under the Plant Protection Act because it would not be “noxious” as APHIS 

ordinarily interprets that term. APHIS has made clear that “noxious” weeds refer to 

weeds that are “likely to be aggressively invasive, have significant negative impacts,” and 

are “extremely difficult to manage or control once established.” 73 Fed. Reg. 60,008, 

60,013 (Oct. 9, 2008). Volunteer MON 87708—like volunteer conventional soybean—

would compete poorly with any succeeding crops and soon be killed, making it extremely 

unlikely to have any prolonged negative effects. Volunteer MON 87708 would also not 

be “extremely difficult to manage” because it can be controlled easily with numerous 

alternative herbicides and other mechanical means. (Carpenter et al., 2002; Dillon et al., 

2006; OECD, 2000; York et al., 2005; Zollinger, 2009). The conclusion that 

MON 87708 does not pose noxious weed risks is consistent with APHIS’s historical 

interpretations of the Plant Protection Act, as APHIS has never before considered a 

genetically engineered crop to be a noxious weed. 

IX.D. Potential for Pollen Mediated Gene Flow 

Gene introgression is a process whereby one or more genes successfully integrate into the 

genome of a recipient plant population. Introgression is affected by many factors, 

including the frequency of the initial pollination event, environmental factors, sexual 

compatibility of pollen donor and recipient plants, pollination biology, flowering 

phenology, hybrid stability and fertility, selection, and the ability to backcross repeatedly. 

Because gene introgression is a natural biological process, it does not constitute an 

environmental risk in and of itself (Sutherland and Poppy, 2005). Gene introgression 

must be considered in the context of the transgene(s) inserted into the biotechnologyderived plant, and the likelihood that the presence of the transgene(s) and their 

subsequent transfer to recipient plants will result in increased plant pest potential. The 

potential for gene introgression from MON 87708 is discussed below. 

The assessment for gene introgression from MON 87708 with other cultivated or wild 

relatives of soybean, discussed in detail below, indicates that MON 87708 is no more 

likely to become a weed than conventional soybean, and MON 87708 is expected to be 

similar to conventional soybean regarding its potential for and impacts from gene flow. 
Monsanto Company 10-SY-210U 255 of 721 

Soybean lacks sexually-compatible relatives in the U.S.; therefore, the only pollenmediated gene flow would be within cultivated soybean. 

IX.D.1. Hybridization with Cultivated Soybean 

Although soybean is largely a self-pollinated species, low levels of natural crosspollination can occur (Caviness, 1966; OECD, 2000; Ray et al., 2003; Yoshimura et al., 

2006). In studies with cultivated soybean, where conditions have been optimized to 

ensure close proximity and flowering synchrony, natural cross-pollination generally has 

been found to be very low. Most outcrossing occurred with surrounding plants, and 

cross-pollination frequencies varied depending on growing season and genotype. Insect 

activity does increase the outcrossing rate, but soybean generally is not a preferred plant 

for pollinators (Abrams et al, 1978; Erickson 1975; Jaycox 1970a, b). 

Numerous studies on soybean cross-pollination have been conducted, and the published 

results, with and without supplemental pollinators, are summarized in Table IX-1. Under 

natural conditions, cross-pollination among adjacent plants in a row or among plants in 

adjacent rows ranged from 0 to 6.3%. In experiments where supplemental pollinators 

(usually bees) were added to the experimental area, cross-pollination ranged from 0.5 to 

7.74% in adjacent plants or adjacent rows. However, cross-pollination does not occur at 

these levels over long distances. Cross-pollination rates decrease to less than 1.5% 

beyond one meter from the pollen source, and rapidly decrease with greater distances 

from the source. The following cross-pollination rates at extended distances have been 

reported: 0.05% at 5.4 meters (Ray et al., 2003), 0% at 6.5 meters (Abud et al., 2003), 

0% at 10.5 m (Yoshimura et al., 2006), and 0.004% at 13.7 meters of separation 

(Caviness, 1966). 

The potential for cross-pollination in soybean is limited. This is recognized in certified 

seed regulations for foundation seed in the U.S., which permit any distance between 

different soybean cultivars in the field as long as the distance is adequate to prevent 

mechanical mixing (USDA-APHIS, 2006). 

The consequence of introgression of the dicamba tolerance trait from MON 87708 into 

other soybean is negligible since soybean gene flow is naturally low; therefore the 

dicamba tolerance trait confers no increased plant pest potential to cultivated soybean. 

IX.D.2. Hybridization with Wild Annual Species within Subgenus Soja

The subgenus Soja includes the cultivated soybean Glycine max and the wild annual 

species Glycine soja. Glycine soja is found in China, Taiwan, Japan, Korea, and Russia 

and can hybridize naturally with the cultivated soybean, G. max (Hymowitz, 2004; Lu, 

2004). Hybridization between female G. soja and male G. max was less successful than 

hybridization in the opposite direction (Dorokhov et al., 2004), where frequency of 

spontaneous cross pollination in reciprocal combinations of G. max and G. soja varied 

from 0.73 (♀ G. soja × ♂ G. max) to 12.8% (♀ G. max × ♂ G. soja). Species 

relationships in the subgenus soja indicated that F1 hybrids of G. max and G. soja carry 

similar genomes and are fertile (Singh and Hymowitz, 1989). Abe et al. (1999) note that 
Monsanto Company 10-SY-210U 256 of 721 

“natural hybrids between G. max and G. soja are rare and hybrid swarms involving both 

species have never been reported.” This is also supported by work from Kuroda et al. 

(2008) in which molecular markers were used and no gene flow from G. max to G. soja

was detected. Many barriers to natural hybridization exist between soybean and wild 

relatives, including the highly selfing nature of both plants, required proximity of wild 

soybean to cultivated soybean, synchrony of flowering, and presence of pollinators. As 

such, it is highly unlikely that naturally occurring, pollen-mediated gene flow and 

transgene introgression into wild soybean relatives from incidentally released 

biotechnology-derived soybean will occur at any meaningful frequency. 

The subgenus Soja also contains an unofficial species, G. gracilis (Hymowitz, 2004). 

Glycine gracilis is known only from Northeast China, and is considered to be a weedy or 

semi-wild form of G. max, with some phenotypic characteristics intermediate to those of 

G. max and G. soja. Glycine gracilis may be a hybrid between G. soja and G. max

(Hymowitz, 1970; Lu, 2004). Interspecific fertile hybrids formed by intentional crosses 

between G. max and G. soja and between G. max and G. gracilis have been easily 

obtained (Dorokhov et al., 2004; Singh and Hymowitz, 1989). Although hybridization 

between G. max and members of the subgenus Soja can take place, G. soja is not found in 

North or South America, and it is highly unlikely that gene transfer will occur. 

IX.D.3. Hybridization with the Wild Perennial Species of Glycine Subgenus 

Wild perennial species of the Glycine subgenus occur in Australia; West, Central and 

South Pacific Islands; China; Papua New Guinea; Philippines; and Taiwan (Hymowitz et 

al., 1992). Therefore, the only opportunities for inter-subgeneric hybridization would 

occur in areas where those species are endemic. Nonetheless, the likelihood of 

interspecific hybridization between G. max and the wild perennial Glycine species is 

extremely low because they are genomically dissimilar (Hymowitz, 2004; Lu, 2004) and 

pod abortion is common. From time to time, immature seeds of the crosses have been 

germinated aseptically in vitro, but the resulting F1 hybrids are slow-growing, 

morphologically weak, and completely sterile. Their sterility is due to poor chromosome 

pairing. Furthermore, species distantly related usually produce nonviable F1 seeds that 

either have premature death of the germinating seedlings or suffer from seedling and 

vegetative lethality (Kollipara et al., 1993). In North and South America, it is not 

possible for gene transfer to occur between cultivated soybean and wild perennial species 

of Glycine subgenera because these wild species do not exist in these regions. 

IX.D.4. Transfer of Genetic Information to Species with Which Soybean Cannot 

Interbreed (Horizontal Gene Flow) 

Monsanto is unaware of any reports regarding the unaided transfer of genetic material 

from soybean species to other sexually-incompatible plant species. The likelihood for 

horizontal gene flow to occur is exceedingly small. Therefore, potential ecological risk 

associated with horizontal gene flow from MON 87708 due to the presence of the 

dicamba tolerance trait is not expected. The consequence of horizontal gene flow of the 

dicamba tolerance trait into other plants that are sexually-incompatible is negligible since, 

as data presented in this petition confirm, the gene and trait confer no increased plant pest 
Monsanto Company 10-SY-210U 257 of 721 

potential to soybean. Thus in the highly unlikely event that horizontal gene transfer were 

to occur, the presence of the dicamba tolerance trait would not be expected to increase 

pest potential in the recipient species. 

IX.E. Potential Impact on Soybean Agronomic Practices 

An assessment of current soybean agronomic practices was conducted to determine 

whether the cultivation of MON 87708 has the potential to impact current soybean and 

weed management practices (Section VIII). Soybean fields are typically highly managed 

agricultural areas that are dedicated to crop production. MON 87708 is likely to be used 

in common rotations on land previously used for agricultural purposes. Certified seed 

production will continue to use well-established industry practices to deliver high quality 

seed containing MON 87708 to growers. Cultivation of MON 87708 is not expected to 

differ from typical soybean cultivation, with the exception of an expanded window of 

dicamba applications. Due to the excellent crop safety of MON 87708 to dicamba, 

growers will have a new herbicide mode-of-action for in-crop control of glyphosate’s 

hard-to-control and resistant broadleaf weeds that are present in U.S. soybean production. 

As a result of cultivation of MON 87708 integrated into the Roundup Ready soybean 

system, the number of dicamba-treated soybean acres is expected to double from historic 

peak levels. Additionally, due to the expanded timing of in-crop applications to soybean, 

dicamba treatments will be later in the growing season than most current labeled dicamba 

uses. These later applications of dicamba to MON 87708 are not expected to impact 

dicamba-sensitive crops from drift (spray or volatility) because, in its application to U.S. 

EPA, Monsanto is registering the use on MON 87708 on a low volatility dicamba (DGA 

salt) formulation, not allowing aerial application on MON 87708, and will consult with 

U.S. EPA to develop any necessary additional measures to protect against offsite impacts. 

MON 87708 is similar to conventional soybean in its agronomic, phenotypic, ecological, 

and compositional characteristics and has levels of resistance to insects and diseases 

comparable to conventional soybean. Therefore, no significant impacts on current 

cultivation and management practices for soybean are expected following the 

introduction of MON 87708. Based on this assessment, the introduction of MON 87708 

will not impact current U.S. soybean cultivation practices or weed management practices, 

other than intended weed control benefits. 

IX.F. Summary of Plant Pest Assessments 

Plant pests are defined in the Plant Protection Act as certain living organisms that can 

directly or indirectly injure, cause damage to, or cause disease to any plant or plant 

product (7 U.S.C. § 7702[14]). Characterization data presented in Sections III through 

VII of this petition confirm that although MON 87708 contains the dicamba-tolerant trait, 

it is not different from conventional soybean in terms of pest potential in its phenotypic, 

agronomic, and environmental interaction characteristics. Monsanto is not aware of any 

study results or observations associated with MON 87708 that would suggest an 

increased plant pest risk would result from its introduction. 
Monsanto Company 10-SY-210U 258 of 721 

The plant pest assessment was based on multiple lines of evidence developed from a 

detailed characterization of MON 87708 compared to conventional soybean, followed by 

a risk assessment on detected differences. The risk assessment considered various factors 

including: 1) insertion of a single functional copy of the dmo expression cassette, 2) 

characterization of the MON 87708 DMO, 3) safety of the MON 87708 DMO, 4) 

compositional equivalence of harvested MON 87708 soybean seed and forage as 

compared to conventional soybean, 5) phenotypic, agronomic, and environmental 

interaction characteristics demonstrating no increased plant pest potential compared to 

conventional soybean, 6) negligible risk to NTOs and threatened or endangered species, 

7) familiarity with soybean as a cultivated crop and the inherently low plant pest potential 

of soybean, and 8) no greater likelihood to impact agronomic practices, including land 

use, cultivation practices, or the management of weeds, diseases, and insects than 

conventional soybean, with the exception of the expanded window of dicamba 

application. 

Based on the data and information presented in this petition, it is concluded that, like 

conventional soybean and currently deregulated biotechnology-derived soybean, 

MON 87708 is highly unlikely to be a plant pest. Therefore, Monsanto Company 

requests a determination from APHIS that MON 87708 and any progeny derived from 

crosses between MON 87708 and other commercial soybean be granted nonregulated 

status under 7 CFR Part 340. 
Monsanto Company 10-SY-210U 259 of 721 

Table IX-1. Summary of Published Literature on Soybean Cross Pollination 

Distance from 

Pollen Source 

(meters) 

CrossPollination (%) Comments Reference 

0.3 0.04 (estimated 

per pod) 

Interspaced plants within a row. Experiment 

conducted in a single year. Single male and female 

parental varieties. Percent outcrossing calculated 

per pod rather than per seed. 

(Woodworth, 

1922) 

0.8 0.07 to 0.18 Adjacent rows. Experiment conducted over two 

years. Several male and female parental varieties. 

(Garber and 

Odland, 1926) 

0.1 0.38 to 2.43 Adjacent plants within a row. Experiment 

conducted in a single year. Several male and 

female parental varieties. 

(Cutler, 1934) 

0.1 0.2 to 1.2 Adjacent plants within a row. Experiment 

conducted in single year at two locations. Several 

male and female parental varieties. 

(Weber and 

Hanson, 1961) 

0.9 

2.7–4.6 

6.4–8.2 

10–15.5 

0.03 to 0.44 

0.007 to 0.06 

0 to 0.02 

0 to 0.01 

Frequency by distance was investigated. 

Experiment conducted over three years. Single 

male and female parental varieties. 

(Caviness, 

1966) 

0.8 m 0.3 to 3.62 Various arrangements within and among adjacent 

rows. Experiment conducted over three years. 

Several male and female parental varieties. 

(Beard and 

Knowles, 1971) 

One row 

(undefined) 

1.15 to 7.74 Bee pollination of single-row, small-plots of pollen 

receptor surrounded by large fields (several acres) 

of pollen donor soybean. Soybean is not a preferred 

flower for alfalfa leafcutting bees. 

(Abrams et al., 

1978) 

0.1–0.6 0.5 to 1.03 

(depending on 

planting design) 

Bee pollination of soybean grown in various spatial 

arrangements. Experiment conducted over four 

years. Several soybean cultivars. 

(Chiang and 

Kiang, 1987) 

1.0 0.09 to 1.63 Adjacent rows. Experiment conducted over two 

years. Several male and female parental varieties. 

(Ahrent and 

Caviness, 1994) 

0.5 

1.0 

6.5 

0.44 to 0.45 

0.04 to 0.14 

none detected 

Frequency by distance was investigated. 

Experiment conducted in a single year. 

Single male and female parental varieties. 

(Abud et al., 

2003) 

0.9 

5.4 

0.29 to 0.41 

0.03 to 0.05 

Frequency by distance was investigated. 

Experiment conducted in a single year. Single male 

and female parental varieties. 

(Ray et al., 

2003) 

0.15 0.65 to 6.32 

(avg. 1.8) 

Interspaced plants within a row. Experiment 

conducted in a single year. Single male and female 

parental varieties. 

(Ray et al., 

2003) 

0.7 

1.4 

2.1 

2.8 

3.5 

7.0 

10.5 

0 to 0.19 

0 to 0.04 

0 to 0.05 

0 to 0.08 

0 to 0.04 

0 to 0.04 

0 

Interspaced plants within a row arranged in small 

plots. Experiment conducted in a four year period. 

Single male and two female parental varieties. 

(Yoshimura et 

al., 2006) 
Monsanto Company 10-SY-210U 260 of 721 

X. ADVERSE CONSEQUENCES OF INTRODUCTION 

Monsanto knows of no results or observations associated with MON 87708 or the 

MON 87708 DMO indicating that there would be an adverse environmental consequence 

from the introduction of MON 87708. MON 87708 contains DMO that renders the 

soybean plant tolerant to the herbicide dicamba. As demonstrated by field results and 

laboratory tests, the only phenotypic difference between MON 87708 and conventional 

soybean is tolerance to dicamba. 

The data and information presented in this petition demonstrate that MON 87708 is 

unlikely to pose an increased plant pest risk or to have an adverse environmental 

consequence compared to conventional soybean. This conclusion is based on multiple 

lines of evidence developed from a detailed characterization of the product compared to 

conventional soybean, followed by risk assessment on detected differences: 1) 

characterization evaluations included molecular analyses, which confirmed the insertion 

of a single functional copy of the dmo expression cassette at a single locus within the 

soybean genome; 2) measurement of the MON 87708 DMO levels in various soybean 

tissues; 3) characterization of the MON 87708 DMO confirming it is not novel and is 

structurally and functionally similar to oxygenase homologs widely present in bacteria 

and plants, where a history of safe use is established; and 4) extensive characterization of 

the plant phenotype, including compositional analysis of key nutrients and antinutrients, 

and environmental interactions. Therefore, based on the lack of increased pest potential 

or adverse environmental consequences compared to conventional soybean, the risks for 

humans, animals, and other NTOs from MON 87708 are negligible under the conditions 

of use. Additionally the introduction of MON 87708 will not adversely impact 

cultivation practices or the management of weeds, diseases, and insects in soybean 

production systems, other than the use of dicamba postemergence in soybean. 

Successful integration of MON 87708 into the Roundup Ready soybean system will 

provide growers with an opportunity for an efficient, effective weed management system 

for the management of glyphosate’s hard-to-control and resistant broadleaf weeds; 

provide an easy system for inclusion of a second herbicide mode-of-action in soybean 

production practices as recommended by weed science experts to manage weed 

resistance development; and continue to provide soybean growers with effective weed 

control systems necessary for production yields to meet the growing needs of the food, 

feed and industrial markets. 
Monsanto Company 10-SY-210U 261 of 721 

REFERENCES 

Abe, J., A. Hasegawa, H. Fukushi, T. Mikami, M. Ohara and Y. Shimamoto. 1999. 

Introgression between wild and cultivated soybeans of Japan revealed by RFLP analysis 

for chloroplast DNAs. Economic Botany 53:285-291. 

Abrams, R.I., C.R. Edwards and T. Harris. 1978. Yields and cross-pollination of 

soybeans as affected by honey bees and alfalfa leafcutting bees. American Bee Journal

118:555-558. 

Abud, S., P.I. Mello de Souza, C.T. Moreira, S.R.M. Andrade, A.V. Ulbrich, G.R. 

Vianna, E.L. Rech and F.J. Lima Aragão. 2003. Gene flow in transgenic soybean in the 

Cerrado region, Brazil. Pesquisa Agropecuária Brasileira 38:1229-1235. 

Adrian-Romero, M., G. Blunden, B.G. Carpenter and E. Tyihák. 1999. HPLC 

quantification of formaldehyde, as formaldemethone, in plants and plant-like organisms. 

Chromatographia 50:160-166. 

Agranova. 2010. Herbicide synopsis - A chronological listing of all herbicide, safener 

and PGR compounds to 1995. Agranova, Kent, United Kingdom. 

http://www.agranova.co.uk/herbhist.asp Accessed June 17, 2010. 

AgroTrak. 2009. Unpublished 2008 agricultural survey data. Gfk Kynetec marketing 

Research, Inc., Berkshire, United Kingdom. 

Ahrens, W.H. 1994. Dicamba. 3,6-dichloro-2-methoxybenzoic acid. Pages 91-94 in 

Herbicide Handbook. Seventh Edition. Weed Science Society of America, Champaign, 

Illinois. 

Ahrent, D.K. and C.E. Caviness. 1994. Natural cross-pollination of twelve soybean 

cultivars in Arkansas. Crop Science 34:376-378. 

Alexander, M.P. 1980. A versatile stain for pollen fungi, yeast and bacteria. Stain 

Technology 55:13-18. 

Al-Kaisi, M. 2001. Value of crop rotation in nitrogen management. Iowa State University, 

Ames, Iowa. http://www.ipm.iastate.edu/ipm/icm/2001/4-23-2001/valuen.html Accessed 

May 12, 2010. 

Al-Kaisi, M., M. Hanna and M. Tidman. 2003. Crop rotation considerations for 2004 

management season rotation. Iowa State University, Ames, Iowa. 

http://www.ipm.iastate.edu/ipm/icm/2003/12-15-2003/croprotation.html Accessed June 

19, 2006. 

Anderson, W.P. 1996. Weed ecology. Pages 27-38 in Weed Science: Principles and 

Applications. Third Edition. West Publishing Company, St. Paul, Minnesota. 
Monsanto Company 10-SY-210U 262 of 721 

AOSA. 2000. Tetrazolium testing handbook. Contribution no. 29. Association of Official 

Seed Analysts, Lincoln, Nebraska. 

AOSA. 2007. Rules for testing seeds. Association of Official Seed Analysis, Ithaca, New 

York. 

AOSCA. 2009. Seed certification handbook: Including genetic and crop standards, 

procedures, and AOSCA service programs. Association of Official Seed Certifying 

Agencies, Moline, Illinois. 

Appunu, C. and B. Dhar. 2006. Differential symbiotic response of phage-typed strains of 

Bradyrhizobium japonicum with soybean cultivars. Journal of Microbiology 44:363-368. 

Aref, S. and D.R. Pike. 1998. Midwest farmers’ perceptions of crop pest infestation. 

Agronomy Journal 90:819-825. 

ASA. 2009. Soybean exports. American Soybean Association, St. Louis, Missouri. 

http://www.soystats.com Accessed March 15, 2010. 

ASA. 2011. Soy stats 2011. American Soybean Association, St. Louis, Missouri. 

http://www.soystats.com/2011/Default-frames.htm [Accessed September 22, 2011]. 

Baker, H.G. 1965. Characteristics and modes of origin of weeds. Pages 147-172 in The 

Genetics of Colonizing Species. H.G. Baker and G.L. Stebbins (eds.). Academic Press, 

New York, New York. 

Barker, R.F., K.B. Idler, D.V. Thompson and J.D. Kemp. 1983. Nucleotide sequence of 

the T-DNA region from the Agrobacterium tumefaciens octopine Ti plasmid pTi15955. 

Plant Molecular Biology 2:335-350. 

Barry, G.F., G.M. Kishore, S.R. Padgette and W.C. Stallings. 1997. Glyphosate-tolerant 

5-enolpyruvylshikimate-3-phosphate synthases. Patent 5,633,435, U.S. Patent Office, 

Washington, D.C. 

Beard, B.H. and P.F. Knowles. 1971. Frequency of cross-pollination of soybeans after 

seed irradiation. Crop Science 11:489-492. 

Beckie, H.J. 2006. Herbicide-resistant weeds: Management tactics and practices. Weed 

Technology 20:793-814. 

Beckie, H.J. and X. Reboud. 2009. Selecting for weed resistance: Herbicide rotation and 

mixture. Weed Technology 23:363-370. 
Monsanto Company 10-SY-210U 263 of 721 

Behrens, M.R., N. Mutlu, S. Chakraborty, R. Dumitru, W.Z. Jiang, B.J. LaVallee, P.L. 

Herman, T.E. Clemente and D.P. Weeks. 2007. Dicamba resistance: Enlarging and 

preserving biotechnology-based weed management strategies. Science 316:1185-1188. 

Berg, G., P. Marten and G. Ballin. 1996. Stenotrophomonas maltophilia in the 

rhizosphere of oilseed rape - Occurrence, characterization and interaction with 

phytopathogenic fungi. Microbiological Research 151:19-27. 

Berg, G., N. Roskot and K. Smalla. 1999. Genotypic and phenotypic relationships 

between clinical and environmental isolates of Stenotrophomonas maltophilia. Journal of 

Clinical Microbiology 37:3594-3600. 

Berg, G., N. Roskot, A. Steidle, L. Eberl, A. Zock and K. Smalla. 2002. Plant-dependent 

genotypic and phenotypic diversity of antagonistic rhizobacteria isolated from different 

Verticillium host plants. Applied and Environmental Microbiology 68:3328-3338. 

Berg, R.D. 1996. The indigenous gastrointestinal microflora. Trends in Microbiology

4:430-435. 

Berman, K.H., G.G. Harrigan, S.G. Riordan, M.A. Nemeth, C. Hanson, M. Smith, R. 

Sorbet, E. Zhu and W.P. Ridley. 2009. Compositions of seed, forage, and processed 

fractions from insect-protected soybean MON 87701 are equivalent to those of 

conventional soybean. Journal of Agricultural and Food Chemistry 57:11360-11369. 

Boerma, H.R. and J.E. Specht. 2004. World distribution and trade of soybean and 

managing inputs for peak production; soybean production in the U.S.A. Pages 3, 502, 507 

and 522 in Soybeans: Improvement, Production, and Uses. Boerma H.R. and J.E. Specht, 

eds. American Society of Agronomy, Inc., Crop Science Society of America, Inc., Soil 

Science Society of America, Inc., Madison, Wisconsin. 

Bond, J.A. and T.W. Walker. 2009. Control of volunteer glyphosate-resistant soybean in 

rice. Weed Technology 23:225-230. 

Bonny, S. 2008. Genetically modified glyphosate-tolerant soybean in the USA: Adoption 

factors, impacts and prospects. A review. Agronomy for Sustainable Development 28:21-

32. 

Bradford, K.J. 2006. Methods to maintain genetic purity of seed stocks. Publication 8189. 

University of California, Oakland, California. 

Breyton, C. 2000. The cytochrome b6f complex: Structural studies and comparison with 

the bc1 complex. Biochimica et Biophysica Acta 1459:467-474. 

Britz, S.J., D.F. Kremer and W.J. Kenworthy. 2008. Tocopherols in soybean seeds: 

Genetic variation and environmental effects in field-grown crops. Journal of the 

American Oil Chemists' Society 85:931-936. 
Monsanto Company 10-SY-210U 264 of 721 

Brookes, G. and P. Barfoot. 2009. GM crops: Global socio-economic and environmental 

impacts 1996-2007. PG Economics, Ltd, Dorchester, United Kingdom. 

www.pgeconomics.co.uk/pdf/2009globalimpactstudy.pdf Accessed May 12, 2010. 

Carpenter, J.E. and L.P. Gianessi. 2001. Agricultural biotechnology: Updated benefits 

estimates. National Center for Food and Agricultural Policy, Washington, D.C. 

Carpenter, J., A. Felsot, T. Goode, M. Hammig, D. Onstad, and S. Sankula. 2002. 

Comparative environmental impacts of biotechnology-derived and traditional soybean, 

corn, and cotton crops. Council for Agricultural Science and Technology, Ames, Iowa. 

CAST. 2007. Implications of gene flow in the scale-up and commercial use of 

biotechnology-derived crops: Economic and policy considerations. Issue paper 37. 

Council for Agricultural Science and Technology, Ames, Iowa. 

Caviness, C.E. 1966. Estimates of natural cross-pollination in Jackson soybeans in 

Arkansas. Crop Science 6:211-212. 

Chakraborty, S., M. Behrens, P.L. Herman, A.F. Arendsen, W.R. Hagen, D.L. Carlson, 

X.-Z. Wang and D.P. Weeks. 2005. A three-component dicamba O-demethylase from 

Pseudomonas maltophilia, strain DI-6: Purification and characterization. Archives of 

Biochemistry and Biophysics 437:20-28. 

Chiang, Y.C. and Y.T. Kiang. 1987. Geometric position of genotypes, honeybee foraging 

patterns and outcrossing in soybean. Botanical Bulletin of Academia Sinica 28:1-11. 

Chung, G. and R.J. Singh. 2008. Broadening the genetic base of soybean: A 

multidisciplinary approach. Critical Reviews in Plant Sciences 27:295-341. 

Clemente, T.E. and E.B. Cahoon. 2009. Soybean oil: Genetic approaches for 

modification of functionality and total content. Plant Physiology 151:1030-1040. 

Codex Alimentarius. 2003. Guideline for the conduct of food safety assessment of foods 

derived from recombinant-DNA plants. CAC/GL 45-2003. Codex Alimentarius 

Commission, Joint FAO/WHO Food Standards Programme, Food and Agriculture 

Organisation, Rome, Italy. 

ftp://ftp.fao.org/codex/Publications/Booklets/Biotech/Biotech_2003e.pdf Accessed May 

12, 2010. 

Cojocariu, C., J. Kreuzwieser and H. Rennenberg. 2004. Correlation of short-chained 

carbonyls emitted from Picea abies with physiological and environmental parameters. 

New Phytologist 162: 717-727. 

Cojocariu, C., P. Escher, K.-H. Häberle, R. Matyssek, H. Rennenberg and J. Kreuszeiser. 

2005. The effect of ozone on the emission of carbonyls from leaves of adult Fagus 

sylvatica. Plant, Cell and Environment 28: 603-611. 
Monsanto Company 10-SY-210U 265 of 721 

Comai, L., N. Larson-Kelly, J. Kiser, C.J.D. Mau, A.R. Pokalsky, C.K. Shewmaker, K. 

McBride, A. Jones and D.M. Stalker. 1988. Chloroplast transport of a ribulose 

bisphosphate carboxylase small subunit-5-enolpyruvyl 3-phosphoshikimate synthase 

chimeric protein requires part of the mature small subunit in addition to the transit 

peptide. The Journal of Biological Chemistry 263:15104-15109. 

Coruzzi, G., R. Broglie, C. Edwards and N.-H. Chua. 1984. Tissue-specific and lightregulated expression of a pea nuclear gene encoding the small subunit of ribulose-1, 5-

bisphosphate carboxylase. EMBO Journal 3:1671-1679. 

Crockett, L. 1977. Contents. Pages vii-ix in Wildly Successful Plants: A Handbook of 

North American Weeds. Macmillan Publishing Co., Inc., New York, New York. 

CTIC. 2004. National survey shows more farmers choose conservation. Conservation 

Technology Information Center, West Lafayette, Indiana. http://www.ctic.purdue.edu/ 

Accessed November 9, 2004. 

CTIC. 2007. 2007 amendment to the national crop residue management survey summary. 

Conservation Technology Information Center, West Lafayette, Indiana. 

http://www.conservationinformation.org/pdf/National_Summary_2007_Amendment.pdf 

Accessed June 14, 2010. 

CTIC. 2011. Top 10 conservation tillage benefits: Conservation for agriculture's future. 

Conservation Technology Information Center, West Lafayette, Indiana. 

http://www.ctic.purdue.edu/resourcedisplay/293/ Accessed September 21, 2011. 

Cunha, B.A. 2010. Stenotrophomonas maltophilia. WebMD, LLC, New York, New York. 

http://www.emedicine.com/med/topic3457.htm Accessed January 2, 2010. 

Cutler, G.H. 1934. A simple method for making soybean hybrids. Journal of the 

American Society of Agronomy 26:252-254. 

Dalley, C.D., K.A. Renner and J.J. Kells. 2001. Weed competition in Roundup Ready 

soybean and corn. Michigan State University, Department of Crop and Soil Science, East 

Lansing, Michigan. http://www.ipm.msu.edu/cat06field/pdf/weedfactsheet.pdf Accessed 

June 15, 2010. 

Darrouzet, E., J.W. Cooley and F. Daldal. 2004. The cytochrome bc1 complex and its 

homologue the b6f complex: Similarities and differences. Photosynthesis Research 79:25-

44. 

Davis, A., K. Renner, C. Sprague, L. Dyer and D. Mutch. 2005. Integrated weed 

management: One year's seeding. Extension Bulletin E-2931. Michigan State University, 

East Lansing, Michigan. 
Monsanto Company 10-SY-210U 266 of 721 

Denton, M., N.J. Todd, K.G. Kerr, P.M. Hawkey and J.M. Littlewood. 1998. Molecular 

epidemiology of Stenotrophomonas maltophilia isolated from clinical specimens from 

patients with cystic fibrosis and associated environmental samples. Journal of Clinical 

Microbiology 36:1953-1958. 

Depicker, A., S. Stachel, P. Dhaese, P. Zambryski and H.M. Goodman. 1982. Nopaline 

synthase: Transcript mapping and DNA sequence. Journal of Molecular and Applied 

Genetics 1:561-573. 

Dillon, T.W., R.C. Scott, N.D. Pearrow and K.A. Meins. 2006. Effect of sulfonylurea rice 

herbicides on soybeans. Page 1 in Southern Weed Science Society Proceedings, Little 

Rock, Arkansas. 

D'Ordine, R.L., T.J. Rydel, M.J. Storek, E.J. Sturman, F. Moshiri, R.K. Bartlett, G.R. 

Brown, R.J. Eilers, C. Dart, Y. Qi, S. Flasinski and S.J. Franklin. 2009. Dicamba 

monooxygenase: Structural insights into a dynamic Rieske oxygenase that catalyzes an 

exocyclic monooxygenation. Journal of Molecular Biology 392:481-497. 

Dorokhov, D., A. Ignatov, E. Deineko, A. Serjapin, A. Ala and K. Skryabin. 2004. 

Potential for gene flow from herbicide-resistant GM soybeans to wild soya in the Russian 

Far East. Pages 151-161 in Introgression from Genetically Modified Plants into Wild 

Relatives. H.C.M. den Nijs, D. Bartsch, and J. Sweet (eds.). CAB International, 

Wallingford, United Kingdom. 

Dorrance, A.E., M.A. Draper and D. Hershman. 2007. Using foliar fungicides to manage 

soybean rust. The Ohio State University, Colombus, Ohio. 

Dumitru, R., W.Z. Jiang, D.P. Weeks and M.A. Wilson. 2009. Crystal structure of 

dicamba monooxygenase: A Rieske nonheme oxygenase that catalyzes oxidative 

demethylation. Journal of Molecular Biology 392:498-510. 

Echemendia, Y. 2010. Microorganism of the month: Stenotrophomonas maltophilia. 

Environmental Microbiology Laboratory, Inc, Cherry Hill, New Jersey. 

http://www.emlab.com/s/sampling/env-report-07-2007.html Accessed January 2, 2007. 

Erickson, E.H. 1975. Honey bees and soybeans. American Bee Journal 115:351-353. 

European Commission. 2008. Review report for the active substance dicamba, Finalised 

in the Standing Committee on the Food Chain and Animal Health at its meeting on 14 

March 2008 in view of the inclusion of dicamba in Annex I of Directive 91/414/EEC. 

European Commission, Health and Consumer Protection Directorate-General, Paris, 

France. 

Fehr, W.R. 2007. Breeding for modified fatty acid composition in soybean. Crop Science

47:S72-S87. 
Monsanto Company 10-SY-210U 267 of 721 

Ferraro, D.J., L. Gakhar and S. Ramaswamy. 2005. Rieske business: Structure-function 

of Rieske non-heme oxygenases. Biochemical and Biophysical Research and 

Communications 338:175-190. 

Fling, M.E., J. Kopf and C. Richards. 1985. Nucleotide sequence of the transposon Tn7

gene encoding an aminoglycoside-modifying enzyme, 3"(9)-O-nucleotidyltransferase. 

Nucleic Acids Research 13:7095-7106. 

Fluhr, R., P. Moses, G. Morelli, G. Coruzzi and N.-H. Chua. 1986. Expression dynamics 

of the pea rbcS multigene family and organ distribution of the transcripts. EMBO Journal

5:2063-2071. 

Frisvold, G.B., T.M. Hurley and P.D. Mitchell. 2009. Adoption of best management 

practices to control weed resistance by corn, cotton, and soybean growers. AgBioForum

12:370-381. 

Gage, D.J. 2004. Infection and invasion of roots by symbiotic, nitrogen-fixing Rhizobia 

during nodulation of temperate legumes. Microbiology and Molecular Biology Reviews

68:280-300. 

Garber, R.J. and T.E. Odland. 1926. Natural crossing in soybeans. Journal of the 

American Society of Agronomy 18:967-970. 

Gianessi, L.P., C.S. Silvers, S. Sankula and J.E. Carpenter. 2002. Plant biotechnology: 

Current and potential impact for improving pest management in U.S. agriculture. An 

analysis of 40 case studies. National Center for Food & Agricultural Policy, Washington, 

D.C. 

Giese, M., U. Bauer-Doranth, C. Langebartels and H. Sandermann. 1994. Detoxification 

of formaldehyde by the spider plant (Chlorophytum comosum L.) and by soybean 

(Glycine max L.) cell-suspension cultures. Plant Physiology 104: 1301-1309 

Givens, W.A., D.R. Shaw, W.G. Johnson, S.C. Weller, B.G. Young, R.G. Wilson, M.D.K. 

Owen and D. Jordan. 2009. A grower survey of herbicide use patterns in glyphosateresistant cropping systems. Weed Technology 23:156-161. 

Giza, P.E. and R.C.C. Huang. 1989. A self-inducing runaway-replication plasmid 

expression system utilizing the Rop protein. Gene 78:73-84. 

Gray, J., E. Wardzala, M. Yang, S. Reinbothe, S. Haller and F. Pauli. 2004. A small 

family of LLS1-related non-heme oxygenases in plants with an origin amongst oxygenic 

photosynthesizers. Plant Molecular Biology 54:39-54. 

Gupta, M., W. Nirunsuksiri, G. Schulenberg, T. Hartl, S. Novak, J. Bryan, N. Vanopdorp, 

J. Bing and S. Thompson. 2008. A non-PCR-based Invader ® assay quantitatively detects 

single-copy genes in complex plant genomes. Molecular Breeding 21:173-181. 
Monsanto Company 10-SY-210U 268 of 721 

Hanson, A.D. and S. Roje. 2001. One-carbon metabolism in higher plants. Annual 

Review of Plant Physiology and Plant Molecular Biology 52: 119-137. 

Harayama, S., M. Kok and E.L. Neidle. 1992. Functional and evolutionary relationships 

among diverse oxygenases. Annual Review of Microbiology 46:565-601. 

Harrigan, G.G., D. Lundry, S. Drury, K. Berman, S.G. Riordan, M.A. Nemeth, W.P. 

Ridley and K.C. Glenn. 2010. Natural variation in crop composition and the impact of 

transgenesis. Nature Biotechnology 28:402-404. 

Heap, I. 2009. International survey of herbicide resistant weeds. Weed Science Society of 

America, Corvallis, Oregon. http://www.weedscience.org/in.asp Accessed November 11, 

2009. 

Heap, I. 2011. Herbicide resistant weeds summary table. WeedScience.com, International 

Survey of Herbicide Resistant Weeds, Corvallis, Oregon. 

http://www.weedscience.org/summary/MOASummary.asp Accessed May 26, 2011.

Heatherly, L.G., and H.F. Hodges. 1999. Soybean production in the Midsouth. CRC 

Press, Boca Raton, Florida. 

Helsel, Z.R. and H.C. Minor. 1993. G4410: Soybean production in Missouri. University 

of Missouri Extension, Columbia, Missouri. 

http://extension.missouri.edu/explore/agguides/crops/g04410.htm Accessed November 

11, 2009. 

Herman, P.L., M. Behrens, S. Chakraborty, B.M. Chrastil, J. Barycki and D.P. Weeks. 

2005. A three-component dicamba -demethylase from Pseudomonas maltophilia, strain 

DI-6. Journal of Biological Chemistry 280:24759-24767. 

Herrmann, K.M. 1995. The shikimate pathway: Early steps in the biosynthesis of 

aromatic compounds. Plant Cell 7:907-919. 

Hibino, T., R. Waditee, E. Araki, H. Ishikawa, K. Aoki, Y. Tanaka and T. Takabe. 2002. 

Functional characterization of choline monooxygenase, an enzyme for betaine synthesis 

in plants. Journal of Biological Chemistry 277:41352-41360. 

Higley, L.G. 1994. Insect injury to soybean. Pages 11-13 in Handbook of Soybean Insect 

Pests. L.G. Higley and D.J. Boethel (eds.). Entomological Society of America, Lanham, 

Maryland. 

Hock, S.M., S.Z. Knezevic, A.R. Martin and J.L. Lindquist. 2006. Soybean row spacing 

and weed emergence time influence weed competitiveness and competitive indices. 

Weed Science 54: 38-46. 
Monsanto Company 10-SY-210U 269 of 721 

Hoeft, R.G., E.D. Nafziger, R.R. Johnson and S.R. Aldrich. 2000. Soybean as a crop. 

Pages 31, 36, 38, 39, 40, 41, 47, 86, 87, 89, 90, 93, 94, 96, 97,120,121, 208, 209 in 

Modern Corn and Soybean Production. MCSP Publications, Champaign, Illinois. 

Holm, L., J.V. Pancho, J.P. Herberger, and D.L. Plucknett. 1979. A geographical atlas of 

world weeds. John Wiley and Sons., Inc., New York, New York. 

Holm, L., J. Doll, E. Holm, J.V. Pancho, and J.P. Herberger. 1997. World weeds: natural 

histories and distribution. John Wiley and Sons., Inc., New York, New York. 

HRAC. 2009. Classification of herbicides according to mode of action. Herbicide 

Resistance Action Committee, Corvallis, Oregon. 

http://www.hracglobal.com/Publications/ClassificationofHerbicideModeofAction/tabid/2

22/Default.aspx Accessed June 26, 2010. 

Hurley, T.M., P.D. Mitchell and G.B. Frisvold. 2009. Weed management costs, weed 

best management practices, and the Roundup Ready weed management program. 

AgBioForum 12:281-290. 

Hymowitz, T. 1970. On the domestication of the soybean. Economic Botany 24:408-421. 

Hymowitz, T. 2004. Speciation and cytogenetics. Pages 97-136 in Soybeans: 

Improvement, Production, and Uses. Third Edition. H.R. Boerma and J.E. Specht (eds.). 

American Society of Agronomy, Inc., Crop Science Society of America, Inc., Soil 

Science Society of America, Inc., Madison, Wisconsin. 

Hymowitz, T., R.G. Palmer and R.J. Singh. 1992. Cytogenetics of the genus Glycine. 

Pages 53-63 in Chromosome Engineering in Plants: Genetics, Breeding and Evolution. T. 

Tsuchiya and P.K. Gupta (eds.). Elsevier Science Publishers, Amsterdam, Netherlands. 

ILSI. 2006. Crop Composition Database, Version 3.0. International Life Sciences 

Institute, Washington, D.C. http://www.cropcomposition.org/. Accessed October 1, 

2009. 

ISO. 2009. Selection and use of the ISO 9000 family of standards. International 

Standards Organization, Geneva, Switzerland. 

ISU. 2004. Soybean growth and development. Iowa State University, Ames, Iowa. 

Israel, D.W., J.N. Mathis, W.M. Barbour and G.H. Elkan. 1986. Symbiotic effectiveness 

and host-strain interactions of Rhizobium fredii USDA 191 on different soybean cultivars. 

Applied and Environmental Microbiology 51:898-903. 

Janas, K.M., M. Cvikrová, A. Paągiewicz and J. Eder. 2000. Alterations in 

phenylpropanoid content in soybean roots during low temperature acclimation. Plant 

Physiology and Biochemistry 38:587-593. 
Monsanto Company 10-SY-210U 270 of 721 

Janda, T., E. Horvath, G. Szalai and E. Paldi. 2007. Role of salicylic acid in the induction 

of abiotic stress tolerance. Pages 91-150 in Salicylic Acid: A Plant Hormone. S. Hayat 

and A. Ahmad (eds.). Springer, Dordrecht, The Netherlands. 

Jaycox, E.R. 1970a. Ecological relationships between honey bees and soybeans: II the 

plant factors. American Bee Journal 110:343-345. 

Jaycox, E.R. 1970b. Ecological relationships between honey bees and soybeans: III the 

honey-bee factors. American Bee Journal 110:383-385. 

Johnson, W.G., M.D.K. Owen, G.R. Kruger, B.G. Young, D.R. Shaw, R.G. Wilson, J.W. 

Wilcut, D.L. Jordan and S.C. Weller. 2009. US farmer awareness of glyphosate-resistant 

weeds and resistance management strategies. Weed Technology 23:308-312. 

Johnson, B., B. Young, J. Matthews, P. Marquardt, C. Slack, K. Bradley, A. York, S. 

Culpepper, A. Hager, K. Al-Khatib, L. Steckel, M. Moechnig, M. Loux, M. Bernards and 

R. Smeda. 2010. Weed control in dicamba-resistant soybeans. Plant Management 

Network, St. Paul, Minnesota. 

http://www.plantmanagementnetwork.org/sub/cm/research/2010/dicamba/ Accessed 

September 20, 2010. 

Jordan, N., D.A. Mortensen, D.M. Prenzlow and K.C. Cox. 1995. Simulation analysis of 

crop rotation effects on weed seedbanks. American Journal of Botany 82:390-398. 

Jordan, T., G. Nice, B. Johnson and T. Bauman. 2009. Reducing spray drift from 

glyphosate and growth regulator herbicide drift caution. Purdue Extension, Purdue 

University, West Lafayette, Indiana. 

http://www.ag.purdue.edu/btny/weedscience/Documents/ReducingDrift09.pdf Accessed 

December 18, 2009. 

Juhnke, M.E. and E. des Jardin. 1989. Selective medium for isolation of Xanthomonas 

maltophilia from soil and rhizosphere environments. Applied and Environmental 

Microbiology 55:747-750. 

Juhnke, M.E., D.E. Mathre and D.C. Sands. 1987. Identification and characterization of 

rhizosphere-competent bacteria of wheat. Applied and Environmental Microbiology

53:2793-2799. 

Klee, H.J., Y.M. Muskopf and C.S. Gasser. 1987. Cloning of an Arabidopsis thaliana

gene encoding 5-enolpyruvylshikimate-3-phosphate synthase: Sequence analysis and 

manipulation to obtain glyphosate-tolerant plants. Molecular and General Genetics

210:437-442. 
Monsanto Company 10-SY-210U 271 of 721 

Klingman, G.C., F.M. Ashton and L.J. Noordhoff. 1975. Methods of weed control. Pages 

13-28 in Weed Science: Principles and Practices. John Wiley & Sons, Inc., New York, 

New York. 

Kollipara, K.P., R.J. Singh and T. Hymowitz. 1993. Genomic diversity in aneudiploid 

(2n=38) and diploid (2n=40) Glycine tomentella revealed by cytogenetic and biochemical 

methods. Genome 36:391-396. 

Krueger, J.P., R.G. Butz, Y.H. Atallah and D.J. Cork. 1989. Isolation and identification 

of microorganisms for the degradation of dicamba. Journal of Agricultural and Food 

Chemistry 37:534-538. 

Kruger, G.R., W.G. Johnson, S.C. Weller, M.D.K. Owen, D.R. Shaw, J.W. Wilcut, D.L. 

Jordan, R.G. Wilson, M.L. Bernards and B.G. Young. 2009. U.S. grower views on 

problematic weeds and changes in weed pressure in glyphosate-resistant corn, cotton, and 

soybean cropping systems. Weed Technology 23:162-166. 

Kuroda, Y., A. Kaga, N. Tomooka and D.A. Vaughan. 2008. Gene flow and genetic 

structure of wild soybean (Glycine soja) in Japan. Crop Science 48:1071-1079. 

Lambert, B., F. Leyns, L. Van Rooyen, F. Gossele, Y. Papon and J. Swings. 1987. 

Rhizobacteria of maize and their antifungal activities. Applied and Environmental 

Microbiology 53:1866-1871. 

Lingenfelter, D.D. and N.L. Hartwig. 2003. Introduction to weeds and herbicides. Penn 

State University Agricultural Research and Cooperative Extension, University Park, 

Pennsylvania. 

Loux, M.M., D. Doohan, A.F. Dobbels, W.G. Johnson, G.R.W. Nice, T.N. Jordan, and 

T.T. Bauman. 2009. Weed control guide for Ohio and Indiana. Purdue University and 

Ohio State. Publication WS16. (http://www.btny.purdue.edu/Pubs/ws/ws-16/) Accessed 

on June 14, 2010. 

LSU. 2009. Asian soybean rust: Background. Louisiana State University, Baton Rouge, 

Louisiana. 

http://www.agctr.lsu.edu/en/crops_livestock/crops/soybeans/Asian_Soybean_Rust/ 

Accessed June 17, 2010. 

Lu, B.-R. 2004. Conserving biodiversity of soybean gene pool in the biotechnology era. 

Plant Species Biology 19:115-125. 

Lundry, D.R., W.P. Ridley, J.J. Meyer, S.G. Riordan, M.A. Nemeth, W.A. Trujillo, M.L. 

Breeze and R. Sorbet. 2008. Composition of grain, forage, and processed fractions from 

second-generation glyphosate-tolerant soybean, MON 89788, is equivalent to that of 
Monsanto Company 10-SY-210U 272 of 721 

conventional soybean (Glycine max L.). Journal of Agricultural and Food Chemistry

56:4611-4622. 

Maiti, I.B. and R.J. Shepherd. 1998. Isolation and expression analysis of peanut chlorotic 

streak caulimovirus (PCISV) full-length transcript (FLt) promoter in transgenic plants. 

Biochemical and Biophysical Research Communications 244:440-444. 

Martinell, B.J., L.S. Julson, C.A. Emler, Y. Huang, D.E. McCabe and E.J. Williams. 

2002. Soybean agrobacterium transformation method. Patent 6,384,301, U.S. Patent 

Office, Washington, D.C. 

Montgomery, R.F., R.M. Hayes, C.H. Tingle and J.A. Kendig. 2002. Control of 

glyphosate-tolerant soybeans (Glycine max) in no-till Roundup Ready® cotton 

(Gossypium hirsutum L.). Proceedings of the Beltwide Cotton Conference, National 

Cotton Council, Atlanta, Georgia. 

MSU. 2010. 2010 weed control guidelines for Mississippi. Mississippi State University 

Extension Service, Mississippi State, Mississippi. 

http://msucares.com/pubs/publications/p1532.pdf Accessed June 14, 2010. 

Mullin, W.J. and W. Xu. 2001. Study of soybean seed coat components and their 

relationship to water absorption. Journal of Agricultural and Food Chemistry 49:5331-

5335. 

Murdock, E.C., M.A. Jones and R.F. Graham. 2002. Control of volunteer glyphosate 

(Roundup)-tolerant cotton and soybean in Roundup Ready cotton. Proceedings of the 

Beltwide Cotton Conference, National Cotton Council, Atlanta, Georgia. 

Naeve, S. 2005. Minnesota soybean production – growth and development. Department 

of Agronomy, University of Minnesota. 

http://www.soybeans.umn.edu/crop/growth/index.htm Accessed December 17, 2009. 

Nam, J.-W., H. Nojiri, T. Yoshida, H. Habe, H. Yamane and T. Omori. 2001. New 

classification system for oxygenase components involved in ring-hydroxylating 

oxygenations. Bioscience, Biotechnology, and Biochemistry 65:254-263. 

NatureServe. 2007. NatureServe explorer: An online encyclopedia of life. Version 6.2. 

NatureServe, Arlington, Virginia. http://www.natureserve.org/explorer. Accessed July 

6, 2007. 

NDSU. 2002. Soybean production field guide for North Dakota and northwestern 

Minnesota. North Dakota State University Extension Service and North Dakota Soybean 

Council, Fargo, North Dakota. http://www.ag.ndsu.edu/pubs/plantsci/rowcrops/a1172.pdf 

Accessed November 11, 2009. 
Monsanto Company 10-SY-210U 273 of 721 

Nemecek-Marshall, M., R.C. Macdonald, J.J. Franzen, C.L. Wojciechowski and R. Fall. 

1995. Methanol emission from leaves: Enzymatic detection of gas-phase methanol and 

relation of methanol fluxes to stomatal conductance and leaf development. Plant 

Physiology 108: 1359-1368. 

Nice, G. and B. Johnson. 2005. Indiana's top ten most problematic weeds. Purdue 

Extension, West Lafayette, Indiana. 

http://www.btny.purdue.edu/WeedScience/2005/topten05.pdf Accessed November 11, 

2009. 

Nielsen, R.L. 2008. Use thermal time to predict leaf stage development in corn. Purdue 

University, West Lafayette, Indiana. 

http://www.agry.purdue.edu/ext/corn/news/timeless/vstageprediction.html Accessed 

December 18, 2009. 

Niepel, M. and D.R. Gallie. 1999. Identification and characterization of the functional 

elements within the tobacco etch virus 5' leader required for cap-independent translation. 

Journal of Virology 73:9080-9088. 

NOAA-ESRL. 2011. Average mean temperature index by month: Climatology by state 

based on climate division data: 1971-2000. National Oceanic and Atmospheric 

Administration, Earth System Research Laboratory, Washington, D.C. 

http://www.esrl.noaa.gov/psd/data/usclimate/tmp.state.19712000.climo Accessed 

September 28, 2011. 

NOAA-NCDC. 2011. Data through 2009. National Oceanic and Atmospheric 

Administration, National Climatic Data Center, Washington, D.C. 

http://www.ncdc.noaa.gov/oa/climate/online/ccd/relhum.txt Accessed September 28, 

2011. 

Norsworthy, J.K. and L.R. Oliver. 2002. Hemp sesbania interference in drill-seeded 

glyphosate-resistant soybean. Weed Science 50: 34-41. 

Nunes, F.V. and I.S. de Melo. 2006. Isolation and characterization of endophytic bacteria 

of coffee plants and their potential in caffeine degradation. Environmental Toxicology

10:293-297. 

OECD. 1993. Safety considerations for biotechnology: Scale-up of crop plants. 

Organisation for Economic Co-operation and Development, Paris, France. 

OECD. 2000. Consensus document on the biology of Glycine max (L.) merr. (soybean). 

ENV/JM/MONO(2000)9. Series on harmonization of regulatory oversight in 

biotechnology No.15. Organisation for Economic Co-operation and Development, Paris, 

France. 
Monsanto Company 10-SY-210U 274 of 721 

OECD. 2001. Consensus document on compositional considerations for new varieties of 

soybean: Key food and feed nutrients and anti-nutrients. ENV/JM/MONO(2001)15. 

Series on the Safety of Novel Foods and Feeds No.2. Organisation for Economic Cooperation and Development, Paris, France. 

Oliver, D.J. 1994. The glycine decarboxylase complex from plant mitochondria. Annual 

Review of Plant Physiology and Plant Molecular Biology 45: 323-337. 

Owen, M.D.K., B.G. Young, D.R. Shaw, R.G. Wilson, D.L. Jordan, S.C. Weller and P. 

Dixon. 2009. Benchmark study: Perspectives on glyphosate-resistant crops and the 

sustainability of chemical weed management. Page 38 in 2009 North Central Weed 

Science Society Proceedings, Las Cruces, New Mexico. 

Padgette, S.R., D.B. Re, G.F. Barry, D.E. Eichholtz, X. Delannay, R.L. Fuchs, G.M. 

Kishore and R.T. Fraley. 1996a. New weed control opportunities: Development of 

soybeans with a Roundup ReadyTM gene. Pages 53-84 in Herbicide-Resistant Crops: 

Agricultural, Environmental, Economic, Regulatory and Technical Aspects. S.O. Duke 

(ed.). CRC Press, Inc., Boca Raton, Florida. 

Padgette, S.R., N.B. Taylor, D.L. Nida, M.R. Bailey, J. MacDonald, L.R. Holden and R.L. 

Fuchs. 1996b. The composition of glyphosate-tolerant soybean seeds is equivalent to that 

of conventional soybeans. Journal of Nutrition 126:702-716. 

Palleroni, N.J. and J.F. Bradbury. 1993. Stenotrophomonas, a new bacterial genus for 

Xanthomonas maltophilia (Hugh 1980) Swings et al. 1983. International Journal of 

Systematic Bacteriology 43:606-609. 

Pedersen, P. 2008a. Soybean production. Iowa State University Extension, Ames, Iowa. 

http://extension.agron.iastate.edu/soybean Accessed November 11, 2009. 

Pedersen, P. 2008b. Soybean - insects and diseases. Iowa State University Extension, 

Ames, Iowa. http://extension.agron.iastate.edu/soybean Accessed November 11, 2009. 

Pedersen, P. 2008c. Managing soybean cyst nematode. Iowa State University Extension, 

Ames, Iowa. http://extension.agron.iastate.edu/soybean/documents/SCN.pdf Accessed 

November 11, 2008. 

Pimentel, D. 1991. Soybean pest management. Pages 341-343 in CRC Handbook of Pest 

Management in Agriculture. Volume 3. Second Edition. CRC Press, Inc., Boca Raton, 

Florida. 

PMRA. 2008. Pest Management Regulatory Agency Re-evaluation – dicamba. Pest 

Management Regulatory Authority, Health Canada. Ottawa, Canada. 

Powles, S.B., C. Preston, I.B. Bryan and A.R. Jutsum. 1996. Herbicide resistance: Impact 

and management. Pages 57-93 in Advances in Agronomy. Volume 58. D.L. Sparks (ed.). 

Academic Press, San Diego, California. 
Monsanto Company 10-SY-210U 275 of 721 

Powles, S.B. 2008. Evolved glyphosate-resistant weeds around the world: Lessons to be 

learnt. Pest Management Science 64:360-365. 

Prather, T.S., J.M. Ditomaso and J.S. Holt. 2000. Herbicide resistance: Definition and 

management strategies. University of California, Division of Agriculture and Natural 

Resources, Oakland, California. 

Price, A.J., K.S. Balkcom, S.A. Culpepper, J.A. Kelton, R.L. Nichols and H. Schomberg. 

2011. Glyphosate-resistant Palmer amaranth: A threat to conservation tillage. Journal of 

Soil and Water Conservation 66:265-275. 

Rathinasabapathi, B., M. Burnet, B.L. Russell, D.A. Gage, P.-C. Liao, G.J. Nye, P. Scott, 

J.H. Golbeck and A.D. Hanson. 1997. Choline monooxygenase, an unusual iron-sulfur 

enzyme catalyzing the first step of Glycine betaine synthesis in plants: Prosthetic group 

characterization and cDNA cloning. Proceedings of the National Academy of Sciences of 

the United States of America 94:3454-3458. 

Ray, J.D., T.C. Kilen, C.A. Abel and R.L. Paris. 2003. Soybean natural cross-pollination 

rates under field conditions. Environmental Biosafety Research 2:133-138. 

Rensing, S.A. and U.-G. Maier. 1994. Phylogenetic analysis of the stress-70 protein 

family. Journal of Molecular Evolution 39:80-86. 

Reynolds, T.L., M.A. Nemeth, K.C. Glenn, W.P. Ridley and J.D. Astwood. 2005. Natural 

variability of metabolites in maize grain: Differences due to genetic background. Journal 

of Agricultural and Food Chemistry 53:10061-10067. 

Ridley, W.P., R.D. Shillito, I. Coats, H.-Y. Steiner, M. Shawgo, A. Phillips, P. Dussold 

and L. Kurtyka. 2004. Development of the International Life Sciences Institute Crop 

Composition Database. Journal of Food Composition and Analysis 17:423-438. 

Rodoni, S., W. Mühlecker, M. Anderl, B. Kräutler, D. Moser, H. Thomas, P. Matile and 

S. Hörtensteiner. 1997. Chlorophyll breakdown in senescent chloroplasts (cleavage of 

pheophorbide a in two enzymic steps). Plant Physiology 115:669-676. 

Rogers, S.G. 2000. Promoter for transgenic plants. Patent 6,018,100, U.S. Patent Office, 

Washington, D.C. 

Rosche, B., B. Tshisuaka, B. Hauer, F. Lingens and S. Fetzner. 1997. 2-oxo-1,2-

dihydroquinoline 8-monooxygenase: Phylogenetic relationship to other multicomponent 

nonheme iron oxygenases. Journal of Bacteriology 179:3549-3554. 

Russell, B.L., B. Rathinasabapathi and A.D. Hanson. 1998. Osmotic stress induces 

expression of choline monooxygenase in sugar beet and amaranth. Plant Physiology

116:859-865. 
Monsanto Company 10-SY-210U 276 of 721 

Ryan, R.P., S. Monchy, M. Cardinale, S. Taghavi, L. Crossman, M.B. Avison, G. Berg, 

D. van der Lelie and J.M. Dow. 2009. The versatility and adaptation of bacteria from the 

genus Stenotrophomonas. Nature Reviews Microbiology 7:514-525. 

Salomon, S. and H. Puchta. 1998. Capture of genomic and T-DNA sequences during 

double-strand break repair in somatic plant cells. EMBO Journal 17:6086-6095. 

Schmidt, C.L. and L. Shaw. 2001. A comprehensive phylogenetic analysis of Rieske and 

Rieske-type iron-sulfur proteins. Journal of Bioenergetics and Biomembranes 33:9-26. 

Scott, W.O. and S.R. Aldrich. 1970. Modern soybean production. The Farm Quarterly: 

16-152. 

SDTF. 1997. A summary of ground application studies. Spray Drift Task Force, St. Louis, 

Missouri. http://www.agdrift.com/PDF_FILES/Ground.pdf Accessed June 7, 2010. 

Seguin, P., P. Turcotte, G. Tremblay, D. Pageau and W. Liu. 2009. Tocopherols 

concentration and stability in early maturing soybean genotypes. Agronomy Journal

101:1153-1159. 

Shurtleff, J.L. and H.D. Coble. 1985. Interference of certain broadleaf weed species in 

soybeans (Glycine max). Weed Science 33: 654-657. 

Singh, R.J. and T. Hymowitz. 1989. The genomic relationships among Glycine soja Sieb. 

and Zucc., G. max (L.) Merr. and 'G. gracilis' Skvortz. Plant Breeding 103:171-173. 

Singh, R.J. and T. Hymowitz. 1999. Soybean genetic resources and crop improvement. 

Genome 42:605-616. 

Soyatech. 2010. Statistics and reference. Pages 314-317 in Soya and Oilseed Bluebook. 

Soyatech, Inc., Southwest Harbor, Maine. 

Stalker, D.M., C.M. Thomas and D.R. Helinski. 1981. Nucleotide sequence of the region 

of the origin of replication of the broad host range plasmid RK2. Molecular and General 

Genetics 181:8-12. 

Steffen, D. 2004. Soybean SN70025 PVPC Number 200400321. U.S. Department of 

Agriculture, Plant Variety Protection Office, Washington, D.C. 

Sundstrom, F.J., J. Williams, A.V. Deynze and K.J. Bradford. 2002. Identity preservation 

of agricultural commodities. Publication 8077. University of California, Division of 

Agricultural and Natural Resources, Oakland, California. 
Monsanto Company 10-SY-210U 277 of 721 

Sutcliffe, J.G. 1979. Complete nucleotide sequence of the Escherichia coli plasmid 

pBR322. Pages 77-90 in Cold Spring Harbor Symposia on Quantitative Biology, Cold 

Spring Harbor, New York. 

Sutherland, J.P. and G.M. Poppy. 2005. Quantifying exposure. Pages 186-212 in Gene 

Flow from GM Plants. G.M. Poppy and M.J. Wilkinson (eds.). Blackwell Publishing, 

Ames, Iowa. 

Swings, J., P. De Vos, M. Van den Mooter and J. De Ley. 1983. Transfer of 

Pseudomonas maltophilia Hugh 1981 to the genus Xanthomonas as Xanthomonas 

maltophilia (Hugh 1981) comb. nov. International Journal of Systematic Bacteriology

33:409-413. 

Szabó, B., E. Tyihák, L.G. Szabó and L. Botz. 2003. Mycotoxin and drought stress 

induced change of alkaloid content of Papaver somniferum plantlets. Acta Botanica 

Hungarica 45: 409-417. 

Szende, B. and E. Tyihák. 2010. Effect of formaldehyde on cell proliferation and death. 

Cell Biology International 34: 1273-1282. 

University of Tennessee. 2010. 2010 weed control manual for Tennessee. University of 

Tennessee, Institute of Agriculture, Knoxville, Tennessee. 

U.S. EPA. 1993. Reregistration eligibility decision (RED): Glyphosate. U.S. 

Environmental Protection Agency, Washington, D.C. 

U.S. EPA. 1996. Residue chemistry test guidelines - Nature of the residue - Plants, 

livestock. OPPTS 860.1300. U.S. Environmental Protection Agency, Office of 

Prevention, Pesticides and Toxic Substances, Washington, D.C. 

U.S. EPA. 2008. Reregistration eligibility decision for formaldehyde and 

paraformaldehyde. EPA 739-R-08-004. U.S. Environmental Protection Agency, Office of 

Pesticide Programs Washington, D.C. 

U.S. EPA. 2009. Reregistration eligibility decision for dicamba and associated salts 

(Amended). U.S. Environmental Protection Agency, Office of Prevention, Pesticides, and 

Toxic Substances, Washington, D.C. 

U.S. FDA. 1992. Statement of policy: Foods derived from new plant varieties. Federal 

Register: 22984-23005. 

USDA-APHIS. 2006. Soybean. U.S. Department of Agriculture, Animal and Plant Health 

Inspection Service, Washington, D.C. http://www.aphis.usda.gov/br/soybean.html 

Accessed January 20, 2006. 

USDA-APHIS. 2007. Environmental assessment, in response to Monsanto petition 06-

178-01p seeking a determination of non-regulated status for Roundup Ready 2 Yield 
Monsanto Company 10-SY-210U 278 of 721 

soybean MON 89788. U.S. Department of Agriculture, Animal and Plant Health 

Inspection Service, Washington, D.C. 

http://www.aphis.usda.gov/brs/aphisdocs2/06_17801p_com.pdf Accessed May 28, 

2010. 

U.S. CPSC. 1997. An update on formaldehyde. U.S. Consumer Product Safety 

Commission, Washington, D.C. 

USDA-ERS. 2001. How much U.S. cropland acreage is under crop residue management. 

Briefing Room. U.S. Department of Agriculture, Economic Research Service, 

Washington, D.C. http://www.ers.usda.gov/Briefing/AgChemicals/soilmangement.htm 

Accessed November 14, 2009. 

USDA-ERS. 2008. Data sets: U.S. and regional cost and return estimates in soybean for 

2008. U.S. Department of Agriculture, Economic Research Service, Washington, D.C. 

http://www.ers.usda.gov/data/CostsandReturns/testpick.htm Accessed January 18, 2010. 

USDA-NASS. 2001. Agricultural chemical usage - 2000 field crops summary. U.S. 

Department of Agriculture, National Agricultural Statistics Service, Washington, D.C. 

USDA-NASS. 2004. Agricultural chemical usage - 2003 field crops summary. U.S. 

Department of Agriculture, National Agricultural Statistics Service, Washington, D.C. 

USDA-NASS. 2006. Agricultural chemical usage - 2005 field crops summary. U.S. 

Department of Agriculture, National Agricultural Statistics Service, Washington, D.C. 

USDA-NASS. 2007a. Acreage - June 2006. U.S. Department of Agriculture, National 

Agricultural Statistics Service, Washington, D.C. 

USDA-NASS. 2007b. Agricultural chemical usage 2006 field crops summary. U.S. 

Department of Agriculture, National Agricultural Statistics Service, Washington, D.C. 

USDA-NASS. 2008. Agricultural chemical usage - 2007 field crops summary. U.S. 

Department of Agriculture, National Agricultural Statistics Service, Washington, D.C. 

USDA-NASS. 2009a. Crop production: 2008 summary. U.S. Department of Agriculture, 

National Agricultural Statistics Service, Washington, D.C. 

USDA-NASS. 2009b. Crop values: 2008 summary. U.S. Department of Agriculture, 

National Agricultural Statistics Service, Washington, D.C. 

USDA-NASS. 2009c. Acreage - June 2009. U.S. Department of Agriculture, National 

Agricultural Statistics Service, Washington, D.C. 
Monsanto Company 10-SY-210U 279 of 721 

USDA-NASS. 2009d. Crop progress reports. U.S. Department of Agriculture, National 

Agricultural Statistics Service, Washington, D.C. 

USDA-NASS. 2010. Charts and maps: Soybeans: Planted acreage by country. U.S. 

Department of Agriculture, National Agricultural Statistics Service, Washington, D.C. 

USDA-NRCS. 2010. The PLANTS database. U.S. Department of Agriculture, Natural 

Resources Conservation Service, Washington, D.C. 

http://www.plants.usda.gov/index.html Accessed March 23, 2010

USGS. 2004. Pesticide national synthesis project: Pesticide use. United States Geological 

Survey, Washington, D.C. http://water.usgs.gov/nawqa/pnsp/usage/maps/ Accessed May 

28, 2010. 

USHHS-ATSDR. 1999. Toxicological profile for formaldehyde. U.S. Department of 

Health and Human Services, Agency for Toxic Substances and Disease Registry, 

Washington, D.C. 

USHHS-NTP. 2011. Report on carcinogens. U.S. Department of Health and Human 

Services, National Toxicology Program, Washington, D.C. 

Vail, G.D. and L.R. Oliver. 1993. Barnyardgrass (Echinochloa crus-galli) interference in 

soybeans (Glycine max). Weed Technology 7: 220-225. 

Wang, X.-Z., B. Li, P.L. Herman and D.P. Weeks. 1997. A three-component enzyme 

system catalyzes the O demethylation of the herbicide dicamba in Pseudomonas 

maltophilia DI-6. Applied and Environmental Microbiology 63:1623-1626. 

Weber, C.R. and W.D. Hanson. 1961. Natural hybridization with and without ionizing 

radiation in soybeans. Crop Science 1:389-392. 

Webster, T.M., M. Patterson, J. Everest, J. Ferrell, B. Brecke, A.S. Culpepper, E.P. 

Prostko, J.D. Green, J.R. Martin, E. Webster, S. Kelly, J. Griffin, D. Sanders, J. Byrd, A. 

Kendig, A. York, D. Jordan, L. Fisher, C. Medlin, D. Murray, J. Norsworthy, J. Chapin, 

L. Nelson, and L. Steckel. 2005. Weed survey: Southern states 2005: Broadleaf crops 

section (cotton, peanut, soybean, tobacco, and forestry). Pages 291-306, Southern Weed 

Science Society Proceedings, Las Cruces, New Mexico. 

Webster, T.M., M. Patterson, J. Everest, K. Smith, J. Norsworthy, B. Scott, C. Fristoe, D. 

Oliver, N. Burgos, J. Boyd, J. Ferrell, P. Minogue, B. Becke, A.S. Culpepper, D. 

Moorhead, E. Prostko, J.M. Moore, J.D. Green, J.R. Martin, B. Miller, D. Miller, J. 

Griffin, D. Stephenson, D. Sanders, K. Bradley, A. York, L. Fisher, S. Jordan, D. Murray, 

M. Marshall, S. Hagood, and C. Johnson. 2009. Weed survey: Southern states 2009: 

Broadleaf crops subsection (cotton, peanut, soybean, tobacco, and forestry). Pages 517-

520 in Southern Weed Science Society Proceedings, Las Cruces, New Mexico. 
Monsanto Company 10-SY-210U 280 of 721 

Werlen, C., H.-P.E. Kohler and J.R. van der Meer. 1996. The broad substrate 

chlorobenzene dioxygenase and cis-chlorobenzene dihydrodiol dehydrogenase of 

Pseudomonas sp. strain P51 are linked evolutionarily to the enzymes for benzene and 

toluene degradation. Journal of Biological Chemistry 271:4009-4016. 

WHO. 2002. Concise International Chemical Assessment Document 40: Formaldehyde. 

World Health Organization, Geneva. 

WHO-IPCS. 1989. Environmental health criteria 89: Formaldehyde. World Health 

Organization, International Programme on Chemical Safety, Geneva, Switzerland. 

http://www.inchem.org/documents/ehc/ehc/ehc89.htm Accessed February 2, 2011. 

Wishart, D.S. 2010. Human Metabolome Database. www.hmdb.ca Accessed June, 2 

2010.

Wishart, D.S., C. Knox, A.C. Guo, R. Eisner, N. Young, B. Gautam, D.D. Hau, N. 

Psychogios, E. Dong, S. Bouatra, R. Mandal, I. Sinelnikov, J. Xia, L. Jia, J.A. Cruz, E. 

Lim, C.A. Sobsey, S. Shrivastava, P. Huang, P. Liu, L. Fang, J. Peng, R. Fradette, D. 

Cheng, D. Tzur, M. Clements, A. Lewis, A. De Souza, A. Zuniga, M. Dawe, Y. Xiong, D. 

Clive, R. Greiner, A. Nazyrova, R. Shaykhutdinov, L. Li, H.J. Vogel and I. Forsythe. 

2009. HMDB: A knowledgebase for the human metabolome. Nucleic Acids Research

37:D603-D610. 

Woodworth, C.M. 1922. The extent of natural cross-pollination in soybeans. Journal of 

the American Society of Agronomy 14:278-283. 

Wrather, J.A., W.C. Stienstra and S.R. Koenning. 2001. Soybean disease loss estimates 

for the United States from 1996 to 1998. Canadian Journal of Plant Pathology-Revue 

Canadienne De Phytopathologie 23:122-131. 

Wrather, J.A. and S. Koenning. 2011. Soybean disease loss estimates for the United 

States, 1996-2010. University of Missouri Delta Research Center, Agricultural 

Experiment Station, Portageville, Missouri. 

http://aes.missouri.edu/delta/research/soyloss.stm Accessed February 22, 2012

Wrather, J.A., W.C. Stienstra and S.R. Koenning. 2001. Soybean disease loss estimates 

for the United States from 1996 to 1998. Canadian Journal of Plant Pathology-Revue 

Canadienne De Phytopathologie 23:122-131. 

WSSA. 1992. Tables. Pages 88, 103, 107, 118, 123, 130, 145, 393-401 in Crop Losses 

Due to Weeds in the United States. D.C. Bridges (ed.). Weed Science Society of America, 

Champaign, Illinois. 

WSSA. 2010. WSSA supports NRC findings on weed control. Weed Science Society of 

America, Champaign, Illinois. Accessed June 9, 2010. 
Monsanto Company 10-SY-210U 281 of 721 

Yang, M., E. Wardzala, G.S. Johal and J. Gray. 2004. The wound-inducible Lls1 gene 

from maize is an orthologue of the Arabidopsis Acd1 gene, and the LLS1 protein is 

present in non-photosynthetic tissues. Plant Molecular Biology 54:175-191. 

York, A.C., J.B. Beam and A.S. Culpepper. 2005. Control of volunteer glyphosateresistant soybean in cotton. Journal of Cotton Science 9:102-109. 

Yoshimura, Y., K. Matsuo and K. Yasuda. 2006. Gene flow from GM glyphosatetolerant to conventional soybeans under field conditions in Japan. Environmental 

Biosafety Research 5:169-173. 

Zambryski, P., A. Depicker, K. Kruger and H.M. Goodman. 1982. Tumor induction by 

Agrobacterium tumefaciens: Analysis of the boundaries of T-DNA. Journal of Molecular 

and Applied Genetics 1:361-370. 

Zhang, L.X., J. Zhang, S. Kye-Boahen and C.E. Watson. 2004. Redefining zones of 

adaptation of soybean maturity groups in the US. 2004 Annual Meeting Abstracts, CDROM. American Society of Agronomy, Inc., Crop Science Society of America, Inc., Soil 

Science Society of America, Inc., Madison, Wisconsin. 

Zhu, J.-K. 2009. Active DNA demethylation mediated by DNA glycosylases. Annual 

Review of Genetics 43: 143-166. 

Zollinger, R.K. 2000. Survey of weeds in North Dakota. North Dakota State University, 

Extension Weed Specialist, Fargo, North Dakota. 

http://www.ag.ndsu.nodak.edu/weeds/ER83/ER83.htm Accessed November 11, 2009. 

Zollinger, R.K. 2009. 2009 North Dakota weed control guide – relative herbicide 

effectiveness on weeds-volunteer Roundup Ready crops. Circular W-253, North Dakota 

State University, Extension Weed Specialist, Fargo, North Dakota 

http://www.ag.ndsu.edu/weeds/w253/w253-4d.htm Accessed November 11, 2009. 

 
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APPENDICES 
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Appendix A: USDA Notifications 

Field trials of MON 87708 have been conducted in the U.S. since 2005. The protocols 

for these trials included field performance, breeding and observation, agronomics, and 

generation of field materials and data necessary for this petition. In addition to 

MON 87708 phenotypic assessment data, observational data on pest and disease stressors 

were collected from these product development trials. The majority of the final reports 

have been submitted to the USDA. However, some final reports, mainly from the 

2008-2009 seasons, are still in preparation. A list of trials conducted under USDA 

notifications and the status of the final reports for these trials are provided in Table A-1. 
Monsanto Company 10-SY-210U 284 of 721 

Table A-1. USDA Notifications Approved for MON 87708 and Status of Trials 

Conducted under These Notifications and Permits 

USDA Number Effective Date Release Site (State) Trial Status 

2005 

05-269-02n 11/16/2005 PR Submitted to USDA 

2006 

06-045-15n 5/18/2006 HI(5) Submitted to USDA 

06-045-17n 5/18/2006 PR(3) Submitted to USDA 

06-052-01n 3/20/2006 IL(7), KS(5) Submitted to USDA 

06-052-02n 4/24/2006 IA(7), IL(5), IN(2) Submitted to USDA 

06-052-09n 4/24/2006 IA(2), IL(6), IN(2) Submitted to USDA 

06-067-05n 4/24/2006 IL(2) Submitted to USDA 

06-090-03n 5/5/2006 IL(2) Submitted to USDA 

06-275-102n 11/14/2006 PR Submitted to USDA 

06-345-101n 1/10/2007 PR(3) Submitted to USDA 

2007 

07-018-103n 2/17/2007 IL(10), IN(3), MO, PR Submitted to USDA 

07-018-106n 2/17/2007 IA(7), KS(6) Submitted to USDA 

07-018-109n 2/17/2007 IA, IL(10), IN(3), MO Submitted to USDA 

07-024-101n 3/18/2007 IA(7), KS(6) Submitted to USDA 

07-039-101n 3/18/2007 IA(4), IL(5), IN(3), KS(3) Submitted to USDA 

07-043-102n 4/10/2007 IA, IL(2), KS, MD, WI Submitted to USDA 

07-050-107n 4/9/2007 IA, IL(2), IN, KS, KY, MN, 

NE, SD 

Submitted to USDA 

07-057-109n 4/6/2007 AL, IA(3), IL, IN, LA, MN, 

MO(2), MS, NE, SD, TN 

Submitted to USDA 

07-094-104n 5/4/2007 IA(2) Submitted to USDA 

07-094-116n 5/4/2007 MN Submitted to USDA 

07-113-103n 6/4/2007 PR(2) Submitted to USDA 

07-241-103n 9/28/2007 PR Submitted to USDA 

07-250-102n 10/7/2007 PR(2) Submitted to USDA 

07-261-101n 10/18/2007 PR Submitted to USDA 

07-271-101n 10/28/2007 PR(2) Submitted to USDA 

07-312-101n 12/5/2007 PR Submitted to USDA 

 
Monsanto Company 10-SY-210U 285 of 721 

Table A-1 (continued). USDA Notifications Approved for MON 87708 and Status of 

Trials Conducted under These Notifications and Permits

USDA Number Effective Date Release Site (State) Trial Status 

2008 

07-352-101rm 3/26/2008 IA(8), IL(16), IN(4), KS(7), 

MO 

Submitted to USDA 

08-030-103n 2/29/2008 PR(2) Submitted to USDA 

08-031-105n 3/13/2008 IA(5), IL(4), KS(5) Submitted to USDA 

08-031-106n 3/1/2008 IA(2), IL(5), IN(3) Submitted to USDA 

08-039-107n 3/9/2008 IA(5), IL, IN(3), KS(5), MO Submitted to USDA 

08-043-107n 3/13/2008 IA(3), IL(10), IN, OH Submitted to USDA 

08-049-101n 3/19/2008 IL, MD, WI Submitted to USDA 

08-058-101n 3/28/2008 IA(3), IL(2), IN, MO, PA, 

WI(2) 

Submitted to USDA 

08-059-109n 3/29/2008 IA Submitted to USDA 

08-059-110n 3/29/2008 IL Submitted to USDA 

08-059-112n 3/29/2008 IN Submitted to USDA 

08-060-103n 4/2/2008 MN Submitted to USDA 

08-063-112n 4/2/2008 IA(4), IL(2), IN, MI, MO, 

NE(2) 

Submitted to USDA 

08-063-113n 4/4/2008 MN(2), ND, SD(5), WI(5) Submitted to USDA 

08-065-101n 4/4/2008 IL(2), IN Submitted to USDA 

08-064-102n 4/3/2008 PA Submitted to USDA 

08-064-103n 4/3/2008 IL Submitted to USDA 

08-064-104n 4/3/2008 AR, GA, KS(5), LA, MO, SC Submitted to USDA 

08-064-105n 4/3/2008 AR, IL(2), IN, KS(3), MD, 

MN(3), NC, SD, WI, ND 

Submitted to USDA 

08-072-110n 4/25/2008 AR, IA, IN(3), KS, MI, MO(2), 

NE 

Submitted to USDA 

08-079-101n 4/17/2008 IA(3) Submitted to USDA 

08-084-102n 4/24/2008 IA, NE Submitted to USDA 

08-182-101n 8/1/2008 PR(2) Submitted to USDA 

08-219-101n 9/5/2008 PR Submitted to USDA 

 

08-263-101n 10/19/2008 AR, IA, IL, MO Submitted to USDA 

08-266-105n 10/22/2008 PR Submitted to USDA 

 

08-323-101n 12/18/2008 PR Submitted to USDA 

08-352-108n 1/26/2009 PR Submitted to USDA 

08-357-101rm 3/17/2009 IA(8), IL(7), IN(3), KS(5), NE Submitted to USDA 
Monsanto Company 10-SY-210U 286 of 721 

Table A-1 (continued). USDA Notifications Approved for MON 87708 and Status of 

Trials Conducted under These Notifications and Permits 

USDA Number Effective Date Release Site (State) Trial Status 

2009 

09-007-106n 2/25/2009 PR Submitted to USDA 

09-036-103n 3/7/2009 IA(2), IL(2), IN, NE Submitted to USDA 

09-042-103n 3/19/2009 MS Submitted to USDA 

09-049-110n 3/20/2009 IA Submitted to USDA 

09-050-136n 4/3/2009 IA(2), IL(2), IN(2), MD, 

MN, OH, PR, SD 

Submitted to USDA 

09-061-108n 4/1/2009 AR, IA, IL(3), KS Submitted to USDA 

09-061-117n 4/1/2009 IL, MO(2) Submitted to USDA 

09-068-111n 4/8/2009 IL(2), IN(2), MS, NE(2), OH Submitted to USDA 

09-071-102n 4/11/2009 NE, SD, TN(2) Submitted to USDA 

09-082-103n 4/22/2009 IN(2) Submitted to USDA 

09-091-103n 5/1/2009 AR Submitted to USDA 

09-093-120n 5/3/2009 AR Submitted to USDA 

09-124-102n 6/3/2009 PR Submitted to USDA 

09-124-105n 5/13/2009 IA Submitted to USDA 

09-135-104n 6/14/2009 IL Submitted to USDA 

09-162-105n 7/11/2009 PR Submitted to USDA 

09-162-106n 7/11/2009 PR Submitted to USDA 

09-222-101n 9/9/2009 PR(2) Submitted to USDA 

09-237-104n 9/24/2009 PR Submitted to USDA 

09-247-101rm 11/17/2009 PR Submitted to USDA 
Monsanto Company 10-SY-210U 287 of 721 

Appendix B: Materials, Methods, and Results for Molecular Analyses of 

MON 87708 

B.1. Materials 

The genomic DNA used in molecular analyses was isolated from leaf tissue harvested 

from MON 87708 and the near isogenic conventional soybean control A3525 (seed lot: 

GLP-0707-18882-S, and GLP-0707-18884-S, respectively). Additional DNA extracted 

from leaf tissue of various MON 87708 generations was used in generational stability 

analyses. The control was conventional soybean variety A3525 that has a similar genetic 

background as MON 87708. Plasmid vector PV-GMHT4355 (Figure III-1) was used as a 

positive hybridization control in Southern blot analyses. Probe templates generated from 

PV-GMHT4355 were used as additional positive hybridization controls. As additional 

reference standards, the 1 kb DNA extension ladder and λ DNA/Hind III segments from 

Invitrogen (Carlsbad, CA) were used for size estimations on Southern blots and agarose 

gels. The GeneRulerTM 1 kb Plus DNA ladder from Fermentas (Hanover, MD) was used 

for size estimations on agarose gels for polymerase chain reaction (PCR) analyses. 

B.2. Characterization of the Materials 

The identity of the leaf material from MON 87708 and the conventional control was 

verified by event-specific PCR analysis to confirm the presence or absence of the dmo 

expression cassette. The stability of the genomic DNA was confirmed in each Southern 

blot analysis by observation of the digested DNA sample on an ethidium bromide-stained 

agarose gel, and/or interpretable signals on Southern blots, and/or produced specific PCR 

products. 

B.3. DNA Isolation for Southern Blot and PCR Analyses 

Genomic DNA from MON 87708 and the conventional control was isolated from leaf 

tissue. The leaf tissue was ground to a fine powder in liquid nitrogen using a mortar and 

pestle. DNA was extracted using a hexadecyltrimethylammonium bromide 

(CTAB)-based method. Briefly, 20 ml of CTAB buffer (1.5% w/v CTAB, 75 mM Tris 

HCl, 100 mM EDTA, 1.05 M NaCl, and 0.75% w/v PVP) and 10 mg RNase A were 

added to approximately 4 ml of ground leaf tissue and incubated at 60-70°C for 

40-50 minutes with intermittent mixing. Twenty ml of chloroform was added to the 

samples and mixed by hand for 2-3 minutes, then centrifuged at 10,300 × g for 

8-10 minutes. The upper aqueous phase was put into a clean tube and the chloroform 

step was repeated twice. After the last chloroform step, the aqueous phase was put into a 

clean tube and the DNA was precipitated with 20 ml of 100% ethanol. The sample was 

centrifuged for one minute to condense the pellet, and then the precipitated DNA was 

hooked out and put into a tube with 4-6 ml of 70% ethanol to wash the DNA pellet. The 

samples were centrifuged at 5,100 × gravity for 5 minutes to pellet the DNA. DNA 

pellets were air dried, then resuspended in 300 μl of TE buffer (10 mM Tris HCl, 1 mM 

EDTA, pH8.0). All extracted DNA was stored in a 4C refrigerator or a -20C freezer. 

 
Monsanto Company 10-SY-210U 288 of 721 

B.4. Quantification of Genomic DNA 

Extracted genomic DNA was quantified using a Hoefer DyNA Quant 200 Fluorometer. 

Molecular size marker IX (Roche, Indianapolis, IN) was used as the calibration standard. 

B.5. Restriction Enzyme Digestion of Genomic DNA 

Approximately 10 μg of genomic DNA extracted from MON 87708 and the conventional 

control was digested with appropriate combinations of restriction enzymes 

Bsp1286 I/Pvu II or Hpa I/Kpn I (New England Biolabs, Ipswich, MA). All digests were 

conducted in 1 × NEBuffer 4 (New England Biolabs) plus 1 × BSA (New England 

Biolabs) at 37 C in a total volume of ~500 μl with ~50 units of each restriction enzyme. 

For the purpose of running positive hybridization controls, ~10 μg of genomic DNA 

extracted from the conventional control was digested with the restriction enzyme 

combination Bsp1286 I/Pvu II or Hpa I/Kpn I and the appropriate positive hybridization 

control(s) were added to these digests. 

B.6. Agarose Gel Electrophoresis 

Digested genomic DNA was resolved on ~0.8% (w/v) agarose gels. Individual digests of 

MON 87708 and the conventional control genomic DNA were loaded on the same gel in 

a long-run/short-run format. The long-run allows for greater resolution of large 

molecular weight DNA, whereas the short-run allows for the detection of small molecular 

weight DNA. For the insert stability analysis, individual digests of genomic DNA 

extracted from leaf tissue across multiple generations were loaded on the agarose gel in a 

single short-run format. The positive hybridization controls were only run in the shortrun format. 

B.7. DNA Probe Preparation for Southern Blot Analyses 

Probe templates were prepared by PCR amplification from plasmid vector 

PV-GMHT4355. Approximately 25 ng of each probe template were radiolabeled with 

32P-deoxycytidine triphosphate (dCTP) (6000 Ci/mmol) or 32P-deoxyadenosine 

triphosphate (dATP) (6000 Ci/mmol) using the RadPrime DNA Labeling System 

(Invitrogen). Probe locations relative to the genetic elements in plasmid vector 

PV-GMHT4355 are depicted in Figure III-1. 

B.8. Southern Blot Analyses of Genomic DNA 

Digested genomic DNA isolated from MON 87708 and the conventional control was 

evaluated using Southern blot analyses. The plasmid vector PV-GMHT4355 DNA 

digested with the enzyme combination Aat II/Nde I was added to the conventional control 

genomic DNA previously digested with the enzyme combination Bsp1286 I/Pvu II or 

Hpa I/Kpn I to serve as a positive hybridization control. When multiple probes were 

hybridized simultaneously to one Southern blot, the appropriate probe templates 

generated from PV-GMHT4355 were mixed with previously digested conventional 

control genomic DNA to serve as additional positive hybridization controls. The 

digested DNA was then separated by agarose gel electrophoresis and transferred onto a 
Monsanto Company 10-SY-210U 289 of 721 

nylon membrane. Southern blots were hybridized and washed at 55, 60, or 65°C, 

depending on the melting temperature (Tm) of the probes. Table B-1 lists the 

radiolabeling conditions and hybridization temperatures of the probes used in this study. 

Multiple exposures of each blot were then generated using Kodak Biomax MS film in 

conjunction with one or two Kodak Biomax MS intensifying screen(s) in a -80C freezer. 

Table B-1. Hybridization Conditions of Utilized Probes 

Probe DNA Probe 

Element Sequence 

Spanned by DNA Probe 

Probe labeled 

with dNTP 

(

32P) 

Hybridization 

Temperature 

(C) 

1 Backbone Probe Backbone sequence dCTP 60 

2 Backbone Probe Backbone sequence dCTP 60 

3 Backbone Probe Backbone sequence dCTP 60 

4 T-DNA II Probe T-E9, and CS-cp4 epsps

(portion) dATP 55 

5 T-DNA II Probe CS-cp4 epsps (portion), 

and TS-CTP2 (portion) dCTP 60 

6 T-DNA II Probe TS-CTP2 (portion), 

L-DnaK, P-FMV dATP 55 

7 Backbone Probe Backbone sequence dCTP 60 

8 T-DNA I Probe B-Right Border, P-PC1SV, 

L-TEV, TS-RbcS dATP 60 

9 T-DNA I Probe TS-RbcS (portion), 

CS-dmo, T-E9 (portion) dCTP 65 

10 T-DNA I Probe T-E9, and B-Left Border dATP 55 
Monsanto Company 10-SY-210U 290 of 721 

B.9. DNA Sequence Analyses of the Insert 

Overlapping PCR products that span the insert and adjacent 5′ and 3′ flanking DNA 

sequences in MON 87708 (Figure B-1) were generated. These products were sequenced 

to determine the nucleotide sequence of the insert in MON 87708 as well as the 

nucleotide sequence of the DNA flanking the 5′ and 3′ ends of the insert. 

The PCR analyses were conducted using 50 ng of genomic DNA template in a 25 μl 

reaction volume containing a final concentration of 1 M betaine, 1 mM MgSO4, 0.8 μM 

of each primer, 0.2 mM of each dNTP, and 0.5 units of KOD Hot Start DNA polymerase 

(Novagen, Madison, WI). The amplification of Product A (Figure B-1) was performed 

under the following cycling conditions: one cycle at 94C for 2 minutes; 35 cycles at 

94C for 45 seconds, 60.2C for 45 seconds, 72C for 5 minutes; one cycle at 72C for 

10 minutes. The amplification of Product B (Figure B-1) was performed under the 

following cycling conditions: one cycle at 94C for 2 minutes; 35 cycles at 94C for 

45 seconds, 60.8C for 45 seconds, 72C for 5 minutes; one cycle at 72C for 10 minutes. 

Following PCR amplification, exonuclease I (Exo; US Biochemicals, Cleveland, OH)/ 

shrimp alkaline phosphatase (SAP; US Biochemicals) purification of the PCR products 

used for sequencing was performed in a 21 μl reaction volume containing 15 μl of the 

PCR product and a final concentration of 0.1 units/μl of Exo and 0.1 units/μl of SAP. 

The reaction was incubated at 37oC for 15 minutes, followed by 80oC for an additional 

15 minutes. 

Prior to sequencing, aliquots of untreated and Exo/SAP treated PCR product were 

separated on 0.8% (w/v) agarose E-gels (Invitrogen) and visualized by ethidium bromide 

staining to verify that the products were of the expected size. The PCR products were 

sequenced using multiple primers including primers used for PCR amplification and 

primers designed internal to the amplified sequences. All sequencing was performed by 

the Monsanto Genomics Sequencing Center using BigDye terminator chemistry (ABI, 

Foster City, CA). 

B.10. PCR and DNA Sequence Analysis to Examine the MON 87708 Insertion Site 

To examine the insertion site of conventional soybean and MON 87708, PCR analysis 

was performed on genomic DNA from both MON 87708 and the conventional control 

(Figure B-2). The primers used in this analysis were designed from the genomic DNA 

sequences flanking the insert in MON 87708. One primer designed from the genomic 

DNA sequence flanking the 5′ end of the insert was paired with a second primer located 

in the genomic DNA sequence flanking the 3′ end of the insert. 

The PCR analysis was conducted using approximately 50 ng of genomic DNA template 

in a 25 μl reaction volume containing a final concentration of 1 M betaine, 1 mM MgSO4, 

0.8 μM of each primer, 0.2 mM of each dNTP, and 0.5 units of KOD Hot Start DNA 

polymerase (Novagen). The amplification of the product was performed under the 

following cycling conditions: one cycle at 94C for 2 minutes; 35 cycles at 94C for 

45 seconds, 60.2C for 45 seconds, 72C for 5 minutes; one cycle at 72C for 10 minutes. 
Monsanto Company 10-SY-210U 291 of 721 

Following PCR amplification, Exo/SAP purification of the PCR products used for 

sequencing was performed in a 21 μl reaction volume containing 15 μl of the PCR 

product and a final concentration of 0.1 units/μl of Exo and 0.1 units/μl of SAP (U.S. 

Biochemicals). The reaction was incubated at 37oC for 15 minutes, followed by 80oC for 

an additional 15 minutes. 

Prior to sequencing, aliquots of untreated and Exo/SAP treated PCR product were 

separated on 0.8 % (w/v) agarose E-gels (Invitrogen) and visualized by ethidium bromide 

staining to verify that the products were of the expected size prior to sequencing. The 

PCR products were sequenced using multiple primers, including primers used for PCR 

amplification and primers designed internal to the amplified sequences. All sequencing 

was performed by the Monsanto Genomics Sequencing Center using BigDye terminator 

chemistry. 
Monsanto Company 10-SY-210U 292 of 721 

Figure B-1. Overlapping PCR Analysis Across the Insert in MON 87708 

PCR analyses were performed on MON 87708 genomic DNA extracted from leaf (Lanes 3 and 

6). Lanes 2 and 5 contain reactions with conventional control DNA while lanes 4 and 7 are 

reactions containing no template DNA. Lanes 1 and 8 contain Fermentas GeneRulerTM 1 kb Plus 

DNA Ladder. Lanes are marked to show which product has been loaded and is visualized on the 

agarose gel. The expected product size for each amplicon is provided in the illustration of the 

insert in MON 87708 that appears at the bottom of the figure. Five microliters of each of the 

PCR products was loaded on the gel. This figure is representative of the data generated; however, 

the specific bands from this gel were not excised and sequenced. 

Lane: 

1. GeneRulerTM 1 kb Plus DNA Ladder 5. Conventional control DNA 

2. Conventional control DNA 6. MON 87708 genomic DNA 

3. MON 87708 genomic DNA 7. No template DNA control 

4. No template DNA control 8. GeneRulerTM 1 kb Plus DNA Ladder 

Arrows denote sizes of DNA, in kilobase pairs, obtained from molecular weight markers on 

ethidium bromide stained gel. 

1 2 3 4 5 6 7 8 

5.0

4.0

3.0

2.0

5.0

4.0

3.0

2.0B-Right Border

P-PC1SV

CS-dmo

T-E9

B-Left Border

TS-RbcS

5′ Flank 3′ Flank

Product B: ~ 3.0 kb

Product A: ~ 3.2 kb

Product A Product B

L-TEV
Monsanto Company 10-SY-210U 293 of 721 

Figure B-2. PCR Amplification of the MON 87708 Insertion Site 

Depiction of the MON 87708 insertion locus in conventional control and MON 87708. PCR 

amplification was performed using Primer A in the 5′ flanking sequence and Primer B in the 3′

flanking sequence of the insert in MON 87708 to verify to examine the insertion site in 

conventional soybean and MON 87708. 

Lane Description 

 1 GeneRulerTM 1 kb Plus DNA Ladder 

2 Conventional control DNA 

 3 MON 87708 genomic DNA 

 4 No template DNA control 

5 GeneRulerTM 1 kb Plus DNA Ladder 

Arrows denote sizes of DNA, in kilobase pairs, obtained from molecular weight markers on 

ethidium bromide stained gel. 
Monsanto Company 10-SY-210U 294 of 721 

Appendix C: Materials, Methods, and Results for the Characterization of 

MON 87708 DMO and Substrate Specificity of DMO 

C.1. Characterization of MON 87708 DMO 

C.1.1. Materials 

MON 87708 DMO (lot 11261646) was purified from defatted soybean flour as described 

in C.3. As described in Section V, MON 87708 produces two forms of DMO. In 

MON 87708, the DMO trimer can be comprised of DMO, DMO+27, or a combination of 

both. Therefore, this document will refer to both forms of the protein and all forms of the 

trimer as the MON 87708 DMO. The identity of the MON 87708 harvested seed 

processed to make the defatted soybean flour was confirmed by event-specific 

polymerase chain reaction (PCR) and a copy of the verification of identity is archived in 

the Monsanto Regulatory archives with the records documenting protein isolation. The 

purified MON 87708 DMO was stored in a -80 oC freezer in a buffer solution containing 

50 mM potassium phosphate, pH 8.0, 100 mM NaCl, 1 mM dithiothreitol (DTT) and 

5% glycerol. The records describing the purification of the MON 87708 DMO are 

archived in the Monsanto Regulatory archives under Orion lot 11261646. 

C.1.2. Description of Assay Control 

Protein molecular weight standards (SeeBlue® Plus2 Pre-stained, Invitrogen,) were used 

to calibrate SDS-PAGE gels and verify protein transfer to polyvinylidene difluoride 

(PVDF) membranes. The broad range SDS-PAGE molecular weight standards (Bio-Rad, 

Hercules, CA) were used to determine the apparent molecular weight of both forms of the 

MON 87708 DMO protein. A peptide mixture (SequazymeTM Peptide Mass Standards 

kit, Applied Biosystems, Foster City, CA) was used to calibrate the MALDI-TOF mass 

spectrometer for tryptic mass and intact mass analysis. Transferrin provided with the kit 

(GE Healthcare, Piscataway, NJ) was used as a positive control for glycosylation 

analysis. 

C.1.3. MON 87708 DMO Purification 

MON 87708 DMO was purified from defatted flour processed from harvested seed of 

MON 87708. MON 87708 DMO was purified using a combination of extraction, 

filtration and diafiltration, and various chromatographic separations. A brief description 

of the purification process is below. 

Defatting of seed from MON 87708 was completed at Pilot Plant Corporation in 

Saskatoon, Canada. The seed was cracked, dehulled, and ground to meal in the presence 

of dry ice. The meal was then solvent extracted, dried, and shipped to Monsanto and 

stored in a -20°C cold room. 

Aliquots of the defatted flour were used as starting material in the purification process. 

Approximately 7.5 kg of defatted MON 87708 flour were extracted with 75 liters (L) of 

extraction buffer (25 mM potassium phosphate, pH 7.2, 10 mM MgCl2, 1 mM DTT, 
Monsanto Company 10-SY-210U 295 of 721 

1 mM benzamidine-HCl, 1 mM phenylmethylsulfonyl fluoride (PMSF), 1 μM E-64, and 

0.1 μM bestatin). The extraction was conducted at room temperature (RT) for 2 hours 

using a Lightnin® mixer with slow stirring (Graham Transmissions Inc, Menomonee 

Falls, WI). The resulting slurry was filtered using an Ertel Alsop filter press (Kingston, 

NY) with Die 42 micro media filter pads and a Cuno filter (Hagedorn and Gannon Co., 

Inc) after the addition of 7.5 kg of diatomaceous earth (5.6 kg fine hy-flo (Celite 

Corporation, Lompoc, CA) and 1.9 kg Celite 560 coarse (Sigma-Aldrich, St Louis, MO)). 

The pads on the press were pre-coated with 1.8 kg of fine hy-flo prior to the filtration of 

the extract. After washing the press with an additional volume of extraction buffer, the 

filtrate was collected (final volume: 150 L). 

The filtrate was then concentrated at RT to 75 L using a hollow fiber cartridge with a 

30,000 kDa molecular weight cutoff (MWCO) (GE Healthcare) at RT to remove small 

molecules. Solid KCl was added to a final concentration of 0.15 M. The concentrated 

filtrate was dialfiltered with four exchanges of 25 L each of a phenyl sepharose 

equilibration buffer (25 mM potassium phosphate, pH 8.0, 1 mM DTT, 

1 mM benzamidine-HCl, 1 mM PMSF, 1 μM E-64, 0.1 μM bestatin, and 0.15 M KCl). 

The first chromatographic step was performed at RT. A 30 L phenyl sepharose (GE 

Healthcare) column equilibrated with phenyl sepharose equilibration buffer was charged 

with the dialfiltered extract and then washed with three column volumes (CV) of the 

phenyl sepharose equilibration buffer. A single CV of elution buffer 

(50 mM triethanolamine, pH 8.0, 1 mM DTT, 1 mM benzamidine-HCl, 1 mM PMSF, 

1 μM E-64, 0.1 μM bestatin, and 100 μM dicamba) was loaded onto the column, the flow 

stopped and the column incubated for 1 hour. The released proteins were eluted with an 

additional CV of elution buffer and stored at 4°C. 

Solid potassium phosphate was added to the phenyl column elution to a final 

concentration of 25 mM and the pH adjusted to 8.0, followed by the addition of 

1 mM DTT, 1 mM benzamidine-HCl, 1 mM PMSF, 1 μM E-64, and 0.1 μM bestatin. A 

3 L ceramic hydroxyapatite column (CHT) (Bio-Rad) was packed at 4 °C and 

equilibrated in a buffer containing 25 mM potassium phosphate, pH 8.0, 1 mM DTT, 

1 mM benzamidine-HCl, 1 mM PMSF, 1 μM E-64, 0.1 μM bestatin, and 

100 μM dicamba. Half of the adjusted phenyl elution was charged on the CHT column. 

The column was washed with two CV of the CHT equilibration buffer. The bound 

proteins were then eluted with 400 mM potassium phosphate, pH 8.0. The flow-through 

containing the MON 87708 DMO, detected by immunoblot analysis, was collected. The 

eluted fractions without the MON 87708 DMO were discarded and the column was reequilibrated. The second half of the phenyl elution was processed with the CHT column 

in the same manner as the first half. The flow-through collected from each CHT column 

run was combined into a single pool. 

Before charging onto the next column, fresh, solid DTT and protease inhibitors were 

added to the CHT column flow-through pool. The flow-through pool from the CHT step 

was then charged on a 5 L DEAE macroprep (Bio-Rad) column at 4°C and equilibrated 

in a buffer containing 25 mM Tris-HCl, pH 8.0, 1 mM DTT, 1 mM benzamidine-HCl, 

1 mM PMSF, 1 μM E-64, 0.1 μM bestatin, and 100 μM dicamba. The DEAE column 
Monsanto Company 10-SY-210U 296 of 721 

was then washed with five CV of the DEAE equilibration buffer followed by five CV of 

the equilibration buffer plus 100 mM NaCl. The MON 87708 DMO was eluted with a 

20 CV linear NaCl gradient from 100 mM to 350 mM in the equilibration buffer. The 

fractions collected throughout the gradient were analyzed by immunoblot and those 

fractions containing the MON 87708 DMO were pooled. 

To concentrate the DEAE macroprep pool, it was first diluted with the DEAE 

equilibration buffer (to reduce the conductivity) and then charged onto a 1 L DEAE 

macroprep column. After charging, the column was washed with three CV of 

equilibration buffer and then eluted with minimal volume of the equilibration buffer plus 

1 M NaCl. This concentrated the DEAE macroprep pool from 16 to 1.6 L. 

The concentrated DEAE macroprep pool was mixed with 1 L of concanavalin A (Con A) 

sepharose 4B (Sigma-Aldrich) that was previously equilibrated with the DEAE 

equilibration buffer with fresh, solid DTT and protease inhibitors added. The purification 

step was run in batch mode at RT and was intended to remove contaminants that bind to 

Con A, while not binding MON 87708 DMO. The concentrated DEAE macroprep pool 

was stirred Con A resin for 1 hour, the resin was filtered out using a Büchner funnel and 

Whatman® filter paper (GE Healthcare). The resin was washed with 3 L of equilibration 

buffer. All filtrates containing MON 87708 DMO were combined. 

The Con A filtrate pool was concentrated on ice for approximately 4 hours using a 

tangential flow membrane (Sartorius-Stedim, Goettingen, Germany) with a 

100 kDa MWCO. After a 10× concentration step, the retentate containing MON 87708 

DMO was dialfiltered with 10 volume exchanges of DEAE macroprep equilibration 

buffer containing 1 mM DTT, 1 mM benzamidine-HCl, 1 mM PMSF, 1 μM E-64, and 

0.1 μM bestatin. 

The concentrated and dialfiltered Con A pool was further purified on CHT at RT, this 

time in a binding mode where MON 87708 DMO was bound to the resin. This is 

achieved in the complete absence of phosphate where MON 87708 DMO binds to the 

CHT column and is then eluted. A 1 L CHT column was packed and equilibrated with 

the DEAE macroprep equilibration buffer with fresh DTT and protease inhibitors. The 

column was washed with three CV of equilibration buffer. The protein was eluted with a 

linear phosphate gradient using an elution buffer (400 mM potassium phosphate, pH 8.0, 

1 mM DTT, 1 mM benzamidine-HCl, 1 mM PMSF, 1 μM E-64, 0.1 μM bestatin, and 

100 μM dicamba) increasing from 0% to 50% over 10 CV. The fractions were collected 

and analyzed by SDS-PAGE. Those containing at least 80% pure MON 87708 DMO as 

estimated by gel densitometry were pooled. 

This entire purification procedure was repeated with two additional batches of 7.5 kg of 

defatted flour from MON 87708. After analysis, all final CHT pools were combined into 

a single final pool that was concentrated on ice for approximately 2 hours to 370 ml with 

a tangential flow membrane with a 30 kDa MWCO. The concentrated pool was dialyzed 

against enzyme storage buffer (50 mM potassium phosphate, pH 8.0, 100 mM NaCl, 

5% glycerol, and 1 mM DTT). Four liters of storage buffer were used and exchanged 
Monsanto Company 10-SY-210U 297 of 721 

twice over two days and the dialysis was conducted at 4°C. The dialysate was aliquoted, 

assigned APS lot 11261646 and stored at -80°C. 

C.1.4. Molecular Weight and Purity Estimation using SDS-PAGE Method 

SDS-PAGE analysis was performed to determine the molecular weight and purity of 

MON 87708 DMO. 

An aliquot of MON 87708 DMO was mixed with 5 × loading buffer (LB) to a final total 

protein concentration of 0.09 μg/μl and heated at 99°C for three minutes. A molecular 

weight marker (Broad Range MW Marker, Bio-Rad) was diluted to a final total protein 

concentration of 0.9 μg/μl. MON 87708 DMO was loaded in duplicate at 0.5, 1.0 and 

1.5 μg of total protein per lane onto a pre-cast Tris glycine 4-20% polyacrylamide 

gradient 10-well gel (Invitrogen). The molecular weight markers were loaded in parallel 

at 4.5 μg protein per lane. Electrophoresis was performed at a constant 125 V for 

90 minutes. Proteins were fixed by placing the gel in a solution of 40% (v/v) methanol 

and 7% (v/v) acetic acid for 30 minutes, stained for 16 hours with Brilliant Blue G 

Colloidal stain (Sigma-Aldrich), destained 30 seconds with a solution containing 10% 

(v/v) acetic acid and 25% (v/v) methanol, and finally destained with 25% (v/v) methanol 

for 6 hours. Analysis of the gel was performed using a Bio-Rad GS-800 densitometer 

with the supplied Quantity One® software (version 4.4.0). Molecular weight markers 

were used to estimate the apparent molecular weight of each observed band. All visible 

bands within each lane were quantified using Quantity One software. Apparent 

molecular weight was obtained for both forms of the MON 87708 DMO protein while the 

purity was calculated based on the addition of the average purity of both proteins. The 

results were reported as an average of all six samples loaded onto the gel containing 

MON 87708 DMO. 

C.1.5. Immunoblot Analysis Method 

Immunoblot analysis was performed to confirm the identity of MON 87708 DMO. 

An aliquot of MON 87708 DMO was diluted with water and mixed with 5 × LB 

(312 mM Tris-HCl, 20% (v/v) 2-mercaptoethanol, 10% (w/v) SDS, 0.025% (w/v) 

bromophenol blue, and 50% (v/v) glycerol, pH 6.8), heated at 99°C for 3 minutes, and 

applied on a pre-cast Tris glycine 4- 20% polyacrylamide gradient 10-well gel 

(Invitrogen). Three amounts (20, 30, and 40 ng) of MON 87708 DMO were loaded in 

duplicate on the gel. Electrophoresis was performed at a constant 125 V for 90 minutes. 

Pre-stained molecular weight markers (SeeBlue® Plus2 Pre-stained, Invitrogen) were 

loaded in parallel to verify electrotransfer of the proteins to the membrane and estimate 

the size of the immunoreactive bands observed. Electrotransfer to a 0.45 μm PVDF 

membrane (Invitrogen) was performed for 90 minutes at a constant 25 V. 

For immunodetection, the membrane was blocked for 1 hour with 10% (w/v) Non-Fat 

Dried Milk (NFDM) in 1× Phosphate Buffered Saline containing 0.05% (v/v) Tween-20 

(PBST). The membrane was then probed with a 1:3,000 dilution of goat anti-DMO 

antibody, which is specific for both forms of the MON 87708 DMO protein, in 5% (w/v) 
Monsanto Company 10-SY-210U 298 of 721 

NFDM in PBST for one hour. Excess antibody was removed using three 10 minutes 

washes with PBST. Finally, the membrane was probed with horseradish peroxidase 

(HRP)-conjugated rabbit anti-goat IgG (Thermo, Rockford, IL) at a dilution of 1:10,000 

in 5% (w/v) NFDM in PBST for 1 hour. Excess HRP-conjugate was removed using three 

10 minutes washes with PBST. All incubations were performed at RT. Immunoreactive 

bands were visualized using the ECL (Enhanced Chemiluminescence) detection system 

(GE Healthcare) and exposed to Amersham Hyperfilm (GE, Healthcare). The film was 

developed using a Konica SRX-101A automated film processor (Tokyo, Japan). Three 

exposures (20, 30, and 60 seconds) were taken and the 20 second exposure was scanned 

using a Bio-Rad GS-800 densitometer with the supplied Quantity One software (version 

4.4.0). 

C.1.6. MALDI-TOF Tryptic Mass Map Analysis Method 

MALDI-TOF mass spectrometry was used to confirm the identity of both forms of the 

MON 87708 DMO proteins. The proteins were first separated by SDS-PAGE prior to 

trypsinization. 

An aliquot (89.5 μl) of MON 87708 DMO was mixed with 22.5 μl of 5 × LB, heated at 

99°C for 3 minutes and loaded in four lanes (three lanes each loaded with 4.2 μg and one 

lane with 3.1 μg of total protein) onto a pre-cast Tris glycine 4-20% polyacrylamide 

gradient 10-well gel (Invitrogen). Pre-stained MW markers (SeeBlue Plus2 Pre-stained, 

Invitrogen) were loaded in parallel to estimate the size of the stained bands observed. 

Electrophoresis was carried out at a constant 150 V for 80 minutes. Following 

electrophoresis, the gel was stained with Brilliant Blue G Colloidal (Sigma-Aldrich). The 

bands corresponding to DMO or DMO+27 were excised from four lanes of the gel, 

destained, reduced, and alkylated. Each gel band was destained for 30 minutes by 

incubation in 100 μl of destain solution (40% methanol, 50% water, and 10% glacial 

acetic acid) in a microfuge tube. This step was repeated twice for 60 minutes each, 

removing all visible Brilliant Blue G Colloidal stain. Following destaining, the gel bands 

were incubated in 100 μl per band of 100 mM ammonium bicarbonate buffer for 16 hours 

at RT. The protein was reduced in 100 μl of 10 mM DTT solution for two hours at 

37 °C. After removing the reducing solution, the protein in the gel was alkylated by 

incubating in 100 μl of 20 mM iodoacetic acid. The alkylation reaction was allowed to 

proceed at RT for 20 minutes in the dark. The gel containing the protein band was 

incubated in 200 μl of 25 mM ammonium bicarbonate buffer for 15 minutes at RT. This 

step was repeated two additional times for 15 minutes each; then the gel band was dried 

using a Savant Speed Vac concentrator (Holbrook, NY). Each gel band was rehydrated 

with 20 μl of 0.02 μg/μl trypsin in 25 mM ammonium bicarbonate and 10% acetonitrile, 

and the incubated for approximately one hour at RT. Following incubation, the excess 

solution was removed and the gel/trypsin reaction mixture was incubated overnight at 

37°C in 40 μl of 25 mM ammonium bicarbonate and 10% acetonitrile. The following 

day, the sample was sonicated for 5 minutes, and the supernatant transferred to a new 

tube and dried using a Speed Vac concentrator (Extract 1). The gel band(s) was 

resuspended in 30 μl of a solution consisting of 60% acetonitrile, 0.1% trifluoroacetic 

acid (TFA), and 0.1% octyl-β-D-glucopyranoside, and then sonicated for 5 minutes. 

After transfer of the supernatant to a new microcentrifuge tube, this step was repeated 
Monsanto Company 10-SY-210U 299 of 721 

once and the combined supernatants were dried using the Speed Vac concentrator 

(Extract 2). Extracts 1 and 2 were separately dissolved in 20 μl 0.1% TFA and then dried 

using a Speed Vac concentrator. Finally, Extract 1 was dissolved in 5 μl of 

50% acetonitrile/0.1% TFA, while Extract 2 was dissolved in 10 μl of the same solution. 

To maximize the solubilization, each sample was sonicated for 5 minutes. 

Mass calibration of the mass spectrometer was performed using an external peptide 

mixture (SequazymeTM Peptide Mass Standards Kit, Calibration Mixture 2, Applied 

Biosystems). The samples Extract 1 and Extract 2 (0.3 μl) were co-crystallized with 

0.75 μl each of the following matrix solutions: dihydroxybenzoic acid (DHB), α-cyano-4-

hydroxy cinnamic acid (α-cyano), and 3,5 dimethoxy-4 hydroxycinnamic acid (sinapinic 

acid) at separate locations on the analysis plate. The samples in DHB matrix were 

analyzed in the 300 to 5,000 Dalton (Da) range. The samples in α-cyano matrix were 

analyzed in the 500 to 5,000 Da range. The samples in sinapinic acid matrix were 

analyzed in the 500 to 7,000 Da range. Protonated (MH+) peptide masses were observed 

monoisotopically in reflector mode (Aebersold, 1993), except above 3,000 Da, where 

mass-averaged values were used. GPMAW32® software (Lighthouse data, Denmark) 

was used to generate a theoretical trypsin digestion of the deduced DMO and DMO+27 

amino acid sequences. Masses were calculated for each theoretical peptide and compared 

to the raw experimental mass data. Below 1000 Da, experimental masses (MH+) were 

assigned to peaks when two or more isotopically resolved peaks were observed. Above 

1000 Da, experimental masses (MH+) were assigned to peaks when three or more 

isotopically resolved peaks were observed. Peaks were not assessed if the peak heights 

were less than approximately twice the baseline noise, or when a mass could not be 

assigned due to overlap with a stronger mass signal. Known autocatalytic segments from 

trypsin digestion were identified in the raw data. The list of experimental masses was 

compared to the theoretical list from the GPMAW software. Those experimental masses 

within 1 Da of a theoretical mass were matched. All matching masses were tallied and a 

coverage map was generated. The tryptic mass map coverage was considered 

acceptable if ≥ 40% of the protein sequence was identified by matching experimental 

masses observed for the tryptic peptide segments to the expected masses for the 

segments. 

C.1.7. N-Terminal Sequencing Method 

N-terminal sequencing using automated Edman degradation chemistry (Hunkapiller et 

al., 1983) was used to confirm the identity of both forms of the MON 87708 DMO 

proteins. 

Ninety microliters of MON 87708 DMO were mixed with 22.5 μl of 5× LB, heated at 

99°C for 3 minutes and loaded in four lanes (10 μl/lane) onto a pre-cast Tris glycine 

4-20% polyacrylamide gradient 10-well gel (Invitrogen). Electrophoresis was carried out 

at a constant voltage of 150 V for 80 minutes. Proteins in the gel were electrotransferred 

to a PVDF (Invitrogen) membrane for 90 minutes in a buffer containing 10 mM CAPS, 

pH 11 and 10% methanol at a constant voltage of 25 V. Pre-stained molecular weight 

markers (SeeBlue Plus2 Pre-stained, Invitrogen) were loaded in parallel to verify the 
Monsanto Company 10-SY-210U 300 of 721 

electrotransfer of protein to the membrane and estimate the size of the stained bands 

observed. The blot was stained with Ponceau S (Sigma-Aldrich). 

Following electrotransfer and staining, the bands corresponding to DMO and DMO+27 

were excised based on apparent molecular weight from the blot and N-terminal sequence 

analyses were performed for 15 cycles using automated Edman degradation chemistry 

(Hunkapiller et al., 1983). An Applied Biosystems 494 ProciseTM Protein Sequencing 

System with 140C Microgradient HPLC pump, ABI 785A Programmable Absorbance 

Detector and ProciseTM Control Software (version 2.1) were used. Chromatographic data 

were collected using AtlasTM 2003 software (Thermo Fisher Scientific Inc, Waltham, 

MA). A PTH (Phenylthiohydantoin) -amino acid standard mixture (Applied Biosystems) 

was used as the calibration standard in the chromatographic analysis. This mixture 

served to verify system suitability criteria such as percent peak resolution and relative 

amino acid chromatographic retention times. A control protein, 10 pmol β lactoglobulin 

(Applied Biosystems), was analyzed before and after the analysis to verify that the 

sequencer met performance criteria for repetitive yield and sequence identity. 

C.1.8. Glycosylation Analysis Method 

Glycosylation analysis was used to determine whether either form of the MON 87708 

DMO proteins were post-translationally modified with covalently bound carbohydrate 

moieties. 

An aliquot of MON 87708 DMO and the positive control, transferrin (GE Healthcare) 

were each diluted with water and mixed with 5 × LB. These samples were heated at 

101.0 °C for three minutes, cooled, and loaded on a Tris glycine 4-20% polyacrylamide 

gradient 10-well mini-gel (Invitrogen). Three amounts of transferrin (50, 100, and 

200 ng) and two amounts (100 and 200 ng) of the purity-corrected DMO enzyme was 

loaded in the gel. SeeBlue® Plus2 Pre-stained protein molecular weight markers 

(Invitrogen) were loaded to verify electrotransfer of the proteins to the membrane. 

Electrophoresis was performed at a constant 150 V for 87 minutes. Electrotransfer to a 

0.45 μm PVDF membrane (Invitrogen) was performed for 60 minutes at a constant 25 V, 

followed by 30 minutes at 30 V. 

Carbohydrate detection was performed directly on the PVDF membrane using the GE 

Healthcare Glycosylation Detection Module (Cat. No. RPN 2190). The manufacturer’s 

protocol was followed and all the reagents except phosphate buffered saline (PBS) were 

provided with the kit. All steps were performed at RT. Following electrotransfer to 

PVDF membrane, the blot was incubated in 30 ml of PBS for 10 minutes, followed by 

incubation with 20 ml of 10 mM NaIO4 for 20 minutes in the dark. The membrane was 

rinsed twice with PBS and washed three times with 20 ml PBS for 10 minutes each. The 

membrane was incubated with 20 ml solution consisting of 0.125 mM biotin-hydrazide, 

100 mM acetate, pH 5.5 for 60 minutes followed by two PBS rinses and three 10 minute 

washes with PBS. The membrane was blocked for 60 minutes using 5% blocking reagent 

in PBS followed by two PBS rinses and three 10 minute washes with PBS. The 

membrane was incubated with strepavidin-HRP at 1:6000 dilution for 30 minutes. After 

two PBS rinses and three 10 minute washes with PBS, the membrane was developed with 
Monsanto Company 10-SY-210U 301 of 721 

ECL detection reagents by mixing 1 ml of Reagent 1 and 1 ml of Reagent 2. After 

one minute incubation, the excess detection solution was removed by blotting with paper 

towels and the blot was exposed to Hyperfilm ECL (GE Healthcare). The film was 

developed using a Konica SRX-101A automated film processor (Tokyo, Japan). Three 

exposures (30 seconds, 1 and 2 minutes) were performed. The image was captured using 

a Bio-Rad GS-800 densitometer with the supplied Quantity One® software (version 

4.4.0). 

C.1.9. Functional Activity Analysis Method 

The specific activity of MON 87708 DMO was determined by quantifying the 

conversion of 3,6-dichloro-2-methoxybenzoic acid (dicamba) to 3,6 dichlorosalicylic acid 

(DCSA) via HPLC (Agilent Technologies 1100 series, Santa Clara, CA) separation and 

fluorescence detection (Agilent Technologies 1200 series, G1321A). The standard assays 

were conducted in 200 μl solutions consisting of 25 mM potassium phosphate, pH 7.2, 

3.4 μg Ferredoxin, 3.4 μg Reductase, 0.5 mM FeSO4, 10 mM MgCl2, 0.7 mM NADH, 

0.3 mM dicamba, 2 μl (42.48 U/ml) of formaldehyde dehydrogenase and either 2 μg 

MON 87708 DMO or 1 μg HIS-DMO as an assay positive control. The reactions were 

performed in PCR tubes (Sorenson, Salt Lake City, UT) and incubated at 30°C for 

15 minutes. Reactions were initiated by the addition of dicamba and quenched with the 

addition of 50 μl of 5% H2SO4. Reactions were then filtered using Whatman Anotop 10 

filters (0.2 μm, GE healthcare), and 40 μl was transferred to a HPLC sample vial (200 μl, 

Agilent) for analysis. Twenty-five microliters of the filtered reaction was injected onto a 

Phenomenex® Synergi 4 μm C18/ODS Hydro-RP column (150 x 4.6 mm ID, Torrance, 

CA). The mobile phase consisted of solvent A (21.5 mM phosphoric acid) and solvent B 

(100% acetonitrile) running at 1.5 ml/min. DCSA was eluted from the column using a 

linear gradient from 90% to 40% solvent A for the first 14 minutes, followed by a step to 

10% solvent A for 1 minute and then re-equilibration at 90% solvent A for 10 minutes 

before the next injection. DCSA was monitored by the detection of fluorescent emission 

at 424 nm (excitation 306 nm) and quantified relative to a standard curve of DCSA 

generated using 0.1, 0.3, 0.6, 0.9, 1.2, 2.4, and 4.8 nmol/250 μl. Chromatographic data 

were collected using AtlasTM 2003 software (Thermo Fisher Scientific Inc). The specific 

activity was calculated based on the amount of purity corrected MON 87708 DMO added 

to the reaction mixture and expressed as nmol of DCSA produced per minute per mg of 

MON 87708 DMO (nmol/min/mg). 

C.1.10. Results of MON 87708 DMO Proteins Molecular Weight and Purity 

The apparent molecular weights of the MON 87708 DMO proteins were determined by 

using SDS-PAGE and the gel stained using Brilliant Blue G Colloidal stain (SigmaAldrich). Purity and apparent molecular weight of DMO and DMO+27 were determined 

using densitometric analysis of the gel (Figure C-1). As summarized in Table C-1, 

apparent molecular weight values were averaged from duplicated loads of 0.5, 1.0, and 

1.5 μg of total protein (Figure C-1, lanes 2-7). The predominant bands identified as 

DMO and DMO+27 were estimated to have apparent molecular weights of 39.8 kDa and 

42.0 kDa, respectively. The average purity of the combined DMO proteins was 81%.
Monsanto Company 10-SY-210U 302 of 721 

Table C-1. Molecular Weight and Purity of the MON 87708-Produced DMO 

Proteins 

Total Protein 

Loaded 

Apparent Molecular 

Weight (kDa) Purity (%) 

DMO DMO+27 DMO DMO+27 DMO 

Proteins

0.5 g in lane 3

0.5 g in lane 4

1.0 g in lane 5

1.0 g in lane 6

1.5 g in lane 7

1.5 g in lane 8

Average 























































 

The apparent molecular weight and the purity of DMO and DMO+27 were determined by 

densitometric analysis of SDS PAGE shown in Figure C-1. Final molecular weight was 

rounded to one decimal place and purity was reported as a whole number percentage. 

Purity of the MON 87708 DMO proteins equals the average purity of DMO plus the 

average purity of DMO+27. 
Monsanto Company 10-SY-210U 303 of 721 

Figure C-1. Molecular Weight and Purity Analysis of MON 87708 DMO Proteins 

An aliquot of MON 87708 DMO was separated on a 4 - 20% Tris glycine polyacrylamide 

gradient gel and then stained with Brilliant Blue G Colloidal stain. Approximate 

apparent molecular weights (kDa) are shown on the left and correspond to the markers 

loaded in Lanes 1 and 8. Empty lanes were cropped. 

Lane Sample Amount (g)

 1 Broad Range MW markers 4.5 

 2 MON 87708 DMO proteins 0.5 

 3 MON 87708 DMO proteins 0.5 

 4 MON 87708 DMO proteins 1.0 

 5 MON 87708 DMO proteins 1.0 

 6 MON 87708 DMO proteins 1.5 

 7 MON 87708 DMO proteins 1.5 

 8 Broad Range MW markers 4.5 
Monsanto Company 10-SY-210U 304 of 721 

C.11. Results of Immunoblot Analysis 

On the immunoblot, the goat anti-DMO antibody recognized two bands migrating at the 

expected apparent molecular weights of approximately 39.8 kDa (DMO) and 42.0 kDa 

(DMO+27), respectively (Figure C-2). As expected, the intensity of the immunoreactive 

bands increased with increasing amount of total protein loaded. No additional bands 

were observed. This immunoblot analysis confirmed the identity of both forms of the 

MON 87708 DMO protein. 

Figure C-2. Immunoblot Analysis of MON 87708 DMO Proteins 

An aliquot of MON 87708 DMO and molecular weight markers were separated by SDSPAGE and electrotransferred to a PVDF membrane. The membrane was incubated with 

goat anti-DMO antibody and immunoreactive bands were visualized using an ECL 

system. Approximate MWs (kDa) are shown on the left and correspond to the markers 

loaded in Lane 1. The 20 second exposure is shown. 

 Lane Sample Amount (ng)

 1 See Blue Plus2 Pre-Stained MW markers 

 2 empty 

 3 MON 87708 DMO proteins 20 

 4 MON 87708 DMO proteins 20 

 5 empty 

 6 MON 87708 DMO proteins 30 

 7 MON 87708 DMO proteins 30 

 8 empty 

 9 MON 87708 DMO proteins 40 

 10 MON 87708 DMO proteins 40 
Monsanto Company 10-SY-210U 305 of 721 

C.12. Results of MALDI-TOF Tryptic Mass Map Analysis 

The identity of both forms of the MON 87708 DMO proteins was confirmed by tryptic 

mapping using MALDI-TOF MS analysis of the fragments. Prior to analysis, DMO and 

DMO+27 were separated by SDS-PAGE, reduced, alkylated and digested with trypsin. 

The ability to identify a protein using this method is dependent upon matching a 

sufficient number of observed tryptic peptide fragment masses with predicted tryptic 

peptide fragment masses. In general, protein identification made by peptide mapping is 

considered to be reliable if the measured coverage of the sequence is 15% or higher with 

a minimum of five matched peptides (Jensen et al., 1997). 

There were 26 unique peptides identified from DMO that corresponded to the expected 

masses of the DMO trypsin-digested peptides while 29 fragments from DMO+27 were 

found to match the expected peptides (Tables C-2 and C-3, respectively). The identified 

peptides were used to assemble a coverage map indicating the matched peptide sequences 

for the entire DMO and DMO+27 protein sequences, resulting in 77.4% (263/340) and 

82.0% (301/367) coverage of the amino acid sequence, respectively (Figures C-3 and 

C-4). N-terminal peptides of both DMO and DMO+27 were identified and consistent 

with the N-terminal sequencing data that determined the N-terminal methionine was 

missing in DMO and methylated in DMO+27 (Tables C-2 and C-3; Figures C-3 and 

C-4). These results confirm the identity of both forms of the MON 87708 DMO proteins. 
Monsanto Company 10-SY-210U 

Table C-2. Summary of the Tryptic Masses Identified for the MON 87708 DMO Protein Using MALDI-T

Matrix 

Expected 

Mass1

Difference2 AA 

position3  Fragme

-Cyano DHB Sinapinic acid Ext.1 Ext.2 Ext.1 Ext.2 Ext.1 Ext.2 

593.61 720.67 833.78 856.77 914.89 

1030.96 1108.93 1276.17 1287.14 1429.18 1507.27 1578.24 1745.42 1762.48 1994.65 2143.78 2294.97 2398.86 2581.87 2700.08 

593.61 720.68 833.80 856.78 914.91 

1030.97 1108.95 1276.20 1429.20 1507.27 1578.27 1745.51 1762.48 1994.67 2143.84 2398.77 

391.34 435.38 593.51 720.60 833.74 856.72 914.84 

1030.92 1108.89 1171.08 1276.19 1287.19 1429.23 1502.35 1507.32 1578.32 1745.59 1762.62 1994.76 2143.94 

331.20 435.38 593.53 720.60 833.77 856.75 914.88 

1030.92 1108.94 1276.21 1429.26 1502.37 1578.33 

1276.19 1429.20 1502.34 1578.30 1745.56 1762.54 1994.78 2143.96 2294.93 2399.15 2582.22 2700.31 

4218.474

2143.95 

331.22 391.18 435.27 593.34 720.37 833.45 856.43 914.53 

1030.57 1108.50 1170.63 1275.73 1286.70 1428.69 1501.79 1506.73 1577.73 1744.93 1761.90 1994.03 2143.12 2294.09 2398.08 2582.34 2699.25 

4217.774

0.02 0.16 0.11 0.27 0.30 0.33 0.34 0.36 0.39 0.43 0.45 0.44 0.44 0.49 0.56 0.54 0.51 0.49 0.58 0.62 0.66 0.88 0.78 0.47 0.83 0.70 

304-305 293-295 206-208 

2-6 

131-136 

99-105 

242-248 296-303 284-292 167-176 194-205 

26-36 

293-303 209-221 180-193 167-179 270-283 225-241 

37-52 

150-166 

7-25 

306-326 249-269 225-248 106-130 

99-136 

RR EDK FLR ATFVR VDPAYR SFPVVER EQSIHSR VVVEAIER SWQAQALVK ANAQTDAFDR IPGGTPSVLMAK TILDTPLALYR EDKVVVEAIER GANTPVDAWNDIR EVIVGDGEIQALMK ANAQTDAFDRLER NFGIDDPEMDGVLR VSAMLNFIAVAPEGTPK QPDGVVAALLDICPHR LLVDNLMDLGHAQYVHR NAWYVAALPEELSEKPLGR AYVEANGIRPAMLSCDEAAVR GTHILTPETEASCHYFFGSSR VSAMLNFIAVAPEGTPKEQSIHSR DALIWICPGDPALADPGAIPDFGCSFPVVERDALIWICPGDPALADPG

1Only experimental masses that matched expected masses are listed in the table. 2The number represents the difference between the expected mass and the first column, which has the correspond3AA position refers to amino acid position within the predicted DMO sequence as depicted in Figure C-3. 4Mass average. 
Monsanto Company 10-SY-210U 

Table C-3. Summary of the Tryptic Masses Identified for the MON 87708 DMO+27 Protein Using MALD

Matrix 

Expected 

Mass1

Difference2 AA 

position3  Fragme

-Cyano DHB Sinapinic acid Ext.1 Ext.2 Ext.1 Ext.2 Ext.1 Ext.2 

720.55 795.61 833.65 856.64 914.74 

1030.80 1069.78 1108.75 1275.97 1286.95 1428.95 1507.01 1565.13 1578.02 1693.26 1745.17 1762.17 1994.34 2143.46 

720.64 795.71 833.73 856.72 

1030.89 1069.92 1108.90 1276.12 1429.10 1507.18 1565.30 1578.14 1693.38 1745.36 1762.37 1994.55 2143.63 2398.72 

435.30 720.47 795.54 833.59 856.58 914.69 

1030.75 1108.71 1275.98 1286.97 1429.00 1470.93 1507.03 1565.22 1578.06 1693.29 1745.28 1762.29 1994.48 2143.57 2398.49 2699.73 

331.19 720.55 795.66 833.69 856.64 

1170.98 1276.14 1429.26 1565.34 1578.29 1745.51 2143.98 

1275.97 1428.93 1469.94 1502.10 1578.04 1745.22 1994.42 2143.57 2294.62 2398.52 2581.78 2699.84 4215.70 

331.22 435.27 720.37 795.42 833.45 856.43 914.53 

1030.57 1069.57 1108.50 1170.66 1275.73 1286.70 1428.69 1470.63 1501.79 1506.73 1564.87 1577.73 1692.97 1744.93 1761.90 1994.03 2143.12 2294.09 2398.08 2582.34 2699.25 4215.03 

0.03 0.03 0.18 0.19 0.20 0.21 0.21 0.23 0.21 0.25 0.32 0.24 0.25 0.26 0.30 0.31 0.28 0.26 0.29 0.29 0.24 0.27 0.31 0.34 0.53 0.64 0.56 0.48 0.67 

331-332 233-235 158-163 

27-33 

126-132 269-275 323-330 311-319 

1-9 

194-203 221-232 

53-63 

320-330 236-248 164-176 207-220 194-206 

11-23 

297-310 

10-23 

252-268 

64-79 

177-193 

34-52 

333-353 276-296 252-275 133-157 126-163 

RR FLR VDPAYR AMATFVR SFPVVER EQSIHSR VVVEAIER SWQAQALVK M*QVWPPIGK ANAQTDAFDR IPGGTPSVLMAK TILDTPLALYR EDKVVVEAIER GANTPVDAWNDIR TVGGYGHVDCNYK EVIVGDGEIQALMK ANAQTDAFDRLER KFETLSYLPPLTR NFGIDDPEMDGVLR KKFETLSYLPPLTR VSAMLNFIAVAPEGTPK QPDGVVAALLDICPHR LLVDNLMDLGHAQYVHR NAWYVAALPEELSEKPLGR AYVEANGIRPAMLSCDEAAVR GTHILTPETEASCHYFFGSSR VSAMLNFIAVAPEGTPKEQSIHSR DALIWICPGDPALADPGAIPDFGCSFPVVERDALIWICPGDPALADPG

1Only experimental masses that matched expected masses are listed in the table. 2The number represents the difference between the expected mass and the first column, which has the corresponding numb3AA position refers to amino acid position within the predicted DMO+27 sequence as depicted in Figure C-4. 

*Methylated methionine.
Monsanto Company 10-SY-210U 308 of 721 

Figure C-3. MALDI-TOF MS Coverage Map of the MON 87708 DMO Protein 

The amino acid sequence of DMO was deduced from the dmo coding region present in 

MON 87708. Boxed regions correspond to tryptic peptides that were identified from DMO using 

MALDI-TOF MS. In total, 77.4% (263 of 340 total amino acids) of the expected protein 

sequence was identified. 

Figure C-4. MALDI-TOF MS Coverage Map of the MON 87708 DMO+27 Protein 

The amino acid sequence of DMO+27 was deduced from the dmo coding region and RbcS

present in MON 87708. Boxed regions correspond to tryptic peptides that were identified from 

DMO+27 using MALDI-TOF MS. In total, 82.0% (301 of 367 total amino acids) of the expected 

protein sequence was identified. 

001 MATFVRNAWY VAALPEELSE KPLGRTILDT PLALYRQPDG VVAALLDICP 

051 HRFAPLSDGI LVNGHLQCPY HGLEFDGGGQ CVHNPHGNGA RPASLNVRSF 

101 PVVERDALIW ICPGDPALAD PGAIPDFGCR VDPAYRTVGG YGHVDCNYKL 

151 LVDNLMDLGH AQYVHRANAQ TDAFDRLERE VIVGDGEIQA LMKIPGGTPS 

201 VLMAKFLRGA NTPVDAWNDI RWNKVSAMLN FIAVAPEGTP KEQSIHSRGT 

251 HILTPETEAS CHYFFGSSRN FGIDDPEMDG VLRSWQAQAL VKEDKVVVEA 

301 IERRRAYVEA NGIRPAMLSC DEAAVRVSRE IEKLEQLEAA

001 MQVWPPIGKK KFETLSYLPP LTRDSRAMAT FVRNAWYVAA LPEELSEKPL

051 GRTILDTPLA LYRQPDGVVA ALLDICPHRF APLSDGILVN GHLQCPYHGL

101 EFDGGGQCVH NPHGNGARPA SLNVRSFPVV ERDALIWICP GDPALADPGA

151 IPDFGCRVDP AYRTVGGYGH VDCNYKLLVD NLMDLGHAQY VHRANAQTDA

201 FDRLEREVIV GDGEIQALMK IPGGTPSVLM AKFLRGANTP VDAWNDIRWN

251 KVSAMLNFIA VAPEGTPKEQ SIHSRGTHIL TPETEASCHY FFGSSRNFGI

301 DDPEMDGVLR SWQAQALVKE DKVVVEAIER RRAYVEANGI RPAMLSCDEA

351 AVRVSREIEK LEQLEAA
Monsanto Company 10-SY-210U 309 of 721 

C.13. Results of N-terminal Sequencing 

N-terminal sequence analysis of the two major protein bands on the PVDF membrane 

(Figure C-2) returned a sequence of 15 amino acids per band that matched the expected 

N-terminal sequences of DMO and DMO+27 (Figures C-5 and C-6, respectively), which 

were deduced from the dmo coding region present in the seed of MON 87708 

(Section IV). The N-terminal methionine residue in DMO was not observed, indicating 

that it was removed during posttranslational processing of the protein. Removal of the 

N-terminal methionine occurs through methionine aminopeptidase (Arfin and Bradshaw, 

1988; Bradshaw et al., 1998; Polevoda and Sherman, 2000) and is common in many 

organisms. 

In the case of DMO+27, the first cycle of N-terminal sequence analysis resulted in a 

PTH-amino acid derivative that corresponds to a methylated modification of the 

N-terminal methionine. It is well-known that the amino-terminal methionine of the 

Rubisco small subunit is post-translationally modified to N-methyl-methionine in vivo in 

pea and other plant species (Grimm et al., 1997; Whitney and Andrews, 2001). 

The N-terminal sequencing results clearly confirm the identity of the DMO and DMO+27 

isolated from the seed of MON 87708. 
Monsanto Company 10-SY-210U 310 of 721 

Figure C-5. N-Terminal Sequence of the MON 87708 DMO Protein 

The expected amino acid sequence of the N-terminus of DMO was deduced from the dmo coding 

region present in MON 87708. The experimental sequence obtained from DMO was compared to 

the expected sequence. (-) indicates the residue was not observed. 

Figure C-6. N-Terminal Sequence of the MON 87708 DMO+27 Protein 

The expected amino acid sequence of the N-terminus of DMO+27 was deduced from the dmo

coding region present in MON 87708. The experimental sequence obtained from DMO+27 was 

compared to the expected sequence. M*, methylated methionine. 

Amino acid 

residue # from 

the N-terminus → 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 

Expected Sequence M A T F V R N A W Y V A A L P E 

 | | | | | | | | | | | | | | | | 

Experimental Sequence - A T F V R N A W Y V A A L P E 

Amino acid 

residue # from 

the N-terminus → 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 

Expected Sequence M Q V W P P I G K K K F E T L 

 | | | | | | | | | | | | | | | 

Experimental Sequence M*Q V W P P I G K K K F E T L 
Monsanto Company 10-SY-210U 311 of 721 

C.1.14. Results of Glycosylation Analysis 

To test whether DMO or DMO+27 was glycosylated when expressed in the seed of 

MON 87708, the MON 87708 DMO proteins were analyzed for glycosylation using a GE 

Glycoprotein Detection Module (GE healthcare). Transferrin, a naturally glycosylated 

protein, was used as a positive control in the assay. The results of this analysis are 

presented in Figure C-7. The positive control was clearly detected at the expected 

molecular weight and the bands increased with increasing protein concentration 

(Figure C-7, lanes 2-4). No bands were observed for DMO or DMO+27 at their expected 

molecular weight positions (39.8 and 42.0 kDa) (Figure C-7, lanes 5 and 6) indicating 

that both forms of the DMO protein are not glycosylated. 

Figure C-7. Glycosylation Analysis of the MON 87708 DMO Proteins 

Molecular weight markers, transferrin (positive control) and an aliquot of the MON 87708 DMO 

were separated by SDS-PAGE (4 - 20%) and electrotransferred to a PVDF membrane. The 

image was captured using a Bio-Rad GS800 with Quantity One software (version 4.4.0). The 

30 second exposure is shown. 

Lane Sample Amount (ng)

 1 See Blue Plus2 Pre-Stained MW markers 

 2 Transferrin 50 

 3 Transferrin 100 

 4 Transferrin 200 

 5 MON 87708 DMO proteins 100 

 6 MON 87708 DMO proteins 200 

 7 Empty lane 

 8 Empty lane 

Lane # 1 2 3 4 5 6 7 8

kDa

250

148

98

64

50

36

22

16

6

4

Expected position 

for DMO+27/DMO

Transferrin 
Monsanto Company 10-SY-210U 312 of 721 

C.15. Results of Functional Activity 

MON 87708 DMO activity was determined by measuring the production of DCSA. The 

specific activity was 62.21 nmol/min/mg of MON 87708 DMO (Table C-4). The value 

represents an average of three independent assays. This result demonstrates that 

MON 87708 DMO isolated from the seed of MON 87708 is functionally active. 

Table C-4. MON 87708 DMO Functional Activity Assay 

Assay# Specific activity 

(DCSA nmol/min/mg) 

Average (nmol/min/mg) 

±Standard Deviation 

1 

2 

3 

61.92 

51.33 

73.39 

62.21 ± 11.03 

C.2. Substrate Specificity of DMO 

C.2.1. Materials 

MON 87708 (lot 11225299-114) and the near isogenic conventional soybean control 

A3525 (lot 11225301-104) were used for the exogenous specificity greenhouse tests 

conducted in 2009 and 2010. At the V2-V3 growth stage, MON 87708 and the 

conventional control plants were sprayed with different herbicides. The herbicides tested 

are listed in Table C-5. 

The DMO protein used in the endogenous and exogenous specificity in vitro experiments 

was generated in Escherichia coli with a histidine-tag at the N-terminus and has an 

identical amino acid sequence to MON 87708 DMO with the exception of a single amino 

acid change at position 112 (W112C) and the lack of alanine at the second position. The 

compounds tested and standards used in the in vitro experiments are listed in Table C-6. 
Monsanto Company 10-SY-210U 313 of 721 

Table C-5. Herbicides Tested in Exogenous Specificity Herbicide Tolerance 

Greenhouse Trials 

Manufacturer/ 

Retailer 

Chemistry/ 

Compound Formulation Lot Number 

 

Agriliance MCPA MCPA L.V.Ester 4 RUD-0102-11027-F 

Albaugh 2,4-DB Butyrac® 200 HPR-0404-14987-F

BASF dicamba Clarity® KIH-0702-18134-F 

BASF imazapyr Arsenal® KIH-0408-15423-F 

Dow picloram Tordon® 22K ABR-9912-99121-F 

Dow clopyralid Stinger® AGD-0104-11295 

Dow triclopyr Garlon® 3A AGD-0205-12641 

Dow atrazine Atrazine® AGT-0804-19336-F 

Dow trifluralin Treflan® MB231656T7 

Dow oxyfluorfen Goal® 2XL EWP-0107-11628-F 

DuPont chlorimuron Classic® MPO-9304-19319 

DuPont chlorsulfuron Glean® MPO-9910-9869 

DuPont linuron Lorox® NIF-0103-1173-F 

Helena 2,4-D 2,4-D Amine 4 RUD-0502-15805-F 

Monsanto alachlor Lasso® MUS-0905-19849-F 

Monsanto acetochlor Harness® MUS-0704-18520-F 

Monsanto glyphosate Roundup WeatherMax® MUS-0905-19887-F 

Monsanto halosulfuron Permit® MUS-0405-15154-F

Syngenta paraquat Gramoxone® GTA-0606-17421 

Valent lactofen Cobra® LVT-0905-19884-F 

 

® Roundup WeatherMax is a trademark of Monsanto Technology LLC. All other trademarks are the 

property of their respective owners. 
Monsanto Company 10-SY-210U 314 of 721 

Table C-6. Compounds Used in Specificity In Vitro Experiments 

C.2.2. Exogenous Specificity Herbicide Tolerance Greenhouse Method 

MON 87708 and the conventional control were planted in pots containing Redi-earth®

and Osmocote® 14-14-14 slow release fertilizer or Peters® 20-20-20 fertilizer. There 

were ten replicate pots each (MON 87708 and the conventional control) with one plant 

per replicate. The pots were placed in a greenhouse and grown under normal agronomic 

conditions for soybean (relative humidity 10-70%, temperature 21-29 ̊C, 14 hour 

photoperiod, and watering as needed). When the plants were at the V2-V3 growth stage, 

the replicates were sprayed with an herbicide (10 replicates of MON 87708 and the 

conventional control per each treatment of each tested herbicide; Table C-5). Two 

different application rates of each herbicide were applied to different replicate sets. 

Twenty to 21 days after treatment, all plants were rated for percent of injury. Ratings 

were based on visual assessment of chlorosis, necrosis, malformation, stunting, and 

biomass reduction with 100% equaling completely dead and 0% equaling no visual 

adverse effects. All ten replicate ratings were averaged. 

C.2.3. In Vitro Specificity Experiments Enzymatic Reaction Mixture Method 

The reaction of E. coli-produced DMO with different compounds evaluated as potential 

substrates was carried out using similar reaction conditions described in the 

characterization portion of this appendix (Appendix C.1.9.). The compounds (Table C-6) 

were combined with E. coli-produced DMO at 0.2 and/or 0.012 mM. The concentrations 

tested ensured adequate reaction conditions in terms of the substrate for the detection of 

product formation or disappearance of substrate. 

Manufacturer/ 

Retailer Compound 

Common 

Name 

Lot/Product 

Number 

 

Compounds Tested: 

Aldrich 2-methoxybenzoic acid o-anisic acid A0230443

Chem Service 3,6-dichloro-2-methoxybenzoic acid dicamba 341-9143

Sigma 2,4-dichlorophenoxyacetic acid 2,4-D D7299-100G

Sigma 3-(4-hydroxy-3,5-

dimethoxyphenyl)prop-2-enoic acid 

sinapic acid D7927-1G

Fluka 3,5-dimethoxy-4-hydroxybenzoic acid syringic acid 86230

Fluka 4-hydroxy-3-methoxybenzoic acid vanillic acid 94770

Fluka 3-(4-hydroxy-3-methoxy-phenyl)prop2-enoic acid ferulic acid 46278 

Compounds Used as Standards:

Monsanto 3,6-dichlorosalicylic acid DCSA GLP-0603-16959-T 

Riedel-de Haen 2,4-dichlorophenol 2,4-DCP 35811 
Monsanto Company 10-SY-210U 315 of 721 

C.2.4. In Vitro Experiments Liquid Chromatography Separation Method 

The reaction mixture was separated by Ultra Performance Liquid Chromatography 

(UPLC) using an ACQUITY UPLC BEH C18 Column containing 1.7 μm Bridged Ethyl 

Hybrid (BEH) particles and an ACQUITY BEH C18 VanGuard Pre-column. The 

column was heated to 40°C. The tested substrates and potential oxidative by-products 

were monitored by ACQUITY UPLC photodiode array (PDA) with wavelength range 

from 200nm to 320nm with 1.2nm resolution. The chromatography was performed at 

0.25ml/min and directed to the mass spectrometer following the separation. Both mobile 

phase A (water) and solvent B (acetonitrile) contained 0.1% v/v formic acid. Gradients 

used were substrate specific: 

 The gradient for dicamba was run from 40 to 50% solvent B in 3min, 50 to 100% 

solvent B in 0.1 min and then kept at 100% solvent B for 1min before returning to 

40% solvent B in 0.1 min. 

 The gradient for 2,4-D was run from 40 to 45% solvent B in 6min, held at 45% 

solvent B for 1min, 45 to 100% solvent B in 0.1 min, and then held at 100% 

solvent B for 0.5 min before returning to 40% solvent B in 0.1 min. 

 The gradient for ferulic acid, o-anisic acid, sinapic acid, syringic acid, and vanillic 

acid were run from 0 to 100% solvent B in 4 min and then held at 100% solvent B 

for 1 min before returning to 0% solvent B in 0.1 min.

Five microliters injection of each sample was used for UPLC analysis where the 

disappearance of the potential substrate was monitored, and a 50 μl injection was used for 

UPLC analysis where formation of potential oxidative by-products was monitored. 

C.2.5. In Vitro Experiments Mass Spectrometry Detection Method 

Elution from the UPLC column (C.2.4) flowed directly to a Waters Micro Q-TOF mass 

spectrometer. The parameters used for the mass determination were: negative mode, 

capillary voltage of 2800 V, sample cone voltage of 26 V for all analytes with the 

exception of 2,4-D and 2,4-DCP, which was 10 V. The extraction cone was 1.5 V. The 

source temperature was 150 °C and the desolvation temperature was 390 °C. The 

desolvation gas flow was 500 L/hour. Scan time was 0.76 seconds and inter scan delay 

was 0.1 seconds. The m/z range used was specific to each substrate and product. The 

m/z range for dicamba and DCSA was from 160 to 225 from 0 to 4 minutes. The m/z at 

175, which is the fragment ion of dicamba, was used as a detection method for dicamba. 

This fragment ion of dicamba gave better sensitivity, than the parent ion. The m/z at 205 

or 207was used to detect DCSA. The m/z range for 2,4-D and 2,4-DCP was from 160-

164 or 160-225 dependent on the specific experiment from 0 to 6 minutes. The m/z 

range for all other acids is from 120 to 230 within 4 minutes. 
Monsanto Company 10-SY-210U 316 of 721 

C.2.6. Results of Herbicide Tolerance Greenhouse Trials 

MON 87708 plants demonstrated similar levels of tolerance as the conventional control 

for 16 of the 19 herbicides tested (Table C-7). When 2,4-D, 2,4-DB, and MCPA were 

applied, the MON 87708 plants showed higher tolerance (expressed as injury rating in 

Table C-8) than the conventional control. Based on these results 2,4-D was further 

examined as a potential substrate for DMO in in vitro experiments. 2,4-D was selected as 

representative of the three auxin herbicides (2,4-D, 2,4-DB, and MCPA) as it is the most 

structurally similar to dicamba. 

C.2.7. Results of In Vitro Experiments with 2,4-D 

The reaction of dicamba with E. coli-produced DMO has been well characterized 

utilizing an in vitro enzymatic assay that monitors the formation of DCSA by LC-MS, 

which allows for the detection of the product with high sensitivity. Both the substrate 

and reaction products can be detected by LC-UV and LC-MS after separation by UPLC 

(Figure C-8). 

The same enzymatic assay using LC-MS as the detection method was used to look for the 

potential conversion of 2,4-D to 2,4-DCP by E. coli-produced DMO. 2,4-DCP is 

predicted to be the product formed from the possible oxidative reaction of 2,4-D and 

DMO, based on the mechanism of action for dicamba conversion to DCSA by DMO. 

Both the amount of substrate and product were monitored in reactions using dicamba and 

2,4-D as the substrate (Figure C-9). Using dicamba as a substrate, the formation of 

DCSA is clearly observed in the presence of E. coli-produced DMO (Figure C-9A). 

When 2,4-D is utilized as a possible substrate, there is no significant formation of 

product, as minimal amounts of 2,4-DCP are observed with and without E. coli-produced 

DMO as well as in a substrate only control (Figure C-9B). 

C.2.8. Results of In Vitro Experiments with Endogenous Soybean Compounds 

Compounds structurally similar to dicamba and found in soybean were used as potential 

substrates to determine if these compounds could be metabolized by DMO (Table C-6). 

The compounds tested were: syringic acid, o-anisic acid, vanillic acid, ferulic acid, and 

sinapic acid. Mass spectrometry scans were taken from 120 m/z to 250 m/z to cover the 

range of all potential oxidation products formed by DMO. Standard reaction conditions 

of dicamba with E. coli-produced DMO were used as a positive control. LC-MS data 

demonstrated that there are no additional peaks formed when reactions of each compound 

incubated with E. coli-produced DMO and without E. coli-produced DMO are compared 

(Figure C-10) (dicamba m/z 205, 2, 4-D m/z 163, ferulic acid m/z 175, o-anisic acid m/z 

137, sinapic acid m/z 209, syringic acid m/z 183, and vanillic acid m/z 153). There were 

no peaks observed at the respective masses for the predicted reaction products of each 

compound incubated with E. coli-produced DMO, indicating these compounds are not 

catabolized by DMO. 
Monsanto Company 10-SY-210U 

Table C-7. Herbicide Tolerance Trials Injury Ratings 

Injury 

Observations 

(days after 

application) 

Injury r

Formulation Manufacturer 

Active 

Ingredient 

Labeled Rate Range 

(g/ha)1

Rates Applied 

(g/ha)1

Control3

Average 

(Range) 

Clarity BASF dicamba 0.13-2.00 (a.e.) 

0.50 (a.e.) 1.00 (a.e.) 20 

100a

100a 

2,4-D Amine 4 Helena 2,4-D 0.13-2.00 (a.e.) 

0.25 (a.e.) 0.50 (a.e.) 20 

96 (90-100) 98 (90-100) 

Butyrac 200 Albaugh 2,4-DB 0.12-1.50 (a.e.) 

0.25 (a.e.) 0.50 (a.e.) 20 

92 (85-100) 93 (85-100) 

MCPA L.V.Ester 4 Agriliance MCPA 0.12-1.50 (a.e.) 

0.25 (a.e.) 0.50 (a.e.) 20 

95 (90-100) 97 (95-100) 

Stinger Dow clopyralid 0.09-0.49 (a.e.) 

0.25 (a.e.) 0.50 (a.e.) 20 

98 (95-100) 

100a 

Garlon 3A Dow triclopyr 0.25-9.01 (a.e.) 

0.25 (a.e.) 0.50 (a.e.) 20 

97 (90-100) 99 (95-100) 

Tordon 22K Dow picloram 0.06-1.00 (a.e.) 

0.25 (a.e.) 0.50 (a.e.) 20 

98 (95-100) 

100 (95-100) 

Classic DuPont chlorimuron 0.02-0.08 (a.i.) 

0.07 (a.i.) 0.18 (a.i.) 20 

88 (85-98) 90 (85-98) 

Glean DuPont chlorsulfuron 0.008-0.046 (a.i.) 

0.07 (a.i.) 0.18 (a.i.) 20 

89 (85-95) 94 (90-98) 

Gramoxone Syngenta paraquat 0.25-1.00 (a.e.) 

0.50 (a.e.) 0.75 (a.e.) 20 

98 (95-99) 99 (98-99) 

Lasso Monsanto alachlor 0.15-4.00 (a.i.) 

4.00 (a.i.) 6.01 (a.i.) 21 

21 (10-35) 45 (30-60) 

Harness Monsanto acetochlor 0.83-4.00 (a.i.) 

4.00 (a.i.) 6.01 (a.i.) 21 

71 (50-85) 92 (80-98) 
Monsanto Company 10-SY-210U 

Table C-7 (continued). Herbicide Tolerance Trials Injury Ratings 

Formulation Manufacturer 

Active 

Ingredient 

Labeled Rate Range 

(lb/acre)1

Rates Applied 

(lb/acre)1

Injury 

Observations 

(days after 

application) 

Injury r

Control3

Average 

(Range) 

Atrazine® Dow atrazine 0.98-3.39 (a.i.) 

1.50 (a.i.) 3.00 (a.i.) 21 

80 (50-100) 97 (85-100) 

Lorox® DuPont linuron 0.25-2.50 (a.i.) 

4.00 (a.i.) 6.01 (a.i.) 21 

58 (30-100) 70 (45-100) 

Treflan® Dow trifluralin 0.50-2.00 (a.i.) 

4.00 (a.i.) 6.01 (a.i.) 21 

9 (5-15) 

11 (5-15) 

Roundup 

WeatherMax® Monsanto glyphosate 0.25-3.72 (a.e.) 

0.07 (a.e.) 0.21 (a.e.) 21 

47 (40-55) 64 (60-70) 

Goal® 2XL Dow oxyfluorfen 0.25-2.00 (a.i.) 

0.50 (a.i.) 0.75 (a.i.) 21 

54 (40-70) 67 (50-80) 

Cobra® Valent lactofen 0.06-0.39 (a.i.) 

0.50 (a.i.) 0.75 (a.i.) 21 

33 (25-50) 41 (25-50) 

Arsenal® BASF imazapyr 0.21-1.50 (a.i.) 

0.07 (a.i.) 0.18 (a.i.) 21 

58 (50-65) 67 (60-75) 

Permit® Monsanto halosulfuron 0.032-0.13 (a.i.) 

0.07 (a.i.) 0.18 (a.i.) 21 

43 (30-50) 52 (40-60) 

1a.e. = acid equivalent; a.i. = active ingredient. Each herbicide contains the active ingredient directly or the salt form of the active ingredient. When d

application, the salt form is calculated back to the acid that is the active ingredient and therefore called acid equivalent. Each labeled rate is for cereal 

since these herbicides are not labeled to be sprayed on soybeans or are labeled for soybeans only as a pre-plant treatment. Based on the labeled rates, 

were chosen and then adjusted for use in-crop on soybeans and for the optimal growing conditions in the greenhouse. 2Injury ratings were determined by visual inspection of each plant. Ratings were based on visual assessment of chlorosis, necrosis, malformation, stun

100 percent = completely dead and 0 percent = no visual adverse effects. 3Control plants were near isogenic conventional soybean control A3525. Reported average and range of 10 replicate plants. 4Reported average and range of 10 replicate plants.
Monsanto Company 10-SY-210U 

Figure C-8. UPLC separation of dicamba (DCB) and DCSA Dicamba and DCSA were separated by UPLC and detected by UV absorbance using a Photo Diode Array (PDA) and mass 
Monsanto Company 10-SY-210U 

Figure C-9. E. coli-produced DMO Conversion of Dicamba (DCB) to DCSA and 2,4-D to 2,4-DichloropheThe formation of DCSA from dicamba (Panel A) and 2,4-DCP from 2,4-D (Panel B) due to E. coli-produced DMO wer

on the lefthand side of each graph represents the concentration of the potential substrate, and the scale on the righth

represents the concentration of the predicted oxidative product. Each bar represents a replicate assay. Substrate only 2,4-D in the 25 mM KPi, 10 mM MgCl2 buffer. The concentration of the dicamba or 2,4-D used in each experiment was 1
Monsanto Company 10-SY-210U 

Figure C-10. E. coli-produced DMO Conversion of Endogenous Substrates Endogenous substrates, as well as dicamba and 2,4-D, were incubated with E. coli-produced DMO and the formdisappearance of substrate was monitored by LC-MS (top two chromatograms) and LC-UV (bottom to chromatogram

(dicamba (a)), (2,4-D (b)) and each endogenous compound:, sinapic acid(c), ferulic acid (d), anisic acid (e), syringic acid For each experiment the reaction mixture was made with (+E. coli-produced DMO, upper) and without (-E. coli-produced 

line indicates the migration of the substrates (and DCSA in the case of dicamba) in each chromatogram.

8,107,108 G4 Vanillic

2.50 2.75 3.00 3.25 3.50 

% 0

100

2.50 2.75 3.00 3.25 3.50 

% 0

100

Dicamba-02-12-09-1126 1: TO

2.48 2.77 3.28 3.44 3.57 3

3.17

2.96

Dicamba-02-12-09-1130 1: TO

2.48 3.61 3

2.77 3.49

2.99

2.50 2.75 3.00 3.25 3.50 

AU

0.0

2.0e+2

2.50 2.75 3.00 3.25 3.50 

AU

0.0

2.0e+2

Dicamba-02-12-09-1126 3: 

2.68 Rang

Dicamba-02-12-09-1130 3: 

2.68 Rang

2.38

+DMO+DMO-DMO-DMO

8,107,108 G5 Syringic

2.50 2.75 3.00 3.25 3.50 3.75

% 0

100

2.50 2.75 3.00 3.25 3.50 3.75

% 0

100

Dicamba-02-12-09-1175 1: TOF MS ES183

1.24e3

2.76

2.51

2.68 3.03 3.47

Dicamba-02-12-09-1179 1: TOF MS ES183

1.05e3

2.51 2.77

2.30 2.68 3.01 3.52

Time

2.50 2.75 3.00 3.25 3.50 3.75

AU

0.0

2.0e+2

2.50 2.75 3.00 3.25 3.50 3.75

AU

0.0

2.0e+2

Dicamba-02-12-09-1175 3: Diode Array 

2.67 Range: 2.417e+2

2.78

Dicamba-02-12-09-1179 3: Diode Array 

2.67 Range: 2.419e+2

2.42 2.78

+DMO+DMO-DMO-DMO

8,107,108 G8 Anistic

2.50 2.75 3.00 3.25 3.50 3.75

% 0

100

2.50 2.75 3.00 3.25 3.50 3.75

% 0

100

Dicamba-02-12-09-1148 1: TOF MS ES137

1.54e3

3.45

3.19

2.78

3.33 3.71

Dicamba-02-12-09-1152 1: TOF MS ES137

2.12e3

3.45

3.19

2.65

3.32

Time

2.50 2.75 3.00 3.25 3.50 3.75

AU

0.0

1.0e+22.0e+2

2.50 2.75 3.00 3.25 3.50 3.75

AU

0.0

1.0e+2

Dicamba-02-12-09-1148 3: Diode Array 

3.09 Range: 1.661e+2

2.44 2.94

Dicamba-02-12-09-1152 3: Diode Array 

3.09 Range: 2.097e+2

2.44

+DMO

+DMO

-DMO

-DMO

8,107,108 G7 Ferrulic 

2.50 2.75 3.00 3.25 3.50 3.75

% 0

100

2.50 2.75 3.00 3.25 3.50 3.75

% 0

100

Dicamba-02-12-09-1103 1: TOF MS ES179

1.89e3

2.70

3.02

2.86

Dicamba-02-12-09-1107 1: TOF MS ES179

2.20e3

2.70

3.00

Di b 02 12 09 1103 3 Di d A

Time

2.50 2.75 3.00 3.25 3.50 3.75

AU

0.0

2.0e+2

2.50 2.75 3.00 3.25 3.50 3.75

AU

0.0

2.0e+2

Dicamba-02-12-09-1103 3: Diode Array

2.91 Range: 2.866e+

2.99

Dicamba-02-12-09-1107 3: Diode Array

2.91 Range: 2.978e+

2.39

2.99

+DMO+DMO-DMO-DMO

8,107,108 B2 DCB

2.00 2.50 3.00

% 0

100

2.00 2.50 3.00

% 0

100

D icam ba-02-12-09-1023 1: TO F M S ES205

4.56e3

2.54

D icam ba-02-12-09-1027 1: TO F M S ES205

9 9

2.64

2.04

2.38

2.81 2.87 3.38 3.09

Tim e

2.00 2.50 3.00

AU

0.0

1.0e+1

2.00 2.50 3.00

AU

0.0

1.0e+12.0e+1

D icam ba-02-12-09-1023 3: D iode Array 

2.66 Range: 2.288e+1

2.44

D icam ba-02-12-09-1027 3: D iode Array 

2.65 Range: 1.837e+1

+DMO-DMO

+DMO-DMO

3,6‐DCSA

3,5‐DCSA

DCB

T im e

3.00 4.00 5.00

AU

0.0

5.0e+1

3.00 4.00 5.00

AU

0.0

5.0e+1

x 1 6 3 .7 3 R ange: 9 .014 e + 1

3.38 4.33

D ic a m b a -0 2 -1 2 -0 9 -1 0 7 0 3 : D io d e A rra y 

x16 3.73 R ange: 8.997e+1

3.38 4.33

8,107,108 C 3 2,4-D

3.00 4.00 5.00

% 0

100

3.00 4.00 5.00

% 0

100

D ic a m b a -0 2 -1 2 -0 9 -1 0 7 0 1: T O F M S E S - 

163

9 4

3.83

3.49 3.55

3.96 4.43

4.51 5.26

D ic a m b a -0 2 -1 2 -0 9 -1 0 7 4 1: T O F M S E S - 

163

8 9

3.86

3.07 3.30

3.90 4.43

5.12 5.23

+DMO

+DMO

-DMO

-DMO

2,4‐DCP

8,107,108 H6 Sinapic 

2.00 2.50 3.00 

% 0

100

2.00 2.50 3.00 

% 0

100

Dicamba-02-12-09-1197 1: TO

2.38

2.19 2.13

1.93

2.49 2.71 3.12 3.17 3

Dicamba-02-12-09-1202 1: TO

2.36 2.52

2.29

2.26

1.94 1.83

2.78

3.063.25

3

2.00 2.50 3.00 

AU

0.0

2.0e+2

2.00 2.50 3.00 

AU

0.0

2.0e+2

Dicamba-02-12-09-1197 3: D

2.15 Rang

1.69

2.82

2.32

Dicamba-02-12-09-1202 3: D

2.15 Range

1.78

2.82

2.33

2.66 3.10 3.20

+DMO

+

-DMO

-

e

a. Dicamba (DCB) b. 2,4-D c. Sinapic Acid

d. Ferulic Acid e. Anisic Acid f. Syringic Acid g. Vanillic Acid
Monsanto Company 10-SY-210U 322 of 721 

References to Appendix C 

Aebersold, R. 1993. Mass spectrometry of proteins and peptides in biotechnology. 

Currrent Opinion in Biotechnology 4:412-419. 

Arfin, S.M., and R.A. Bradshaw. 1988. Cotranslational processing and protein turnover 

in eukaryotic cells. Biochemistry 27:7979-7984. 

Bradshaw, R.A., W.W. Brickey, and K.W. Walker. 1998. N-terminal processing: the 

methionine aminopeptidase and N alpha-acetyl transferase families. Trends in 

Biochemical Sciences 23:263-267. 

Grimm, R., M. Grimm, C. Eckerskorn, K. Pohlmeyer, T. Rohl, and J. Soll. 1997. 

Postimport methylation of the small subunit of ribulose-1,5-bisphosphate carboxylase in 

chloroplasts. FEBS Letters 408:350-354. 

Hunkapiller, M.W., R.M. Hewick, W.J. Dreyer, and L.E. Hood. 1983. High-sensitivity 

sequencing with a gas-phase sequenator. Methods in Enzymology 91:399-413. 

Jensen, O.N., A.V. Podtelejnikov, and M. Mann. 1997. Identification of the components 

of simple protein mixtures by high-accuracy peptide mass mapping and database 

searching. Analytical Chemistry 69:4741-4750. 

Polevoda, B., and F. Sherman. 2000. N alpha terminal acetylation of eukaryotic proteins. 

The Journal of Biological Chemistry 275:36479-36482. 

Whitney, S.M., and T.J. Andrews. 2001. The gene for the ribulose-1,5-bisphosphate 

carboxylase/oxygenase (Rubisco) small subunit relocated to the plastid genome of 

tobacco directs the synthesis of small subunits that assemble into Rubisco. Plant Cell 

13:193-205. 
Monsanto Company 10-SY-210U 323 of 721 

Appendix D: Materials and Methods Used for the Analysis of the Levels of 

MON 87708 DMO 

D.1. Materials 

Over-season leaf, root, forage, and seed tissue samples from MON 87708 and the near 

isogenic conventional soybean control A3525 were harvested from five field sites in the 

U.S. during 2008 from plants grown from starting seed lot 10001256 and 10001257, 

respectively. An E. coli-produced DMO protein (lot 11247247) was used as the 

analytical reference standard. 

The identities of MON 87708 and the conventional control samples were confirmed by 

verifying the chain of custody documentation prior to analysis. To further confirm the 

identities of MON 87708 and the conventional control samples, event specific 

polymerase chain reaction (PCR) analyses were conducted on the seed tissue samples 

from each site to confirm the presence or absence of the dmo expression cassette. The 

PCR analyses and the resulting verification of identities were archived under the starting 

seed lot numbers. 

D.2. Field Design and Tissue Collection 

Field trials were initiated during the 2008 planting season to generate MON 87708 and 

the conventional control samples at the following locations in the U.S.: Jefferson County, 

Iowa; Stark County, Illinois; Clinton County, Illinois; Parke County, Indiana; and Berks 

County, Pennsylvania. The field sites were representative of soybean producing regions 

suitable for commercial soybean production. At each site, three replicated plots 

containing MON 87708, as well as the conventional control, were planted using a 

randomized complete block field design. Over-season leaf (OSL), root, forage, and seed 

samples were collected from each replicated plot at each field site (except for the 

conventional control from Berks County, Pennsylvania where only two replicates were 

tested). 

The OSL tissue samples were collected from the youngest set of fully expanded trifoliate 

leaves at the following growth stages: OSL-1 at V3-V4; OSL-2 at V5-V8; OSL-3 at 

R2-V12; and OSL-4 at R5-V16. The root and forage tissue samples were collected at 

approximately the R6 growth stage. Seed tissue samples were collected at the R8 growth 

stage. 

D.3. Tissue Processing and Protein Extraction 

All tissue samples harvested were shipped to Monsanto’s processing facility and were 

prepared by the Monsanto Sample Management Team. The prepared tissue samples were 

stored in a -80°C freezer until transferred on dry ice to the analytical facility. 

MON 87708 DMO was extracted from the seed tissue samples at a tissue to buffer ratio 

of 1:100 with a Tris-borate buffer (0.1 M Tris, 0.1 M Na2B4O7 • 10H2O, 0.01 M MgCl2, 

0.05% (v/v) Tween-20 at pH 7.8). MON 87708 DMO was extracted from OSL, forage, 
Monsanto Company 10-SY-210U 324 of 721 

and root tissues samples at a tissue to buffer ratio of 1:100, 1:50, and 1:50, respectively, 

using phosphate buffered saline (PBS) with Tween 20 and 0.5% (w/v) bovine serum 

albumin (BSA) (1 × PBST with 0.5% (w/v) BSA). Extractions were done using 8 1/4′′

chrome-steel beads, and shaking in a Harbil mixer (Fluid Management, Wheeling, 

Illinois). Insoluble material was removed from all tissue extracts using a serum filter 

(Fisher Scientific, Pittsburgh, PA). The extracts were aliquotted and stored frozen in 

a -80°C freezer until enzyme-linked immunosorbent assay (ELISA) analysis. 

D.4. DMO Antibodies 

Goat polyclonal anti-DMO antibodies were purified using Protein-G Agarose affinity 

chromatography. The concentration of the purified IgG was determined to be 8.1 mg/ml 

by spectrophotometric methods. The purified antibody was stored in 1× PBS, pH 7.4. 

The purified anti-DMO antibodies were coupled with biotin (Pierce, Rockford, IL) 

according to the manufacturer’s instructions. The detection reagent was NeutrAvidin 

(Pierce, Rockford, IL) conjugated to horseradish peroxidase (HRP). The goat polyclonal 

anti-DMO antibodies react with the DMO and DMO+27 present in the MON 87708 

DMO and were used as capture antibodies for the DMO ELISA method. 

D.5. DMO ELISA Method 

Goat polyclonal anti-DMO antibodies were diluted in coating buffer (15 mM Na2CO3, 

35 mM NaHCO3, and 150 mM NaCl, pH 9.6) to a final concentration of 5.0 μg/ml and 

then immobilized onto 96-well microtiter plates followed by incubation in a 4 °C 

refrigerator for ≥8 hours. Plates were washed with 1 × PBS containing 

0.05% (v/v) Tween-20 (1 × PBST). The plates were blocked using 10% Casein in tris 

buffered saline (TBS) blocking buffer (Pierce, Rockford, IL) at 200 μl per well for 1 hour 

at room temperature. The blocking buffer was aspirated and DMO standard or tissue 

sample extract was added at 100 μl per well and incubated for 1 hour at 37°C. Prior to 

the addition of biotinylated antibody, NuetrAvidin-HRP and 3,3',5,5' tetramethylbenzidine (TMB; Kirkegaard & Perry, Gaithersburg, MD) reagents, plates were washed 

with 1 × PBST. The captured MON 87708 DMO was detected by the addition of 100 μl 

per well of biotinylated goat anti-DMO antibodies and NeutrAvidin-HRP. Plates were 

developed by adding 100 μl per well of TMB. The enzymatic reaction was terminated by 

the addition of 100 μl per well of 3 M H3PO4. Quantification of MON 87708 DMO was 

accomplished by interpolation on a DMO standard curve that ranged from 

0.313-20 ng/ml. 

D.6. Moisture Analysis 

Tissue moisture content was determined using an IR-200 Moisture Analyzer (Denver 

Instrument Company, Arvada, CO). A homogeneous site and tissue-specific pool (TSSP) 

was prepared using MON 87708 and the conventional control samples from each tissue 

type grown at each site. The average percent moisture for each TSSP was calculated 

from triplicate analyses. A TSSP Dry Weight Conversion Factor (DWCF) was calculated 

as follows: 
Monsanto Company 10-SY-210U 325 of 721 

 

  100

Mean % TSSP Moisture DWCF  1  

The DWCF was used to convert protein levels from a μg/g fresh weight (fwt) basis into a 

μg/g dry weight (dwt) basis using the following calculation: 

 

  DWCF

Protein Level Fresh Weight Protein Level in Dry Weight  

The protein levels that were reported on a fwt basis to be less than or equal to the limit of 

detection or less than the limit of quantification (LOQ) were not reported on a dwt basis. 

D.7. Data Analyses 

All MON 87708 DMO ELISA plates were analyzed on a SPECTRAmax Plus 384 

(Molecular Devices, Sunnyvale, CA) microplate spectrophotometer, using a dual 

wavelength detection method. All protein concentrations were determined by optical 

absorbance at a wavelength of 450 nm with a simultaneous reference reading of 

620-650 nm. Data reduction analyses were performed using Molecular Devices 

SOFTmax PRO GxP version 5.0.1 software. Absorbance readings and protein standard 

concentrations were fitted with a four parameter logistic curve fit. Following the 

interpolation from the standard curve, the amount of protein (ng/ml) in the tissue was 

reported on a μg/g fwt basis for data that were greater than or equal to the LOQ. For 

MON 87708 DMO, this conversion utilized a sample dilution factor and a tissue-tobuffer ratio. The enzyme values in μg/g fwt were converted to μg/g dwt by applying the 

DWCF. Microsoft Excel 2007 (Version 12.0.6514.5000 SP2 Microsoft, Redmond, WA) 

was used to calculate MON 87708 DMO levels in tissue samples. The sample mean, 

standard deviations, and ranges were also calculated using Microsoft Excel 2007. 
Monsanto Company 10-SY-210U 326 of 721 

Appendix E: Materials, Methods, and Individual-Site Results for Compositional 

Analysis of MON 87708 Soybean Seed and Forage 

Compositional comparisons between MON 87708 and the near isogenic conventional 

soybean control A3525 were performed using the principles and analytes outlined in the 

OECD consensus documents for soybean composition (OECD, 2001). These principles 

are accepted globally and have been employed previously in assessments of soybean 

products derived through biotechnology. The compositional assessment was conducted 

on seed and forage samples harvested from a single growing season conducted in the U.S. 

during 2008 under typical agronomic practices. 

The materials and methods for compositional analysis, as well as the individual-site 

results (Tables E-4 to E-18), are provided below. 

E.1. Materials 

Forage and harvested seed from MON 87708, a near isogenic conventional soybean 

control A3525 that has similar genetic background to that of MON 87708, and 

commercial reference varieties were compositionally assessed. The commercial 

reference varieties are listed in Table E-1. 

Table E-1. Commercial Reference Varieties 

Material Name Seed Lot# Field Site Code 

CST3461 10000890 IARL 

Wilken 3316 10001505 IARL 

Midland 363 10001570 IARL 

Stine 3300-0 10001312 IARL 

Croplan HT3596STS 10001450 ILWY 

FS 3591 10001448 ILWY 

Garst 3585N 10000883 ILWY 

Pioneer 93M52 10001311 ILWY 

Stine 3608-0 10001392 ILCY 

Quality Plus 365C 10001608 ILCY 

Crows C37003N 10001508 ILCY 

NK S38-T8 10001509 ILCY 

Lewis 372 10001475 INRC 

Pioneer 93M50 10000888 INRC 

Dekalb DKB34-51 10000889 INRC 

Stewart SB3454 10000887 INRC 

Dekalb DKB31-51 10001285 PAHM 

NK 32Z3 10001607 PAHM 

Hoegemeyer 333 10001590 PAHM 

Pioneer 93B15 10001304 PAHM 
Monsanto Company 10-SY-210U 327 of 721 

E.2. Characterization of the Materials 

The identities of the forage and seed samples from MON 87708, the conventional control, 

and the commercial reference varieties were verified by the Study Director prior to the 

study by confirming the chain-of-custody documentation supplied with the forage and 

seed harvested from the field sites. The seed of MON 87708, the conventional control, 

and the commercial reference varieties were characterized by event-specific polymerase 

chain reaction (PCR) analysis to confirm the presence or absence of the dmo expression 

cassette. 

E.3. Field Production of the Samples 

Harvested seed and forage of MON 87708, the conventional control, and the commercial 

reference varieties were collected from five replicated sites in the U.S. during the 2008 

growing season. These sites are Jefferson County, Iowa (IARL); Stark County, Illinois 

(ILWY); Clinton County, Illinois (ILCY); Parke County, Indiana (INRC); and Berks 

County, Pennsylvania (PAHM). Starting seeds were planted in a randomized complete 

block design with three plots for each of MON 87708, the conventional control, and the 

commercial reference varieties. The production was conducted under normal agronomic 

field conditions. All soybean plants including MON 87708, the conventional control, and 

commercial reference varieties were treated with maintenance pesticides as necessary 

throughout the growing season. In addition, MON 87708 plots were treated at the V2-V3 

growth stage with dicamba herbicide at the target label rate (0.5 lb/Acre a.e.). Seed and 

forage samples were harvested from all plots and shipped on dry ice (forage) or at 

ambient temperature (harvested seed) to Monsanto Company, St. Louis, MO. A 

subsample for compositional analysis was obtained for each tissue sample collected. 

These subsamples were ground and stored in a freezer set to maintain -20°C until their 

shipment on dry ice to Covance Laboratories Inc. (Madison, WI) for analysis. 

E.4. Summary of Analytical Methods 

Nutrients assessed in this study included proximates (ash, carbohydrates by calculation, 

moisture, protein, and fat), fiber, amino acids (18 components), fatty acids (FA, C8-C22), 

and vitamin E (α-tochopherol) in seed, and proximates (ash, carbohydrates by calculation, 

moisture, protein, and fat) and fiber in forage. Anti-nutrients assessed in seed included 

raffinose, stachyose, lectin, phytic acid, trypsin inhibitors, and isoflavones (daidzein, 

genistein, and glycitein). 

All compositional analyses were performed at Covance Laboratories, Inc. (Madison, WI). 

Methods for analysis were based on internationally-recognized procedures and literature 

publications. Brief descriptions of the methods utilized for the analyses are described 

below. 
Monsanto Company 10-SY-210U 328 of 721 

E.5. Analytical Method Summaries and Reference Standards 

E.5.1. Acid Detergent Fiber 

The sample was placed in a fritted vessel and washed with an acidic boiling detergent 

solution that dissolved the protein, carbohydrate, and ash. An acetone wash removed the 

fats and pigments. The lignocellulose fraction was collected on the frit and determined 

gravimetrically (Goering and Van Soest, 1970). The limit of quantitation (LOQ) for this 

analysis was 0.100%. 

E.5.2. Amino Acid Composition 

The sample was assayed by three methods to obtain the full profile. Tryptophan required 

a base hydrolysis with sodium hydroxide. The sulfur-containing amino acids required an 

oxidation with performic acid prior to hydrolysis with hydrochloric acid. Analysis of the 

samples for the remaining amino acids was accomplished through direct acid hydrolysis 

with hydrochloric acid. Once hydrolyzed, the individual amino acids were then 

quantitated using an automated amino acid analyzer (AOAC-International, 2005a). The 

LOQ for this analysis was 0.100 mg/g. 

Reference Standards: 

 ThermoScientific K18, 2.5 mol/mL per constituent except cystine 

(1.25 mol/mL), Lot Number JK126327 

 Sigma, L-Tryptophan, 100%, Lot Number 076K0075 

 Sigma/BioChemika, L-Cysteic Acid Monohydrate, 99.5% (used as 100%), Lot 

Number 1305674 

 Sigma, L-Methionine Sulfone, 100%, Lot Number 047K1321 

E.5.3. Ash 

The sample was placed in an electric furnace at 550 C and ignited to drive off all volatile 

organic matter. The nonvolatile matter remaining was quantitated gravimetrically and 

calculated to determine percent ash (AOAC-International, 2005b). The LOQ for this 

analysis was 0.100%. 

E.5.4. Carbohydrates 

The total carbohydrate level was calculated by difference using the fwt-derived data and 

the following equation (USDA, 1973): 

% carbohydrates = 100 % - (% protein + % fat + % moisture + % ash) 

The LOQ for this analysis was 0.100%. 

 
Monsanto Company 10-SY-210U 329 of 721 

E.5.5. Crude Fiber 

Crude fiber was quantitated as the loss on ignition of dried residue remaining after 

digestion of the sample with 1.25% sulfuric acid and 1.25% sodium hydroxide solutions 

under specific conditions (AOAC-International, 2005c). The limit of quantitation for this 

study was 0.100%. 

 E.5.6. Fat by Acid Hydrolysis 

The sample was hydrolyzed with hydrochloric acid at an elevated temperature. The fat 

was extracted with ether and hexane. The extract was evaporated on a steambath, redissolved in hexane and filtered through a sodium sulfate column. The hexane extract 

was then evaporated again on a steambath under nitrogen, dried, and weighed (AOACInternational, 2005d). The LOQ for this analysis was 0.100%. 

E.5.7. Fat by Soxhlet Extraction 

The sample was weighed into a cellulose thimble containing sodium sulfate and dried to 

remove excess moisture. Pentane was dripped through the sample to remove the fat. The 

extract was then evaporated, dried, and weighed (AOAC-International, 2005e). The 

LOQ for this analysis was 0.100%. 

E.5.8. Fatty Acids 

The lipid was extracted and saponified with 0.5 N sodium hydroxide in methanol. The 

saponification mixture was methylated with 14% boron trifluoride in methanol. The 

resulting methyl esters were extracted with heptane containing an internal standard. The 

methyl esters of the fatty acids were analyzed by gas chromatography using external 

standards for quantitation (AOAC-International, 2005f; AOCS, 1997a, c). The limit of 

quantitation was 0.0200%. 

Reference Standards: 

 Nu Chek Prep GLC Reference Standard Hazleton No. 1, *, Lot Number AU18-S 

 Nu Chek Prep GLC Reference Standard Hazleton No. 2, *, Lot Number M13-O 

 Nu Chek Prep GLC Reference Standard Hazleton No. 3, *, Lot Number MA18-S 

 Nu Chek Prep GLC Reference Standard Hazleton No. 4, *, Lot Number JA16-T 

 Nu Chek Prep Methyl Gamma Linolenate, used as 100%, Lot Number U-63MJY12-R 

 Nu Chek Prep Methyl Tridecanoate, used as 100%, Lot Number N-13M-JA16-T 

* Overall purity of the sum of the mixture of components is used as 100%. 

 E.5.9. Isoflavones 

The sample was extracted using a solution of hydrochloric acid and reagent alcohol 

heated on steam baths or hot plates. The extract was brought to volume, diluted, and 

centrifuged. An aliquot of the supernatant was placed onto a C18 solid-phase extraction 

column. Unwanted components of the matrix were rinsed off with 20% methanol and 

then the isoflavones were eluted with 80% methanol. The sample was analyzed on a 
Monsanto Company 10-SY-210U 330 of 721 

high-performance liquid chromatography system with ultraviolet detection and was 

compared to an external standard curve of known standards for quantitation (Pettersson 

and Kiessling, 1984; Seo and Morr, 1984). The LOQ for each component was 

10.0 ppm (μg/g). 

Reference Standards: 

 Chromadex, Daidzein, 96.5%, Lot Number 04007-120 

 Chromadex, Glycitein, 96.3%, Lot Number 07344-571 

 Indofine, Genistein, 99% (100% used in calculations), Lot Number 0309074 

E.5.10. Lectin 

The sample was suspended in phosphate buffered saline (PBS), shaken, and filtered. An 

aliquot of the resulting extract was serially diluted in 10 cuvettes containing PBS. A 10% 

hematocrit of lyophilized rabbit blood in PBS was added to each dilution. After 

2.5 hours, the absorbance of each dilution of the sample and lectin control was measured 

on a spectrophotometer at 620 nm, using PBS to zero the instrument. One 

hemagglutinating unit (H.U.) was defined as the level that caused 50% of the standard 

cell suspension to sediment in 2.5 hours (Klurfeld and Kritchevsky, 1987; Liener, 1955). 

The LOQ for this analysis was 0.10 H.U./mg. 

Reference Standard: 

 Sigma-Aldrich, Red Blood Cells, Rabbit, Product #R1629, Lot Number 

105K6042 

E.5.11. Moisture 

The sample was dried in a vacuum oven at approximately 100C to a constant weight. 

The moisture weight loss was determined and converted to percent moisture (AOACInternational, 2005g). The LOQ for this analysis was 0.100%. 

E.5.12. Neutral Detergent Fiber, Enzyme Method 

The sample was placed in a fritted vessel and washed with a neutral boiling detergent 

solution that dissolved the protein, carbohydrate, enzyme, and ash. An acetone wash 

removed the fats and pigments. Hemicellulose, cellulose, and lignin fractions were 

collected on the frit and determined gravimetrically (AACC, 1998; Goering and Van 

Soest, 1970). The LOQ for this analysis was 0.100%. 

E.5.13. Phytic Acid 

The sample was extracted using 0.5 M HCl with ultrasonication. Purification and 

concentration were accomplished on a silica-based anion-exchange column. The sample 

was analyzed on a polymer high-performance liquid chromatography column PRP-1, 

5 μm (150 × 4.1 mm) with a refractive index detector (Lehrfeld, 1989; Lehrfeld, 1994). 

The LOQ for this analysis was 0.100%. 
Monsanto Company 10-SY-210U 331 of 721 

Reference Standard: 

 Aldrich, Phytic Acid, Dodecasodium Salt Hydrate, 98%, Lot Number 068K0755 

E.5.14. Protein 

Nitrogenous compounds in the sample were reduced in the presence of boiling sulfuric 

acid and a mercury catalyst mixture to form ammonia. The acid digest was made alkaline. 

The ammonia was distilled and then titrated with a previously standardized acid. The 

percent nitrogen was calculated and converted to equivalent protein using the factor 6.25 

(AOAC-International, 2005h; Bradstreet, 1965; Kolthoff and Sandell, 1948). The LOQ 

for this analysis was 0.100%. 

E.5.15. Raffinose and Stachyose 

The sample was extracted with deionized water and the extract treated with a 

hydroxylamine hydrochloride solution in pyridine, containing phenyl-β-D-glucoside as 

an internal standard. The resulting oximes were converted to silyl derivatives by 

treatment with hexamethyldisilazane and trifluoracetic acid and analyzed by gas 

chromatography using a flame ionization detector (Brobst, 1972; Mason and Slover, 

1971). The LOQ for this analysis was 0.0500%. 

Reference Standards: 

 Sigma, D-(+)-Raffinose Pentahydrate, 95.5% after correction for degree of 

hydration, Lot Number 037K1059 

 Sigma, Stachyose, 97.1% after correction for degree of hydration, Lot Number 

078K3802 

E.5.16. Trypsin Inhibitor 

The sample was ground and defatted with petroleum ether. A sample of matrix was 

extracted with 0.01 N sodium hydroxide. Varying aliquots of the sample suspension 

were exposed to a known amount of trypsin and benzoy1-DL-arginine~p~nitroanilide 

hydrochloride. The sample was allowed to react for 10 minutes at 37°C. After 

10 minutes, the reaction was halted by the addition of acetic acid. The solution was 

centrifuged and then the absorbance was determined at 410 nm. Trypsin inhibitor 

activity was determined by photometrically measuring the inhibition of trypsin’s reaction 

with benzoyl-DL-arginine~p~nitroanilide hydrochloride (AOCS, 1997b). The LOQ for 

this analysis was 1.00 Trypsin Inhibitor Units (TIU)/mg. 

E.5.17. Vitamin E 

The sample was saponified to break down any fat and release vitamin E. The saponified 

mixture was extracted with ethyl ether and then quantitated by high-performance liquid 

chromatography using a silica column (Cort et al., 1983; McMurray et al., 1980; Speek et 

al., 1985). The LOQ for this analysis was 0.500 mg/100g. 
Monsanto Company 10-SY-210U 332 of 721 

Reference Standard: 

 USP, Alpha Tocopherol, 100%, Lot Number M 

E.6. Data Processing and Statistical Analysis 

After compositional analyses were performed, data spreadsheets were forwarded to 

Monsanto Company. The data were reviewed, formatted, and sent to Certus International, 

Inc. for statistical analysis. 

The following formulas were used for re-expression of soybean composition data for 

statistical analysis (Table E-2): 

Table E-2. Re-Expression Formulas for Statistical Analysis of Composition Data 

Component From (X) To Formula1

Proximates (excluding Moisture), 

Fiber, Phytic Acid, Raffinose, 

Stachyose

% fwt % dwt X/d

Isoflavones μg/g fwt μg/g dwt X/d

Lectin H.U./fwt H.U./dwt X/d

Trypsin Inhibitor TIU/mg fwt TIU/mg dwt X/d

Vitamin E mg/100g fwt mg/100g dwt X/d

Amino Acids (AA) mg/g fwt % dwt X/(10d)

Fatty Acids (FA) % fwt % Total FA

(100)Xj/X, for 

each FAj where 

X is over all 

the FA

1

‘X’ is the individual sample value; ‘d’ is the fraction of the sample that is dry matter. 

In order to complete a statistical analysis for a compositional analyte, at least 50% of the 

values for an analyte had to be greater than the assay LOQ. The following 14 analytes 

with more than 50% of observations below the assay LOQ were excluded from statistical 

analysis: 8:0 caprylic acid, 10:0 capric acid, 12:0 lauric acid, 14:0 myristic acid, 

14:1 myristoleic acid, 15:0 pentadecanoic acid, 15:1 pentadecenoic acid, 16:1 palmitoleic 

acid, 17:0 heptadecanoic acid, 17:1 heptadecenoic acid, 18:3 gamma-linolenic acid, 

20:3 eicosatrienoic acid, 20:2 eicosadienoic acid, and 20:4 arachidonic acid. 

If less than 50% of the observations for a component were below the LOQ, individual 

analyses that were below the LOQ were assigned a value equal to one-half the LOQ. The 

following analyte was assigned a value (Table E-3): 

Table E-3. Component with Observations Below the Assay Limit of Quantitation 

Not Excluded from Statistical Analysis 

 Obs. Below LOQ Total

N LOQ 

Value 

Component Units N (%) Assigned 

Seed Fatty Acid

20:1 Eicosenoic % fwt 45 42.9 105 0.020 0.010 
Monsanto Company 10-SY-210U 333 of 721 

The data were assessed for potential outliers using a studentized PRESS residuals 

calculation. A PRESS residual is the difference between any value and its predicted 

value from a statistical model that excludes the data point. The studentized version scales 

these residuals so that the values tend to have a standard normal distribution when 

outliers are absent. Thus, most values are expected to be between  3. Extreme data 

points that are also outside of the  6 studentized PRESS residual range are considered 

for exclusion, as outliers, from the final analyses. No results had PRESS residual values 

outside of the  6 range. 

All soybean compositional components were statistically analyzed using a mixed model 

analysis of variance. The five replicated sites were analyzed both separately and 

combined. Individual replicated site analyses used model (1). 

(1) Yij = U + Ti + Bj + eij, 

where Yij = unique individual observation, U = overall mean, Ti = substance effect, 

Bj = random block effect, and eij = residual error. 

Combined-site analyses used model (2). 

(2) Yijk = U + Ti + Lj + B(L)jk + LTij + eijk, 

where Yijk = unique individual observation, U = overall mean, Ti = substance effect, 

Lj = random site effect, B(L)jk = random block within site effect, 

LTij = random site by substance interaction effect, and eijk = residual error. 

A range of observed values from the reference varieties was determined for each 

analytical component. Additionally, data from the reference varieties were used to 

develop tolerance intervals. A tolerance interval is an interval that one can claim, with a 

specified degree of confidence, contains at least a specified proportion, p, of an entire 

sampled population for the parameter measured. 

For each compositional component, 99% tolerance intervals were calculated that are 

expected to contain, with 95% confidence, 99% of the quantities expressed in the 

population of commercial reference varieties. Because negative quantities are not 

possible, negative calculated lower tolerance bounds were set to zero. 

SAS® (Version 9.2) software was used to generate all summary statistics and perform all 

analyses. 

Report tables present p-values from SAS as either <0.001 or the actual value truncated to 

three decimal places. 
Monsanto Company 10-SY-210U 

Table E-4. Statistical Summary of Site IARL Soybean Seed Nutrients for MON 87708 (Treated) vs. Conve

Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 877082 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Signific

(p-Val

Proximate (% dwt)Ash 5.30 (0.070) 5.29 (0.070) 0.0063 (0.098) -0.27, 0.28 0.95

(5.20 - 5.37) (5.19 - 5.47) (-0.27 - 0.17) 

Carbohydrates 38.55 (0.56) 37.97 (0.56) 0.57 (0.55) -0.96, 2.11 0.35

(38.03 - 39.11) (37.17 - 38.45) (-0.27 - 1.33) 

Moisture (% fwt) 7.13 (0.28) 6.07 (0.28) 1.06 (0.39) -0.033, 2.15 0.05

(6.92 - 7.27) (5.84 - 6.36) (0.56 - 1.43) 

Protein 40.92 (0.27) 41.09 (0.27) -0.18 (0.16) -0.63, 0.28 0.34

(40.40 - 41.41) (40.69 - 41.74) (-0.33 - 0.088) 

Total Fat 15.22 (0.47) 15.61 (0.47) -0.40 (0.66) -2.23, 1.43 0.57

(14.77 - 15.62) (15.38 - 15.82) (-1.05 - -0.017) 

Fiber (% dwt)Acid Detergent Fiber 13.03 (0.38) 12.60 (0.38) 0.43 (0.54) -1.08, 1.94 0.47

(12.68 - 13.58) (11.92 - 13.17) (-0.34 - 1.66) 
Monsanto Company 10-SY-210U 

Table E-4 (continued). Statistical Summary of Site IARL Soybean Seed Nutrients for MON 87708 (TreaControl 

Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 877082 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Signific

(p-Val

Fiber (% dwt)Crude Fiber 7.71 (0.33) 7.41 (0.33) 0.30 (0.46) -0.98, 1.58 0.54

(7.26 - 8.48) (7.17 - 7.60) (-0.34 - 1.03) 

Neutral Detergent Fiber 14.25 (0.89) 13.27 (0.89) 0.98 (1.25) -2.51, 4.46 0.47

(13.11 - 16.38) (11.81 - 14.42) (-1.31 - 4.57) 

Amino Acid (% dwt)Alanine 1.72 (0.026) 1.74 (0.026) -0.027 (0.036) -0.13, 0.074 0.50

(1.66 - 1.76) (1.69 - 1.77) (-0.11 - 0.042) 

Arginine 3.28 (0.055) 3.45 (0.055) -0.17 (0.061) -0.33, 0.0040 0.05

(3.26 - 3.30) (3.27 - 3.56) (-0.26 - -0.013) 

Aspartic Acid 4.55 (0.060) 4.63 (0.060) -0.076 (0.084) -0.31, 0.16 0.41

(4.44 - 4.63) (4.46 - 4.74) (-0.29 - 0.12) 

Cystine 0.61 (0.0094) 0.59 (0.0094) 0.018 (0.013) -0.019, 0.054 0.25

(0.60 - 0.62) (0.56 - 0.62) (-0.0056 - 0.053) 
Monsanto Company 10-SY-210U 

Table E-4 (continued). Statistical Summary of Site IARL Soybean Seed Nutrients for MON 87708 (TreaControl 

Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 877082 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Amino Acid (% dwt)Glutamic Acid 7.24 (0.10) 7.41 (0.10) -0.16 (0.15) -0.57, 0.25 0.332 

(7.05 - 7.38) (7.12 - 7.58) (-0.53 - 0.17) 

Glycine 1.72 (0.024) 1.76 (0.024) -0.037 (0.034) -0.13, 0.057 0.331 

(1.67 - 1.76) (1.70 - 1.79) (-0.12 - 0.042) 

Histidine 1.04 (0.014) 1.06 (0.014) -0.018 (0.019) -0.071, 0.036 0.408 

(1.02 - 1.06) (1.02 - 1.08) (-0.066 - 0.031) 

Isoleucine 1.84 (0.038) 1.88 (0.038) -0.036 (0.054) -0.19, 0.11 0.540 

(1.75 - 1.90) (1.79 - 1.94) (-0.16 - 0.11) 

Leucine 3.01 (0.041) 3.07 (0.041) -0.055 (0.058) -0.22, 0.11 0.401 

(2.93 - 3.07) (2.96 - 3.14) (-0.21 - 0.072) 

Lysine 2.60 (0.030) 2.62 (0.030) -0.017 (0.042) -0.13, 0.10 0.710 

(2.53 - 2.64) (2.54 - 2.66) (-0.12 - 0.090) 
Monsanto Company 10-SY-210U 

Table E-4 (continued). Statistical Summary of Site IARL Soybean Seed Nutrients for MON 87708 (TreaControl 

Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 877082 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Amino Acid (% dwt)Methionine 0.58 (0.013) 0.56 (0.013) 0.016 (0.019) -0.037, 0.068 0.452 

(0.56 - 0.60) (0.53 - 0.60) (-0.022 - 0.071) 

Phenylalanine 1.98 (0.036) 2.03 (0.036) -0.052 (0.050) -0.19, 0.087 0.357 

(1.92 - 2.04) (1.95 - 2.09) (-0.17 - 0.023) 

Proline 1.92 (0.035) 1.98 (0.035) -0.063 (0.050) -0.20, 0.076 0.274 

(1.90 - 1.96) (1.89 - 2.07) (-0.17 - 0.065) 

Serine 1.98 (0.044) 2.00 (0.044) -0.024 (0.036) -0.12, 0.076 0.545 

(1.92 - 2.03) (1.95 - 2.08) (-0.087 - 0.047) 

Threonine 1.53 (0.023) 1.54 (0.023) -0.011 (0.023) -0.075, 0.053 0.653 

(1.48 - 1.56) (1.51 - 1.58) (-0.051 - 0.052) 

Tryptophan 0.45 (0.0092) 0.45 (0.0092) 0.00085 (0.013) -0.035, 0.037 0.950 

(0.45 - 0.46) (0.44 - 0.47) (-0.0079 - 0.0069) 
Monsanto Company 10-SY-210U 

Table E-4 (continued). Statistical Summary of Site IARL Soybean Seed Nutrients for MON 87708 (TreaControl 

Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 877082 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Amino Acid (% dwt)Tyrosine 1.34 (0.013) 1.38 (0.013) -0.042 (0.016) -0.087, 0.0035 0.062 

(1.32 - 1.37) (1.37 - 1.42) (-0.079 - -0.00044) 

Valine 1.95 (0.046) 1.99 (0.046) -0.042 (0.065) -0.22, 0.14 0.552 

(1.82 - 2.03) (1.90 - 2.05) (-0.20 - 0.13) 

Fatty Acid (% Total FA)16:0 Palmitic 11.49 (0.051) 11.00 (0.051) 0.49 (0.062) 0.32, 0.66 0.001 

(11.44 - 11.54) (10.92 - 11.08) (0.39 - 0.62) 

18:0 Stearic 4.06 (0.067) 4.00 (0.067) 0.059 (0.095) -0.21, 0.32 0.568 

(3.99 - 4.19) (3.99 - 4.01) (-0.016 - 0.20) 

18:1 Oleic 19.38 (0.20) 21.67 (0.20) -2.29 (0.28) -3.07, -1.52 0.001 

(19.07 - 19.73) (21.48 - 21.78) (-2.71 - -1.75) 

18:2 Linoleic 53.85 (0.33) 52.70 (0.33) 1.16 (0.46) -0.13, 2.44 0.066 

(53.42 - 54.07) (52.66 - 52.73) (0.68 - 1.41) 
Monsanto Company 10-SY-210U 

Table E-4 (continued). Statistical Summary of Site IARL Soybean Seed Nutrients for MON 87708 (TreaControl 

Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 877082 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Fatty Acid (% Total FA)18:3 Linolenic 10.64 (0.13) 10.04 (0.13) 0.60 (0.19) 0.074, 1.12 0.033 

(10.58 - 10.74) (10.00 - 10.12) (0.46 - 0.74) 

20:0 Arachidic 0.25 (0.0038) 0.25 (0.0038) 0.0016 (0.0054) -0.013, 0.017 0.777 

(0.25 - 0.26) (0.25 - 0.26) (-0.0021 - 0.0080) 

20:1 Eicosenoic 0.073 (0.0020) 0.070 (0.0020) 0.0030 (0.0028) -0.0048, 0.011 0.348 

(0.071 - 0.075) (0.069 - 0.071) (0.0011 - 0.0062) 

22:0 Behenic 0.26 (0.0035) 0.28 (0.0035) -0.015 (0.0042) -0.027, -0.0036 0.022 

(0.25 - 0.27) (0.27 - 0.28) (-0.023 - -0.0038) 

Vitamin (mg/100g dwt)Vitamin E 1.15 (0.058) 0.94 (0.058) 0.21 (0.066) 0.027, 0.39 0.033 

(1.10 - 1.22) (0.89 - 0.97) (0.18 - 0.24) 

1dwt = dry weight; fwt = fresh weight; FA = fatty acid. 2MON 87708 was treated with dicamba. 3Mean (S.E.) = least-square mean (standard error). 4Control refers to the near isogenic conventional soybean control A3525. 5With 95% confidence, interval contains 99% of the values expressed in the population of commercial reference soybean varieties. Negative limits set to zero.
Monsanto Company 10-SY-210U 

Table E-5. Statistical Summary of Site IARL Soybean Seed Anti-nutrients for MON 87708 (Treated) vs. C

Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 877082 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Anti-nutrient (% dwt)Lectin (H.U./mg dwt) 2.04 (0.64) 1.53 (0.64) 0.52 (0.90) -1.98, 3.02 0.595 

(0.80 - 3.44) (0.46 - 2.70) (-0.81 - 2.98) 

Phytic Acid 1.36 (0.032) 1.53 (0.032) -0.17 (0.045) -0.30, -0.048 0.018 

(1.33 - 1.38) (1.47 - 1.62) (-0.29 - -0.10) 

Raffinose 0.38 (0.022) 0.43 (0.022) -0.051 (0.020) -0.11, 0.0034 0.059 

(0.34 - 0.42) (0.40 - 0.45) (-0.10 - -0.018) 

Stachyose 3.61 (0.20) 3.89 (0.20) -0.29 (0.28) -1.06, 0.49 0.365 

(3.15 - 4.02) (3.76 - 4.15) (-1.00 - 0.26) 

Trypsin Inhibitor (TIU/mg dwt) 33.59 (2.63) 29.67 (2.63) 3.92 (3.19) -4.94, 12.79 0.286 

(29.66 - 37.60) (25.64 - 32.71) (-3.05 - 7.87) 

Isoflavone (μg/g dwt)Daidzein 1489.23 (145.92) 1447.97 (145.92) 41.27 (206.36) -531.67, 614.20 0.851 

(1175.46 - 1654.49) (1404.40 - 1505.77) (-258.27 - 233.35) 
Monsanto Company 10-SY-210U 

Table E-5 (continued). Statistical Summary of Site IARL Soybean Seed Anti-nutrients for MON Conventional Control 

Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 877082 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Isoflavone (μg/g dwt)Genistein 1050.87 (83.29) 954.99 (83.29) 95.88 (117.79) -231.16, 422.92 0.461 

(1019.09 - 1070.05) (900.10 - 984.62) (38.84 - 163.37) 

Glycitein 108.39 (5.35) 106.40 (5.35) 1.98 (2.24) -4.23, 8.20 0.425 

(97.06 - 117.44) (97.49 - 111.71) (-0.44 - 5.73) 

1dwt= dry weight; H.U. = Hemagglutinating Units; TIU = Trypsin Inhibitor Units.2MON 87708 was treated with dicamba.3Mean (S.E.) = least-square mean (standard error).4Control refers to the near isogenic conventional soybean control A3525 5With 95% confidence, interval contains 99% of the values expressed in the population of commercial reference soybean vset to zero.
Monsanto Company 10-SY-210U 

Table E-6. Statistical Summary of Site IARL Soybean Forage Nutrients for MON 87708 (Treated) vs. C

Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 877082 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Proximate (% dwt)Ash 9.44 (0.62) 8.87 (0.62) 0.57 (0.52) -0.88, 2.03 0.334 

(9.14 - 9.65) (7.58 - 10.46) (-0.81 - 1.56) 

Carbohydrates 63.02 (0.80) 65.57 (0.80) -2.55 (1.13) -5.69, 0.60 0.087 

(62.21 - 63.79) (63.58 - 67.74) (-3.95 - -1.37) 

Moisture (% fwt) 82.60 (0.33) 81.97 (0.33) 0.63 (0.44) -0.59, 1.86 0.223 

(82.40 - 82.80) (81.40 - 82.70) (0.10 - 1.20) 

Protein 25.21 (0.54) 23.00 (0.54) 2.21 (0.76) 0.098, 4.33 0.043 

(24.71 - 25.52) (22.15 - 24.07) (1.33 - 2.75) 

Total Fat 2.30 (0.32) 2.52 (0.32) -0.22 (0.46) -1.49, 1.06 0.662 

(2.00 - 2.59) (2.01 - 3.27) (-0.68 - 0.042) 

Fiber (% dwt)Acid Detergent Fiber 40.04 (2.83) 32.62 (2.83) 7.42 (3.30) -1.75, 16.59 0.088 

(32.90 - 45.11) (28.87 - 38.15) (2.07 - 16.24) 
Monsanto Company 10-SY-210U 

Table E-6 (continued). Statistical Summary of Site IARL Soybean Forage Nutrients for MON 87708 (TreaConventional Control 

Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 877082 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Fiber (% dwt)Neutral Detergent Fiber 35.16 (2.66) 35.01 (2.66) 0.15 (3.76) -10.30, 10.60 0.969 

(30.00 - 38.51) (27.47 - 39.42) (-8.13 - 11.03) 

1dwt = dry weight; fwt = fresh weight.2MON 87708 was treated with dicamba.3Mean (S.E.) = least-square mean (standard error).4Control refers to the near isogenic conventional soybean control A3525. 5With 95% confidence, interval contains 99% of the values expressed in the population of commercial reference soybean vaNegative limits set to zero.
Monsanto Company 10-SY-210U 

Table E-7. Statistical Summary of Site ILCY Soybean Seed Nutrients for MON 87708 (Treated) vs. C

Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 877082 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Proximate (% dwt)Ash 5.10 (0.080) 5.05 (0.080) 0.050 (0.089) -0.20, 0.30 0.602 

(5.07 - 5.13) (4.88 - 5.22) (-0.14 - 0.23) 

Carbohydrates 36.35 (0.68) 35.77 (0.68) 0.58 (0.97) -2.11, 3.26 0.583 

(35.65 - 36.91) (34.11 - 37.16) (-0.38 - 2.37) 

Moisture (% fwt) 5.73 (0.20) 6.44 (0.20) -0.71 (0.28) -1.49, 0.062 0.062 

(5.17 - 6.25) (6.20 - 6.63) (-1.46 - -0.24) 

Protein 40.17 (0.40) 41.72 (0.40) -1.55 (0.55) -3.08, -0.024 0.047 

(39.44 - 40.96) (40.81 - 42.67) (-2.56 - -0.73) 

Total Fat 18.39 (0.45) 17.49 (0.45) 0.89 (0.63) -0.87, 2.65 0.231 

(18.25 - 18.56) (16.81 - 18.39) (-0.047 - 1.74) 

Fiber (% dwt)Acid Detergent Fiber 15.13 (0.45) 14.04 (0.45) 1.10 (0.61) -0.59, 2.78 0.144 

(14.86 - 15.57) (13.47 - 14.57) (0.41 - 2.10) 
Monsanto Company 10-SY-210U 

Table E-7 (continued). Statistical Summary of Site ILCY Soybean Seed Nutrients for MON 87708 (TreaControl 

Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 877082 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Fiber (% dwt)Crude Fiber 9.13 (0.29) 8.38 (0.29) 0.76 (0.41) -0.38, 1.90 0.139 

(8.84 - 9.39) (8.20 - 8.64) (0.55 - 0.97) 

Neutral Detergent Fiber 16.62 (0.63) 16.85 (0.63) -0.23 (0.73) -2.27, 1.81 0.766 

(16.34 - 16.77) (15.19 - 17.99) (-1.23 - 1.56) 

Amino Acid (% dwt)Alanine 1.80 (0.014) 1.81 (0.014) -0.0030 (0.020) -0.060, 0.054 0.891 

(1.77 - 1.82) (1.78 - 1.84) (-0.028 - 0.034) 

Arginine 3.22 (0.053) 3.30 (0.053) -0.081 (0.049) -0.22, 0.056 0.174 

(3.14 - 3.28) (3.19 - 3.43) (-0.20 - 0.0059) 

Aspartic Acid 4.67 (0.037) 4.76 (0.037) -0.089 (0.034) -0.18, 0.0051 0.058 

(4.59 - 4.75) (4.73 - 4.82) (-0.15 - -0.043) 

Cystine 0.60 (0.0078) 0.59 (0.0078) 0.010 (0.011) -0.021, 0.041 0.416 

(0.58 - 0.62) (0.58 - 0.60) (-0.0071 - 0.034) 
Monsanto Company 10-SY-210U 

Table E-7 (continued). Statistical Summary of Site ILCY Soybean Seed Nutrients for MON 87708 (TreaControl

Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 877082 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Amino Acid (% dwt)Glutamic Acid 7.43 (0.073) 7.61 (0.073) -0.18 (0.056) -0.34, -0.024 0.032 

(7.27 - 7.54) (7.52 - 7.76) (-0.25 - -0.067) 

Glycine 1.79 (0.013) 1.81 (0.013) -0.021 (0.014) -0.059, 0.018 0.213 

(1.75 - 1.81) (1.79 - 1.83) (-0.049 - 0.0022) 

Histidine 1.06 (0.0071) 1.08 (0.0071) -0.020 (0.0055) -0.035, -0.0046 0.022 

(1.04 - 1.07) (1.07 - 1.09) (-0.030 - -0.0050) 

Isoleucine 1.89 (0.013) 1.97 (0.013) -0.078 (0.017) -0.13, -0.031 0.010 

(1.87 - 1.93) (1.97 - 1.97) (-0.10 - -0.037) 

Leucine 3.09 (0.022) 3.17 (0.022) -0.077 (0.012) -0.11, -0.044 0.002 

(3.04 - 3.14) (3.14 - 3.19) (-0.10 - -0.051) 

Lysine 2.66 (0.017) 2.67 (0.017) -0.013 (0.013) -0.048, 0.022 0.366 

(2.62 - 2.69) (2.65 - 2.69) (-0.041 - 0.0088) 
Monsanto Company 10-SY-210U 

Table E-7 (continued). Statistical Summary of Site ILCY Soybean Seed Nutrients for MON 87708 (TreaControl 

Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 877082 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Amino Acid (% dwt)Methionine 0.55 (0.012) 0.57 (0.012) -0.017 (0.017) -0.064, 0.030 0.370 

(0.53 - 0.59) (0.56 - 0.58) (-0.039 - 0.024) 

Phenylalanine 2.11 (0.022) 2.13 (0.022) -0.016 (0.018) -0.067, 0.035 0.429 

(2.08 - 2.14) (2.09 - 2.19) (-0.048 - 0.011) 

Proline 2.02 (0.019) 2.06 (0.019) -0.044 (0.019) -0.096, 0.0082 0.079 

(1.98 - 2.04) (2.04 - 2.09) (-0.063 - -0.016) 

Serine 2.10 (0.027) 2.08 (0.027) 0.023 (0.025) -0.046, 0.093 0.404 

(2.06 - 2.12) (2.01 - 2.15) (-0.027 - 0.054) 

Threonine 1.59 (0.015) 1.59 (0.015) 0.0058 (0.019) -0.046, 0.058 0.772 

(1.55 - 1.62) (1.58 - 1.59) (-0.035 - 0.046) 

Tryptophan 0.51 (0.0090) 0.48 (0.0090) 0.030 (0.0084) 0.0064, 0.053 0.024 

(0.49 - 0.53) (0.47 - 0.50) (0.015 - 0.045) 
Monsanto Company 10-SY-210U 

Table E-7 (continued). Statistical Summary of Site ILCY Soybean Seed Nutrients for MON 87708 (TreaControl 

Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 877082 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Amino Acid (% dwt)Tyrosine 1.44 (0.033) 1.43 (0.033) 0.014 (0.045) -0.11, 0.14 0.773 

(1.43 - 1.46) (1.39 - 1.51) (-0.079 - 0.078) 

Valine 1.96 (0.015) 2.05 (0.015) -0.090 (0.021) -0.15, -0.031 0.013 

(1.94 - 2.01) (2.05 - 2.06) (-0.12 - -0.048) 

Fatty Acid (% Total FA)16:0 Palmitic 12.05 (0.073) 11.95 (0.073) 0.096 (0.089) -0.15, 0.34 0.340 

(11.95 - 12.16) (11.73 - 12.08) (-0.082 - 0.29) 

18:0 Stearic 3.91 (0.044) 3.93 (0.044) -0.025 (0.062) -0.20, 0.15 0.705 

(3.88 - 3.93) (3.86 - 4.02) (-0.086 - 0.018) 

18:1 Oleic 19.74 (0.29) 21.57 (0.29) -1.83 (0.41) -2.96, -0.69 0.011 

(19.44 - 19.94) (21.07 - 22.44) (-2.51 - -1.21) 

18:2 Linoleic 54.54 (0.25) 53.26 (0.25) 1.28 (0.35) 0.31, 2.26 0.021 

(54.45 - 54.70) (52.77 - 53.74) (0.73 - 1.68) 
Monsanto Company 10-SY-210U 

Table E-7 (continued). Statistical Summary of Site ILCY Soybean Seed Nutrients for MON 87708 (TreaControl 

Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 877082 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Fatty Acid (% Total FA)18:3 Linolenic 9.07 (0.074) 8.58 (0.074) 0.50 (0.099) 0.22, 0.77 0.007 

(8.99 - 9.16) (8.42 - 8.71) (0.36 - 0.56) 

20:0 Arachidic 0.26 (0.0035) 0.26 (0.0035) -0.0048 (0.0050) -0.019, 0.0091 0.393 

(0.26 - 0.26) (0.26 - 0.27) (-0.012 - 0.00087) 

20:1 Eicosenoic 0.16 (0.0016) 0.16 (0.0016) -0.0064 (0.0023) -0.013, 0.00005 0.051 

(0.16 - 0.16) (0.16 - 0.17) (-0.010 - -0.0034) 

22:0 Behenic 0.28 (0.0030) 0.29 (0.0030) -0.011 (0.0042) -0.023, 0.00033 0.054 

(0.27 - 0.28) (0.28 - 0.30) (-0.019 - -0.0066) 

Vitamin (mg/100g dwt)Vitamin E 2.13 (0.077) 1.86 (0.077) 0.27 (0.089) 0.022, 0.52 0.038 

(2.10 - 2.17) (1.71 - 2.11) (0.059 - 0.42) 

1dwt = dry weight; fwt = fresh weight; FA = fatty acid.2MON 87708 was treated with dicamba.3Mean (S.E.) = least-square mean (standard error).4Control refers to the near isogenic conventional soybean control A3525. 5With 95% confidence, interval contains 99% of the values expressed in the population of commercial reference soybean va Negative limits set to zero.
Monsanto Company 10-SY-210U 

Table E-8. Statistical Summary of Site ILCY Soybean Seed Anti-nutrients for MON 87708 (Treated) vs. C

Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 877082 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Anti-nutrient (% dwt)Lectin (H.U./mg dwt) 6.89 (1.81) 5.54 (1.81) 1.34 (2.55) -5.75, 8.43 0.626 

(4.28 - 10.27) (2.14 - 10.38) (-4.27 - 8.13) 

Phytic Acid 1.12 (0.032) 1.16 (0.032) -0.037 (0.045) -0.16, 0.088 0.456 

(1.08 - 1.20) (1.10 - 1.22) (-0.14 - 0.10) 

Raffinose 0.58 (0.013) 0.57 (0.013) 0.011 (0.018) -0.039, 0.061 0.575 

(0.57 - 0.59) (0.54 - 0.60) (-0.027 - 0.046) 

Stachyose 3.33 (0.10) 3.64 (0.10) -0.31 (0.14) -0.70, 0.085 0.095 

(3.25 - 3.46) (3.43 - 3.83) (-0.58 - 0.023) 

Trypsin Inhibitor (TIU/mg dwt) 31.75 (1.10) 32.78 (1.10) -1.02 (1.56) -5.35, 3.30 0.546 

(30.61 - 33.32) (31.06 - 34.22) (-2.43 - 0.26) 

Isoflavone (μg/g dwt)Daidzein 925.54 (72.66) 922.21 (72.66) 3.32 (102.76) -281.99, 288.63 0.975 

(899.83 - 974.38) (762.49 - 1098.08) (-198.25 - 139.91) 
Monsanto Company 10-SY-210U 

Table E-8 (continued). Statistical Summary of Site ILCY Soybean Seed Anti-nutrients for MON Conventional Control 

Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 877082 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Isoflavone (μg/g dwt)Genistein 655.78 (45.60) 653.27 (45.60) 2.51 (64.49) -176.54, 181.56 0.970 

(594.13 - 712.01) (588.17 - 770.79) (-58.78 - 60.35) 

Glycitein 98.02 (8.11) 113.29 (8.11) -15.27 (11.47) -47.13, 16.59 0.254 

(77.67 - 112.00) (96.25 - 122.09) (-43.86 - 15.75) 

1dwt = dry weight; H.U. = Hemagglutinating Units; TIU = Trypsin Inhibitor Units. 2MON 87708 was treated with dicamba. 3Mean (S.E.) = least-square mean (standard error). 4Control refers to the near isogenic conventional soybean control A3525. 5With 95% confidence, interval contains 99% of the values expressed in the population of commercial reference soybean varieties. Negative limits set to zero.
Monsanto Company 10-SY-210U 

Table E-9. Statistical Summary of Site ILCY Soybean Forage Nutrients for MON 87708 (Treated) vs. C

Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 877082 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Proximate (% dwt)Ash 6.45 (0.22) 6.27 (0.22) 0.18 (0.32) -0.70, 1.05 0.601 

(6.12 - 6.70) (6.10 - 6.49) (-0.37 - 0.48) 

Carbohydrates 65.26 (0.61) 66.38 (0.61) -1.12 (0.87) -3.53, 1.29 0.265 

(64.42 - 65.78) (65.53 - 67.94) (-3.52 - 0.11) 

Moisture (% fwt) 73.13 (0.25) 73.53 (0.25) -0.40 (0.35) -1.37, 0.57 0.316 

(72.40 - 73.70) (73.20 - 73.80) (-1.20 - 0.50) 

Protein 22.20 (0.51) 21.64 (0.51) 0.56 (0.68) -1.33, 2.46 0.454 

(21.25 - 23.11) (20.88 - 23.03) (-0.78 - 2.23) 

Total Fat 6.11 (0.38) 5.77 (0.38) 0.34 (0.52) -1.09, 1.77 0.549 

(5.62 - 6.88) (5.15 - 6.72) (0.17 - 0.47) 

Fiber (% dwt)Acid Detergent Fiber 30.24 (1.09) 28.46 (1.09) 1.78 (1.54) -2.49, 6.05 0.310 

(28.75 - 32.14) (27.05 - 30.90) (-2.15 - 5.09) 
Monsanto Company 10-SY-210U 

Table E-9 (continued). Statistical Summary of Site ILCY Soybean Forage Nutrients for MON 87708 (TreaConventional Control 

Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 877082 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Fiber (% dwt)Neutral Detergent Fiber 27.72 (1.65) 28.06 (1.65) -0.35 (2.33) -6.82, 6.13 0.889 

(27.34 - 27.98) (23.66 - 32.73) (-4.90 - 3.68) 

1dwt = dry weight; fwt = fresh weight. 2MON 87708 was treated with dicamba. 3Mean (S.E.) = least-square mean (standard error). 4Control refers to the near isogenic conventional soybean control A3525. 5With 95% confidence, interval contains 99% of the values expressed in the population of commercial reference soybean varieties. Negative limits set to zero.
Monsanto Company 10-SY-210U 

Table E-10. Statistical Summary of Site ILWY Soybean Seed Nutrients for MON 87708 (Treated) vs. C

Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 877082 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Proximate (% dwt)Ash 5.43 (0.084) 5.23 (0.084) 0.20 (0.084) -0.031, 0.44 0.073 

(5.24 - 5.69) (5.13 - 5.29) (0.097 - 0.40) 

Carbohydrates 38.84 (0.44) 38.29 (0.44) 0.54 (0.45) -0.71, 1.79 0.292 

(38.13 - 39.21) (38.19 - 38.42) (-0.064 - 0.96) 

Moisture (% fwt) 6.96 (0.16) 6.16 (0.16) 0.80 (0.22) 0.18, 1.42 0.022 

(6.80 - 7.17) (5.79 - 6.41) (0.63 - 1.01) 

Protein 40.88 (0.28) 41.99 (0.28) -1.11 (0.38) -2.15, -0.062 0.042 

(40.56 - 41.37) (41.72 - 42.25) (-1.54 - -0.63) 

Total Fat 14.83 (0.38) 14.49 (0.38) 0.34 (0.44) -0.89, 1.57 0.487 

(14.00 - 15.90) (14.40 - 14.54) (-0.53 - 1.49) 

Fiber (% dwt)Acid Detergent Fiber 13.43 (0.43) 12.96 (0.43) 0.47 (0.61) -1.21, 2.15 0.480 

(12.71 - 14.61) (12.48 - 13.69) (-0.71 - 2.13) 
Monsanto Company 10-SY-210U 

Table E-10 (continued). Statistical Summary of Site ILWY Soybean Seed Nutrients for MON 87708 (TreaControl 

Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 877082 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Fiber (% dwt)Crude Fiber 8.22 (0.52) 7.58 (0.52) 0.64 (0.74) -1.40, 2.69 0.430 

(7.39 - 9.07) (7.39 - 7.82) (-0.14 - 1.68) 

Neutral Detergent Fiber 15.62 (0.72) 13.78 (0.72) 1.84 (0.94) -0.77, 4.45 0.122 

(13.84 - 17.83) (13.44 - 14.00) (-0.064 - 4.39) 

Amino Acid (% dwt)Alanine 1.74 (0.0076) 1.75 (0.0076) -0.014 (0.0093) -0.039, 0.012 0.219 

(1.73 - 1.76) (1.75 - 1.76) (-0.024 - -0.0071) 

Arginine 3.30 (0.037) 3.57 (0.037) -0.27 (0.040) -0.38, -0.16 0.002 

(3.24 - 3.33) (3.55 - 3.60) (-0.32 - -0.23) 

Aspartic Acid 4.59 (0.017) 4.67 (0.017) -0.089 (0.020) -0.14, -0.034 0.011 

(4.55 - 4.61) (4.67 - 4.68) (-0.13 - -0.065) 

Cystine 0.62 (0.0090) 0.61 (0.0090) 0.014 (0.0092) -0.012, 0.039 0.205 

(0.62 - 0.63) (0.58 - 0.62) (-0.0029 - 0.036) 
Monsanto Company 10-SY-210U 

Table E-10 (continued). Statistical Summary of Site ILWY Soybean Seed Nutrients for MON 87708 (TreaControl 

Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 877082 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Amino Acid (% dwt)Glutamic Acid 7.29 (0.030) 7.51 (0.030) -0.21 (0.031) -0.30, -0.13 0.002 

(7.20 - 7.35) (7.49 - 7.53) (-0.29 - -0.16) 

Glycine 1.75 (0.0053) 1.77 (0.0053) -0.021 (0.0075) -0.042, 0.00033 0.052 

(1.74 - 1.76) (1.76 - 1.77) (-0.034 - -0.0096) 

Histidine 1.05 (0.0032) 1.07 (0.0032) -0.017 (0.0046) -0.030, -0.0046 0.019 

(1.05 - 1.05) (1.06 - 1.07) (-0.027 - -0.0057) 

Isoleucine 1.87 (0.012) 1.90 (0.012) -0.023 (0.0040) -0.034, -0.012 0.004 

(1.85 - 1.89) (1.88 - 1.91) (-0.033 - -0.017) 

Leucine 3.02 (0.011) 3.10 (0.011) -0.077 (0.0062) -0.094, -0.060 <0.00

(3.00 - 3.04) (3.09 - 3.13) (-0.086 - -0.061) 

Lysine 2.63 (0.0062) 2.63 (0.0062) 0.0011 (0.0088) -0.023, 0.026 0.904 

(2.63 - 2.64) (2.62 - 2.64) (-0.011 - 0.0071) 
Monsanto Company 10-SY-210U 

Table E-10 (continued). Statistical Summary of Site ILWY Soybean Seed Nutrients for MON 87708 (TreaControl 

Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 877082 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Amino Acid (% dwt)Methionine 0.59 (0.0074) 0.58 (0.0074) 0.0079 (0.010) -0.021, 0.037 0.492 

(0.57 - 0.60) (0.56 - 0.59) (-0.016 - 0.029) 

Phenylalanine 2.01 (0.019) 2.07 (0.019) -0.061 (0.021) -0.12, -0.0015 0.046 

(1.96 - 2.06) (2.05 - 2.10) (-0.085 - -0.047) 

Proline 1.94 (0.020) 2.05 (0.020) -0.10 (0.028) -0.18, -0.027 0.020 

(1.93 - 1.96) (2.01 - 2.09) (-0.16 - -0.048) 

Serine 1.98 (0.013) 2.06 (0.013) -0.079 (0.013) -0.11, -0.044 0.003 

(1.97 - 2.00) (2.02 - 2.09) (-0.095 - -0.048) 

Threonine 1.52 (0.0042) 1.55 (0.0042) -0.026 (0.0049) -0.040, -0.013 0.005 

(1.51 - 1.53) (1.54 - 1.55) (-0.030 - -0.022) 

Tryptophan 0.44 (0.0083) 0.47 (0.0083) -0.025 (0.011) -0.055, 0.0061 0.089 

(0.44 - 0.45) (0.45 - 0.48) (-0.035 - -0.015) 
Monsanto Company 10-SY-210U 

Table E-10 (continued). Statistical Summary of Site ILWY Soybean Seed Nutrients for MON 87708 (TreaControl 

Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 877082 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Amino Acid (% dwt)Tyrosine 1.35 (0.024) 1.36 (0.024) -0.013 (0.027) -0.090, 0.063 0.651 

(1.32 - 1.41) (1.34 - 1.40) (-0.032 - 0.0095) 

Valine 1.99 (0.013) 2.01 (0.013) -0.022 (0.012) -0.057, 0.012 0.149 

(1.95 - 2.01) (2.01 - 2.02) (-0.053 - 0.0028) 

Fatty Acid (% Total FA)16:0 Palmitic 11.26 (0.041) 11.04 (0.041) 0.22 (0.057) 0.063, 0.38 0.017 

(11.25 - 11.27) (10.97 - 11.12) (0.15 - 0.28) 

18:0 Stearic 4.32 (0.063) 4.25 (0.063) 0.076 (0.089) -0.17, 0.32 0.439 

(4.23 - 4.40) (4.16 - 4.31) (0.067 - 0.085) 

18:1 Oleic 19.52 (0.25) 21.14 (0.25) -1.62 (0.36) -2.62, -0.62 0.010 

(19.34 - 19.64) (20.78 - 21.55) (-1.97 - -1.43) 

18:2 Linoleic 53.74 (0.32) 52.90 (0.32) 0.85 (0.46) -0.43, 2.12 0.139 

(53.55 - 54.06) (52.33 - 53.20) (0.47 - 1.22) 
Monsanto Company 10-SY-210U 

Table E-10 (continued). Statistical Summary of Site ILWY Soybean Seed Nutrients for MON 87708 (TreaControl 

Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 877082 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Fatty Acid (% Total FA)18:3 Linolenic 10.54 (0.10) 10.05 (0.10) 0.49 (0.14) 0.095, 0.89 0.026 

(10.51 - 10.59) (9.89 - 10.14) (0.39 - 0.65) 

20:0 Arachidic 0.27 (0.0041) 0.27 (0.0041) -0.00093 (0.0058) -0.017, 0.015 0.879 

(0.26 - 0.27) (0.26 - 0.27) (-0.0057 - 0.0027) 

20:1 Eicosenoic 0.075 (0.0020) 0.076 (0.0020) -0.0014 (0.0023) -0.0076, 0.0049 0.583 

(0.070 - 0.079) (0.075 - 0.077) (-0.0071 - 0.0019) 

22:0 Behenic 0.26 (0.0032) 0.28 (0.0032) -0.019 (0.0040) -0.030, -0.0079 0.008 

(0.26 - 0.27) (0.28 - 0.29) (-0.021 - -0.017) 

Vitamin (mg/100g dwt)Vitamin E 1.18 (0.037) 0.94 (0.037) 0.23 (0.053) 0.086, 0.38 0.011 

(1.08 - 1.26) (0.89 - 0.99) (0.19 - 0.31) 

1dwt = dry weight; fwt = fresh weight; FA = fatty acid. 2MON 87708 was treated with dicamba. 3Mean (S.E.) = least-square mean (standard error). 4Control refers to the near isogenic conventional soybean control A3525. 5With 95% confidence, interval contains 99% of the values expressed in the population of commercial reference soybean varieties. Negative limits set to zero.
Monsanto Company 10-SY-210U 

Table E-11. Statistical Summary of Site ILWY Soybean Seed Anti-nutrients for MON 87708 (TreatControl

Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 877082 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Anti-nutrient (% dwt)Lectin (H.U./mg dwt) 1.10 (0.44) 2.33 (0.44) -1.23 (0.43) -2.43, -0.035 0.045 

(0.59 - 1.51) (1.34 - 3.68) (-2.17 - -0.75) 

Phytic Acid 1.40 (0.033) 1.55 (0.033) -0.14 (0.044) -0.27, -0.023 0.030 

(1.33 - 1.46) (1.47 - 1.61) (-0.22 - -0.054) 

Raffinose 0.37 (0.026) 0.41 (0.026) -0.037 (0.036) -0.14, 0.065 0.371 

(0.32 - 0.45) (0.41 - 0.41) (-0.086 - 0.040) 

Stachyose 3.44 (0.19) 3.76 (0.19) -0.33 (0.27) -1.08, 0.43 0.294 

(3.07 - 4.02) (3.68 - 3.85) (-0.78 - 0.34) 

Trypsin Inhibitor (TIU/mg dwt) 34.32 (2.07) 29.73 (2.07) 4.59 (2.89) -3.43, 12.62 0.187 

(29.54 - 39.27) (25.43 - 32.22) (-2.68 - 8.72) 

Isoflavone (μg/g dwt)Daidzein 1458.08 (35.08) 1271.60 (35.08) 186.48 (31.48) 99.09, 273.88 0.004 

(1416.31 - 1535.98) (1196.71 - 1354.96) (153.17 - 225.25) 
Monsanto Company 10-SY-210U 

Table E-11 (continued). Statistical Summary of Site ILWY Soybean Seed Anti-nutrients for MON Conventional Control 

Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 877082 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Isoflavone (μg/g dwt)Genistein 898.89 (28.15) 860.58 (28.15) 38.31 (39.82) -72.23, 148.86 0.390 

(873.39 - 913.50) (784.27 - 913.26) (-10.80 - 129.23) 

Glycitein 111.77 (2.15) 79.70 (2.15) 32.07 (2.23) 25.87, 38.27 <0.00

(109.88 - 113.86) (77.14 - 81.62) (31.24 - 32.73) 

1dwt = dry weight; H.U. = Hemagglutinating Units; TIU = Trypsin Inhibitor Units. 2MON 87708 was treated with dicamba. 3Mean (S.E.) = least-square mean (standard error). 4Control refers to the near isogenic conventional soybean control A3525. 5With 95% confidence, interval contains 99% of the values expressed in the population of commercial reference soybean varieties. Negative limits set to zero.
Monsanto Company 10-SY-210U 

Table E-12. Statistical Summary of Site ILWY Soybean Forage Nutrients for MON 87708 (Treated) vs. C

Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 877082 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Proximate (% dwt)Ash 6.61 (0.23) 6.88 (0.23) -0.27 (0.30) -1.10, 0.56 0.416 

(6.02 - 7.21) (6.82 - 6.99) (-0.79 - 0.37) 

Carbohydrates 67.99 (0.77) 66.26 (0.77) 1.73 (1.08) -1.28, 4.74 0.185 

(66.09 - 69.50) (65.71 - 66.67) (-0.30 - 3.78) 

Moisture (% fwt) 75.17 (0.55) 75.40 (0.55) -0.23 (0.65) -2.04, 1.57 0.737 

(74.10 - 76.70) (75.10 - 75.60) (-1.40 - 1.10) 

Protein 20.93 (0.58) 21.97 (0.58) -1.03 (0.82) -3.32, 1.25 0.278 

(19.27 - 22.70) (21.81 - 22.13) (-2.69 - 0.57) 

Total Fat 4.44 (0.52) 4.91 (0.52) -0.47 (0.35) -1.44, 0.50 0.252 

(3.92 - 5.10) (4.50 - 5.63) (-0.67 - -0.19) 

Fiber (% dwt)Acid Detergent Fiber 29.18 (1.73) 27.77 (1.73) 1.41 (2.44) -5.36, 8.19 0.593 

(26.72 - 31.00) (25.12 - 31.00) (-4.28 - 4.71) 
Monsanto Company 10-SY-210U 

Table E-12 (continued). Statistical Summary of Site ILWY Soybean Forage Nutrients for MON Conventional Control 

Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 877082 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Fiber (% dwt)Neutral Detergent Fiber 28.64 (1.83) 33.25 (1.83) -4.61 (2.59) -11.80, 2.58 0.149 

(27.22 - 31.26) (31.89 - 34.06) (-6.63 - -0.62) 

1dwt = dry weight; fwt = fresh weight. 2MON 87708 was treated with dicamba. 3Mean (S.E.) = least-square mean (standard error). 4Control refers to the near isogenic conventional soybean control A3525. 5With 95% confidence, interval contains 99% of the values expressed in the population of commercial reference soybean varieties. Negative limits set to zero.
Monsanto Company 10-SY-210U 

Table E-13. Statistical Summary of Site INRC Soybean Seed Nutrients for MON 87708 (Treated) vs. Conv

Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 877082 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Proximate (% dwt)Ash 5.03 (0.11) 4.95 (0.11) 0.075 (0.15) -0.35, 0.50 0.644 

(4.94 - 5.18) (4.73 - 5.23) (-0.28 - 0.45) 

Carbohydrates 37.60 (0.78) 35.95 (0.78) 1.65 (1.10) -1.41, 4.71 0.208 

(37.15 - 38.07) (35.27 - 37.04) (1.03 - 2.05) 

Moisture (% fwt) 6.74 (0.20) 6.53 (0.20) 0.21 (0.29) -0.59, 1.01 0.513 

(6.48 - 7.13) (6.32 - 6.84) (-0.23 - 0.69) 

Protein 42.11 (0.53) 43.58 (0.53) -1.48 (0.76) -3.57, 0.62 0.122 

(41.33 - 42.53) (43.50 - 43.69) (-2.36 - -0.97) 

Total Fat 15.30 (0.45) 15.55 (0.45) -0.25 (0.56) -1.82, 1.32 0.678 

(14.54 - 15.95) (14.52 - 16.10) (-0.63 - 0.025) 

Fiber (% dwt)Acid Detergent Fiber 13.22 (0.35) 12.77 (0.35) 0.46 (0.46) -0.81, 1.73 0.374 

(12.64 - 13.79) (12.28 - 13.25) (-0.14 - 1.52) 
Monsanto Company 10-SY-210U 

Table E-13 (continued). Statistical Summary of Site INRC Soybean Seed Nutrients for MON 87708 (TreaControl 

Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 877082 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Fiber (% dwt)Crude Fiber 8.06 (0.18) 6.89 (0.18) 1.17 (0.25) 0.47, 1.88 0.009 

(7.76 - 8.47) (6.59 - 7.12) (0.64 - 1.51) 

Neutral Detergent Fiber 15.98 (0.67) 14.66 (0.67) 1.32 (0.95) -1.32, 3.96 0.237 

(14.97 - 16.49) (13.24 - 15.56) (0.93 - 1.73) 

Amino Acid (% dwt)Alanine 1.81 (0.021) 1.84 (0.021) -0.032 (0.022) -0.093, 0.029 0.223 

(1.80 - 1.83) (1.80 - 1.87) (-0.072 - 0.013) 

Arginine 3.44 (0.044) 3.72 (0.044) -0.27 (0.062) -0.45, -0.10 0.011 

(3.39 - 3.50) (3.64 - 3.81) (-0.38 - -0.15) 

Aspartic Acid 4.76 (0.062) 4.89 (0.062) -0.13 (0.078) -0.34, 0.087 0.173 

(4.70 - 4.80) (4.80 - 4.95) (-0.23 - 0.0034) 

Cystine 0.61 (0.0067) 0.59 (0.0067) 0.020 (0.0094) -0.0067, 0.046 0.107 

(0.61 - 0.61) (0.59 - 0.60) (0.0087 - 0.029) 
Monsanto Company 10-SY-210U 

Table E-13 (continued). Statistical Summary of Site INRC Soybean Seed Nutrients for MON 87708 (TreaControl 

Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 877082 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Amino Acid (% dwt)Glutamic Acid 7.66 (0.11) 7.93 (0.11) -0.26 (0.14) -0.66, 0.13 0.138 

(7.57 - 7.73) (7.75 - 8.02) (-0.45 - -0.018) 

Glycine 1.82 (0.020) 1.85 (0.020) -0.035 (0.027) -0.11, 0.039 0.257 

(1.80 - 1.83) (1.82 - 1.87) (-0.072 - 0.013) 

Histidine 1.09 (0.012) 1.11 (0.012) -0.028 (0.016) -0.073, 0.018 0.166 

(1.07 - 1.10) (1.10 - 1.12) (-0.056 - -0.0026) 

Isoleucine 1.94 (0.042) 2.00 (0.042) -0.060 (0.057) -0.22, 0.099 0.354 

(1.90 - 1.97) (1.95 - 2.03) (-0.11 - 0.025) 

Leucine 3.16 (0.041) 3.24 (0.041) -0.082 (0.052) -0.23, 0.061 0.186 

(3.11 - 3.19) (3.20 - 3.26) (-0.14 - -0.0086) 

Lysine 2.70 (0.030) 2.72 (0.030) -0.030 (0.035) -0.13, 0.068 0.446 

(2.66 - 2.71) (2.68 - 2.76) (-0.070 - 0.031) 
Monsanto Company 10-SY-210U 

Table E-13 (continued). Statistical Summary of Site INRC Soybean Seed Nutrients for MON 87708 (TreaControl 

Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 877082 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Amino Acid (% dwt)Methionine 0.58 (0.0086) 0.59 (0.0086) -0.0055 (0.012) -0.039, 0.028 0.674 

(0.58 - 0.59) (0.58 - 0.60) (-0.017 - 0.00070) 

Phenylalanine 2.15 (0.030) 2.20 (0.030) -0.049 (0.040) -0.16, 0.062 0.289 

(2.12 - 2.18) (2.13 - 2.23) (-0.11 - 0.048) 

Proline 2.07 (0.010) 2.06 (0.010) 0.0045 (0.015) -0.036, 0.045 0.771 

(2.06 - 2.09) (2.05 - 2.07) (-0.0071 - 0.016) 

Serine 2.09 (0.028) 2.13 (0.028) -0.038 (0.029) -0.12, 0.043 0.260 

(2.06 - 2.12) (2.13 - 2.14) (-0.070 - -0.016) 

Threonine 1.60 (0.019) 1.61 (0.019) -0.014 (0.027) -0.089, 0.060 0.622 

(1.57 - 1.62) (1.60 - 1.62) (-0.040 - 0.012) 

Tryptophan 0.47 (0.013) 0.45 (0.013) 0.019 (0.018) -0.032, 0.069 0.365 

(0.44 - 0.50) (0.43 - 0.46) (-0.015 - 0.064) 
Monsanto Company 10-SY-210U 

Table E-13 (continued). Statistical Summary of Site INRC Soybean Seed Nutrients for MON 87708 (TreaControl 

Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 877082 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Amino Acid (% dwt)Tyrosine 1.38 (0.024) 1.44 (0.024) -0.065 (0.022) -0.13, -0.0024 0.044 

(1.35 - 1.43) (1.38 - 1.47) (-0.12 - -0.036) 

Valine 2.05 (0.045) 2.12 (0.045) -0.074 (0.063) -0.25, 0.10 0.305 

(2.01 - 2.09) (2.06 - 2.16) (-0.14 - 0.026) 

Fatty Acid (% Total FA)16:0 Palmitic 11.42 (0.068) 11.19 (0.068) 0.23 (0.097) -0.036, 0.50 0.074 

(11.39 - 11.46) (11.14 - 11.21) (0.19 - 0.26) 

18:0 Stearic 4.18 (0.047) 4.25 (0.047) -0.064 (0.066) -0.25, 0.12 0.387 

(4.11 - 4.27) (4.16 - 4.31) (-0.19 - 0.11) 

18:1 Oleic 18.78 (0.085) 20.19 (0.085) -1.41 (0.093) -1.66, -1.15 <0.00

(18.58 - 18.95) (20.12 - 20.23) (-1.64 - -1.27) 

18:2 Linoleic 54.98 (0.10) 54.43 (0.10) 0.54 (0.14) 0.15, 0.94 0.019 

(54.80 - 55.14) (54.32 - 54.64) (0.16 - 0.82) 
Monsanto Company 10-SY-210U 

Table E-13 (continued). Statistical Summary of Site INRC Soybean Seed Nutrients for MON 87708 (TreaControl 

Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 877082 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Fatty Acid (% Total FA)18:3 Linolenic 10.03 (0.050) 9.31 (0.050) 0.71 (0.070) 0.52, 0.91 <0.00

(9.89 - 10.10) (9.26 - 9.40) (0.60 - 0.84) 

20:0 Arachidic 0.26 (0.0029) 0.27 (0.0029) -0.0053 (0.0041) -0.017, 0.0062 0.270 

(0.26 - 0.27) (0.26 - 0.27) (-0.013 - 0.0051) 

20:1 Eicosenoic 0.072 (0.0020) 0.071 (0.0020) 0.0010 (0.0025) -0.0059, 0.0079 0.701 

(0.069 - 0.076) (0.069 - 0.076) (-0.00013 - 

0.0025)

22:0 Behenic 0.28 (0.0033) 0.29 (0.0033) -0.014 (0.0047) -0.027, -0.0012 0.038 

(0.27 - 0.29) (0.29 - 0.30) (-0.023 - 0.00015) 

Vitamin (mg/100g dwt)Vitamin E 1.28 (0.054) 1.16 (0.054) 0.12 (0.067) -0.065, 0.31 0.146 

(1.25 - 1.30) (1.10 - 1.23) (0.018 - 0.18) 

1dwt = dry weight; fwt = fresh weight; FA = fatty acid. 2MON 87708 was treated with dicamba. 3Mean (S.E.) = least-square mean (standard error). 4Control refers to the near isogenic conventional soybean control A3525. 5With 95% confidence, interval contains 99% of the values expressed in the population of commercial reference varieties. Negative limits set to zero.
Monsanto Company 10-SY-210U 

Table E-14. Statistical Summary of Site INRC Soybean Seed Anti-nutrients for MON 87708 (TreatControl 

Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 877082 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Anti-nutrient (% dwt)Lectin (H.U./mg dwt) 3.03 (0.32) 2.56 (0.32) 0.47 (0.45) -0.79, 1.74 0.357 

(2.75 - 3.45) (2.33 - 3.02) (-0.26 - 1.13) 

Phytic Acid 1.27 (0.071) 1.19 (0.071) 0.078 (0.087) -0.16, 0.32 0.421 

(1.22 - 1.34) (1.09 - 1.36) (-0.022 - 0.15) 

Raffinose 0.37 (0.023) 0.40 (0.023) -0.032 (0.033) -0.12, 0.060 0.384 

(0.33 - 0.43) (0.36 - 0.43) (-0.098 - 0.069) 

Stachyose 3.14 (0.077) 3.46 (0.077) -0.32 (0.11) -0.62, -0.015 0.043 

(3.12 - 3.17) (3.33 - 3.67) (-0.51 - -0.21) 

Trypsin Inhibitor (TIU/mg dwt) 29.17 (1.79) 29.28 (1.79) -0.12 (1.83) -5.21, 4.97 0.952 

(26.09 - 33.09) (25.22 - 31.77) (-4.76 - 3.09) 

Isoflavone (μg/g dwt)Daidzein 1683.50 (67.03) 1419.40 (67.03) 264.10 (94.79) 0.92, 527.28 0.049 

(1593.24 - 1777.49) (1416.92 - 1421.55) (173.52 - 360.58) 
Monsanto Company 10-SY-210U 

Table E-14 (continued). Statistical Summary of Site INRC Soybean Seed Anti-nutrients for MON Conventional Control 

Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 877082 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Isoflavone (μg/g dwt)Genistein 1033.37 (50.59) 862.03 (50.59) 171.34 (71.54) -27.29, 369.97 0.074 

(963.43 - 1092.19) (840.10 - 890.94) (108.39 - 204.37) 

Glycitein 111.51 (3.23) 98.42 (3.23) 13.10 (3.31) 3.91, 22.29 0.016 

(110.91 - 112.28) (89.42 - 103.14) (7.77 - 21.94) 

1dwt = dry weight; H.U. = Hemagglutinating Units; TIU = Trypsin Inhibitor Units. 2MON 87708 was treated with dicamba. 3Mean (S.E.) = least-square mean (standard error). 4Control refers to the near isogenic conventional soybean control A3525. 5With 95% confidence, interval contains 99% of the values expressed in the population of commercial reference soybean vaNegative limits set to zero.
Monsanto Company 10-SY-210U 

Table E-15. Statistical Summary of Site INRC Soybean Forage Nutrients for MON 87708 (Treated) vs. C

Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 877082 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Proximate (% dwt)Ash 6.45 (0.24) 6.95 (0.24) -0.50 (0.26) -1.23, 0.23 0.128 

(5.94 - 7.05) (6.84 - 7.07) (-0.89 - 0.11) 

Carbohydrates 65.19 (0.69) 63.82 (0.69) 1.38 (0.98) -1.34, 4.10 0.232 

(63.10 - 66.54) (62.91 - 64.44) (-1.34 - 3.03) 

Moisture (% fwt) 73.00 (0.32) 72.27 (0.32) 0.73 (0.39) -0.34, 1.80 0.129 

(72.40 - 73.70) (71.60 - 72.70) (-0.10 - 1.30) 

Protein 21.78 (0.41) 23.33 (0.41) -1.55 (0.41) -2.69, -0.41 0.019 

(20.99 - 22.51) (22.64 - 24.11) (-2.26 - -0.73) 

Total Fat 6.54 (0.27) 5.88 (0.27) 0.66 (0.39) -0.42, 1.73 0.164 

(6.12 - 7.34) (5.39 - 6.19) (-0.069 - 1.96) 

Fiber (% dwt)Acid Detergent Fiber 26.46 (1.50) 23.83 (1.50) 2.63 (2.13) -3.27, 8.54 0.283 

(23.30 - 31.06) (22.93 - 25.53) (-0.51 - 8.13) 
Monsanto Company 10-SY-210U 

Table E-15 (continued). Statistical Summary of Site INRC Soybean Forage Nutrients for MON 87708 TreaConventional Control 

Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 877082 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Fiber (% dwt)Neutral Detergent Fiber 27.20 (1.62) 26.11 (1.62) 1.08 (2.04) -4.59, 6.76 0.623 

(24.21 - 31.27) (23.91 - 29.42) (0.30 - 1.85) 

1dwt = dry weight; fwt = fresh weight. 2MON 87708 was treated with dicamba. 3Mean (S.E.) = least-square mean (standard error). 4Control refers to the near isogenic conventional soybean control A3525. 5With 95% confidence, interval contains 99% of the values expressed in the population of commercial reference soybean varieties. Negative limits set to zero.
Monsanto Company 10-SY-210U 

Table E-16. Statistical Summary of Site PAHM Soybean Seed Nutrients for MON 87708 (Treated) vs. C

Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 877082 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Proximate (% dwt)Ash 5.33 (0.11) 5.05 (0.11) 0.28 (0.16) -0.16, 0.72 0.148 

(5.27 - 5.39) (4.96 - 5.17) (0.15 - 0.44) 

Carbohydrates 38.30 (0.51) 35.23 (0.51) 3.07 (0.64) 1.29, 4.84 0.008 

(37.69 - 38.65) (34.49 - 35.75) (2.23 - 4.07) 

Moisture (% fwt) 7.84 (0.22) 10.50 (0.22) -2.66 (0.22) -3.27, -2.04 <0.00

(7.38 - 8.47) (10.40 - 10.60) (-3.12 - -2.13) 

Protein 40.25 (0.41) 43.69 (0.41) -3.43 (0.51) -4.86, -2.01 0.002 

(39.00 - 41.05) (43.46 - 43.85) (-4.84 - -2.70) 

Total Fat 16.10 (0.63) 16.05 (0.63) 0.050 (0.89) -2.41, 2.51 0.957 

(14.95 - 18.03) (15.64 - 16.85) (-1.90 - 2.37) 

Fiber (% dwt)Acid Detergent Fiber 12.91 (0.35) 11.96 (0.35) 0.96 (0.43) -0.24, 2.15 0.089 

(12.45 - 13.21) (11.62 - 12.17) (0.37 - 1.44) 
Monsanto Company 10-SY-210U 

Table E-16 (continued). Statistical Summary of Site PAHM Soybean Seed Nutrients for MON 87708 (TreaConventional Control 

Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 877082 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Fiber (% dwt)Crude Fiber 8.30 (0.63) 6.61 (0.63) 1.68 (0.71) -0.30, 3.66 0.077 

(6.23 - 9.65) (6.05 - 6.98) (0.18 - 2.67) 

Neutral Detergent Fiber 13.97 (0.36) 13.11 (0.36) 0.86 (0.50) -0.54, 2.25 0.164 

(13.43 - 14.97) (12.63 - 13.62) (-0.18 - 1.88) 

Amino Acid (% dwt)Alanine 1.75 (0.017) 1.86 (0.017) -0.11 (0.024) -0.17, -0.042 0.010 

(1.74 - 1.77) (1.82 - 1.90) (-0.16 - -0.054) 

Arginine 3.25 (0.053) 3.88 (0.053) -0.63 (0.074) -0.83, -0.42 0.001 

(3.09 - 3.36) (3.83 - 3.93) (-0.83 - -0.47) 

Aspartic Acid 4.56 (0.041) 4.94 (0.041) -0.38 (0.057) -0.54, -0.22 0.002 

(4.45 - 4.63) (4.90 - 5.01) (-0.56 - -0.27) 

Cystine 0.62 (0.0062) 0.59 (0.0062) 0.028 (0.0081) 0.0061, 0.051 0.024 

(0.60 - 0.63) (0.59 - 0.60) (0.0092 - 0.039) 
Monsanto Company 10-SY-210U 

Table E-16 (continued). Statistical Summary of Site PAHM Soybean Seed Nutrients for MON 87708 (TreaConventional Control 

Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 877082 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Amino Acid (% dwt)Glutamic Acid 7.28 (0.081) 8.00 (0.081) -0.73 (0.11) -1.05, -0.41 0.003 

(7.06 - 7.40) (7.91 - 8.14) (-1.09 - -0.54) 

Glycine 1.73 (0.015) 1.86 (0.015) -0.13 (0.022) -0.19, -0.066 0.004 

(1.69 - 1.75) (1.83 - 1.89) (-0.20 - -0.076) 

Histidine 1.05 (0.0089) 1.13 (0.0089) -0.085 (0.013) -0.12, -0.050 0.002 

(1.02 - 1.06) (1.11 - 1.14) (-0.12 - -0.053) 

Isoleucine 1.85 (0.026) 2.00 (0.026) -0.15 (0.036) -0.25, -0.051 0.014 

(1.79 - 1.90) (1.94 - 2.04) (-0.24 - -0.046) 

Leucine 3.03 (0.027) 3.28 (0.027) -0.24 (0.038) -0.35, -0.14 0.002 

(2.96 - 3.09) (3.24 - 3.32) (-0.36 - -0.15) 

Lysine 2.60 (0.023) 2.75 (0.023) -0.15 (0.032) -0.24, -0.060 0.009 

(2.53 - 2.65) (2.71 - 2.77) (-0.23 - -0.091) 
Monsanto Company 10-SY-210U 

Table E-16 (continued). Statistical Summary of Site PAHM Soybean Seed Nutrients for MON 87708 (TreaConventional Control 

Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 877082 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Amino Acid (% dwt)Methionine 0.58 (0.0090) 0.58 (0.0090) -0.00056 (0.0072) -0.021, 0.020 0.942 

(0.57 – 0.60) (0.57 – 0.60) (-0.0027 – 

0.00085)

Phenylalanine 2.04 (0.027) 2.21 (0.027) -0.18 (0.038) -0.28, -0.069 0.010 

(2.00 – 2.07) (2.16 – 2.27) (-0.27 - -0.083) 

Proline 1.98 (0.024) 2.10 (0.024) -0.13 (0.032) -0.21, -0.038 0.016 

(1.94 – 2.00) (2.08 – 2.13) (-0.14 - -0.11) 

Serine 2.04 (0.035) 2.16 (0.035) -0.12 (0.050) -0.26, 0.018 0.073 

(2.01 – 2.06) (2.06 – 2.21) (-0.19 – 0.0063) 

Threonine 1.55 (0.015) 1.62 (0.015) -0.069 (0.021) -0.13, -0.011 0.029 

(1.52 – 1.57) (1.60 – 1.64) (-0.10 - -0.032) 

Tryptophan 0.47 (0.012) 0.46 (0.012) 0.010 (0.017) -0.036, 0.057 0.567 

(0.44 – 0.48) (0.45 – 0.46) (-0.0098 – 0.023) 
Monsanto Company 10-SY-210U 

Table E-16 (continued). Statistical Summary of Site PAHM Soybean Seed Nutrients for MON 87708 (TreaConventional Control 

Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 877082 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Amino Acid (% dwt)Tyrosine 1.35 (0.027) 1.49 (0.027) -0.14 (0.032) -0.23, -0.052 0.011 

(1.28 – 1.43) (1.48 – 1.52) (-0.20 - -0.088) 

Valine 1.95 (0.029) 2.13 (0.029) -0.17 (0.041) -0.29, -0.061 0.012 

(1.89 – 2.00) (2.05 – 2.17) (-0.27 - -0.050) 

Fatty Acid (% Total FA)16:0 Palmitic 11.74 (0.12) 11.49 (0.12) 0.25 (0.15) -0.17, 0.67 0.169 

(11.39 – 12.07) (11.38 – 11.55) (-0.15 – 0.52) 

18:0 Stearic 3.85 (0.12) 3.76 (0.12) 0.093 (0.14) -0.30, 0.48 0.544 

(3.60 – 4.12) (3.67 – 3.91) (-0.078 – 0.42) 

18:1 Oleic 18.58 (0.31) 20.01 (0.31) -1.43 (0.35) -2.40, -0.45 0.015 

(17.85 – 19.42) (19.60 – 20.32) (-1.74 - -0.90) 

18:2 Linoleic 54.89 (0.41) 54.68 (0.41) 0.21 (0.53) -1.25, 1.67 0.708 

(53.59 – 55.67) (54.18 – 54.99) (-0.59 – 0.68) 
Monsanto Company 10-SY-210U 

Table E-16 (continued). Statistical Summary of Site PAHM Soybean Seed Nutrients for MON 87708 (TreaConventional Control 

Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 877082 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Fatty Acid (% Total FA)18:3 Linolenic 10.33 (0.21) 9.47 (0.21) 0.85 (0.16) 0.40, 1.31 0.006 

(9.91 - 10.88) (9.13 - 9.68) (0.59 - 1.20) 

20:0 Arachidic 0.25 (0.0063) 0.24 (0.0063) 0.0034 (0.0071) -0.016, 0.023 0.652 

(0.23 - 0.26) (0.24 - 0.25) (-0.0079 - 0.020) 

20:1 Eicosenoic 0.091 (0.015) 0.072 (0.015) 0.018 (0.022) -0.042, 0.078 0.445 

(0.073 - 0.12) (0.068 - 0.075) (-0.0014 - 0.050) 

22:0 Behenic 0.27 (0.0046) 0.27 (0.0046) -0.0067 (0.0034) -0.016, 0.0028 0.121 

(0.26 - 0.28) (0.27 - 0.28) (-0.012 - 0.0024) 

Vitamin (mg/100g dwt)Vitamin E 1.32 (0.10) 1.23 (0.10) 0.097 (0.022) 0.037, 0.16 0.010 

(1.21 - 1.54) (1.11 - 1.40) (0.049 - 0.14) 

1dwt = dry weight; fwt = fresh weight; FA = fatty acid. 2MON 87708 was treated with dicamba. 3Mean (S.E.) = least-square mean (standard error) 4Control refers to the near isogenic conventional soybean control A3525. 5With 95% confidence, interval contains 99% of the values expressed in the population of commercial reference soybean varieties. Negative limits set to zero.
Monsanto Company 10-SY-210U 

Table E-17. Statistical Summary of Site PAHM Soybean Seed Anti-nutrients for MON 87708 (TreatControl 

Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 877082 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Anti-nutrient (% dwt)Lectin (H.U./mg dwt) 2.81 (0.80) 3.85 (0.80) -1.03 (1.03) -3.88, 1.81 0.370 

(2.08 - 3.34) (3.28 - 4.45) (-1.73 - 0.051) 

Phytic Acid 1.35 (0.077) 1.50 (0.077) -0.15 (0.092) -0.40, 0.11 0.179 

(1.13 - 1.51) (1.41 - 1.62) (-0.28 - -0.054) 

Raffinose 0.47 (0.047) 0.54 (0.047) -0.072 (0.066) -0.26, 0.11 0.339 

(0.32 - 0.55) (0.49 - 0.57) (-0.24 - 0.058) 

Stachyose 3.29 (0.18) 3.36 (0.18) -0.075 (0.25) -0.77, 0.62 0.777 

(3.19 - 3.47) (3.07 - 3.90) (-0.69 - 0.40) 

Trypsin Inhibitor (TIU/mg dwt) 32.53 (4.33) 30.39 (4.33) 2.14 (5.59) -13.38, 17.66 0.721 

(27.64 - 36.16) (26.59 - 33.33) (1.05 - 2.83) 

Isoflavone (μg/g dwt)Daidzein 1918.51 (144.27) 1642.38 (144.27) 276.14 (204.03) -290.35, 842.62 0.247 

(1565.54 - 2305.26) (1510.07 - 1729.91) (-121.61 - 795.19) 
Monsanto Company 10-SY-210U 

Table E-17 (continued). Statistical Summary of Site PAHM Soybean Seed Anti-nutrients for MON 87708 (Conventional Control 

Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 877082 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Isoflavone (μg/g dwt)Genistein 1196.16 (107.80) 1101.98 (107.80) 94.18 (152.45) -329.09, 517.45 0.570 

(976.03 - 1496.78) (983.22 - 1162.01) (-185.98 - 513.56) 

Glycitein 110.37 (7.95) 81.44 (7.95) 28.93 (11.25) -2.30, 60.16 0.061 

(93.26 - 119.09) (68.68 - 90.51) (2.75 - 50.41) 

1dwt = dry weight; H.U. = Hemagglutinating Units; TIU = Trypsin Inhibitor Units. 2 MON 87708 was treated with dicamba. 3Mean (S.E.) = least-square mean (standard error). 4Control refers to the near isogenic conventional soybean control A3525. 5With 95% confidence, interval contains 99% of the values expressed in the population of commercial reference soybean varieties. Negative limits set to zero.
Monsanto Company 10-SY-210U 

Table E-18. Statistical Summary of Site PAHM Soybean Forage Nutrients for MON 87708 (Treated) vs. C

Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 877082 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Proximate (% dwt)Ash 7.51 (0.26) 7.88 (0.32) -0.37 (0.41) -1.67, 0.94 0.438 

(7.18 – 8.13) (7.67 – 8.09) (-0.86 - -0.49) 

Carbohydrates 70.95 (1.04) 65.81 (1.16) 5.14 (1.03) 1.85, 8.43 0.015 

(69.23 – 73.31) (65.74 – 66.41) (3.49 – 6.90) 

Moisture (% fwt) 74.27 (0.63) 74.91 (0.63) -0.64 (0.15) -1.11, -0.18 0.021 

(73.40 – 75.40) (73.80 – 74.90) (-0.90 - -0.40) 

Protein 17.47 (1.26) 21.96 (1.45) -4.49 (1.45) -9.10, 0.12 0.053 

(15.23 – 19.58) (21.49 – 21.91) (-6.26 - -2.34) 

Total Fat 3.97 (0.26) 4.18 (0.31) -0.20 (0.31) -1.19, 0.78 0.553 

(3.82 – 4.21) (4.30 – 4.35) (-0.42 - -0.14) 

Fiber (% dwt)Acid Detergent Fiber 26.97 (1.95) 26.02 (2.39) 0.96 (3.08) -8.86, 10.77 0.776 

(25.46 – 29.89) (21.79 – 30.24) (-4.78 – 8.09) 
Monsanto Company 10-SY-210U 

Table E-18 (continued). Statistical Summary of Site PAHM Soybean Forage Nutrients for MON 87708 (TConventional Control 

Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 877082 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Fiber (% dwt)Neutral Detergent Fiber 29.45 (1.01) 30.20 (1.24) -0.75 (1.60) -5.83, 4.34 0.672 

(27.00 - 32.07) (30.00 - 30.40) (-3.40 - -0.71) 

1dwt = dry weight; fwt = fresh weight. 2 MON 87708 was treated with dicamba. 3Mean (S.E.) = least-square mean (standard error) 4Control refers to the near isogenic conventional soybean control A3525. 5With 95% confidence, interval contains 99% of the values expressed in the population of commercial reference soybean varieties. Negative limits set to zero.
Monsanto Company 10-SY-210U 

Table E-19. Statistical Summary of Site IARL Soybean Seed Nutrients for MON 87708 (Untreated) vs. Co

 Difference (MON 87708minus Control)

Analytical Component (Units)1

MON 877082 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Proximate (% dw)Ash 5.35 (0.070) 5.29 (0.070) 0.058 (0.098) -0.22, 0.33 0.589 

(5.24 - 5.46) (5.19 - 5.47) (-0.0028 - 0.16) 

Carbohydrates 37.68 (0.56) 37.97 (0.56) -0.29 (0.55) -1.83, 1.25 0.628 

(36.27 - 39.10) (37.17 - 38.45) (-0.90 - 0.65) 

Moisture (% fw) 7.84 (0.28) 6.07 (0.28) 1.77 (0.39) 0.68, 2.86 0.010 

(7.39 - 8.73) (5.84 - 6.36) (1.05 - 2.89) 

Protein 40.95 (0.27) 41.09 (0.27) -0.15 (0.16) -0.60, 0.31 0.420 

(40.72 - 41.20) (40.69 - 41.74) (-0.54 - 0.069) 

Total Fat 16.03 (0.47) 15.61 (0.47) 0.42 (0.66) -1.41, 2.25 0.562 

(14.69 - 17.31) (15.38 - 15.82) (-0.69 - 1.49) 

Fiber (% dw)Acid Detergent Fiber 13.78 (0.38) 12.60 (0.38) 1.18 (0.54) -0.33, 2.69 0.096 

(12.82 - 14.36) (11.92 - 13.17) (-0.35 - 2.45) 
Monsanto Company 10-SY-210U 

Table E-19 (continued). Statistical Summary of Site IARL Soybean Seed Nutrients for MON 87708 (UntreatedControl 

 Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 877082 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Fiber (% dw)Crude Fiber 7.63 (0.33) 7.41 (0.33) 0.23 (0.46) -1.05, 1.50 0.650 

(6.85 - 8.07) (7.17 - 7.60) (-0.60 - 0.90) 

Neutral Detergent Fiber 14.50 (0.89) 13.27 (0.89) 1.22 (1.25) -2.26, 4.71 0.384 

(13.26 - 15.98) (11.81 - 14.42) (-0.34 - 4.17) 

Amino Acid (% dw)Alanine 1.75 (0.026) 1.74 (0.026) 0.0049 (0.036) -0.096, 0.11 0.900 

(1.72 - 1.79) (1.69 - 1.77) (-0.028 - 0.030) 

Arginine 3.35 (0.055) 3.45 (0.055) -0.099 (0.061) -0.27, 0.070 0.177 

(3.30 - 3.41) (3.27 - 3.56) (-0.19 - 0.037) 

Aspartic Acid 4.60 (0.060) 4.63 (0.060) -0.033 (0.084) -0.27, 0.20 0.711 

(4.57 - 4.62) (4.46 - 4.74) (-0.11 - 0.10) 

Cystine 0.61 (0.0094) 0.59 (0.0094) 0.018 (0.013) -0.019, 0.054 0.256 

(0.61 - 0.61) (0.56 - 0.62) (-0.0068 - 0.044) 
Monsanto Company 10-SY-210U 

Table E-19 (continued). Statistical Summary of Site IARL Soybean Seed Nutrients for MON 87708 (UntreatedControl 

 Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 877082 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Amino Acid (% dw)Glutamic Acid 7.33 (0.10) 7.41 (0.10) -0.074 (0.15) -0.48, 0.34 0.644 

(7.25 - 7.43) (7.12 - 7.58) (-0.19 - 0.12) 

Glycine 1.75 (0.024) 1.76 (0.024) -0.013 (0.034) -0.11, 0.081 0.720 

(1.74 - 1.76) (1.70 - 1.79) (-0.049 - 0.041) 

Histidine 1.05 (0.014) 1.06 (0.014) -0.0055 (0.019) -0.059, 0.048 0.786 

(1.05 - 1.06) (1.02 - 1.08) (-0.026 - 0.029) 

Isoleucine 1.87 (0.038) 1.88 (0.038) -0.011 (0.054) -0.16, 0.14 0.851 

(1.86 - 1.88) (1.79 - 1.94) (-0.068 - 0.085) 

Leucine 3.05 (0.041) 3.07 (0.041) -0.020 (0.058) -0.18, 0.14 0.746 

(3.01 - 3.08) (2.96 - 3.14) (-0.065 - 0.055) 

Lysine 2.62 (0.030) 2.62 (0.030) -0.00013 (0.042) -0.12, 0.12 0.997 

(2.61 - 2.62) (2.54 - 2.66) (-0.047 - 0.083) 
Monsanto Company 10-SY-210U 

Table E-19 (continued). Statistical Summary of Site IARL Soybean Seed Nutrients for MON 87708 (UntreatedControl 

 Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 877082 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Amino Acid (% dw)Methionine 0.58 (0.013) 0.56 (0.013) 0.019 (0.019) -0.034, 0.072 0.376 

(0.57 - 0.59) (0.53 - 0.60) (-0.0030 - 0.056) 

Phenylalanine 2.03 (0.036) 2.03 (0.036) -0.00029 (0.050) -0.14, 0.14 0.995 

(1.98 - 2.09) (1.95 - 2.09) (-0.024 - 0.022) 

Proline 1.94 (0.035) 1.98 (0.035) -0.048 (0.050) -0.19, 0.091 0.390 

(1.89 - 1.98) (1.89 - 2.07) (-0.10 - 0.043) 

Serine 2.02 (0.044) 2.00 (0.044) 0.017 (0.036) -0.082, 0.12 0.653 

(1.91 - 2.10) (1.95 - 2.08) (-0.043 - 0.073) 

Threonine 1.53 (0.023) 1.54 (0.023) -0.0028 (0.023) -0.067, 0.061 0.909 

(1.49 - 1.57) (1.51 - 1.58) (-0.026 - 0.033) 

Tryptophan 0.44 (0.0092) 0.45 (0.0092) -0.012 (0.013) -0.048, 0.024 0.405 

(0.41 - 0.46) (0.44 - 0.47) (-0.053 - 0.018) 
Monsanto Company 10-SY-210U 

Table E-19 (continued). Statistical Summary of Site IARL Soybean Seed Nutrients for MON 87708 (UntreatedControl 

 Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 877082 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Amino Acid (% dw)Tyrosine 1.40 (0.013) 1.38 (0.013) 0.012 (0.016) -0.033, 0.057 0.494 

(1.38 - 1.40) (1.37 - 1.42) (-0.011 - 0.032) 

Valine 1.98 (0.046) 1.99 (0.046) -0.0086 (0.065) -0.19, 0.17 0.900 

(1.97 - 2.00) (1.90 - 2.05) (-0.077 - 0.097) 

Fatty Acid (% Total FA)16:0 Palmitic 11.51 (0.051) 11.00 (0.051) 0.51 (0.062) 0.34, 0.68 0.001 

(11.39 - 11.63) (10.92 - 11.08) (0.41 - 0.59) 

18:0 Stearic 3.95 (0.067) 4.00 (0.067) -0.052 (0.095) -0.32, 0.21 0.613 

(3.75 - 4.06) (3.99 - 4.01) (-0.23 - 0.044) 

18:1 Oleic 19.28 (0.20) 21.67 (0.20) -2.39 (0.28) -3.17, -1.61 0.001 

(18.77 - 19.68) (21.48 - 21.78) (-2.71 - -2.07) 

18:2 Linoleic 54.30 (0.33) 52.70 (0.33) 1.60 (0.46) 0.32, 2.89 0.025 

(53.70 - 55.34) (52.66 - 52.73) (1.04 - 2.61) 
Monsanto Company 10-SY-210U 

Table E-19 (continued). Statistical Summary of Site IARL Soybean Seed Nutrients for MON 87708 (UntreatedControl 

 Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 877082 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Fatty Acid (% Total FA)18:3 Linolenic 10.38 (0.13) 10.04 (0.13) 0.34 (0.19) -0.18, 0.86 0.144 

(9.94 - 10.60) (10.00 - 10.12) (-0.18 - 0.61) 

20:0 Arachidic 0.25 (0.0038) 0.25 (0.0038) -0.0024 (0.0054) -0.017, 0.013 0.680 

(0.24 - 0.26) (0.25 - 0.26) (-0.014 - 0.0033) 

20:1 Eicosenoic 0.071 (0.0020) 0.070 (0.0020) 0.00087 (0.0028) -0.0070, 0.0087 0.773 

(0.065 - 0.076) (0.069 - 0.071) (-0.0039 - 0.0043) 

22:0 Behenic 0.26 (0.0035) 0.28 (0.0035) -0.014 (0.0042) -0.025, -0.0021 0.030 

(0.26 - 0.27) (0.27 - 0.28) (-0.017 - -0.010) 

Vitamin (mg/100g dw)Vitamin E 1.26 (0.058) 0.94 (0.058) 0.32 (0.066) 0.13, 0.50 0.008 

(1.11 - 1.42) (0.89 - 0.97) (0.23 - 0.46) 

1dw = dry weight; fw = fresh weight; FA = fatty acid. 2MON 87708 plants were not sprayed with dicamba, but received another conventional treatment as was done for the conve3Mean (S.E.) = least-square mean (standard error). 4Control refers to the non-biotechnology derived, conventional control (A3525). 5With 95% confidence, interval contains 99% of the values expressed in the population of conventional substances. Negativ
Monsanto Company 10-SY-210U 

Table E-20. Statistical Summary of Site IARL Soybean Seed Anti-Nutrients for MON 87708 (Untreated) vControl 

Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 877082 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Anti-nutrientLectin (H.U./mg dw) 2.98 (0.64) 1.53 (0.64) 1.46 (0.90) -1.04, 3.96 0.180 

(2.35 - 3.87) (0.46 - 2.70) (-0.35 - 2.46) 

Phytic Acid (% dw) 1.43 (0.032) 1.53 (0.032) -0.10 (0.045) -0.23, 0.024 0.088 

(1.38 - 1.47) (1.47 - 1.62) (-0.18 - -0.040) 

Raffinose (% dw) 0.41 (0.022) 0.43 (0.022) -0.024 (0.020) -0.079, 0.030 0.284 

(0.38 - 0.45) (0.40 - 0.45) (-0.063 - 0.00015) 

Stachyose (% dw) 3.30 (0.20) 3.89 (0.20) -0.59 (0.28) -1.37, 0.19 0.101 

(2.94 - 3.59) (3.76 - 4.15) (-0.82 - -0.18) 

Trypsin Inhibitor (TIU/mg dw) 29.97 (2.63) 29.67 (2.63) 0.30 (3.19) -8.57, 9.17 0.929 

(23.32 - 33.42) (25.64 - 32.71) (-2.32 - 2.51) 

Isoflavone (μg/g dw)Daidzein 1734.44 (145.92) 1447.97 (145.92) 286.47 (206.36) -286.47, 859.41 0.237 

(1522.84 - 2125.56) (1404.40 - 1505.77) (17.08 - 691.83) 
Monsanto Company 10-SY-210U 

Table E-20 (continued). Statistical Summary of Site IARL Soybean Seed Anti-Nutrients for MON 87708 (UntrConventional Control 

 Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 877082 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Isoflavone (μg/g dw)Genistein 1099.12 (83.29) 954.99 (83.29) 144.13 (117.79) -182.91, 471.17 0.288 

(954.54 - 1380.52) (900.10 - 984.62) (-22.32 - 400.27) 

Glycitein 110.70 (5.35) 106.40 (5.35) 4.30 (2.24) -1.92, 10.52 0.127 

(99.92 - 117.72) (97.49 - 111.71) (2.43 - 7.73) 

1dw = dry weight; H.U. = Hemagglutinating Units; TIU = Trypsin Inhibitor Units. 2 MON 87708 plants were not sprayed with dicamba, but received another conventional treatment as was done for the conve3Mean (S.E.) = least-square mean (standard error). 4Control refers to the non-biotechnology derived, conventional control (A3525). 5With 95% confidence, interval contains 99% of the values expressed in the population of conventional substances. Negativ
Monsanto Company 10-SY-210U 

Table E-21. Statistical Summary of Site IARL Soybean Forage Nutrients for MON 87708 (Untreated) vs. Control 

 Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 877082 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Proximate (% dw)Ash 8.37 (0.62) 8.87 (0.62) -0.50 (0.52) -1.95, 0.95 0.395 

(7.23 - 9.45) (7.58 - 10.46) (-1.01 - -0.13) 

Carbohydrates 62.49 (0.80) 65.57 (0.80) -3.08 (1.13) -6.22, 0.067 0.053 

(61.64 - 63.41) (63.58 - 67.74) (-6.11 - -0.17) 

Moisture (% fw) 83.47 (0.33) 81.97 (0.33) 1.50 (0.44) 0.28, 2.72 0.027 

(82.70 - 84.10) (81.40 - 82.70) (0.90 - 2.70) 

Protein 25.76 (0.54) 23.00 (0.54) 2.76 (0.76) 0.65, 4.88 0.022 

(24.63 - 27.04) (22.15 - 24.07) (1.54 - 4.89) 

Total Fat 3.29 (0.32) 2.52 (0.32) 0.77 (0.46) -0.50, 2.04 0.169 

(2.61 - 3.91) (2.01 - 3.27) (-0.66 - 1.63) 

Fiber (% dw)Acid Detergent Fiber 31.48 (2.83) 32.62 (2.83) -1.14 (3.30) -10.31, 8.04 0.748 

(29.42 - 34.63) (28.87 - 38.15) (-3.52 - 1.51) 
Monsanto Company 10-SY-210U 

Table E-21 (continue). Statistical Summary of Site IARL Soybean Forage Nutrients for MON 87708 (UntreateControl 

 Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 877082 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Fiber (% dw)Neutral Detergent Fiber 37.35 (2.66) 35.01 (2.66) 2.34 (3.76) -8.11, 12.79 0.567 

(37.11 - 37.80) (27.47 - 39.42) (-2.29 - 9.63) 

1dw = dry weight; fw = fresh weight. 2MON 87708 plants were not sprayed with dicamba, but received another conventional treatment as was done for the conve3Mean (S.E.) = least-square mean (standard error). 4Control refers to the non-biotechnology derived, conventional control (A3525). 5With 95% confidence, interval contains 99% of the values expressed in the population of conventional substances. Negativ
Monsanto Company 10-SY-210U 

Table E-22. Statistical Summary of Site ILCY Soybean Seed Nutrients for MON 87708 (Untreated) vs. Co

 Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 877082 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Proximate (% dw)Ash 5.10 (0.080) 5.05 (0.080) 0.046 (0.089) -0.20, 0.29 0.632 

(4.94 - 5.27) (4.88 - 5.22) (0.025 - 0.059) 

Carbohydrates 35.98 (0.68) 35.77 (0.68) 0.21 (0.97) -2.48, 2.90 0.839 

(35.27 - 37.36) (34.11 - 37.16) (-1.89 - 3.25) 

Moisture (% fw) 5.78 (0.20) 6.44 (0.20) -0.66 (0.28) -1.43, 0.11 0.077 

(5.68 - 5.88) (6.20 - 6.63) (-0.75 - -0.52) 

Protein 41.07 (0.40) 41.72 (0.40) -0.65 (0.55) -2.17, 0.88 0.305 

(40.92 - 41.22) (40.81 - 42.67) (-1.60 - 0.42) 

Total Fat 17.87 (0.45) 17.49 (0.45) 0.37 (0.63) -1.39, 2.13 0.588 

(16.66 - 18.66) (16.81 - 18.39) (-1.73 - 1.46) 

Fiber (% dw)Acid Detergent Fiber 14.54 (0.45) 14.04 (0.45) 0.50 (0.61) -1.18, 2.19 0.452 

(13.36 - 15.72) (13.47 - 14.57) (-0.71 - 1.16) 
Monsanto Company 10-SY-210U 

Table E-22 (continued). Statistical Summary of Site ILCY Soybean Seed Nutrients for MON 87708 (UntreatedControl 

 Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 877082 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Fiber (% dw)Crude Fiber 8.08 (0.29) 8.38 (0.29) -0.30 (0.41) -1.44, 0.84 0.508 

(7.30 - 8.89) (8.20 - 8.64) (-1.33 - 0.60) 

Neutral Detergent Fiber 17.05 (0.63) 16.85 (0.63) 0.20 (0.73) -1.84, 2.24 0.797 

(16.33 - 18.38) (15.19 - 17.99) (-1.05 - 1.27) 

Amino Acid (% dw)Alanine 1.78 (0.014) 1.81 (0.014) -0.027 (0.020) -0.084, 0.030 0.259 

(1.77 - 1.80) (1.78 - 1.84) (-0.067 - -0.0037) 

Arginine 3.21 (0.053) 3.30 (0.053) -0.094 (0.049) -0.23, 0.043 0.129 

(3.12 - 3.27) (3.19 - 3.43) (-0.21 - -0.0074) 

Aspartic Acid 4.69 (0.037) 4.76 (0.037) -0.072 (0.034) -0.17, 0.022 0.099 

(4.63 - 4.74) (4.73 - 4.82) (-0.12 - 0.0057) 

Cystine 0.60 (0.0078) 0.59 (0.0078) 0.013 (0.011) -0.018, 0.043 0.322 

(0.59 - 0.61) (0.58 - 0.60) (-0.0016 - 0.030) 
Monsanto Company 10-SY-210U 

Table E-22 (continued). Statistical Summary of Site ILCY Soybean Seed Nutrients for MON 87708 (UntreatedControl 

 Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 877082 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Amino Acid (% dw)Glutamic Acid 7.44 (0.073) 7.61 (0.073) -0.17 (0.056) -0.33, -0.014 0.039 

(7.33 - 7.51) (7.52 - 7.76) (-0.28 - -0.042) 

Glycine 1.78 (0.013) 1.81 (0.013) -0.027 (0.014) -0.066, 0.012 0.126 

(1.77 - 1.78) (1.79 - 1.83) (-0.046 - -0.0099) 

Histidine 1.06 (0.0071) 1.08 (0.0071) -0.014 (0.0055) -0.029, 0.00090 0.059 

(1.05 - 1.07) (1.07 - 1.09) (-0.019 - -0.0059) 

Isoleucine 1.94 (0.013) 1.97 (0.013) -0.035 (0.017) -0.083, 0.013 0.110 

(1.91 - 1.95) (1.97 - 1.97) (-0.058 - -0.021) 

Leucine 3.11 (0.022) 3.17 (0.022) -0.058 (0.012) -0.090, -0.025 0.008 

(3.08 - 3.13) (3.14 - 3.19) (-0.060 - -0.056) 

Lysine 2.66 (0.017) 2.67 (0.017) -0.012 (0.013) -0.047, 0.024 0.413 

(2.62 - 2.67) (2.65 - 2.69) (-0.032 - 0.017) 
Monsanto Company 10-SY-210U 

Table E-22 (continued). Statistical Summary of Site ILCY Soybean Seed Nutrients for MON 87708 (UntreatedControl 

 Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 877082 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Amino Acid (% dw)Methionine 0.57 (0.012) 0.57 (0.012) -0.0026 (0.017) -0.049, 0.044 0.886 

(0.56 - 0.58) (0.56 - 0.58) (-0.0052 - -

0.00024)

Phenylalanine 2.07 (0.022) 2.13 (0.022) -0.064 (0.018) -0.12, -0.014 0.024 

(2.05 - 2.08) (2.09 - 2.19) (-0.11 - -0.038) 

Proline 1.95 (0.019) 2.06 (0.019) -0.11 (0.019) -0.16, -0.058 0.004 

(1.90 - 1.98) (2.04 - 2.09) (-0.14 - -0.058) 

Serine 2.06 (0.027) 2.08 (0.027) -0.015 (0.025) -0.084, 0.055 0.593 

(2.05 - 2.08) (2.01 - 2.15) (-0.069 - 0.037) 

Threonine 1.58 (0.015) 1.59 (0.015) -0.00051 (0.019) -0.052, 0.051 0.979 

(1.56 - 1.61) (1.58 - 1.59) (-0.023 - 0.020) 

Tryptophan 0.48 (0.0090) 0.48 (0.0090) -0.0026 (0.0084) -0.026, 0.021 0.770 

(0.46 - 0.48) (0.47 - 0.50) (-0.015 - 0.017) 
Monsanto Company 10-SY-210U 

Table E-22 (continued). Statistical Summary of Site ILCY Soybean Seed Nutrients for MON 87708 (UntreatedControl 

 Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 877082 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Amino Acid (% dw)Tyrosine 1.42 (0.033) 1.43 (0.033) -0.0065 (0.045) -0.13, 0.12 0.892 

(1.35 - 1.48) (1.39 - 1.51) (-0.075 - 0.098) 

Valine 2.01 (0.015) 2.05 (0.015) -0.043 (0.021) -0.10, 0.016 0.113 

(1.99 - 2.04) (2.05 - 2.06) (-0.070 - -0.011) 

Fatty Acid (% Total FA)16:0 Palmitic 12.12 (0.073) 11.95 (0.073) 0.17 (0.089) -0.076, 0.42 0.127 

(12.10 - 12.15) (11.73 - 12.08) (0.067 - 0.37) 

18:0 Stearic 3.94 (0.044) 3.93 (0.044) 0.0064 (0.062) -0.16, 0.18 0.922 

(3.87 - 4.05) (3.86 - 4.02) (-0.13 - 0.19) 

18:1 Oleic 19.85 (0.29) 21.57 (0.29) -1.72 (0.41) -2.86, -0.58 0.013 

(19.62 - 20.22) (21.07 - 22.44) (-2.82 - -0.85) 

18:2 Linoleic 54.31 (0.25) 53.26 (0.25) 1.05 (0.35) 0.074, 2.02 0.040 

(53.67 - 54.63) (52.77 - 53.74) (-0.064 - 1.85) 
Monsanto Company 10-SY-210U 

Table E-22 (continued). Statistical Summary of Site ILCY Soybean Seed Nutrients for MON 87708 (UntreatedControl 

 Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 877082 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Fatty Acid (% Total FA)18:3 Linolenic 9.09 (0.074) 8.58 (0.074) 0.51 (0.099) 0.23, 0.79 0.006 

(8.94 - 9.23) (8.42 - 8.71) (0.23 - 0.67) 

20:0 Arachidic 0.26 (0.0035) 0.26 (0.0035) -0.0013 (0.0050) -0.015, 0.013 0.802 

(0.26 - 0.27) (0.26 - 0.27) (-0.014 - 0.014) 

20:1 Eicosenoic 0.15 (0.0016) 0.16 (0.0016) -0.0078 (0.0023) -0.014, -0.0013 0.028 

(0.15 - 0.16) (0.16 - 0.17) (-0.013 - -0.0010) 

22:0 Behenic 0.28 (0.0030) 0.29 (0.0030) -0.0065 (0.0042) -0.018, 0.0052 0.199 

(0.28 - 0.28) (0.28 - 0.30) (-0.019 - -0.00002) 

Vitamin (mg/100g dw)Vitamin E 2.21 (0.077) 1.86 (0.077) 0.35 (0.089) 0.10, 0.60 0.016 

(2.11 - 2.27) (1.71 - 2.11) (0.16 - 0.49) 

1dw = dry weight; fw = fresh weight; FA = fatty acid. 2 MON 87708 plants were not sprayed with dicamba, but received another conventional treatment as was done for the conve3Mean (S.E.) = least-square mean (standard error). 4Control refers to the non-biotechnology derived, conventional control (A3525). 5With 95% confidence, interval contains 99% of the values expressed in the population of conventional substances. Negativ
Monsanto Company 10-SY-210U 

Table E-23. Statistical Summary of Site ILCY Soybean Seed Anti-Nutrients for MON 87708 (Untreated) vControl 

 Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 877082 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Anti-nutrientLectin (H.U./mg dw) 3.52 (1.81) 5.54 (1.81) -2.02 (2.55) -9.11, 5.07 0.473 

(2.56 - 4.82) (2.14 - 10.38) (-7.83 - 1.05) 

Phytic Acid (% dw) 1.08 (0.032) 1.16 (0.032) -0.077 (0.045) -0.20, 0.048 0.163 

(1.05 - 1.11) (1.10 - 1.22) (-0.17 - 0.015) 

Raffinose (% dw) 0.56 (0.013) 0.57 (0.013) -0.0071 (0.018) -0.057, 0.043 0.711 

(0.55 - 0.58) (0.54 - 0.60) (-0.045 - 0.049) 

Stachyose (% dw) 3.56 (0.10) 3.64 (0.10) -0.086 (0.14) -0.48, 0.31 0.576 

(3.35 - 3.74) (3.43 - 3.83) (-0.32 - 0.30) 

Trypsin Inhibitor (TIU/mg dw) 34.71 (1.10) 32.78 (1.10) 1.93 (1.56) -2.39, 6.26 0.282 

(33.01 - 37.61) (31.06 - 34.22) (-0.72 - 6.55) 

Isoflavone (μg/g dw)Daidzein 1061.06 (72.66) 922.21 (72.66) 138.85 (102.76) -146.46, 424.16 0.247 

(910.73 - 1156.87) (762.49 - 1098.08) (-187.35 - 394.38) 
Monsanto Company 10-SY-210U 

Table E-23 (continued). Statistical Summary of Site ILCY Soybean Seed Anti-Nutrients for MON 87708 (UntrConventional Control 

 Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 877082 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Isoflavone (μg/g dw)Genistein 721.38 (45.60) 653.27 (45.60) 68.12 (64.49) -110.93, 247.17 0.350 

(654.16 - 792.83) (588.17 - 770.79) (-116.63 - 204.65) 

Glycitein 125.94 (8.11) 113.29 (8.11) 12.65 (11.47) -19.20, 44.51 0.332 

(120.06 - 133.73) (96.25 - 122.09) (-2.04 - 37.48) 

1dw = dry weight; H.U. = Hemagglutinating Units; TIU = Trypsin Inhibitor Units. 2 MON 87708 plants were not sprayed with dicamba, but received another conventional treatment as was done for the conve3Mean (S.E.) = least-square mean (standard error). 4Control refers to the non-biotechnology derived, conventional control (A3525). 5With 95% confidence, interval contains 99% of the values expressed in the population of conventional substances. Negativ
Monsanto Company 10-SY-210U 

Table E-24. Statistical Summary of Site ILCY Soybean Forage Nutrients for MON 87708 (Untreated) vs. Control 

 Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 877082 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Proximate (% dw)Ash 5.45 (0.22) 6.27 (0.22) -0.82 (0.32) -1.70, 0.052 0.059 

(4.92 - 6.04) (6.10 - 6.49) (-1.57 - -0.054) 

Carbohydrates 66.21 (0.61) 66.38 (0.61) -0.17 (0.87) -2.58, 2.24 0.855 

(65.30 - 67.29) (65.53 - 67.94) (-1.89 - 1.62) 

Moisture (% fw) 73.27 (0.25) 73.53 (0.25) -0.27 (0.35) -1.24, 0.70 0.488 

(73.20 - 73.40) (73.20 - 73.80) (-0.60 - 0.20) 

Protein 22.53 (0.51) 21.64 (0.51) 0.89 (0.68) -1.00, 2.79 0.261 

(22.29 - 22.65) (20.88 - 23.03) (-0.38 - 1.77) 

Total Fat 5.73 (0.38) 5.77 (0.38) -0.040 (0.52) -1.47, 1.39 0.942 

(5.34 - 6.04) (5.15 - 6.72) (-1.38 - 0.67) 

Fiber (% dw)Acid Detergent Fiber 28.43 (1.09) 28.46 (1.09) -0.027 (1.54) -4.30, 4.24 0.986 

(27.35 - 30.49) (27.05 - 30.90) (-0.41 - 0.31) 
Monsanto Company 10-SY-210U 

Table E-24 (continued). Statistical Summary of Site ILCY Soybean Forage Nutrients for MON 87708 (UntreatControl 

 Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 877082 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Fiber (% dw)Neutral Detergent Fiber 32.83 (1.65) 28.06 (1.65) 4.76 (2.33) -1.72, 11.24 0.110 

(30.68 - 34.48) (23.66 - 32.73) (0.59 - 10.81) 

1dw = dry weight; fw = fresh weight. 2MON 87708 plants were not sprayed with dicamba, but received another conventional treatment as was done for the conve3Mean (S.E.) = least-square mean (standard error). 4Control refers to the non-biotechnology derived, conventional control (A3525). 5With 95% confidence, interval contains 99% of the values expressed in the population of conventional substances. Negativ
Monsanto Company 10-SY-210U 

Table E-25. Statistical Summary of Site ILWY Soybean Seed Nutrients for MON 87708 (Untreated) vs. C

 Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 877082 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Proximate (% dw)Ash 5.30 (0.084) 5.23 (0.084) 0.068 (0.084) -0.17, 0.30 0.465 

(5.24 - 5.34) (5.13 - 5.29) (0.039 - 0.11) 

Carbohydrates 38.39 (0.44) 38.29 (0.44) 0.10 (0.45) -1.15, 1.35 0.835 

(37.03 - 39.08) (38.19 - 38.42) (-1.17 - 0.83) 

Moisture (% fw) 6.66 (0.16) 6.16 (0.16) 0.51 (0.22) -0.11, 1.12 0.084 

(6.34 - 6.83) (5.79 - 6.41) (-0.070 - 1.03) 

Protein 41.96 (0.28) 41.99 (0.28) -0.022 (0.38) -1.07, 1.02 0.956 

(41.53 - 42.72) (41.72 - 42.25) (-0.46 - 0.47) 

Total Fat 14.36 (0.38) 14.49 (0.38) -0.14 (0.44) -1.37, 1.10 0.776 

(13.95 - 15.03) (14.40 - 14.54) (-0.59 - 0.62) 

Fiber (% dw)Acid Detergent Fiber 13.71 (0.43) 12.96 (0.43) 0.75 (0.61) -0.93, 2.43 0.283 

(13.31 - 14.17) (12.48 - 13.69) (-0.39 - 1.68) 
Monsanto Company 10-SY-210U 

Table E-25 (continued). Statistical Summary of Site ILWY Soybean Seed Nutrients for MON 87708 (UntreateControl 

 Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 877082 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Fiber (% dw)Crude Fiber 7.37 (0.52) 7.58 (0.52) -0.21 (0.74) -2.25, 1.83 0.787 

(6.14 - 8.72) (7.39 - 7.82) (-1.25 - 1.19) 

Neutral Detergent Fiber 15.79 (0.72) 13.78 (0.72) 2.00 (0.94) -0.61, 4.62 0.100 

(15.13 - 16.53) (13.44 - 14.00) (1.23 - 3.09) 

Amino Acid (% dw)Alanine 1.77 (0.0076) 1.75 (0.0076) 0.020 (0.0093) -0.0057, 0.046 0.096 

(1.76 - 1.79) (1.75 - 1.76) (0.0086 - 0.043) 

Arginine 3.52 (0.037) 3.57 (0.037) -0.049 (0.040) -0.16, 0.062 0.288 

(3.41 - 3.60) (3.55 - 3.60) (-0.14 - 0.051) 

Aspartic Acid 4.72 (0.017) 4.67 (0.017) 0.050 (0.020) -0.0047, 0.11 0.064 

(4.68 - 4.75) (4.67 - 4.68) (-0.0019 - 0.082) 

Cystine 0.63 (0.0090) 0.61 (0.0090) 0.021 (0.0092) -0.0042, 0.047 0.081 

(0.61 - 0.64) (0.58 - 0.62) (0.0091 - 0.034) 
Monsanto Company 10-SY-210U 

Table E-25 (continued). Statistical Summary of Site ILWY Soybean Seed Nutrients for MON 87708 (UntreateControl 

 Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 877082 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Amino Acid (% dw)Glutamic Acid 7.53 (0.030) 7.51 (0.030) 0.023 (0.031) -0.064, 0.11 0.506 

(7.49 - 7.56) (7.49 - 7.53) (-0.0030 - 0.066) 

Glycine 1.78 (0.0053) 1.77 (0.0053) 0.013 (0.0075) -0.0078, 0.034 0.155 

(1.77 - 1.79) (1.76 - 1.77) (-0.0019 - 0.032) 

Histidine 1.08 (0.0032) 1.07 (0.0032) 0.011 (0.0046) -0.0016, 0.024 0.071 

(1.07 - 1.08) (1.06 - 1.07) (0.00005 - 0.024) 

Isoleucine 1.90 (0.012) 1.90 (0.012) 0.0031 (0.0040) -0.0081, 0.014 0.486 

(1.88 - 1.91) (1.88 - 1.91) (-0.00070 - 

0.0093)

Leucine 3.12 (0.011) 3.10 (0.011) 0.017 (0.0062) -0.00030, 0.034 0.052 

(3.10 - 3.13) (3.09 - 3.13) (0.0081 - 0.030) 

Lysine 2.68 (0.0062) 2.63 (0.0062) 0.046 (0.0088) 0.022, 0.071 0.006 

(2.66 - 2.69) (2.62 - 2.64) (0.029 - 0.069) 
Monsanto Company 10-SY-210U 

Table E-25 (continued). Statistical Summary of Site ILWY Soybean Seed Nutrients for MON 87708 (UntreateControl 

 Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 877082 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Amino Acid (% dw)Methionine 0.59 (0.0074) 0.58 (0.0074) 0.0067 (0.010) -0.022, 0.036 0.553 

(0.58 - 0.59) (0.56 - 0.59) (-0.0018 - 0.017) 

Phenylalanine 2.07 (0.019) 2.07 (0.019) -0.0031 (0.021) -0.063, 0.056 0.892 

(2.06 - 2.08) (2.05 - 2.10) (-0.044 - 0.023) 

Proline 2.02 (0.020) 2.05 (0.020) -0.028 (0.028) -0.11, 0.049 0.368 

(1.97 - 2.05) (2.01 - 2.09) (-0.12 - 0.041) 

Serine 2.10 (0.013) 2.06 (0.013) 0.043 (0.013) 0.0084, 0.078 0.026 

(2.09 - 2.11) (2.02 - 2.09) (0.023 - 0.073) 

Threonine 1.59 (0.0042) 1.55 (0.0042) 0.044 (0.0049) 0.031, 0.058 <0.00

(1.58 - 1.60) (1.54 - 1.55) (0.031 - 0.052) 

Tryptophan 0.48 (0.0083) 0.47 (0.0083) 0.016 (0.011) -0.015, 0.046 0.227 

(0.46 - 0.50) (0.45 - 0.48) (-0.014 - 0.032) 
Monsanto Company 10-SY-210U 

Table E-25 (continued). Statistical Summary of Site ILWY Soybean Seed Nutrients for MON 87708 (UntreateControl 

 Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 877082 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Amino Acid (% dw)Tyrosine 1.40 (0.024) 1.36 (0.024) 0.032 (0.027) -0.044, 0.11 0.305 

(1.35 - 1.44) (1.34 - 1.40) (-0.0023 - 0.084) 

Valine 2.00 (0.013) 2.01 (0.013) -0.011 (0.012) -0.045, 0.024 0.443 

(1.97 - 2.02) (2.01 - 2.02) (-0.031 - 0.012) 

Fatty Acid (% Total FA)16:0 Palmitic 11.13 (0.041) 11.04 (0.041) 0.085 (0.057) -0.074, 0.24 0.212 

(11.02 - 11.21) (10.97 - 11.12) (-0.094 - 0.24) 

18:0 Stearic 4.47 (0.063) 4.25 (0.063) 0.23 (0.089) -0.019, 0.47 0.062 

(4.30 - 4.57) (4.16 - 4.31) (-0.010 - 0.38) 

18:1 Oleic 20.24 (0.25) 21.14 (0.25) -0.89 (0.36) -1.89, 0.11 0.068 

(19.52 - 20.73) (20.78 - 21.55) (-2.03 - -0.045) 

18:2 Linoleic 52.87 (0.32) 52.90 (0.32) -0.024 (0.46) -1.30, 1.25 0.961 

(52.18 - 53.74) (52.33 - 53.20) (-0.98 - 1.41) 
Monsanto Company 10-SY-210U 

Table E-25 (continued). Statistical Summary of Site ILWY Soybean Seed Nutrients for MON 87708 (UntreateControl 

 Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 877082 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Fatty Acid (% Total FA)18:3 Linolenic 10.65 (0.10) 10.05 (0.10) 0.60 (0.14) 0.20, 1.00 0.014 

(10.37 - 10.90) (9.89 - 10.14) (0.25 - 1.01) 

20:0 Arachidic 0.28 (0.0041) 0.27 (0.0041) 0.011 (0.0058) -0.0048, 0.027 0.123 

(0.27 - 0.29) (0.26 - 0.27) (-0.0031 - 0.022) 

20:1 Eicosenoic 0.077 (0.0020) 0.076 (0.0020) 0.00030 (0.0023) -0.0060, 0.0066 0.901 

(0.073 - 0.079) (0.075 - 0.077) (-0.0037 - 0.0039) 

22:0 Behenic 0.28 (0.0032) 0.28 (0.0032) -0.0029 (0.0040) -0.014, 0.0081 0.503 

(0.28 - 0.29) (0.28 - 0.29) (-0.010 - 0.0054) 

Vitamin (mg/100g dw)Vitamin E 1.15 (0.037) 0.94 (0.037) 0.21 (0.053) 0.061, 0.35 0.017 

(1.11 - 1.20) (0.89 - 0.99) (0.12 - 0.31) 

1dw = dry weight; fw = fresh weight; FA = fatty acid. 2 MON 87708 plants were not sprayed with dicamba, but received another conventional treatment as was done for the conve3Mean (S.E.) = least-square mean (standard error). 4Control refers to the non-biotechnology derived, conventional control (A3525). 5With 95% confidence, interval contains 99% of the values expressed in the population of conventional substances. Negativ
Monsanto Company 10-SY-210U 

Table E-26. Statistical Summary of Site ILWY Soybean Seed Anti-Nutrients for MON 87708 (Untreated) Control 

 Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 877082 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Anti-nutrientLectin (H.U./mg dw) 1.36 (0.44) 2.33 (0.44) -0.97 (0.43) -2.17, 0.23 0.087 

(1.18 - 1.63) (1.34 - 3.68) (-2.04 - -0.16) 

Phytic Acid (% dw) 1.43 (0.033) 1.55 (0.033) -0.12 (0.044) -0.24, 0.0051 0.056 

(1.43 - 1.43) (1.47 - 1.61) (-0.19 - -0.045) 

Raffinose (% dw) 0.37 (0.026) 0.41 (0.026) -0.038 (0.036) -0.14, 0.064 0.361 

(0.34 - 0.41) (0.41 - 0.41) (-0.065 - 0.0072) 

Stachyose (% dw) 3.60 (0.19) 3.76 (0.19) -0.17 (0.27) -0.92, 0.59 0.573 

(3.37 - 3.85) (3.68 - 3.85) (-0.39 - 0.0078) 

Trypsin Inhibitor (TIU/mg dw) 38.93 (2.07) 29.73 (2.07) 9.20 (2.89) 1.18, 17.22 0.033 

(37.89 - 39.49) (25.43 - 32.22) (5.67 - 13.97) 

Isoflavone (μg/g dw)Daidzein 1307.12 (35.08) 1271.60 (35.08) 35.52 (31.48) -51.87, 122.92 0.322 

(1291.91 - 1320.17) (1196.71 - 1354.96) (-34.79 - 95.20) 
Monsanto Company 10-SY-210U 

Table E-26 (continued). Statistical Summary of Site ILWY Soybean Seed Anti-Nutrients for MON 87708 (UntConventional Control 

 Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 877082 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Isoflavone (μg/g dw)Genistein 832.43 (28.15) 860.58 (28.15) -28.14 (39.82) -138.69, 82.40 0.518 

(780.21 - 862.94) (784.27 - 913.26) (-103.98 - 69.88) 

Glycitein 96.96 (2.15) 79.70 (2.15) 17.26 (2.23) 11.06, 23.46 0.001 

(92.19 - 103.25) (77.14 - 81.62) (11.83 - 21.63) 

1dw = dry weight; H.U. = Hemagglutinating Units; TIU = Trypsin Inhibitor Units. 2MON 87708 plants were not sprayed with dicamba, but received another conventional treatment as was done for the conve3Mean (S.E.) = least-square mean (standard error). 4Control refers to the non-biotechnology derived, conventional control (A3525). 5With 95% confidence, interval contains 99% of the values expressed in the population of conventional substances. Negativ
Monsanto Company 10-SY-210U 

Table E-27. Statistical Summary of Site ILWY Soybean Forage Nutrients for MON 87708 (Untreated) vs. Control 

 Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 877082 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Proximate (% dw)Ash 6.76 (0.23) 6.88 (0.23) -0.12 (0.30) -0.95, 0.71 0.706 

(6.52 - 7.14) (6.82 - 6.99) (-0.30 - 0.15) 

Carbohydrates 65.94 (0.77) 66.26 (0.77) -0.31 (1.08) -3.32, 2.69 0.786 

(64.34 - 67.08) (65.71 - 66.67) (-1.37 - 0.68) 

Moisture (% fw) 75.13 (0.55) 75.40 (0.55) -0.27 (0.65) -2.07, 1.54 0.702 

(74.10 - 75.70) (75.10 - 75.60) (-1.00 - 0.10) 

Protein 22.27 (0.58) 21.97 (0.58) 0.30 (0.82) -1.98, 2.59 0.730 

(22.05 - 22.54) (21.81 - 22.13) (0.091 - 0.58) 

Total Fat 5.01 (0.52) 4.91 (0.52) 0.11 (0.35) -0.86, 1.08 0.773 

(4.25 - 6.52) (4.50 - 5.63) (-0.31 - 0.88) 

Fiber (% dw)Acid Detergent Fiber 27.49 (1.73) 27.77 (1.73) -0.28 (2.44) -7.06, 6.49 0.912 

(23.32 - 29.63) (25.12 - 31.00) (-7.68 - 4.38) 
Monsanto Company 10-SY-210U 

Table E-27 (continued). Statistical Summary of Site ILWY Soybean Forage Nutrients for MON 87708 (UntreaControl 

 Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 877082 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Fiber (% dw)Neutral Detergent Fiber 29.89 (1.83) 33.25 (1.83) -3.36 (2.59) -10.55, 3.83 0.264 

(26.37 - 35.43) (31.89 - 34.06) (-5.93 - 1.37) 

1dw = dry weight; fw = fresh weight. 2MON 87708 plants were not sprayed with dicamba, but received another conventional treatment as was done for the conve3Mean (S.E.) = least-square mean (standard error). 4Control refers to the non-biotechnology derived, conventional control (A3525). 5With 95% confidence, interval contains 99% of the values expressed in the population of conventional substances. Negativ
Monsanto Company 10-SY-210U 

Table E-28. Statistical Summary of Site INRC Soybean Seed Nutrients for MON 87708 (Untreated) vs. Co

 Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 877082 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Proximate (% dw)Ash 5.13 (0.11) 4.95 (0.11) 0.18 (0.15) -0.24, 0.60 0.295 

(5.05 - 5.30) (4.73 - 5.23) (-0.18 - 0.41) 

Carbohydrates 37.64 (0.78) 35.95 (0.78) 1.69 (1.10) -1.37, 4.75 0.200 

(35.63 - 39.79) (35.27 - 37.04) (0.10 - 4.52) 

Moisture (% fw) 6.67 (0.20) 6.53 (0.20) 0.13 (0.29) -0.67, 0.93 0.667 

(6.26 - 7.11) (6.32 - 6.84) (-0.18 - 0.31) 

Protein 41.47 (0.53) 43.58 (0.53) -2.11 (0.76) -4.21, -0.014 0.049 

(40.22 - 43.06) (43.50 - 43.69) (-3.28 - -0.63) 

Total Fat 15.75 (0.45) 15.55 (0.45) 0.20 (0.56) -1.37, 1.77 0.739 

(14.93 - 16.26) (14.52 - 16.10) (-1.10 - 1.55) 

Fiber (% dw)Acid Detergent Fiber 12.89 (0.35) 12.77 (0.35) 0.12 (0.46) -1.15, 1.39 0.800 

(12.06 - 13.49) (12.28 - 13.25) (-0.72 - 1.22) 
Monsanto Company 10-SY-210U 

Table E-28 (continued). Statistical Summary of Site INRC Soybean Seed Nutrients for MON 87708 (UntreatedControl 

 Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 877082 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Fiber (% dw)Crude Fiber 7.38 (0.18) 6.89 (0.18) 0.49 (0.25) -0.21, 1.19 0.123 

(7.06 - 7.61) (6.59 - 7.12) (0.11 - 0.87) 

Neutral Detergent Fiber 14.43 (0.67) 14.66 (0.67) -0.23 (0.95) -2.86, 2.41 0.822 

(12.91 - 15.21) (13.24 - 15.56) (-2.27 - 1.97) 

Amino Acid (% dw)Alanine 1.79 (0.021) 1.84 (0.021) -0.054 (0.022) -0.12, 0.0067 0.068 

(1.76 - 1.84) (1.80 - 1.87) (-0.10 - -0.027) 

Arginine 3.40 (0.044) 3.72 (0.044) -0.32 (0.062) -0.49, -0.14 0.007 

(3.35 - 3.50) (3.64 - 3.81) (-0.46 - -0.19) 

Aspartic Acid 4.72 (0.062) 4.89 (0.062) -0.17 (0.078) -0.39, 0.044 0.091 

(4.59 - 4.90) (4.80 - 4.95) (-0.34 - -0.050) 

Cystine 0.62 (0.0067) 0.59 (0.0067) 0.031 (0.0094) 0.0050, 0.057 0.029 

(0.60 - 0.63) (0.59 - 0.60) (0.0031 - 0.047) 
Monsanto Company 10-SY-210U 

Table E-28 (continued). Statistical Summary of Site INRC Soybean Seed Nutrients for MON 87708 (UntreatedControl 

 Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 877082 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Amino Acid (% dw)Glutamic Acid 7.56 (0.11) 7.93 (0.11) -0.36 (0.14) -0.76, 0.035 0.064 

(7.36 - 7.88) (7.75 - 8.02) (-0.66 - -0.14) 

Glycine 1.80 (0.020) 1.85 (0.020) -0.054 (0.027) -0.13, 0.020 0.111 

(1.76 - 1.85) (1.82 - 1.87) (-0.11 - -0.016) 

Histidine 1.07 (0.012) 1.11 (0.012) -0.038 (0.016) -0.083, 0.0072 0.079 

(1.05 - 1.11) (1.10 - 1.12) (-0.072 - -0.0075) 

Isoleucine 1.92 (0.042) 2.00 (0.042) -0.079 (0.057) -0.24, 0.080 0.238 

(1.80 - 2.02) (1.95 - 2.03) (-0.22 - -0.0049) 

Leucine 3.12 (0.041) 3.24 (0.041) -0.12 (0.052) -0.26, 0.023 0.080 

(3.03 - 3.24) (3.20 - 3.26) (-0.22 - -0.023) 

Lysine 2.67 (0.030) 2.72 (0.030) -0.053 (0.035) -0.15, 0.045 0.205 

(2.60 - 2.76) (2.68 - 2.76) (-0.13 - -0.0027) 
Monsanto Company 10-SY-210U 

Table E-28 (continued). Statistical Summary of Site INRC Soybean Seed Nutrients for MON 87708 (UntreatedControl 

 Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 877082 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Amino Acid (% dw)Methionine 0.59 (0.0086) 0.59 (0.0086) 0.00055 (0.012) -0.033, 0.034 0.966 

(0.56 - 0.61) (0.58 - 0.60) (-0.036 - 0.021) 

Phenylalanine 2.08 (0.030) 2.20 (0.030) -0.12 (0.040) -0.23, -0.011 0.037 

(2.03 - 2.14) (2.13 - 2.23) (-0.17 - -0.090) 

Proline 2.01 (0.010) 2.06 (0.010) -0.054 (0.015) -0.095, -0.014 0.020 

(1.99 - 2.04) (2.05 - 2.07) (-0.079 - -0.015) 

Serine 2.06 (0.028) 2.13 (0.028) -0.072 (0.029) -0.15, 0.0094 0.070 

(1.97 - 2.11) (2.13 - 2.14) (-0.15 - -0.026) 

Threonine 1.57 (0.019) 1.61 (0.019) -0.040 (0.027) -0.11, 0.034 0.206 

(1.51 - 1.61) (1.60 - 1.62) (-0.088 - -0.0061) 

Tryptophan 0.48 (0.013) 0.45 (0.013) 0.026 (0.018) -0.025, 0.076 0.231 

(0.46 - 0.50) (0.43 - 0.46) (0.0012 - 0.044) 
Monsanto Company 10-SY-210U 

Table E-28 (continued). Statistical Summary of Site INRC Soybean Seed Nutrients for MON 87708 (UntreatedControl 

 Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 877082 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Amino Acid (% dw)Tyrosine 1.39 (0.024) 1.44 (0.024) -0.055 (0.022) -0.12, 0.0073 0.070 

(1.37 - 1.40) (1.38 - 1.47) (-0.077 - -0.018) 

Valine 2.03 (0.045) 2.12 (0.045) -0.097 (0.063) -0.27, 0.078 0.198 

(1.90 - 2.13) (2.06 - 2.16) (-0.25 - -0.015) 

Fatty Acid (% Total FA)16:0 Palmitic 11.45 (0.068) 11.19 (0.068) 0.27 (0.097) -0.00083, 0.54 0.050 

(11.23 - 11.57) (11.14 - 11.21) (0.014 - 0.44) 

18:0 Stearic 4.15 (0.047) 4.25 (0.047) -0.096 (0.066) -0.28, 0.088 0.219 

(4.07 - 4.24) (4.16 - 4.31) (-0.20 - -0.022) 

18:1 Oleic 18.80 (0.085) 20.19 (0.085) -1.39 (0.093) -1.65, -1.14 <0.00

(18.63 - 18.96) (20.12 - 20.23) (-1.60 - -1.26) 

18:2 Linoleic 54.94 (0.10) 54.43 (0.10) 0.51 (0.14) 0.11, 0.90 0.024 

(54.79 - 55.13) (54.32 - 54.64) (0.15 - 0.81) 
Monsanto Company 10-SY-210U 

Table E-28 (continued). Statistical Summary of Site INRC Soybean Seed Nutrients for MON 87708 (UntreatedControl 

 Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 877082 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Fatty Acid (% Total FA)18:3 Linolenic 10.05 (0.050) 9.31 (0.050) 0.74 (0.070) 0.54, 0.93 <0.00

(10.00 - 10.10) (9.26 - 9.40) (0.60 - 0.84) 

20:0 Arachidic 0.26 (0.0029) 0.27 (0.0029) -0.0072 (0.0041) -0.019, 0.0043 0.156 

(0.26 - 0.27) (0.26 - 0.27) (-0.014 - -0.0026) 

20:1 Eicosenoic 0.070 (0.0020) 0.071 (0.0020) -0.0014 (0.0025) -0.0083, 0.0056 0.615 

(0.068 - 0.073) (0.069 - 0.076) (-0.0073 - 0.0035) 

22:0 Behenic 0.28 (0.0033) 0.29 (0.0033) -0.016 (0.0047) -0.029, -0.0032 0.025 

(0.27 - 0.28) (0.29 - 0.30) (-0.023 - -0.0045) 

Vitamin (mg/100g dw)Vitamin E 1.25 (0.054) 1.16 (0.054) 0.091 (0.067) -0.094, 0.28 0.244 

(1.11 - 1.40) (1.10 - 1.23) (0.0086 - 0.17) 

1dw = dry weight; fw = fresh weight; FA = fatty acid. 2 MON 87708 plants were not sprayed with dicamba, but received another conventional treatment as was done for the conve3Mean (S.E.) = least-square mean (standard error). 4Control refers to the non-biotechnology derived, conventional control (A3525). 5With 95% confidence, interval contains 99% of the values expressed in the population of conventional substances. Negativ
Monsanto Company 10-SY-210U 

Table E-29. Statistical Summary of Site INRC Soybean Seed Anti-Nutrients for MON 87708 (Untreated) vControl 

 Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 877082 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Anti-nutrientLectin (H.U./mg dw) 2.94 (0.32) 2.56 (0.32) 0.39 (0.45) -0.88, 1.65 0.443 

(2.24 - 3.81) (2.33 - 3.02) (-0.78 - 1.48) 

Phytic Acid (% dw) 1.24 (0.071) 1.19 (0.071) 0.052 (0.087) -0.19, 0.29 0.585 

(1.09 - 1.37) (1.09 - 1.36) (-0.099 - 0.26) 

Raffinose (% dw) 0.45 (0.023) 0.40 (0.023) 0.049 (0.033) -0.043, 0.14 0.215 

(0.42 - 0.47) (0.36 - 0.43) (0.042 - 0.056) 

Stachyose (% dw) 3.50 (0.077) 3.46 (0.077) 0.044 (0.11) -0.26, 0.35 0.704 

(3.35 - 3.62) (3.33 - 3.67) (-0.32 - 0.28) 

Trypsin Inhibitor (TIU/mg dw) 30.68 (1.79) 29.28 (1.79) 1.40 (1.83) -3.69, 6.49 0.487 

(28.59 - 32.19) (25.22 - 31.77) (0.42 - 3.37) 

Isoflavone (μg/g dw)Daidzein 1671.09 (67.03) 1419.40 (67.03) 251.69 (94.79) -11.48, 514.87 0.056 

(1485.63 - 1842.13) (1416.92 - 1421.55) (68.71 - 422.41) 
Monsanto Company 10-SY-210U 

Table E-29 (continued). Statistical Summary of Site INRC Soybean Seed Anti-Nutrients for MON 87708 (UntrConventional Control 

 Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 877082 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Isoflavone (μg/g dw)Genistein 992.90 (50.59) 862.03 (50.59) 130.88 (71.54) -67.75, 329.51 0.141 

(842.93 - 1103.14) (840.10 - 890.94) (-48.01 - 248.10) 

Glycitein 119.27 (3.23) 98.42 (3.23) 20.86 (3.31) 11.67, 30.05 0.003 

(113.04 - 124.24) (89.42 - 103.14) (17.40 - 23.62) 

1dw = dry weight; H.U. = Hemagglutinating Units; TIU = Trypsin Inhibitor Units. 2MON 87708plants were not sprayed with dicamba, but received another conventional treatment as was done for the conven3Mean (S.E.) = least-square mean (standard error). 4Control refers to the non-biotechnology derived, conventional control (A3525). 5With 95% confidence, interval contains 99% of the values expressed in the population of conventional substances. Negativ
Monsanto Company 10-SY-210U 

Table E-30. Statistical Summary of Site INRC Soybean Forage Nutrients for MON 87708 (Untreated) vs. Control 

 Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 877082 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Proximate (% dw)Ash 6.20 (0.24) 6.95 (0.24) -0.74 (0.26) -1.47, -0.015 0.047 

(5.87 - 6.72) (6.84 - 7.07) (-1.20 - -0.22) 

Carbohydrates 64.35 (0.69) 63.82 (0.69) 0.53 (0.98) -2.18, 3.25 0.614 

(63.90 - 64.94) (62.91 - 64.44) (-0.23 - 0.99) 

Moisture (% fw) 72.70 (0.32) 72.27 (0.32) 0.43 (0.39) -0.64, 1.50 0.323 

(72.30 - 72.90) (71.60 - 72.70) (-0.20 - 1.30) 

Protein 23.46 (0.41) 23.33 (0.41) 0.13 (0.41) -1.01, 1.28 0.760 

(23.10 - 24.15) (22.64 - 24.11) (-0.10 - 0.46) 

Total Fat 5.98 (0.27) 5.88 (0.27) 0.10 (0.39) -0.97, 1.17 0.809 

(5.92 - 6.01) (5.39 - 6.19) (-0.18 - 0.63) 

Fiber (% dw)Acid Detergent Fiber 25.59 (1.50) 23.83 (1.50) 1.77 (2.13) -4.14, 7.67 0.452 

(24.69 - 27.00) (22.93 - 25.53) (-0.44 - 3.99) 
Monsanto Company 10-SY-210U 

Table E-30 (continued). Statistical Summary of Site INRC Soybean Forage Nutrients for MON 87708 (UntreaControl 

 Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 877082 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Fiber (% dw)Neutral Detergent Fiber 26.13 (1.62) 26.11 (1.62) 0.020 (2.04) -5.65, 5.69 0.992 

(25.38 - 27.64) (23.91 - 29.42) (-4.04 - 3.73) 

1dw = dry weight; fw = fresh weight. 2MON 87708 plants were not sprayed with dicamba, but received another conventional treatment as was done for the conve3Mean (S.E.) = least-square mean (standard error). 4Control refers to the non-biotechnology derived, conventional control (A3525). 5With 95% confidence, interval contains 99% of the values expressed in the population of conventional substances. Negativ
Monsanto Company 10-SY-210U 

Table E-31. Statistical Summary of Site PAHM Soybean Seed Nutrients for MON 87708 (Untreated) vs. C

 Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 877082 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Proximate (% dw)Ash 5.20 (0.11) 5.05 (0.11) 0.16 (0.16) -0.28, 0.59 0.378 

(4.89 - 5.51) (4.96 - 5.17) (-0.28 - 0.55) 

Carbohydrates 36.81 (0.51) 35.23 (0.51) 1.58 (0.64) -0.20, 3.35 0.069 

(35.36 - 37.56) (34.49 - 35.75) (0.87 - 2.06) 

Moisture (% fw) 9.53 (0.22) 10.50 (0.22) -0.97 (0.22) -1.58, -0.36 0.011 

(9.21 - 9.91) (10.40 - 10.60) (-1.19 - -0.69) 

Protein 40.38 (0.41) 43.69 (0.41) -3.31 (0.51) -4.73, -1.88 0.002 

(39.96 - 40.97) (43.46 - 43.85) (-3.89 - -2.78) 

Total Fat 17.61 (0.63) 16.05 (0.63) 1.56 (0.89) -0.90, 4.02 0.153 

(17.32 - 18.17) (15.64 - 16.85) (1.32 - 1.70) 

Fiber (% dw)Acid Detergent Fiber 11.96 (0.35) 11.96 (0.35) 0.0073 (0.43) -1.18, 1.20 0.987 

(11.01 - 12.89) (11.62 - 12.17) (-0.61 - 0.72) 
Monsanto Company 10-SY-210U 

Table E-31 (continued). Statistical Summary of Site PAHM Soybean Seed Nutrients for MON 87708 (UntreateControl 

 Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 877082 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Fiber (% dw)Crude Fiber 7.05 (0.63) 6.61 (0.63) 0.43 (0.71) -1.55, 2.41 0.578 

(6.81 - 7.28) (6.05 - 6.98) (0.064 - 0.76) 

Neutral Detergent Fiber 14.44 (0.36) 13.11 (0.36) 1.33 (0.50) -0.064, 2.73 0.056 

(14.21 - 14.87) (12.63 - 13.62) (1.12 - 1.62) 

Amino Acid (% dw)Alanine 1.79 (0.017) 1.86 (0.017) -0.068 (0.024) -0.13, -0.0016 0.046 

(1.76 - 1.81) (1.82 - 1.90) (-0.10 - -0.043) 

Arginine 3.39 (0.053) 3.88 (0.053) -0.49 (0.074) -0.70, -0.29 0.002 

(3.35 - 3.44) (3.83 - 3.93) (-0.55 - -0.45) 

Aspartic Acid 4.66 (0.041) 4.94 (0.041) -0.28 (0.057) -0.44, -0.12 0.008 

(4.64 - 4.69) (4.90 - 5.01) (-0.35 - -0.22) 

Cystine 0.61 (0.0062) 0.59 (0.0062) 0.015 (0.0081) -0.0078, 0.037 0.144 

(0.61 - 0.61) (0.59 - 0.60) (0.0099 - 0.021) 
Monsanto Company 10-SY-210U 

Table E-31 (continued). Statistical Summary of Site PAHM Soybean Seed Nutrients for MON 87708 (UntreateControl 

 Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 877082 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Amino Acid (% dw)Glutamic Acid 7.42 (0.081) 8.00 (0.081) -0.58 (0.11) -0.90, -0.26 0.007 

(7.32 - 7.50) (7.91 - 8.14) (-0.70 - -0.46) 

Glycine 1.77 (0.015) 1.86 (0.015) -0.090 (0.022) -0.15, -0.029 0.014 

(1.76 - 1.78) (1.83 - 1.89) (-0.13 - -0.068) 

Histidine 1.07 (0.0089) 1.13 (0.0089) -0.061 (0.013) -0.096, -0.026 0.008 

(1.07 - 1.08) (1.11 - 1.14) (-0.073 - -0.045) 

Isoleucine 1.89 (0.026) 2.00 (0.026) -0.11 (0.036) -0.21, -0.013 0.035 

(1.88 - 1.91) (1.94 - 2.04) (-0.16 - -0.036) 

Leucine 3.10 (0.027) 3.28 (0.027) -0.18 (0.038) -0.28, -0.074 0.009 

(3.08 - 3.11) (3.24 - 3.32) (-0.23 - -0.15) 

Lysine 2.64 (0.023) 2.75 (0.023) -0.11 (0.032) -0.20, -0.023 0.025 

(2.62 - 2.65) (2.71 - 2.77) (-0.12 - -0.091) 
Monsanto Company 10-SY-210U 

Table E-31 (continued). Statistical Summary of Site PAHM Soybean Seed Nutrients for MON 87708 (UntreateControl 

 Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 877082 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Amino Acid (% dw)Methionine 0.57 (0.0090) 0.58 (0.0090) -0.017 (0.0072) -0.037, 0.0031 0.078 

(0.55 - 0.59) (0.57 - 0.60) (-0.036 - -0.0056) 

Phenylalanine 2.08 (0.027) 2.21 (0.027) -0.13 (0.038) -0.24, -0.027 0.025 

(2.03 - 2.11) (2.16 - 2.27) (-0.18 - -0.047) 

Proline 2.00 (0.024) 2.10 (0.024) -0.11 (0.032) -0.20, -0.019 0.028 

(1.93 - 2.04) (2.08 - 2.13) (-0.17 - -0.060) 

Serine 2.08 (0.035) 2.16 (0.035) -0.078 (0.050) -0.22, 0.060 0.191 

(2.02 - 2.11) (2.06 - 2.21) (-0.19 - 0.054) 

Threonine 1.57 (0.015) 1.62 (0.015) -0.047 (0.021) -0.10, 0.010 0.085 

(1.55 - 1.60) (1.60 - 1.64) (-0.077 - 0.0039) 

Tryptophan 0.48 (0.012) 0.46 (0.012) 0.021 (0.017) -0.026, 0.067 0.287 

(0.44 - 0.50) (0.45 - 0.46) (-0.013 - 0.049) 
Monsanto Company 10-SY-210U 

Table E-31 (continued). Statistical Summary of Site PAHM Soybean Seed Nutrients for MON 87708 (UntreateControl 

 Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 877082 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Amino Acid (% dw)Tyrosine 1.44 (0.027) 1.49 (0.027) -0.057 (0.032) -0.15, 0.033 0.152 

(1.42 - 1.46) (1.48 - 1.52) (-0.086 - -0.028) 

Valine 2.00 (0.029) 2.13 (0.029) -0.13 (0.041) -0.24, -0.014 0.035 

(2.00 - 2.00) (2.05 - 2.17) (-0.17 - -0.049) 

Fatty Acid (% Total FA)16:0 Palmitic 11.79 (0.12) 11.49 (0.12) 0.31 (0.15) -0.11, 0.72 0.112 

(11.74 - 11.89) (11.38 - 11.55) (0.20 - 0.37) 

18:0 Stearic 3.70 (0.12) 3.76 (0.12) -0.056 (0.14) -0.45, 0.33 0.708 

(3.55 - 3.93) (3.67 - 3.91) (-0.12 - 0.023) 

18:1 Oleic 18.45 (0.31) 20.01 (0.31) -1.56 (0.35) -2.53, -0.58 0.011 

(18.26 - 18.80) (19.60 - 20.32) (-2.02 - -1.31) 

18:2 Linoleic 55.44 (0.41) 54.68 (0.41) 0.76 (0.53) -0.70, 2.22 0.223 

(55.13 - 55.62) (54.18 - 54.99) (0.26 - 1.43) 
Monsanto Company 10-SY-210U 

Table E-31 (continued). Statistical Summary of Site PAHM Soybean Seed Nutrients for MON 87708 (UntreateControl 

 Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 877082 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Fatty Acid (% Total FA)18:3 Linolenic 10.03 (0.21) 9.47 (0.21) 0.56 (0.16) 0.11, 1.01 0.026 

(9.74 - 10.18) (9.13 - 9.68) (0.49 - 0.61) 

20:0 Arachidic 0.24 (0.0063) 0.24 (0.0063) -0.0067 (0.0071) -0.026, 0.013 0.396 

(0.23 - 0.25) (0.24 - 0.25) (-0.011 - -0.0013) 

20:1 Eicosenoic 0.086 (0.015) 0.072 (0.015) 0.014 (0.022) -0.046, 0.073 0.565 

(0.066 - 0.13) (0.068 - 0.075) (-0.0088 - 0.058) 

22:0 Behenic 0.26 (0.0046) 0.27 (0.0046) -0.013 (0.0034) -0.022, -0.0036 0.018 

(0.25 - 0.27) (0.27 - 0.28) (-0.018 - -0.0076) 

Vitamin (mg/100g dw)Vitamin E 1.37 (0.10) 1.23 (0.10) 0.15 (0.022) 0.087, 0.21 0.002 

(1.26 - 1.59) (1.11 - 1.40) (0.10 - 0.19) 

1dw = dry weight; fw = fresh weight; FA = fatty acid. 2MON 87708 plants were not sprayed with dicamba, but received another conventional treatment as was done for the conve3Mean (S.E.) = least-square mean (standard error). 4Control refers to the non-biotechnology derived, conventional control (A3525). 5With 95% confidence, interval contains 99% of the values expressed in the population of conventional substances. Negativ
Monsanto Company 10-SY-210U 

Table E-32. Statistical Summary of Site PAHM Soybean Seed Anti-Nutrients for MON 87708 (Untreated) Control 

 Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 877082 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Anti-nutrientLectin (H.U./mg dw) 4.41 (0.80) 3.85 (0.80) 0.57 (1.03) -2.28, 3.41 0.610 

(1.96 - 6.35) (3.28 - 4.45) (-1.85 - 3.07) 

Phytic Acid (% dw) 1.44 (0.077) 1.50 (0.077) -0.057 (0.092) -0.31, 0.20 0.569 

(1.41 - 1.48) (1.41 - 1.62) (-0.21 - 0.034) 

Raffinose (% dw) 0.53 (0.047) 0.54 (0.047) -0.014 (0.066) -0.20, 0.17 0.841 

(0.48 - 0.55) (0.49 - 0.57) (-0.022 - -0.0042) 

Stachyose (% dw) 3.43 (0.18) 3.36 (0.18) 0.067 (0.25) -0.63, 0.76 0.800 

(3.26 - 3.65) (3.07 - 3.90) (-0.64 - 0.53) 

Trypsin Inhibitor (TIU/mg dw) 40.89 (4.33) 30.39 (4.33) 10.50 (5.59) -5.02, 26.01 0.133 

(28.31 - 51.50) (26.59 - 33.33) (-2.94 - 18.17) 

Isoflavone (μg/g dw)Daidzein 2085.21 (144.27) 1642.38 (144.27) 442.84 (204.03) -123.65, 1009.32 0.095 

(1931.40 - 2297.58) (1510.07 - 1729.91) (296.74 - 610.43) 
Monsanto Company 10-SY-210U 

Table E-32 (continued). Statistical Summary of Site PAHM Soybean Seed Anti-Nutrients for MON 87708 (UnConventional Control 

 Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 877082 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Isoflavone (μg/g dw)Genistein 1300.54 (107.80) 1101.98 (107.80) 198.56 (152.45) -224.71, 621.83 0.262 

(1209.90 - 1469.13) (983.22 - 1162.01) (61.89 - 307.12) 

Glycitein 99.12 (7.95) 81.44 (7.95) 17.68 (11.25) -13.55, 48.91 0.191 

(83.25 - 112.67) (68.68 - 90.51) (10.93 - 27.53) 

1dw = dry weight; H.U. = Hemagglutinating Units; TIU = Trypsin Inhibitor Units. 2MON 87708 plants were not sprayed with dicamba, but received another conventional treatment as was done for the conve3Mean (S.E.) = least-square mean (standard error). 4Control refers to the non-biotechnology derived, conventional control (A3525). 5With 95% confidence, interval contains 99% of the values expressed in the population of conventional substances. Negativ
Monsanto Company 10-SY-210U 

Table E-33. Statistical Summary of Site PAHM Soybean Forage Nutrients for MON 87708 (Untreated) vsControl 

 Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 877082 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Proximate (% dw)Ash 8.26 (0.26) 7.88 (0.32) 0.38 (0.41) -0.93, 1.69 0.422 

(7.81 - 8.65) (7.67 - 8.09) (-0.28 - 0.97) 

Carbohydrates 68.76 (1.04) 65.81 (1.16) 2.95 (1.03) -0.35, 6.24 0.065 

(67.19 - 71.05) (65.74 - 66.41) (1.45 - 4.64) 

Moisture (% fw) 74.47 (0.63) 74.91 (0.63) -0.44 (0.15) -0.91, 0.021 0.055 

(73.40 - 75.60) (73.80 - 74.90) (-0.50 - -0.40) 

Protein 19.46 (1.26) 21.96 (1.45) -2.49 (1.45) -7.10, 2.12 0.183 

(16.28 - 21.25) (21.49 - 21.91) (-5.21 - -0.66) 

Total Fat 3.46 (0.26) 4.18 (0.31) -0.72 (0.31) -1.70, 0.27 0.102 

(2.69 - 3.87) (4.30 - 4.35) (-0.49 - -0.48) 

Fiber (% dw)Acid Detergent Fiber 25.62 (1.95) 26.02 (2.39) -0.40 (3.08) -10.21, 9.42 0.905 

(24.22 - 28.01) (21.79 - 30.24) (-2.23 - 2.83) 
Monsanto Company 10-SY-210U 

Table E-33 (continued). Statistical Summary of Site PAHM Soybean Forage Nutrients for MON 87708 (UntreControl 

 Difference (MON 87708 minus Control)

Analytical Component (Units)1

MON 877082 

Mean (S.E.)3 

(Range)

Control4

Mean (S.E.) 

(Range)

Mean (S.E.) 

(Range)

95% 

Confidence Interval

Significa

(p-Valu

Fiber (% dw)Neutral Detergent Fiber 28.70 (1.01) 30.20 (1.24) -1.50 (1.60) -6.59, 3.58 0.415 

(27.46 - 29.45) (30.00 - 30.40) (-0.95 - -0.83) 

1dw = dry weight; fw = fresh weight. 2MON 87708 plants were not sprayed with dicamba, but received another conventional treatment as was done for the conve3Mean (S.E.) = least-square mean (standard error). 4Control refers to the non-biotechnology derived, conventional control (A3525). 5With 95% confidence, interval contains 99% of the values expressed in the population of conventional substances. Negativ
Monsanto Company 10-SY-210U 434 of 721 

References for Appendix E 

AACC. 1999. Official method 32-20. American Association of Cereal Chemists, St. Paul, 

Minnesota. 

AOAC-International. 2005a. AOAC official method 982.30. Official Methods of 

Analysis of AOAC International, Association of Analytical Communities International, 

Gaithersburg, Maryland. 

AOAC-International. 2005b. AOAC official method 923.03. Official Methods of 

Analysis of AOAC International, Association of Analytical Communities International, 

Gaithersburg, Maryland. 

AOAC-International. 2005c. AOAC official method 962.09. Official Methods of 

Analysis of AOAC International, Association of Analytical Communities International, 

Gaithersburg, Maryland. 

AOAC-International. 2005d. AOAC official method 922.06 and 954.02. Official 

Methods of Analysis of AOAC International, Association of Analytical Communities 

International, Gaithersburg, Maryland. 

AOAC-International. 2005e. AOAC official method 960.39 and 948.22. Official Methods 

of Analysis of AOAC International, Association of Analytical Communities International, 

Gaithersburg, Maryland. 

AOAC-International. 2005f. AOAC official method 983.23. Official Methods of 

Analysis of AOAC International, Association of Analytical Communities International, 

Gaithersburg, Maryland. 

AOAC-International. 2005g. AOAC official method 926.08 and 925.09. Official 

Methods of Analysis of AOAC International, Association of Analytical Communities 

International, Gaithersburg, Maryland. 

AOAC-International. 2005h. AOAC official method 955.04 and 979.09. Official 

Methods of Analysis of AOAC International, Association of Analytical Communities 

International, Gaithersburg, Maryland.

AOCS. 1997a. Method Ce 2-66. Official Methods and Recommended Practices of the 

American Oil Chemists Society, 5th Edition. American Oil Chemists Society, 

Champaign. 

AOCS. 1997b. Method Ba 12-75. Official Methods and Recommended Practices of the 

American Oil Chemists Society, 5th Edition. American Oil Chemists Society, 

Champaign. 
Monsanto Company 10-SY-210U 435 of 721 

AOCS. 1997c. Method Ce 1e-62. Official Methods and Recommended Practices of the 

American Oil Chemists Society, 5th Edition American Oil Chemists Society, Champaign. 

Bradstreet, R.B. 1965. The Kjeldahl method for organic nitrogen. Academic Press. New 

York, New York. 

Brobst, K.H. 1972. Gas-liquid chromatography of trimethylsilyl derivatives. Pages 3-8 in 

Methods in Carbohydrate Chemistry. Volume 6. Academic Press, New York, New York.

Cort, W., T.S. Vicente, E.H. Waysek and B.D. Williams. 1983. Vitamin E content of 

feedstuffs determined by high-performance liquid chromatographic fluorescence. Journal 

of Agriculture and Food Chemistry 31:1330-1333. 

Goering, H.K. and P.J. Van Soest. 1970. Forage and fiber analyses (apparatus, reagents, 

procedures, and some applications). Agriculture Handbook No. 379. U.S. Department of 

Agriculture, Agricultural Research Service, Washington, D.C. 

Klurfeld, D.M., and D. Kritchevsky. 1987. Isolation and quantitation of lectins from 

vegetable oils. Lipids 22:667-668. 

Kolthoff, I.M., and E.B. Sandell. 1948. Quantitative inorganic analysis. MacMillan, New 

York, New York. 

Lehrfeld, J. 1989. High-performance liquid chromatography analysis of phytic acid on a 

pH-stable, macroporous polymer column. Cereal Chemistry 66:510-515. 

Lehrfeld, J. 1994. HPLC separation and quantitation of phytic acid and some inositol 

phosphates in foods: Problem and solutions. Journal of Agricultural and Food Chemistry 

42:2726-2731. 

Liener, I.E. 1955. The photometric determination of the hemagglutinating activity of 

soyin and crude soybean extracts. Archives of Biochemistry and Biophysics 54:223-231. 

Mason, B.S., and H.T. Slover. 1971. A gas chromatographic method for the 

determination of sugars in foods. Journal of Agricultural and Food Chemistry 19:551-

554. 

McMurray, C.H., W.J. Blanchflower, and D.A. Rice. 1980. Influence of extraction 

techniques on determination of alpha-tocopherol in animal feedstuffs. JournalAssociation of Official Analytical Chemists 63:1258-1261. 

OECD. 2001. Consensus document on compositional considerations for new varieties of 

soybean: Key food and feed nutrients and anti-nutrients. ENV/JM/MONO(2001)15. 

Series on the Safety of Novel Foods and Feeds No.2. Organisation for Economic Cooperation and Development, Paris, France. 
Monsanto Company 10-SY-210U 436 of 721 

Pettersson, H., and K.-H. Kiessling. 1984. Liquid chromatographic determination of the 

plant estrogen coumestrol and isoflavones in animal feed. Journal of Association of 

Official Analytical Chemists International 67:503-506. 

SAS Software Release 9.2 (TS1M0). ©2002-2008 by SAS Institute Inc., Cary, North 

Carolina. 

Seo, A., and C.V. Morr. 1984. Improved high-performance liquid chromatographic 

analysis of phenolic acids and isoflavonoids from soybean protein products. Journal of 

Agricultural and Food Chemistry 32:530-533. 

Speek, A.J., J. Schrijver, and W.H.P. Schreurs. 1985. Vitamin E composition of some 

seed oils as determined by high-performance liquid chromatography with fluorometric 

detection. Journal of Food Science 50:121-124. 

USDA. 1973. Energy value of foods. Agricultural handbook No. 74:2-11. 
Monsanto Company 10-SY-210U 437 of 721 

Appendix F: Materials, Methods, and Individual Site Results for the Seed 

Dormancy and Germination Assessment of MON 87708 

F.1. Materials 

Seed dormancy and germination characteristics were assessed on seed from MON 87708, 

the near isogenic conventional soybean control A3525, and commercial reference 

varieties produced at the Howard County, Iowa; Stark County, Illinois; and Shelby 

County, Missouri sites in 2008 field trials (Appendix G). The field trial at each site was 

established in a randomized complete block design with three replications. The seed 

from MON 87708, conventional control, and the commercial reference varieties were 

harvested from all three replicated plots at each of the three field sites. In order to provide 

a representative sample from each field site for dormancy and germination testing, the 

replicate samples from each field trial were pooled to produce one seed sample (lot) of 

MON 87708, the conventional control, and each commercial reference variety (Table F1). 

Table F-1. Starting Seed of MON 87708, Conventional Control, and Commercial 

Reference Varieties Used in the Dormancy and Germination Assessment 

Production 

Site1 Material Name2 

Material 

Type Phenotype Seed Lot # 

IA MON 87708 Test Dicamba-Tolerant 11222450-001 

IA A3525 Control Conventional 11222449-001 

IA Garst 3585N Reference Conventional 11222451-001 

IA Pioneer 93B15 Reference Conventional 11222452-001 

IA Crows C37003N Reference Conventional 11222453-001 

IA NK S33-A8 Reference Roundup Ready 11222454-001 

IL MON 87708 Test Dicamba-Tolerant 11222456-001 

IL A3525 Control Conventional 11222455-001 

IL Croplan HT3596STS Reference Conventional 11222457-001 

IL NK S37-N4 Reference Roundup Ready 11222458-001 

IL Stewart SB3454 Reference Conventional 11222459-001 

IL Midland 363 Reference Conventional 11222460-001 

MO MON 87708 Test Dicamba-Tolerant 11222462-001 

MO A3525 Control Conventional 11222461-001 

MO Garst 3585N Reference Conventional 11222463-001 

MO Pioneer 93B15 Reference Conventional 11222464-001 

MO Crows C37003N Reference Conventional 11222465-001 

MO NK S33-A8 Reference Roundup Ready 11222466-001 1

IA = Howard County, IA; IL = Stark County, IL; MO = Shelby County, MO. 

2

MON 87708, the conventional control, and commercial reference varieties seed used to assess dormancy 

and germination characteristics were all produced from replicated field trials conducted in 2008 to assess 

plant phenotypic characteristics and, therefore, were not obtained from commercial sources. The 
Monsanto Company 10-SY-210U 438 of 721 

commercial reference varieties were all conventional soybean varieties with the exception of NK S33-A8 

and NK S37-N4, which were Roundup Ready soybean varieties.

F.2. Characterization of the Materials 

For the MON 87708, conventional control, and the commercial reference varieties 

starting seed lots, the presence or absence of the dmo expression cassette was confirmed 

by event-specific polymerase chain reaction analyses. 

F.3. Germination Testing Facility and Experimental Methods 

Seed dormancy and germination evaluations were conducted at BioDiagnostics, Inc. in 

River Falls, WI. The principal investigator was qualified to conduct seed dormancy and 

germination testing consistent with the standards established by the Association of 

Official Seed Analysts, a seed trade association (AOSA, 2000; AOSA, 2006; AOSA, 

2007). 

Seed lots of MON 87708, the conventional control, and four commercial reference 

varieties were produced from each of three sites and tested under six different 

temperature regimes. Six germination chambers were maintained dark under one of the 

following temperature regimes: constant temperature of approximately 10, 20, or 30°C 

or alternating temperatures of approximately 10/20, 10/30, or 20/30°C. The alternating 

temperature regimes were maintained at the lower temperature for 16 hours and the 

higher temperature for eight hours. The temperature inside each germination chamber 

was monitored and recorded every 15 minutes throughout the duration of the assessment. 

For each seed lot, four replicated paper germination towels were prepared per facility 

SOPs for each temperature regime. Wax coated paper was placed on a large tray 

followed by a water-moistened germination towel. A target of 100 seeds per seed lot 

were placed on the germination towel (i.e., one seed lot per towel) using a vacuum 

planting system. A second water-moistened germination towel was placed on top of the 

seed. The towels were then rolled up and secured with a rubber band. All rolled 

germination towels were placed into appropriately labeled buckets that were then covered 

with ventilated plastic bags attached with rubber bands. The buckets were arranged in 

the germination chambers in a split-plot design, where the whole-plot treatment was seed 

production site and the sub-plot treatment was seed material (i.e., MON 87708, the 

conventional control, or commercial reference varieties). 

A description of each germination characteristic evaluated and the timing of evaluations 

are presented in Table VII-1. The types of data collected depended on the temperature 

regime. Each rolled germination towel in the AOSA-recommended temperature regime 

(i.e., 20/30°C) was evaluated periodically during the study for normal germinated, 

abnormal germinated, hard, dead, and firm-swollen seed as defined by AOSA guidelines 

(AOSA, 2006; AOSA, 2007). AOSA only provides guidelines (AOSA, 2007) for testing 

seed under optimal temperatures (20/30°C); however, additional temperature regimes 

were included to test a range of temperature conditions. Each rolled germination towel in 

 

 Roundup Ready is a registered trademark of Monsanto Technology LLC. All other trademarks are the 

property of their respective owners. 
Monsanto Company 10-SY-210U 439 of 721 

the additional temperature regimes (i.e., 10, 20, 30, 10/20, and 10/30°C) was evaluated 

periodically for germinated, hard, dead, and firm-swollen seed. Emergence and/or 

development of essential structures of seedlings that otherwise would be categorized as 

“normal germinated” under optimal temperature conditions may not be so at non-optimal 

temperatures. Therefore, for the additional temperature regimes, no distinction was made 

between normal and abnormal germinated seed. 

F.4. Statistical Analysis 

Analysis of variance was conducted according to a split-plot design with four replications. 

SAS® (Version 9.2) was used to compare MON 87708 to the conventional control within 

each seed production site (individual-site analyses) and in a combined-site analysis, in 

which the data were pooled across all sites, for the following germination characteristics: 

percent germinated (categorized as percent normal germinated and percent abnormal 

germinated for the AOSA temperature regime), percent viable hard, percent dead, and 

percent viable firm-swollen seed. The level of statistical significance was predetermined 

to be 5% (α = 0.05). MON 87708 was not statistically compared to the commercial 

reference varieties nor were comparisons made across temperature regimes. For each 

assessed characteristic, the minimum and maximum means were determined from among 

the commercial reference varieties to provide a range of values that are representative of 

commercial soybean varieties. The following is a summary of the results from the 

individual-site analyses. Results from the combined-site analysis are presented in Table 

VII-2. 

F.5. Individual-Site Seed Dormancy and Germination Analysis 

In the individual-site analyses, no statistically significant differences were detected 

between MON 87708 and the conventional control for any of the measured 

characteristics (i.e., percent germinated, viable hard, dead, or viable firm-swollen seed) in 

any temperature regime for seed produced at the MO site. Six statistically significant 

differences in total were detected between MON 87708 and the conventional control for 

seed produced at the IA and IL sites (Table F-2). MON 87708 had lower percent 

germinated seed than the conventional control at 10°C for seed produced at the IA 

(97.5% vs. 99.3%) and IL (99.3% vs. 100.0%) sites and at 10/30°C for seed produced at 

the IA site (96.8% vs. 99.0%). MON 87708 had lower percent viable hard seed than the 

conventional control at 10/30°C for seed produced at the IA site (0.0% vs. 0.3%). 

Percent dead seed was higher for MON 87708 than the conventional control at 10°C 

(2.0% vs. 0.3%) and 10/30°C (3.3% vs. 0.8%) for seed produced at the IA site. 

Statistically significant differences between MON 87708 and the conventional control for 

germination characteristics in the individual-site analyses were not consistently detected 

across temperature regimes or seed production sites. While some statistically significant 

differences were detected in the combined-site analysis, the assessed dormancy and 

germination characteristics of MON 87708 were within the range of values expected for 

the commercial reference varieties and therefore are considered not biologically 

meaningful in terms of increased weediness of MON 87708 compared to the 

conventional soybean. 
Monsanto Company 10-SY-210U 

Table F-2. Dormancy and Germination Characteristics of MON 87708 and Conventional Control Seed 

Three Field Sites 

Temperature Regime Germination Category 

Mean % (S.E.)1

IA IL MO 

MON 87708 Control MON 87708 Control MON 87708 

10°C Germinated 97.5 (1.0)* 99.3 (0.5) 99.3 (0.3)* 100.0 (0.0) 100.0 (0.0)  Viable Hard 0.0 (0.0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0)  Dead 2.0 (0.9)* 0.3 (0.3) 0.5 (0.3) 0.0 (0.0) 0.0 (0.0)  Viable Firm-swollen 0.5 (0.3) 0.5 (0.3) 0.3 (0.3) 0.0 (0.0) 0.0 (0.0) 20°C Germinated 98.0 (0.9) 98.3 (0.6) 99.8 (0.3) 99.8 (0.3) 100.0 (0.0)  Viable Hard 0.0 (0.0) 0.0 (0.0) 0.0 (0.0) 0.3 (0.3) 0.0 (0.0)  Dead 2.0 (0.9) 1.8 (0.6) 0.3 (0.3) 0.0 (0.0) 0.0 (0.0)  Viable Firm-swollen 0.0 (0.0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0) 30°C Germinated 96.3 (1.1) 98.3 (0.3) 100.0 (0.0) 100.0 (0.0) 99.8 (0.3)  Viable Hard 0.0 (0.0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0)  Dead 3.8 (1.1) 1.8 (0.3) 0.0 (0.0) 0.0 (0.0) 0.3 (0.3)  Viable Firm-swollen 0.0 (0.0)† 0.0 (0.0) 0.0 (0.0)† 0.0 (0.0) 0.0 (0.0)† 
Monsanto Company 10-SY-210U 

Table F-2 (continued). Dormancy and Germination Characteristics of MON 87708 and Conventional Coat each of Three Field Sites Temperature Regime Germination Category 

Mean % (S.E.)1

IA IL 

MON 87708 Control MON 87708 Control MON 87708 

10/20°C Germinated 98.0 (0.4) 97.8 (0.3) 100.0 (0.0) 100.0 (0.0) 100.0 (0.0 Viable Hard 0.3 (0.3) 0.3 (0.3) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0 Dead 1.8 (0.3) 1.8 (0.3) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0 Viable Firm-swollen 0.0 (0.0) 0.3 (0.3) 0.0 (0.0) 0.0 (0.0) 0.0 (0.010/30°C Germinated 96.8 (0.8)* 99.0 (0.4) 99.0 (0.6) 100.0 (0.0) 100.0 (0.0 Viable Hard 0.0 (0.0)* 0.3 (0.3) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0 Dead 3.3 (0.8)* 0.8 (0.5) 1.0 (0.6) 0.0 (0.0) 0.0 (0.0 Viable Firm-swollen 0.0 (0.0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.020/30°C2 Normal Germinated 91.5 (2.1) 92.0 (0.7) 97.5 (1.0) 99.0 (0.4) 98.8 (0.5 Abnormal Germinated 5.8 (1.0) 6.3 (1.0) 1.8 (0.9) 0.8 (0.5) 1.3 (0.5 Viable Hard 0.0 (0.0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0 Dead 2.8 (1.3) 1.8 (0.8) 0.8 (0.3) 0.3 (0.3) 0.0 (0.0 Viable Firm-swollen 0.0 (0.0)† 0.0 (0.0) 0.0 (0.0)† 0.0 (0.0) 0.0 (0.0Note: Seed for the germination study were produced in Howard County, IA; Stark County, IL; and Shelby County, MO in 2008. Seed 

germination chambers in a split-plot design where the whole-plot treatment was seed production site and the sub-plot treatment was see

MON 87708, the conventional control, or commercial reference soybean varieties). *Indicates a statistically significant difference between MON 87708 and the conventional control (

α=0.05). 

†No statistical comparisons were made due to lack of variability in the data. 

1Means based on four replicates (n = 4) of 100 seeds. The total percentage of all germination characteristics of MON 87708 or the con

temperature regimes is greater than 100.0% due to numerical rounding of the means. S.E. = Standard Error

2Germinated seed in the AOSA temperature regime 20/30°C were categorized as either normal germinated or abnormal germinated see
Monsanto Company 10-SY-210U 442 of 721 

References for Appendix F 

AOSA. 2000. Tetrazolium testing handbook. Association of Official Seed Analysts, 

Lincoln, Nebraska 

AOSA. 2006. Seed evaluation handbook. Contribution No. 35. in Handbook on Seed 

Testing Association of Official Seed Analysts, Lincoln, Nebraska. 

AOSA. 2007. Rules for testing seeds. Association of Official Seed Analysis, Ithaca, New 

York. 

SAS Software Release 9.2 (TS1M0). ©2002-2008 by SAS Institute Inc., Cary, North 

Carolina. 
Monsanto Company 10-SY-210U 443 of 721 

Appendix G: Materials, Methods, and Individual-Site Results from the 

Phenotypic, Agronomic, and Environmental Interaction Assessment of 

MON 87708 under Field Conditions 

G.1. Materials 

The soybean materials for the phenotypic and environmental interactions assessment in 

the 2008 field included untreated MON 87708, the near isogenic conventional soybean 

control A3525, and 18 commercial reference soybean varieties. The references included 

both conventional and Roundup Ready soybean varieties. The list of the soybean 

materials planted at each of 18 field sites is presented in Table G-1. 

The soybean materials for the phenotypic and environmental interactions assessment in 

the 2009 field included dicamba-treated MON 87708, the near isogenic conventional 

soybean control A3525, and 14 conventional, commercial reference soybean varieties. 

The list of the soybean materials planted at each of 8 field sites is presented in Table G-2. 

G.2. Characterization of the Materials 

For the MON 87708 and the conventional control starting seed lots, the presence or 

absence of the dmo expression cassette was confirmed by event-specific polymerase 

chain reaction analyses. 

G.3. Field Sites and Plot Design 

Data were collected at 16 field sites in the U.S. and two sites in Canada during 2008 

(Section VII, Table VII-3). Data were collected at 8 field sites in the U.S. during 2009 

(Section VII, Table VII-4). These 26 locations provided a diverse range of 

environmental and agronomic conditions representative of commercial soybean 

production areas in North America. The researchers at each field site were familiar with 

the growth, production, and evaluation of soybean characteristics. 

The experiment was established at each of the 18 sites in the 2008 field trials in a 

randomized complete block design with three replications. Each plot at the IL2, IN1, MI, 

and MO1 sites consisted of twelve 30 feet long rows spaced approximately 30 inches 

apart. Rows # 2 and 3 were designated for the collection of phenotypic data. Rows # 5 

and 6 were designated for the collection of abiotic stress response, disease damage, and 

arthropod-related damage data. Rows # 8-10 were designated for the collection of 

arthropod samples. Rows # 1, 4, 7, 11, and 12 were used as buffer rows. Each plot was 

surrounded by approximately 5-15 feet of a commercial soybean variety by planting 

border rows in the alleyways between blocks and around the entire perimeter of the plot 

area. The purpose of the planted borders was to create a continuous soybean stand across 

the plot area to ensure collection of more robust arthropod abundance data. 

 

 Roundup Ready is a registered trademark of Monsanto Technology LLC. All other trademarks are the 

property of their respective owners. 
Monsanto Company 10-SY-210U 444 of 721 

Each plot at the AR, Can1, Can2, IA1, IA2, IA3, IL1, IN2, IN3, KS, MO2, NE, PA, and 

WI sites consisted of four 20 feet long rows spaced approximately 30 inches apart. 

Rows # 2 and 3 were designated for the collection of phenotypic, abiotic stress response, 

disease damage, and arthropod-related damage data. Rows # 1 and 4 were used as buffer 

rows. The entire plot area was surrounded by an approximately 10 foot wide, four-row 

border of a commercial soybean variety. 

In the 2009 field trials the experiment was established at each of the sites in a randomized 

complete block design with four replications. Each plot consisted of eight 20 feet long 

rows spaced approximately 30 inches apart. Rows 1 and 2 were designated for the 

collection of plant tissue and harvested seed samples for use in other studies. Rows 4 and 

5 were designated for the collection of phenotypic, abiotic stress response, disease 

damage, and arthropod damage data in this report. Rows 3 and 6 - 8 were used as buffer 

rows. The plots within each replicate were separated by approximately a two-row buffer 

of a commercially-available soybean variety, and the entire plot area was surrounded by 

approximately a four-row border of a commercially-available soybean variety. A 

minimum of a 15 foot fallow area was established around the perimeter of the study area 

at each site to clearly isolate the experiment from other soybean. 

G.4. Planting and Field Operations 

Field and planting information for the 2008 field trials are listed in Table G-3. Field and 

planting information for the 2009 field trials are listed in Table G-4. Agronomic 

practices used to prepare and maintain each study site were characteristic of those used in 

each respective geographic region. All maintenance operations were performed 

uniformly over the entire trial area. 
Monsanto Company 10-SY-210U 

Table G-1. Starting Seed for the Phenotypic, Agronomic, and Environmental Interaction Assessment for 2

Material Name 

Material Type 

Relative Maturity Phenotype 

Monsanto Seed Lot # Sites1

MON 87708 Test 3.5 Dicamba-Tolerant 10001256 All A3525 Control 3.5 Conventional 10001257 All FS 3591 Reference 3.5 Conventional 10001448 AR, IA2, IN1, MI, PA AG3505 Reference 3.5 Roundup Ready2 10001281 AR, IA2, IN1, MI, PA 

Wilken 3316 Reference 3.3 Conventional 10001505 AR, IA2, IN1, MI, PA Stine 3300-0 Reference 3.3 Conventional 10001312 AR, IA2, IN1, MI, PA Garst 3585N Reference 3.5 Conventional 10000883 IA3, IN2, MO1, WI Crows C37003N Reference 3.7 Conventional 10001508 IA3, IN2, MO1, WI Garst S33-A8 Reference 3.3 Roundup Ready2 10001284 Can1, IA3, IN2, MO

Pioneer 93B15 Reference 3.1 Conventional 10001304 Can1, IA3, IN2, MO

Dekalb DKB28-53 Reference 2.8 Roundup Ready2 10001950 Can1, Can2 

Asgrow AG2801 Reference 2.8 Roundup Ready2 10001951 Can1, Can2 Pioneer 93M52 Reference 3.5 Conventional 10001311 Can2, IL1, IN3, MO2 

NK S38-T8 Reference 3.8 Conventional 10001509 IL1, IN3, MO2 Lewis 3716 Reference 3.7 Roundup Ready2 10001278 IL1, IN3, MO2 Hoegemeyer 333 Reference 3.2 Conventional 10001590 Can2, IL1, IN3, MO2 Croplan Reference 3.5 Conventional 10001450 IA1, IL2, KS, NE 

NK S37-N4 Reference 3.7 Roundup Ready2 10001286 IA1, IL2, KS, NE 

Stewart SB3454 Reference 3.4 Conventional 10000887 IA1, IL2, KS, NE Midland 363 Reference 3.3 Conventional 10001570 IA1, IL2, KS, NE 1MON 87708 and the conventional control were planted at all field sites; the commercial reference varieties were site-specific. Site cod

Jackson County, AR; Can1 = Norfolk, Ontario, Canada; Can2 = Kent, Ontario, Canada; IA1 = Jefferson County, IA; IA2 = Benton CouCounty, IA; IL1 = Clinton County, IL; IL2 = Stark County, IL, IN1 = Boone County, IN; IN2 = Clinton County, IN; IN3 = Parke Coun

County, KS; MI = Ottawa County, MI; MO1 = Shelby County, MO; MO2 = Macon County, MO; NE = York County, NE; PA = Berks 

Walworth County, WI. 

2Commercial Roundup Ready soybean variety
Monsanto Company 10-SY-210U 

Table G-2. Starting Seed for the Phenotypic, Agronomic, and Environmental Interaction Assessment for 2Material Name Material Type1 Relative Maturity Phenotype2 Monsanto Seed Lot # A3525 Control 3.5 Conventional 11225301 FS 3591 Reference 3.5 Conventional 10001448 Crows C3908 Reference 3.9 Conventional 10001074 NK S38-T8 Reference 3.8 Conventional 10001509 Croplan HT3596STS Reference 3.5 Conventional 10001117 

Midland 363 Reference 3.3 Conventional 10001570 Stewart SB3454 Reference 3.4 Conventional 10001130 Quality Plus 365C Reference 3.6 Conventional 10001129 Channel Bio 3461 Reference 3.4 Conventional 10001115 Pioneer 93M52 Reference 3.5 Conventional 10001128 FS 3591 Reference 3.5 Conventional 10001448 Stewart SB3454 Reference 3.4 Conventional 10001130 NK 32Z3 Reference 3.2 Conventional 10001126 Garst 3585N Reference 3.5 Conventional 10001119 Channel Bio 37002 Reference 3.7 Conventional 10001116 Quality Plus 365C Reference 3.6 Conventional 10001129 Croplan HT3596STS Reference 3.5 Conventional 10001117 Stewart SB3454 Reference 3.4 Conventional 10001130 Crows C37003N Reference 3.7 Conventional 10001508 Pioneer 93M52 Reference 3.5 Conventional 10001128 NK S38-T8 Reference 3.8 Conventional 10001509 Quality Plus 365C Reference 3.6 Conventional 10001129 Garst 3585N Reference 3.5 Conventional 10001119 

Wilken 3316 Reference 3.3 Conventional 10001505 

Midland 363 Reference 3.3 Conventional 10001570 

MON 877084 Test DT 11225299 
Monsanto Company 10-SY-210U 

1 T = Test, C = Control, and R = reference. 2 Phenotypic abbreviations: NT = non-biotech conventional, Conventional = conventional commercial, DT = dicamba-toler3 Site codes are as follows: ARNE = Jackson County, AR; IARL = Jefferson County, IA; ILCY = Clinton County, IL; ILW

INRC = Parke County, IN; INSH = Boone County, IN; KSLA = Pawnee County, KS; NEYO = York County, NE

4 Received a mandatory dicamba application. 
Monsanto Company 10-SY-210U 

Table G-3. Field and Planting Information for 2008 Field Trials 

Site 

Planting Date1 

Planting 

Rate 

(seeds/ft

)

Planting 

Depth 

(in) 

Plot 

Size 

(ft)2 

Rows/ 

Plot Soil Series, Organic Matter, pH 

Cr

200

AR 05-29-

08 8-9 0.75 10 × 20 4 Bosket sandy loam; 1.2%; 6.4 Cotto

Can1 05-26-

08 

9 1.5 10 × 20 4 Norfolk sandy loam; 1.5%; 6.6 Cor

Can2 05-27-

08 

9 1.5 10 × 20 4 Thames clay loam; 3.4%; 7.4 Cor

IA1 06-07-

08 

9 1.25 10 × 20 4 Mahaska silty clay loam; 3.18%; 5.9 Sorgh

IA2 06-19-

08 

9 2.0 10 × 20 4 Tama Muscatine silty clay loam; 3.9%; 6.1 Soybe

IA3 06-26-

08 

9 1.0 10 × 20 4 Lawler loam; 7.3%; 7.6 Cor

IL1 06-19-

08 

9 1.25 10 × 20 4 Cisne-Huey Complex silt loam; 1.3%; 7.1 Mil

IL2 06-02-

08 

9 1.25 30 × 30 12 Plano silt loam; 3.5%; 6.4 Cor

IN1 05-28-

08 

9 1.5 30 × 30 12 Crosby silt loam; 2.5%; 7.1 Cor

IN2 05-27-

08 

9 1.5 10 × 20 4 Fincastle silt loam; 2.0%; 7.0 Sweet c

IN3 07-01-

08 

9 1.0 10 × 20 4 Silty loam; 3%; 6.5 Soybe

KS 06-04-

08 

9 1

−1.25 10 × 20 4 Farnum loam; 2.6%; 7.6 Sorgh
Monsanto Company 10-SY-210U 

1Month-day-year. 2Width × length. 

MI 05-27-

08 

9 1.5 30 × 30 12 Nester loam; 2.1%; 6.5 Cor

MO1 06-18-

08 

9 1.25 30 × 30 12 Putnam silt loam; 2.1%; 6.6 Cor

MO2 06-19-

08 

9 1.0 10 × 20 4 Gorin silt loam; 4.2%; 6.3 Soybe

NE 06-02-

08 

9 1.0 10 × 20 4 Hastings silt loam; 3.0%; 6.2 Soybe

PA 06-03-

08 

9 1.25 10 × 20 4 Philo/Atkins silt loam; 2.0%; 6.2 Fallo

WI 05-29-

08 

9 1.25 10 × 20 4 Radford silt loam; 2.2%; 5.9 Cor
Monsanto Company 10-SY-210U 

Table G-4. Field and Planting Information for 2009 Field Trials 1 Site codes are as follows: ARNE = Jackson County, AR; IARL = Jefferson County, IA; ILCY = Clinton County, IL; ILW

INRC = Parke County, IN; INSH = Boone County, IN; KSLA = Pawnee County, KS; NEYO = York County, NE 

2 Month-day-year. 3 Width × length.  * Two packets were planted on 7/06/2009 to replace two packets broken on 6/30/2009. 

Site Code1 

Planting 

date2

Planting 

rate 

(seeds/ft)

Planting 

depth 

(in) 

Plot 

size (ft)3 

Rows/ 

plot Soil series, organic matter, pH 

ARNE 6/20/2009 9 0.5 20 × 20 8 Bosket sandy loam, 1.0%, 6.0 IARL 6/26/2009 9 1.3 20 × 20 8 Taintor/Mahaska silty clay loam, 3.5%, 7.0 ILCY 6/29/2009 9 1.4 20 × 20 8 Hoyleton-Darmstadt silt loam, 2.6%, 7.3 ILWY 6/24/2009 9 1.8 20 × 20 8 Flanagan silt loam, 3.8%, 6.5 INRC 6/30/2009 

7/06/2009* 

9 1.0 20 × 20 8 Reeseville silt loam, 1.4%, 5.8 

INSH 6/19/2009 9 1.5 20 × 20 8 Crosby silt loam, 2.3%, 5.6 KSLA 6/23/2009 9 1.0 20 × 20 8 Silt loam, 2.6%, 7.6 NEYO 6/24/2009 9 1.0 20 × 20 8 Hastings silt loam, 3.0%, 6.2 
Monsanto Company 10-SY-210U 451 of 721 

G.5. Phenotypic Observations 

The description of the characteristics measured and the designated developmental stages 

when observations occurred are listed in Table VII-1. 

G.6. Environmental Interaction Observations 

Environmental interactions (i.e., interactions between the crop plants and their receiving 

environment) were used to characterize MON 87708 by evaluating plant response to 

abiotic stress, disease damage, arthropod-related damage, and pest and beneficial 

arthropod abundance in the plots using the methods described in G.7 and G.8. 

G.7. Abiotic Stress Response, Disease Damage, and Arthropod-Related Damage 

MON 87708 and the conventional control were evaluated at all 26 sites for differences in 

plant response to abiotic stress, disease damage, and arthropod-related damage. Three 

abiotic stressors, three diseases, and three arthropod pests were evaluated four times 

during the growing season at the following intervals: 

Observation 1: V2 – V4 growth stage12

Observation 2: R1 – R2 growth stage 

Observation 3: R3 – R5 growth stage 

Observation 4: R6 – R8 growth stage 

The researcher at each field site chose abiotic stressors, diseases, and arthropod pests that 

were either actively causing plant injury in the study area or were likely to occur in 

soybean during the given observation period. Therefore, abiotic stressors, diseases, and 

arthropod pests assessed often varied between observations at a site and between sites. 

Abiotic stress response and disease damage observations were collected from each plot 

using a continuous 0 – 9 scale of increasing severity. Data were collected numerically 

and then placed into one of the following categories for reporting purposes: 

Rating Severity of plant damage 

0 none (no symptoms observed) 

1 – 3 slight (symptoms not damaging to plant development) 

4 – 6 moderate (intermediate between slight and severe) 

7 – 9 severe (symptoms damaging to plant development) 

Arthropod-related damage was assessed from each plot on the upper four nodes of 10 

representative plants using the arthropod-specific 0 – 5 rating scales of increasing 

severity listed below. 

 

 

12 For the 2009 field trials Observation 1 occurred at the V2-V3 growth stage. 
Monsanto Company 10-SY-210U 452 of 721 

Defoliating arthropods (e.g., corn earworm, bean leaf beetle, 

Japanese beetle, soybean looper)

Rating Severity of plant damage 

0 None 

1 1 – 20 % defoliation 

2 21 – 40% defoliation 

3 41 – 60% defoliation 

4 61 – 80% defoliation 

5 > 80% defoliation 

Pod feeding arthropods (e.g., corn earworm, bean leaf beetle, 

stink bug, Lygus bug on reproductive plant parts)

Rating Severity of plant damage 

0 None 

1 1 – 20 % damaged pods 

2 21 – 40% damaged pods 

3 41 – 60% damaged pods 

4 61 – 80% damaged pods 

5 > 80% damaged pods 

Leafhoppers (e.g., potato leafhopper)

Rating Severity of plant damage 

0 None 

1 1 – 50% of foliage with leaf yellowing; no leaf puckering 

or leaf margin necrosis 

2 1 – 50% of foliage with leaf yellowing, leaf puckering 

and/or leaf margin necrosis 

3 > 50% of foliage with leaf yellowing; no leaf puckering 

or leaf margin necrosis 

4 > 50% of foliage with leaf yellowing, leaf puckering, 

and/or leaf margin necrosis 

5 > 50% of foliage with necrotic leaves (leaves dead due to 

leafhopper damage) 

Aphids (e.g., soybean aphid)

Rating Severity of plant damage 

0 None 

1 1 – 100 aphids per plant; no leaf puckering 

2 101 – 250 aphids per plant; no leaf puckering 

3 ≥ 250 aphids per plant with leaf puckering 

4 ≥ 250 aphids per plant with leaf puckering and leaf 

yellowing and/or necrosis 

5 ≥ 250 aphids per plant with plant stunting 
Monsanto Company 10-SY-210U 453 of 721 

G.8. Arthropod Abundance 

Pest and beneficial arthropods were collected at the IL2, IN1, MI, and MO1 sites four 

times during the growing season at the following intervals: 

Collection 1: R1 – R2 growth stage 

Collection 2: Approximately two weeks after collection 1 

Collection 3: Approximately two weeks after collection 2 

Collection 4: Approximately two weeks after collection 3 

Arthropods were collected using a beat sheet sampling method (Kogan and Pitre, 1980). 

The beat sheet was a 36 × 42 inch white, vinyl sheet that was spread between the plants 

of two adjacent rows. Plants were shaken vigorously along the length of each side of the 

beat sheet to dislodge arthropods from the plants. A total of four subsamples were 

collected in this way from each plot. Specifically, two subsamples were collected from 

rows # 8 and 9 of each plot (subsamples 1 and 3) and two subsamples were collected 

from rows # 9 and 10 of each plot (subsamples 2 and 4). The subsamples collected from 

within each pair of rows were at least 10 feet apart and at least 3 feet from the edge of 

each plot. The four subsamples were combined into one pre-labeled container and placed 

on freezer ice packs. The samples were then sent overnight to Monsanto Company, 

St. Louis, MO for arthropod identification and enumeration. 

A maximum of six pest and six beneficial arthropods were evaluated for each collection 

interval. These specific arthropods were then enumerated across all samples from a given 

collection interval at each individual site. Three of the six pest and three of the six 

beneficial arthropods were predetermined prior to the collection of samples, namely bean 

leaf beetle, green cloverworm, and stink bugs for the pests and Araneae (spiders), Nabis

spp. and Orius spp. for the beneficial arthropods, and were evaluated from all collections 

from all sites. For each specific collection interval at each individual site, up to three 

additional pest and three additional beneficial arthropods, which were determined to be 

the most abundant, were evaluated across all samples from the site in addition to the 

predetermined arthropods. The suite of pest and beneficial arthropods assessed often 

varied between collections from a site and between sites due to differences in temporal 

activity and geographical distribution of arthropod taxa. 

No arthropod abundance data were collected from the 2009 field trials. 

G.9. Environmental Interactions Evaluation Criteria 

For the assessments of abiotic stress response and disease damage, MON 87708 and the 

conventional control were considered different in susceptibility or tolerance to an abiotic 

stress or disease on a particular observation date at a site if the range of injury severity to 

MON 87708 did not overlap with the range of injury severity to the conventional control 

across all three replications. These data are categorical and were not subjected to 

statistical analysis. For each observation at a site, the range of injury severity across the 

commercial reference varieties provided data that are representative of commercial 
Monsanto Company 10-SY-210U 454 of 721 

soybean varieties. Arthropod-related damage and abundance data were quantitatively 

evaluated and subjected to statistical analysis as appropriate. 

G.10. Data Assessment 

Experienced scientists familiar with the experimental design and evaluation criteria were 

involved in all components of data collection, summarization, and analysis. Personnel 

assessed that measurements were taken properly, data were consistent with expectations 

based on experience with the crop, and the experiment was carefully monitored. Prior to 

analysis, the overall dataset was evaluated for evidence of biologically relevant changes 

and for possible evidence of an unexpected plant response. Any unexpected observations 

or issues that would impact the evaluation objectives were noted. Data were then 

subjected to statistical analysis as indicated below. 

G.11. Statistical Analysis 

Analysis of variance was conducted according to a randomized complete block design 

using SAS® (Version 9.2). The level of statistical significance was predetermined to be 

5% (α=0.05). MON 87708 was compared to the conventional control within each site 

(individual-site analyses) and in a combined-site analysis, in which the data were pooled 

across sites, for early stand count, seedling vigor, days to 50% flowering, plant height, 

lodging, pod shattering, final stand count, seed moisture, 100 seed weight, seed test 

weight, and yield. Growth stage, flower color, plant pubescence, abiotic stress response, 

and disease damage data were categorical and not statistically analyzed. Arthropodrelated damage and pest and beneficial arthropod abundance data were statistically 

analyzed only within individual observations/collections and sites due to the variation in 

temporal activity and geographical distribution of the taxa. 

No statistical comparisons were made between MON 87708 and the commercial 

reference varieties. The reference range for each measured phenotypic characteristic was 

determined from the minimum and maximum mean values from among the 18 

commercial reference varieties planted among the sites. The reference range for the 

damage from and abundance of each arthropod evaluated from a given 

observation/collection and site was determined from the minimum and maximum mean 

damage or abundance values collected from the commercial reference varieties planted at 

the site. 

G.12. Individual Field Site Plant Growth and Development Results and Discussion 

G.12.1. 2008 Untreated MON 87708 Individual Field Site Plant Growth and 

Development Results and Discussion 

In the individual-site analyses, no statistically significant differences were detected 

between MON 87708 and the conventional control for 153 out of 179 comparisons for 

the assessed phenotypic characteristics (Table G-5). Lack of variability in the data 

precluded statistical comparisons between MON 87708 and the conventional control for 

seedling vigor at the IL2 site; days to 50% flowering at the IN1 site and MI sites; lodging 

at the Can1, IN3, and MO1 sites; and pod shattering at the Can1, Can2, IA1, IA2, IA3, 
Monsanto Company 10-SY-210U 455 of 721 

IL1, IL2, IN1, IN2, IN3, KS, MI, and NE sites. For these data, the means for 

MON 87708 and the conventional control were the same value, indicating no biological 

differences (Table G-5). 

A total of 26 statistically significant differences were detected between MON 87708 and 

the conventional control in the individual-site analyses (Table G-5). These differences 

were distributed among nine phenotypic characteristics. Seedlings of MON 87708 were 

less vigorous than the conventional control at the Can2 site (6.3 vs. 5.0 rating). 

MON 87708 flowered one day later than the conventional control at the MO2 site (214 vs. 

213 days after Jan. 1, 2008), but one day earlier than the conventional control at the WI 

site (210 vs. 211 days after Jan. 1, 2008). Plants of MON 87708 were taller than the 

conventional control at the AR (28.9 vs. 27.1 inches), IA1 (39.1 vs. 34.1 inches), IA2 

(37.1 vs. 34.6 inches), IL1 (26.1 vs. 22.3 inches), IN1 (33.6 vs. 30.5 inches), IN2 (39.6 vs. 

37.3 inches), MO1 (23.4 vs. 21.1 inches), and MO2 (31.4 vs. 28.7 inches) sites. 

MON 87708 had more lodging than the conventional control at the IA1 (2.0 vs. 0.7 

rating), IL2 (1.0 vs. 0.3 rating), and the WI (1.3 vs. 0.0 rating) sites, but less lodging than 

the conventional control at the KS site (1.3 vs. 2.0 rating). Final stand count was higher 

for MON 87708 than the conventional control at the IN3 site (330.3 vs. 298.0 plants/plot). 

Seed moisture was higher for MON 87708 than the conventional control at the WI site 

(11.7 vs. 11.2%). The weight of 100 seeds was lower for MON 87708 than the 

conventional control at the Can1 (15.9 vs. 17.1 g), Can2 (16.9 vs. 18.1 g), IL2 (14.2 vs. 

15.0 g), NE (15.4 vs. 15.9 g), and PA (13.4 vs. 15.4 g) sites. Test weight was higher for 

MON 87708 than the conventional control at the AR site (55.4 vs. 54.0 lb/bu), but lower 

than the conventional control at the IL1 site (54.2 vs. 55.9 lb/bu). Yield was lower for 

MON 87708 than the conventional control at the AR (63.3 vs. 70.4 bu/a) and PA (53.9 vs. 

65.5 bu/a) sites. Since the statistically significant differences detected in the individualsite analyses for seedling vigor, days to 50% flowering, lodging, final stand count, seed 

moisture, test weight, and yield were not detected in the combined--site analysis, this 

suggests these differences were not indicative of a consistent response in the data 

associated with the trait and are considered not biologically meaningful in terms of 

increased weediness of MON 87708 compared to the conventional control. While a 

statistically significant difference was detected for plant height and 100 seed weight in 

the combined-site analysis, the assessed phenotypic values of MON 87708 for both 

characteristics in the combined-site analysis were within the range of values observed for 

the commercial reference varieties. 

G.12.2. 2009 Dicamba-Treated MON 87708 Individual Field Site Plant Growth and 

Development Results and Discussion 

In the individual-site analysis, a total of 88 comparisons were made (Table G-6). Of 

these comparisons, no numerical differences were observed for 9 comparisons for which 

p-values could not be generated due to lack of variability. The eight flower color 

comparisons were categorical and were not statistically analyzed; however, at each site, 

all plants of treated MON 87708 and the control had purple flowers as expected. For the 

remaining comparisons, a total of fourteen statistically significant differences were 

detected out of 71 comparisons between treated MON 87708 and the conventional 
Monsanto Company 10-SY-210U 456 of 721 

control. These differences were distributed among eight out of the 11 phenotypic 

characteristics. Early stand count was lower for treated MON 87708 than the control at 

the ARNE site (263.0 vs. 300.5 plants/plot) and higher for the ILWY site (338.0 vs. 321.0 

plants/plot). Plants of MON 87708 flowered earlier than the control at the ILCY site 

(215.3 vs. 217.3 days after 1 Jan. 2009), and later than the control at the NEYO site 

(214.0 vs. 210.0 days after 1 Jan. 2009. Plants of MON 87708 were taller than the 

control at the ILCY site (26.4 vs. 24.1 inches). MON 87708 had less lodging than the 

control at the ARNE site (5.3 vs. 6.3 rating) and the NEYO site (1.3 vs. 2.0 rating). Pod 

shattering was higher for MON 87708 then the control at the NEYO site (1.8 vs. 1.3 

rating). Treated MON 87708 had a lower final stand count than the control at the ARNE 

site (254.3 vs. 290.8 plants/plot). Seed moisture was lower for treated MON 87708 than 

the control at the ARNE site (10.8 vs. 13.3%). The weight of 100 seeds was lower for 

MON 87708 than the control at the ARNE site (13.8 vs. 15.2 g), INSH site (15.4 vs. 16.9 

g), KSLA site (17.0 vs. 17.8 g), and the NEYO site (14.0 vs. 15.3 g). Considering that 

the statistical differences detected in the individual-site analyses for early stand count, 

days to 50% flowering, plant height, lodging, pod shattering, final stand count, and seed 

moisture were not detected in the combined-site analysis, this suggests these differences 

were not indicative of a consistent plant response associated with the trait and are 

unlikely to be biologically meaningful in terms of increased weed potential of MON 

87708 when treated with dicamba compared to the control. While a statistical difference 

was detected for 100 seed weight at three sites and in the combined-site analysis, the 

assessed phenotypic values of treated MON 87708 was within the expected values for 

commercial soybean (Section 3.8, step 4, “no” answer). Conventional cultivars seed 

weight ranges from 0.12 to 0.18 grams per seed (Heatherly and Elmore, 2004). 
Monsanto Company 10-SY-210U 

Table G-5. 2008 Individual-Site Phenotypic Comparison of Untreated MON 87708 to Conventional Contr

 Phenotypic Characteristic (units)  Early stand count (#/plot) Seedling vigor (1-9 scale) Days to 50% flowering1 

Flower Color/P

pubescence2 

Site 

MON 87708 

Mean (S.E.) 

Control 

Mean (S.E.) 

MON 87708 

Mean (S.E.) 

Control 

Mean (S.E.) 

MON 87708 

Mean (S.E.) 

Control 

Mean (S.E.) MON 87708 C

AR 160.3 (11.3) 142.7 (13.3) 6.0 (0.0) 6.0 (0.0) 183.3 (0.3) 183.0 (0.0) Purple/hairy PurpCan1 224.7 (2.0) 237.3 (15.1) 3.0 (0.0) 3.0 (0.0) 201.0 (0.6) 200.3 (0.3) Purple/hairy Purp

Can2 327.3 (24.5) 363.0 (13.3) 6.3 (0.3)* 5.0 (0.0) 202.0 (0.0) 202.7 (0.7) Purple/hairy PurpIA1 281.0 (11.9) 251.7 (16.8) 1.7 (0.7) 1.7 (0.3) 203.7 (0.7) 202.3 (1.3) Purple/hairy PurpIA2 142.3 (23.0) 111.0 (21.3) 3.3 (0.3) 4.0 (0.6) 221.7 (0.7) 221.7 (0.7) Purple/hairy PurpIA3 328.3 (13.7) 293.7 (2.7) 1.0 (0.0) 1.0 (0.0) 216.0 (0.0) 216.0 (0.0) Purple/hairy PurpIL1 317.3 (4.4) 315.3 (5.0) 4.3 (0.3) 4.3 (0.3) 212.0 (0.0) 212.0 (0.0) Purple/hairy PurpIL2 299.9 (3.5) 309.9 (1.2) 2.0 (0.0) † 2.0 (0.0) 203.3 (0.7) 203.3 (0.3) Purple/hairy PurpIN1 307.9 (3.7) 305.0 (4.2) 5.7 (0.3) 4.7 (0.3) 206.0 (0.0)† 206.0 (0.0) Purple/hairy Purp

IN2 285.7 (55.4) 304.0 (39.3) 5.0 (1.0) 4.3 (0.7) 202.0 (0.0) 202.0 (0.0) Purple/hairy Purp

IN3 338.0 (5.1) 310.3 (15.5) 3.7 (0.9) 3.3 (0.9) 220.0 (0.0) 220.7 (0.7) Purple/hairy PurpKS 198.0 (11.2) 175.3 (29.2) 2.7 (0.3) 3.3 (0.7) 198.7 (0.3) 199.0 (0.0) Purple/hairy PurpMI 333.7 (3.1) 342.8 (3.2) 4.0 (0.0) 3.7 (0.3) 208.0 (0.0)† 208.0 (0.0) Purple/hairy Purp

MO1 195.2 (13.9) 219.4 (14.6) 4.7 (0.3) 4.7 (0.3) 212.7 (0.9) 213.0 (0.0) Purple/hairy Purp

MO2 325.7 (0.9) 318.0 (6.0) 3.3 (0.3) 2.7 (0.3) 214.0 (0.0)* 213.0 (0.0) Purple/hairy Purp

NE 271.3 (9.5) 254.7 (3.3) 2.3 (0.3) 2.3 (0.3) 199.3 (1.3) 198.0 (0.0) Purple/hairy Purp

PA 266.3 (14.3) 262.3 (14.0) 3.0 (0.6) 2.7 (0.3) 201.3 (0.3) 201.0 (0.0) Purple/hairy PurpWI 277.0 (24.2) 254.0 (0.0) 1.3 (0.3) 1.0 (0.0) 210.0 (0.0)* 211.0 (0.0) Purple/hairy Purp
Monsanto Company 10-SY-210U 

Table G-5 (continued). 2008 Individual-Site Phenotypic Comparison of Untreated MON 87708 to Conven

 Phenotypic Characteristic (units)  Plant height (inch) Lodging (0-9 scale) Pod shattering (0-9 scale) Final stand cou

Site 

MON 87708 

Mean (S.E.) 

Control 

Mean (S.E.) 

MON 87708 

Mean (S.E.) 

Control Mean 

(S.E.) 

MON 87708 

Mean (S.E.) 

Control 

Mean (S.E.) 

MON 87708 

Mean (S.E.) 

AR 28.9 (0.2)* 27.1 (0.1) 2.7 (0.3) 2.0 (0.6) 0.3 (0.3) 0.3 (0.3) 140.3 (8.7) Can1 32.1 (0.6) 31.7 (0.5) 0.0 (0.0)† 0.0 (0.0) 0.0 (0.0)† 0.0 (0.0) 240.3 (1.7) Can2 40.1 (0.7) 38.6 (0.8) 4.0 (0.6) 3.3 (0.9) 0.0 (0.0)† 0.0 (0.0) 309.3 (6.9) IA1 39.1 (0.5)* 34.1 (0.3) 2.0 (0.6)* 0.7 (0.3) 0.0 (0.0)† 0.0 (0.0) 242.7 (5.8) IA2 37.1 (0.3)* 34.6 (0.4) 2.0 (0.0) 2.0 (0.0) 0.0 (0.0)† 0.0 (0.0) 123.0 (17.7) IA3 24.6 (0.5) 23.5 (0.8) 0.7 (0.3) 0.7 (0.3) 0.0 (0.0)† 0.0 (0.0) 293.3 (4.7) IL1 26.1 (1.4)* 22.3 (0.2) 1.0 (0.0) 0.3 (0.3) 0.0 (0.0)† 0.0 (0.0) 321.7 (6.1) IL2 39.0 (0.3) 37.4 (0.5) 1.0 (0.0)* 0.3 (0.3) 0.0 (0.0)† 0.0 (0.0) 295.9 (4.5) IN1 33.6 (2.3)* 30.5 (1.9) 1.3 (0.3) 0.7 (0.3) 0.0 (0.0)† 0.0 (0.0) 223.5 (25.1) IN2 39.6 (0.6)* 37.3 (0.4) 1.0 (0.0) 1.0 (0.0) 0.0 (0.0)† 0.0 (0.0) 262.3 (7.0) IN3 25.3 (0.3) 24.3 (0.5) 0.0 (0.0)† 0.0 (0.0) 0.0 (0.0)† 0.0 (0.0) 330.3 (7.7)* KS 40.5 (1.5) 38.2 (1.3) 1.3 (0.3)* 2.0 (0.0) 0.0 (0.0)† 0.0 (0.0) 187.7 (10.5) MI 29.5 (2.6) 30.9 (1.8) 0.7 (0.3) 0.3 (0.3) 0.0 (0.0)† 0.0 (0.0) 304.2 (4.5) MO1 23.4 (1.0)* 21.1 (0.8) 0.0 (0.0)† 0.0 (0.0) 0.0 (0.0) 0.0 (0.0) 186.8 (3.7) MO2 31.4 (1.7)* 28.7 (1.1) 0.7 (0.3) 0.3 (0.3) 1.0 (0.0) 1.0 (0.0) 308.3 (1.7) NE 40.5 (0.9) 38.9 (1.3) 0.3 (0.3) 1.0 (0.0) 0.0 (0.0)† 0.0 (0.0) 254.0 (7.8) PA 32.9 (0.8) 33.3 (0.8) 1.3 (0.3) 0.7 (0.7) 0.0 (0.0) 0.0 (0.0) 260.0 (16.2) WI 39.5 (0.7) 37.3 (0.3) 1.3 (0.3)* 0.0 (0.0) 0.0 (0.0) 0.5 (0.5) 235.7 (20.2) 
Monsanto Company 10-SY-210U 

Table G-5 (continued). 2008 Individual-Site Phenotypic Comparison of Untreated MON 87708 to Conven

 Phenotypic Characteristic (units)  Seed moisture (%) 100 seed weight (g) Test weight (lb/bu) Yield b

Site 

MON 87708 

Mean (S.E.) 

Control 

Mean (S.E.) 

MON 87708 

Mean (S.E.) 

Control Mean 

(S.E.) 

MON 87708 

Mean (S.E.) 

Control Mean 

(S.E.) 

MON 87708 

Mean (S.E.) 

AR 9.5 (0.1) 9.7 (0.3) 16.1 (0.3) 15.1 (1.1) 55.4 (0.4)* 54.0 (0.7) 63.3 (3.7)* Can1 12.4 (0.1) 12.1 (0.3) 15.9 (0.5)* 17.1 (0.0) 55.7 (0.4) 55.5 (0.5) 66.7 (3.0) Can2 13.3 (0.2) 13.2 (0.1) 16.9 (0.2)* 18.1 (0.2) 57.3 (0.2) 57.4 (0.2) 76.4 (0.1) IA1 11.4 (0.2) 11.2 (0.1) 15.0 (0.0) 14.7 (0.3) 55.6 (0.8) 54.5 (0.3) 71.7 (3.3) IA2 12.6 (0.1) 12.1 (0.5) 16.5 (0.3) 16.0 (0.2) 57.3 (0.3) 57.3 (0.3) 54.7 (6.7) IA3 7.5 (0.1) 7.5 (0.1) 12.0 (0.3) 12.5 (0.2) 58.1 (0.6) 57.2 (0.7) 20.5 (1.1) IL1 10.4 (0.0) 10.4 (0.1) 11.7 (0.5) 12.0 (0.5) 54.2 (0.7)* 55.9 (0.2) 46.5 (5.5) IL2 12.3 (0.1) 12.3 (0.3) 14.2 (0.1)* 15.0 (0.2) 54.3 (0.3) 54.3 (0.9) 55.3 (0.8) IN1 9.2 (0.1) 9.3 (0.1) 14.5 (0.0) 14.6 (0.0) 55.4 (0.9) 54.6 (0.9) 56.0 (2.4) IN2 10.9 (0.2) 10.7 (0.3) 14.9 (0.1) 15.0 (0.1) 51.0 (1.1) 50.7 (0.5) 61.9 (4.3) IN3 10.0 (0.1) 10.0 (0.2) 16.0 (0.2) 16.4 (0.3) 59.5 (1.3) 59.6 (1.5) 48.7 (2.6) KS 17.7 (0.2) 17.9 (0.3) 15.0 (0.0) 15.7 (0.3) 56.2 (0.4) 55.4 (0.3) 66.0 (2.9) MI 15.5 (0.0) 14.8 (0.2) 18.2 (0.7) 18.8 (0.3) 56.6 (0.3) 56.2 (0.2) 42.9 (4.7) MO1 11.9 (0.4) 12.1 (0.4) 14.3 (0.3) 15.0 (2.0) 55.8 (0.2) 55.7 (0.3) 35.1 (2.4) MO2 12.0 (0.0) 11.8 (0.2) 15.3 (0.3) 15.7 (0.3) 54.0 (0.3) 53.5 (1.2) 43.5 (1.4) NE 13.1 (0.1) 13.4 (0.3) 15.4 (0.2)* 15.9 (0.3) 59.2 (0.1) 59.1 (0.2) 69.3 (1.3) PA 10.4 (0.2) 10.5 (0.2) 13.4 (0.3)* 15.4 (0.2) 57.2 (0.3) 57.5 (0.3) 53.9 (1.3)* WI 11.7 (0.1)* 11.2 (0.1) 14.2 (0.2) 15.4 (0.4) 58.0 (0.6) 58.3 (0.3) 65.0 (2.7) Note: The experimental design at each site was a randomized complete block with three replications. S.E. = Standard Error 1Calendar day number when approximately 50% of the plants in each plot were flowering. 2Flower color and plant pubescence data were categorical and were not statistically analyzed.*Indicates a statistically significant difference between MON 87708 and the conventional control (

α=0.05). 

†No statistical comparisons were made due to lack of variability in the data. 
Monsanto Company 10-SY-210U 

Table G-6. 2009 Individual-Site Phenotypic Comparison of Dicamba Treated MON 87708 to Conventiona

 Phenotypic Characteristic (units) 

 Early stand count (#/plot)

Seedling vigor 

(1-9 scale)

Days After 1 Jan 2009 to 

50% flowering 

Site1 

MON 87708 

Mean (S.E.) 

Control 

Mean (S.E.) 

 MON 87708 

Mean (S.E.) 

Control 

Mean(S.E.) 

 MON 87708 

Mean (S.E.) 

Control 

Mean (S.E.) 

 M

M

ARNE 263.0* 

(13.73) 300.5 (5.42) 5.8 (0.25) 5.3 (0.48) 203.8 (0.25) 204.0 (0.41) 

IARL 290.8 (3.42) 285.0 (12.88) 4.8 (1.18) 5.8 (0.48) 220.3 (0.48) 220.3 (0.25) ILCY 

337.0 (2.35) 328.3 (6.70) 1.0 (0.00) 1.8 (0.25) 

 215.3* 

(0.48) 217.3 (0.25) 

ILWY 338.0* (5.00) 321.0 (3.03) 1.0† (0.00) 1.0 (0.00) 216.0 (0.00) 216.0 (0.00) 

INRC 276.5 (5.07) 293.3 (7.34) 3.3 (0.75) 2.3 (0.48) 224.3 (0.85) 225.3 (0.25) 

INSH 300.8 (8.19) 287.8 (6.94) 2.0 (0.41) 1.5 (0.29) 217.0 (0.00) 217.0 (0.00) 

KSLA 276.3 (9.41) 285.3 (6.80) 3.3 (0.25) 3.3 (0.25) 207.0 (0.00) 207.3 (0.25) 

NEYO 

309.0 (2.35) 307.8 (4.52) 3.0† (0.00) 3.0 (0.00) 

 214.0* 

(0.00) 210.0 (0.00) 
Monsanto Company 10-SY-210U 

Table G-6 (continued). 2009 Individual-Site Phenotypic Comparison of Dicamba Treated MON 877Control  Phenotypic Characteristic (units)  Plant Height (in) Lodging (1-9 scale)

Pod Shattering 

(1-9 scale) Fin

Site1 

MON 87708 

Mean (S.E.) 

Control 

Mean (S.E.) 

 MON 87708 

Mean (S.E.) 

Control 

Mean (S.E.) 

 MON 87708 

Mean (S.E.) 

Control 

Mean (S.E.) 

 MO

Me

ARNE 32.9 (1.07) 32.3 (0.77) 5.3* (0.48) 6.3 (0.25) 1.0† (0.00) 1.0 (0.00) 254

(12.

IARL 33.6 (0.60) 32.7 (0.43) 3.3 (0.63) 3.5 (0.65) 1.0 (0.00) 1.5 (0.50) 280ILCY 26.4* (0.99) 24.1 (0.26) 1.0† (0.00) 1.0 (0.00) 1.0† (0.00) 1.0 (0.00) 231ILWY 35.2 (0.18) 35.3 (0.33) 2.0 (0.00) 2.0 (0.00) 1.0† (0.00) 1.0 (0.00) 300INRC 23.1 (0.74) 21.7 (0.72) 1.0† (0.00) 1.0 (0.00) 1.0† (0.00) 1.0 (0.00) 226INSH 33.3 (0.97) 33.4 (0.56) 1.5 (0.50) 1.0 (0.00) 1.0† (0.00) 1.0 (0.00) 270KSLA 35.1 (0.82) 35.1 (0.30) 2.3 (0.25) 2.5 (0.29) 1.0 (0.00) 1.0 (0.00) 260NEYO 33.4 (0.42) 35.4 (1.45) 1.3* (0.25) 2.0 (0.00) 1.8* (0.25) 1.3 (0.25) 295
Monsanto Company 10-SY-210U 

Table G-6 (continued). 2009 Individual-Site Phenotypic Comparison of Dicamba Treated MON 877Control  Phenotypic Characteristic (units)  Seed Moisture (%) 100 Seed Weight (g) Yield (bu/ac) Site1 

MON 87708 

Mean (S.E.) 

Control 

Mean (S.E.) 

 MON 87708 

Mean (S.E.) 

Control 

Mean (S.E.) 

 MON 87708 

Mean (S.E.) 

Control 

Mean (S.E

ARNE 10.8* (0.38) 13.3 (0.76) 13.8* (0.23) 15.2 (0.18) 48.5 (2.96) 47.9 (2.33IARL 11.7 (0.05) 11.5 (0.16) 13.0 (0.41) 14.0 (0.41) 37.3 (1.52) 35.6 (1.25ILCY 12.9 (0.14) 13.1 (0.25) 14.9 (0.27) 15.7 (0.25) 52.0 (8.22) 51.6 (1.58ILWY 17.7 (0.14) 17.8 (0.20) 15.1 (0.21) 15.4 (0.24) 49.7 (1.97) 52.9 (2.74INRC 12.5 (0.27) 12.7 (0.31) 13.8 (0.15) 14.2 (0.30) 29.6 (2.75) 31.3 (2.00INSH 12.6 (0.04) 12.5 (0.13) 15.4* (0.31) 16.9 (0.22) 58.2 (2.56) 57.7 (2.48KSLA 11.9 (0.34) 11.5 (0.38) 17.0* (0.00) 17.8 (0.25) 65.5 (1.63) 66.0 (1.54NEYO 15.0 (0.13) 15.0 (0.09) 14.0* (0.41) 15.3 (0.25) 33.2 (1.58) 31.7 (0.55Note: The experimental design was a randomized complete block with four replications. * Statistically significant differences (

α=0.05) between MON 87708 and the conventional soybean control. † No statistical comparisons were made due to lack of variability in the data. 

1 Site codes are as follows: ARNE = Jackson County, AR; IARL = Jefferson County, IA; ILCY = Clinton County, IL; ILW

INRC = Parke County, IN; INSH = Boone County, IN; KSLA = Pawnee County, KS; NEYO = York County, NE 2 Flower color data were categorical and were not statistically analyzed. 
Monsanto Company 10-SY-210U 

Table G-7. Growth Stage Monitoring of Untreated MON 87708, Conventional Control, and the Co

Varieties from 2008 Field Trials 

 Assessment Date and Range of Growth Stages Observed2

Site1 Material Obs. 1 Obs. 2 Obs. 3 Obs. 4 Obs. 5 Obs. 6 Obs. 7 Obs. 8 

AR 06/18/2008 07/6/2008 07/23/2008 08/06/2008 08/23/2008 09/02/2008 9/24/2008 —  MON 87708 V2 R2 R3 R5 R6 R6 R8 —  Control V2 R2 R3 R5 R6 R6 R8 —  References V2-V3 R2 R3 R5 R6 R6 R8 — 

Can1 06/11/2008 06/24/2008 07/08/2008 07/25/2008 08/11/2008 08/25/2008 09/08/2008 09/23/2008 1

 MON 87708 VE V1-V2 V3-V4 R2 R3-R4 R4-R5 R5-R6 R6-R7  Control VE V1-V2 V3-V4 R2 R3-R4 R4-R5 R5-R6 R6-R7  References VE V1-V2 V3-V4 R2 R3-R4 R4-R5 R5-R6 R6-R7 Can2 06/10/2008 06/25/2008 07/10/2008 07/21/2008 08/06/2008 08/23/2008 09/03/2008 09/20/2008 1 MON 87708 VE V2-V3 V5 R2 R3 R5 R6 R7 

 Control VE V2-V3 V5-V6 R2 R3 R5 R6 R7 

 References VE V2-V3 V5-V6 R2 R3 R5 R6 R7 

IA1 07/01/2008 07/16/2008 08/06/2008 08/27/2008 09/15/2008 10/1/2008 

 MON 87708 V1-V2 V6-V7 R2 R4 R6 R7-R8 — —  Control V2 V6 R2-R3 R4 R6 R7-R8 — —  References V1-V2 V6-V7 R2-R3 R4 R6 R7 — — IA2 07/17/2008 08/05/2008 08/20/2008 09/10/2008 09/29/2008 10/15/2008 11/04/2008  MON 87708 V3 R2 R3 R5 R6 R7 R8 —  Control V3 R2 R3 R5 R6 R7 R8 —  References V3 R2 R3 R5 R6 R7 R8 — 
Monsanto Company 10-SY-210U 

Table G-7 (continued). Growth Stage Monitoring of Untreated MON 87708, Conventional Control, a

Reference Varieties from 2008 Field Trials 

 Assessment Date and Range of Growth Stages Observed2

Site1 Material Obs. 1 Obs. 2 Obs. 3 Obs. 4 Obs. 5 Obs. 6 Obs. 7 Obs. 8 

IA3 07/10/2008 07/20/2008 08/04/2008 08/24/2008 09/13/2008 09/30/2008  MON 87708 VC-V1 V1-V2 R2 R4 R5 R6 — —  Control V1 V2 R2 R4 R5 R6 — —  References VC-V1 V1-V2 R2-R3 R3-R4 R4-R6 R6 — — IL1 07/07/2008 07/22/2008 08/12/2008 09/01/2008 09/19/2008 10/06/2008  MON 87708 V1 R1 R2-R3 R6 R6 R8 — —  Control V1 R1 R2 R5-R6 R6-R7 R8 — —  References V1 R1 R2-R3 R5-R6 R6-R7 R8 — — IL2 06/25/2008 07/16/2008 08/07/2008 08/27/2008 09/17/2008 10/08/2008  MON 87708 V2 V6 R3 R5 R7 R7-R8 — —  Control V2 V6 R3 R5 R7 R7-R8 — —  References V2 V6 R3 R5 R7 R7-R8 — — IN1 06/25/2008 07/14/2008 07/25/2008 08/13/2008 08/28/2008 09/18/2008 10/01/2008  MON 87708 V2-V3 R1 R2 R5 R6 R7 R8 —  Control V2-V3 R1 R2 R4-R5 R6 R7 R8 —  References V2-V3 R1 R2 R4-R5 R6 R7-R8 R8 — IN2 06/25/2008 07/14/2008 07/21/2008 08/11/2008 08/26/2008 09/10/2008 10/01/2008  MON 87708 V3 R1 R2 R5 R6 R6 R8 —  Control V2-V3 R1 R2 R5 R6 R6-R7 R8 —  References V2-V3 R1 R2 R4-R5 R6 R6-R7 R8 — 
Monsanto Company 10-SY-210U 

Table G-7 (continued). Growth Stage Monitoring of Untreated MON 87708, Conventional Control, aReference Varieties from 2008 Field Trials 

Assessment Date and Range of Growth Stages Observed2

Site1 Material Obs. 1 Obs. 2 Obs. 3 Obs. 4 Obs. 5 Obs. 6 Obs. 7 Obs. 8 

IN3 07/15/2008 08/08/2008 08/27/2008 09/11/2008 09/25/2008 10/07/2008 10/20/2008  MON 87708 V2 R1-R2 R4 R5 R6 R7 R8 —  Control V2 R1-R2 R4 R5 R6 R7 R8 —  References V2 R1-R2 R4 R5 R6 R7 R8 — KS 06/27/2008 07/08/2008 07/23/2008 08/04/2008 08/18/2008 08/29/2008 09/10/2008 09/26/2008  MON 87708 V2 V5-V6 R2 R3 R5 R5 R6 R8  Control V2 V5-V6 R2 R3 R5 R5 R6 R8  References V2 V5-V6 R2 R3 R4-R5 R5 R6 R7 MI 06/24/2008 07/08/2008 07/22/2008 08/05/2008 08/18/2008 09/02/2008 09/17/2008 10/02/2008  MON 87708 V2 V4-V5 V8-V9 R3 R5 R6 R6 R7 

 Control V2 V4-V5 V8-V9 R3 R5 R6 R6 R7  References V2 V4-V5 V8-V9 R3 R5 R6 R6 R7 

MO1 07/16/2008 07/30/2008 08/19/2008 09/09/2008 09/23/2008 10/07/2008 10/21/2008 10/31/2008 

 MON 87708 V2-V3 V5-R2 R4 R5 R6 R8 R8 R8  Control V3 R1 R4 R5 R6 R8 R8 R8  References V2-V3 R1-R2 R4 R5 R6 R7-R8 R8 R8 MO2 07/14/2008 08/02/2008 08/14/2008 08/20/2008 09/10/2008 09/24/2008 10/07/2008 10/31/2008  MON 87708 V2 R2 R3 R4 R6 R6 R8 R8  Control V2 R2 R3 R4 R6 R6 R8 R8  References V2-V3 R1-R2 R2-R3 R4 R5-R6 R6 R7-R8 R8 
Monsanto Company 10-SY-210U 

Table G-7 (continued). Growth Stage Monitoring of Untreated MON 87708, Conventional Control, aReference Varieties from 2008 Field Trials 

 Assessment Date and Range of Growth Stages Observed2

Site1 Material Obs. 1 Obs. 2 Obs. 3 Obs. 4 Obs. 5 Obs. 6 Obs. 7 Obs. 8 

NE 06/26/2008 07/17/2008 08/07/2008 08/28/2008 09/23/2008 10/08/2008  MON 87708 V2 R1-R2 R4 R6 R7 R8 — —  Control V2 R2 R4 R6 R7 R8 — —  References V2 R1-R2 R4 R6 R7 R8 — — PA 06/20/2008 07/10/2008 07/24/2008 08/12/2008 08/29/2008 09/16/2008 09/30/2008 10/15/2008  MON 87708 V1 V4 R2 R4 R5-R6 R7 R8 R8  Control V1 V4 R2 R4 R5 R7 R7-R8 R8  References V1 V4-V5 R2-R3 R4-R5 R5-R6 R6-R7 R7-R8 R8 WI 07/01/2008 07/16/2008 08/08/2008 08/29/2008 09/16/2008 09/30/2008  MON 87708 V3 V5* R3 R5 R6 R7 — —  Control V3 V4 R3 R5 R6 R7 — —  References V3 V4-V5 R3 R5 R6 R7 — — *Indicates that MON 87708 and the conventional control were not within the same range of plant growth stages on this observation dat1Site codes are as follows: AR = Jackson County, AR; Can1 = Norfolk, Ontario, Canada; Can2 = Kent, Ontario, Canada; IA1 = Jefferso

IA2 = Benton County, IA; IA3 = Howard County, IA; IL1 = Clinton County, IL; IL2 = Stark County, IL; IN1 = Boone County, IN; IN2 IN3 = Parke County, IN; KS = Pawnee County, KS; MI = Ottawa County, MI; MO1 = Shelby County, MO; MO2 = Macon County, MNE; PA = Berks County, PA; WI = Walworth County, WI. 

2Obs. = Observation number; dates in month/day/year format. 

Dash (—) indicates information not available or plants had already reached full maturity (R8).
Monsanto Company 10-SY-210U 

Table G-8. Growth Stage Monitoring of Dicamba Treated MON 87708, Conventional Control, and the CoVarieties from 2009 Field Trials 

 Assessment Date and Range of Growth Stages Observed1

Site 2 Material Obs. 1 Obs. 2 Obs. 3 Obs. 4 Obs. 5 Obs. 6 Obs. 7 Obs. 8 Obs. 9 ObsARNE 7/10/2009 7/25/2009 8/12/2009 9/02/2009 9/22/2009 10/08/2009  MON 877083 V2 R1-R2 R4-R5 R6 R6 R8  Control V2 R1-R2 R5 R6 R6 R8  References V2 V6-R2 R4-R5 R6 R6 R8 IARL 7/17/2009 7/23/2009 7/27/2009 7/29/2009 8/12/2009 8/14/2009 8/21/2009 8/28/2009 9/7/2009 9/14/ MON 877083 V2 V3 V3 V4 R1 R2 R2-R3 R4 R5 R Control V2 V3 V3 V3-V4 R1 R2 R2-R3 R4 R5 R References V2 V3 V3 V3-V4 R1 R2 R2-R4 R4 R5 RINRC 7/30/2009 8/6/2009 8/22/2009 8/26/2009 9/3/2009 9/15/2009 10/5/2009 10/12/2009 10/29/2009 —  MON 877083 V2 V3 R2 R3 R5 R5 R6 R7 R8 —  Control V2 V3 R2 R3 R5 R5 R6 R7 R8 —  References V2 V3 R2 R2-R3 R5 R5 R6 R7 R8 — 
Monsanto Company 10-SY-210U 

Table G-8 (continued). Growth Stage Monitoring of Dicamba Treated MON 87708, Conventiona

Commercial Reference Varieties from 2009 Field Trials 

 Assessment Date and Range of Growth Stages Observed1

Site2 Material Obs. 1 Obs. 2 Obs. 3 Obs. 4 Obs. 5 Obs. 6 Obs. 7 Obs. 8 Obs. 9 INSH 7/14/2009 8/11/2009 9/1/2009 10/5/2009 11/6/2009 — — — —  MON 877083 V3 R2 R4 R7 R8 — — — —  Control V3 R2 R4 R7 R8 — — — —  References V3 V10-R2 R4 R7 R8 — — — — KSLA 7/9/2009 7/27/2009 8/6/2009 8/20/2009 9/3/2009 9/16/2009 10/1/2009 10/16/2009 10/28/2009  MON 877083 V2 R1 R1 R3-R4 R5 R6 R7 R8 R8  Control V2 R1 R1 R4 R5 R6 R7 R8 R8  References V2 R1 R1 R3-R4 R4-R5 R6 R6-R7 R8 R8 NEYO 7/17/2009 8/7/2009 8/26/2009 — — — — — —  MON 877083 V3 R1 R4 — — — — — —  Control V3 R1 R4 — — — — — —  References V3 R1 R4 — — — — — — Obs. = Observation number 1 Month-day-year. 2 Site codes are as follows: ARNE = Jackson County, AR; IARL = Jefferson County, IA; ILCY = Clinton County, IL; ILW

INRC = Parke County, IN; INSH = Boone County, IN; KSLA = Pawnee County, KS; NEYO = York County, NE

3 Received a mandatory dicamba application. 

— Dashes indicate information not provided. Growth stage data from the ILWY and ILCY site were not collected properly 
Monsanto Company 10-SY-210U 469 of 721 

Table G-9. Abiotic Stress Response Evaluations of Untreated MON 87708 and 

Conventional Control from 2008 Field Trials Using an Observational Severity Scale 

Note: The experimental design was a randomized complete block with three replications. 

Observations were conducted at four crop developmental stages: Observation 1 at V2-V4; 

Observation 2 at R1-R2; Observation 3 at R3-R5; Observation 4 at R6-R8. 

*Indicates a difference observed between MON 87708 (slight damage observed) and the 

conventional control (no damage observed) for wind damage during Observation 4 at the WI site. 

Data were not subjected to statistical analysis. 

1

Includes iron deficiency.

Abiotic Stressor Number of 

observations 

across all sites 

Number of observations where no 

differences were observed between 

MON 87708 and conventional control 

Total 194 193 

Cold 6 6 

Compaction 9 9 

Crusting 1 1 

Drought 27 27 

Excess moisture 20 20 

Flooding 18 18 

Frost 3 3 

Hail 30 30 

Heat damage 21 21 

Mineral Toxicity 1 1 

Nutrient deficiency1

 28 28 

Wind 30 29* 
Monsanto Company 10-SY-210U 470 of 721 

Table G-10. Disease Damage Evaluations of Untreated MON 87708 and 

Conventional Control from 2008 Field Trials Using an Observational Severity Scale 

Disease stressor Number of 

observations 

across all sites 

Number observations where no 

differences were observed between 

MON 87708 and conventional control 

Total 215 215 

Alternaria leaf spot 7 7 

Anthracnose 10 10 

Asian rust 4 4 

Bacterial blight 16 16 

Brown stem rot 5 5 

Cercospora1 3 3 

Charcoal rot 4 4 

Downy mildew 7 7 

Frogeye leaf spot 24 24 

Phytophthora2

 13 13 

Pod and stem blight 1 1 

Powdery mildew 20 20 

Pythium 7 7 

Rhizoctonia 10 10 

Septoria brown spot3

 39 39 

Soybean cyst nematode 2 2 

Soybean mosaic virus 1 1 

Soybean rust4 14 14 

Stem canker 2 2 

Sudden death syndrome5 14 14 

White mold 11 11 

Yellow mosaic virus 1 1 

Note: The experimental design was a randomized complete block with three replications. 

Observations were conducted at four crop developmental stages: Observation 1 at V2-V4; 

Observation 2 at R1-R2; Observation 3 at R3-R5; Observation 4 at R6-R8. 

1

Includes Cercospora leaf blight and Cercospora leaf disease. 

2

Includes Phytophthora root rot. 

3

Includes Septoria.

4

Includes rust. 

5

Includes sudden death.
Monsanto Company 10-SY-210U 

Table G-11. Arthropod–Related Damage Evaluations of Untreated MON 87708 and Conventional ConTrials Using an Observational Severity Scale Arthropod 

Number of 

observations 

across sites1

Number of observations 

where no differences were 

detected between 

MON 87708 and 

conventional control2 

Statistically Significant Differe

Site3 Observation Number 

Arthropod-Re

Rating (0

MON 87708 

(S.E.) Contro

Aphid 33 31 IA2 3 0.3 (0.06) 0.5 (

IA3 2 0.8 (0.03) 0.9 (

Armyworm 2 2 

−

−

−

Bean leaf beetle 48 48 

−

−

−

Blister beetle 5 4 MO1 2 0.1 (0.03) 0.4 Cabbage looper 1 1 

−

−

−

Corn rootworm beetle 3 3 

−

−

−

Cutworm 1 1 

−

−

−

Fall armyworm 3 3 

−

−

−

Grasshopper 29 29 

−

−

−

Green cloverworm 10 10 

−

−

−
Monsanto Company 10-SY-210U 

Table G-11 (continued). Arthropod-Related Damage Evaluations of Untreated MON 87708 and Conven

2008 Field Trials Using an Observational Severity Scale 

Arthropod 

Number of 

observations 

across sites1 

Number of observations 

where no differences were 

detected between 

MON 87708 and 

conventional control2

Statistically Significant Differenc

Site3 Observation Number 

Arthropod-Rela

Rating (0-

MON 87708 

(S.E.) Control 

Japanese beetle 27 25 IN1 2 0.6 (0.03) 0.9 (0

PA 4 0.6 (0.07) 0.4 (0

Leafhopper5 5 4 PA 1 1.1 (0.07) 0.6 (0

Seedcorn maggot 1 1 

−

−

−

−

Soybean looper 7 7 

−

−

−

−

Spider mite 8 8 

−

−

−

−

Sting bug 24 24 

−

−

−

−

Thistle caterpillar 3 3 

−

−

−

−

Thrips 2 2 

−

−

−

−

Yellow woollybear 4 4 

−

−

−

−

Note: The experimental design was a randomized complete block design with three replications. Observations were conducted at four stages: Observation 1 = V2-V4, Observation 2 = R1-R2, Observation 3 = R3-R5, and Observation 4 = R6-R8. 1Statistical comparisons were made between MON 87708 and the conventional control for 95 of the observations. Lack of variability i

statistical comparisons for 121 of the observations. 2No statistically significant difference (

α = 0.05) or numerical difference. 3Site codes IA2 = Benton County, IA; IA3 = Howard County, IA; IN1 = Boone County, IN; MO1 = Shelby County, MO; PA = Berks C

4Means, standard errors (S.E.), and reference ranges for differences between MON 87708 and the conventional control that were statist

Reference ranges were determined from the minimum and maximum mean values from among the commercial reference varieties. 5The difference detected in leafhopper damage during the first observation at the PA site was for damage caused by potato leafhoppers.Dash (

−) indicates no statistically significant difference between MON 87708 and the conventional control (

α = 0.05). 
Monsanto Company 10-SY-210U 

Table G-12. Abundance of Pest Arthropods in Beat Sheet Samples Collected from Untreated MON Control, and the Commercial Reference Varieties from 2008 Field Trials  Pest Arthropod2 Aphid Bean Leaf Beetle Grape ColaColl. Site1 MON 87708 (S.E.) Control (S.E.) Reference range MON 87708 (S.E.) Control (S.E.) Reference range MON 87708 (S.E.) Contro

(S.E.) 

1 IL2 

−

−

− 13.7 (4.8) 13.7 (4.1) 6.3 - 13.3 0.7 (0.3) 1.3 (1.3

 IN1 

−

−

− 0.3 (0.3) 0.3 (0.3) 0.7 - 1.0 0.0 (0.0) 0.3 (0.3

 MI 

−

−

− 0.0 (0.0) † 0.0 (0.0) 0.0 - 0.0 

−

−

 MO1 

−

−

− 0.0 (0.0) 0.0 (0.0) 0.3 - 0.7 

−

−

2 IL2 

−

−

− 4.3 (2.6) 5.7 (3.5) 2.7 - 5.0 

−

−

 IN1 

−

−

− 1.3 (0.9) 2.0 (0.6) 0.3 - 1.7 

−

−

 MI 816.7 (252.2) 383.3 (116.7) 600.0 - 1366.7 0.0 (0.0) † 0.0 (0.0) 0.0 - 0.0 

−

−

 MO1 

−

−

− 0.0 (0.0) 0.0 (0.0) 0.0 - 0.3 0.0 (0.0) 0.3 (0.3

3 IL2 

−

−

− 0.3 (0.3) 1.3 (0.9) 0.7 - 1.3 

−

−

 IN1 

−

−

− 0.3 (0.3) 0.7 (0.3) 0.0 - 1.3 

−

−

 MI 93.3 (78.4) 33.3 (16.7) 3.3 - 60.0 0.0 (0.0) 0.0 (0.0) 0.0 - 0.3 

−

−

 MO1 13.3 (3.3) 16.7 (3.3) 6.7 - 33.3 1.0 (1.0) 0.0 (0.0) 0.0 - 1.3 

−

−

4 IL2 

−

−

− 14.0 (3.5) 14.3 (4.7) 5.7 - 10.3 

−

−

 IN1 

−

−

− 2.0 (0.6) 2.3 (1.2) 0.3 - 2.0 

−

−

 MI 

−

−

− 0.0 (0.0) † 0.0 (0.0) 0.0 - 0.0 

−

−

 MO1 

−

−

− 5.0 (2.6) 3.3 (0.9) 3.3 - 4.7 

−

−
Monsanto Company 10-SY-210U 

Table G-12 (continued). Abundance of Pest Arthropods in Beat Sheet Samples Collected from Un

Conventional Control, and the Commercial Reference Varieties from 2008 Field Trials 

 Pest Arthropod2 Garden Flea-hopper Green Clover-worm JapaneColl. Site1 MON 87708 (S.E.) Control (S.E.) Reference range MON 87708 (S.E.) Control (S.E.) Reference range MON 87708 (S.E.) Co

(S

1 IL2 - - - 0.0 (0.0) * 2.0 (0.0) 0.3 - 2.7 0.3 (0.3) 0 IN1 - - - 0.0 (0.0) 0.3 (0.3) 0.0 - 1.3 9.0 (4.6) 12 MI - - - 1.7 (0.9) * 0.0 (0.0) 0.0 - 1.7 2.3 (1.9) * 8 MO1 - - - 2.3 (1.9) 5.0 (2.1) 1.0 - 3.7 

−

2 IL2 - - - 16.0 (5.0) 14.7 (2.8) 7.3 - 17.7 

−

 IN1 - - - 6.3 (0.3) 3.7 (0.9) 3.7 - 7.3 1.3 (0.7) 0 MI - - - 1.7 (0.7) 2.7 (1.7) 1.0 - 3.0 2.7 (0.3) 5 MO1 0.3 (0.3) 1.0 (0.6) 0.0 - 1.0 7.0 (2.3) 7.7 (0.3) 0.7 - 8.3 

−

3 IL2 - - - 6.3 (2.8) 6.3 (2.4) 7.7 - 15.7 

−

 IN1 - - - 7.0 (1.7) * 11.3 (2.2) 8.0 - 13.0 2.3 (0.3) 0 MI - - - 1.0 (0.6) 2.0 (0.6) 0.3 - 2.3 1.3 (0.9) 5 MO1 - - - 1.3 (0.9) 1.3 (0.9) 0.0 - 1.3 

−

4 IL2 - - - 1.0 (1.0) 1.0 (0.0) 0.3 - 2.0 

−

 IN1 - - - 1.7 (0.9) 2.3 (1.2) 1.0 - 3.7 0.0 (0.0) 0 MI - - - 0.0 (0.0) * 0.7 (0.3) 0.0 - 0.0 0.0 (0.0) 0 MO1 0.3 (0.3) 0.0 (0.0) 0.0 - 0.3 0.0 (0.0) 0.7 (0.7) 0.0 - 0.7 

−
Monsanto Company 10-SY-210U 

Table G-12 (continued). Abundance of Pest Arthropods in Beat Sheet Samples Collected from Un

Conventional Control, and the Commercial Reference Varieties from 2008 Field Trials 

 Pest Arthropod2 Potato Leafhopper Stink Bug Tarnished Coll. Site1 MON 87708 (S.E.) Control (S.E.) Reference range MON 87708 (S.E.) Control (S.E.) Reference range MON 87708 (S.E.) Co

(S

1 IL2 13.0 (10.1) 28.7 (12.3) 15.7 - 43.3 0.0 (0.0) 0.0 (0.0) 0.0 - 2.7 

−

 IN1 

−

−

− 0.0 (0.0) 0.3 (0.3) 0.0 - 0.3 

−

 MI 

−

−

− 0.0 (0.0) 0.3 (0.3) 0.0 - 0.3 1.3 (1.3) 0

 MO1 0.0 (0.0) 0.3 (0.3) 0.0 - 1.0 0.3 (0.3) 1.0 (0.6) 0.0 - 0.7 

−

2 IL2 

−

−

− 0.3 (0.3) 0.3 (0.3) 0.3 - 1.3 

−

 IN1 

−

−

− 0.7 (0.3) 0.0 (0.0) 0.3 - 0.7 

−

 MI 

−

−

− 0.3 (0.3) 1.0 (0.6) 1.0 - 6.3 1.3 (1.3) 3

 MO1 

−

−

− 0.3 (0.3) 0.0 (0.0) 0.0 - 1.3 

−

3 IL2 

−

−

− 0.0 (0.0) * 1.3 (0.9) 0.0 - 0.7 0.0 (0.0) 0

 IN1 

−

−

− 2.0 (1.5) * 0.0 (0.0) 0.0 - 0.7 

−

 MI 

−

−

− 2.0 (1.2) 2.3 (0.9) 1.7 - 3.7 6.0 (1.5) 12

 MO1 

−

−

− 0.3 (0.3) 0.7 (0.7) 0.3 - 1.3 

−

4 IL2 

−

−

− 2.3 (0.3) 2.0 (1.5) 1.7 - 5.0 0.7 (0.7) 2

 IN1 

−

−

− 1.3 (0.9) 1.0 (0.6) 1.0 - 5.0 

−

 MI 

−

−

− 3.7 (0.7) 3.0 (0.6) 1.7 - 4.7 2.0 (1.2) 1

 MO1 

−

− - 1.3 (0.9) 1.0 (0.6) 0.3 - 4.0 

−
Monsanto Company 10-SY-210U 

Table G-12 (continued). Abundance of Pest Arthropods in Beat Sheet Samples Collected from Un

Conventional Control, and the Commercial Reference Varieties from 2008 Field Trials

 Pest Arthropod2 Velvet-bean Caterpillar Wooly-bear Caterpillar Coll. Site1 MON 87708 (S.E.) Control (S.E.) Reference range MON 87708 (S.E.) Control (S.E.) Reference range 1 IL2 

−

−

−

−

−

−

 IN1 

−

−

−

−

−

−

 MI 

−

−

−

−

−

−

 MO1 

−

−

−

−

−

−

2 IL2 

−

−

−

−

−

−

 IN1 

−

−

−

−

−

−

 MI 

−

−

−

−

−

−

 MO1 

−

−

−

−

−

−

3 IL2 

−

−

−

−

−

−

 IN1 0.7 (0.7) 1.3 (0.3) 0.0 - 1.5 

−

−

−

 MI 

−

−

−

−

−

−

 MO1 

−

−

−

−

−

−

4 IL2 

−

−

−

−

−

−

 IN1 

−

−

− 2.3 (0.9) 3.0 (1.2) 0.7 - 3.0 

 MI 

−

−

− 0.7 (0.3) 0.3 (0.3) 0.3 - 1.0 

 MO1 

−

−

− 0.0 (0.0) 0.0 (0.0) 0.0 - 0.7 

Note: The experimental design was a randomized complete block with three replications. Arthropod collection (Coll.) 1 was conducte

stage, and the three subsequent collections were conducted at approximately two week intervals thereafter. * Indicates a statistically significant difference between MON 87708 and the conventional control (

α=0.05). 

† No statistical comparisons were made due to lack of variability in the data. 

1Site codes are as follows: IL2 = Stark County, IL; IN1 = Boone County, IN; MI = Ottawa County, MI; MO1 = Shelby County, MO. 2MON 87708 and the conventional control values represent mean number of arthropods collected from three replications. S.E. = Standa

ranges were determined from the minimum and maximum mean values from among the commercial reference varieties at the site. Dash (

−) indicates arthropod not evaluated. 
Monsanto Company 10-SY-210U 

Table G-13. Abundance of Beneficial Arthropods in Beat Sheet Samples Collected from Untreated MON 

Control, and the Commercial Reference Varieties from 2008 Field Trials 

 Beneficial Arthropod2 Araneae (spiders) Big eyed bug Carabidae Coll. Site1 MON 87708 (S.E.) Control (S.E.) Reference range MON 87708 (S.E.) Control (S.E.) Reference range MON 87708 (S.E.) Control (S.E.) 1 IL2 1.0 (1.0) 0.0 (0.0) 0.0 - 1.0 

−

−

−

−

−

 IN1 0.0 (0.0) 0.0 (0.0) 0.0 - 0.7 

−

−

−

−

−

 MI 0.0 (0.0) 0.0 (0.0) 0.0 - 0.3 

−

−

−

−

−

 MO1 0.0 (0.0) 0.0 (0.0) 0.3 - 1.0 0.0 (0.0) 0.0 (0.0) 0.0 - 0.3 

−

−

2 IL2 0.7 (0.3) 1.0 (0.0) 0.0 - 1.3 

−

−

−

−

−

 IN1 0.3 (0.3) 0.7 (0.7) 0.3 - 0.7 

−

−

−

−

−

 MI 1.3 (0.3) 0.7 (0.3) 0.0 - 1.0 

−

−

−

−

−

 MO1 3.7 (2.7) 1.0 (0.0) 0.7 - 2.3 0.0 (0.0) 0.3 (0.3) 0.0 - 0.3 

−

−

3 IL2 0.0 (0.0) 0.3 (0.3) 0.0 - 0.3 

−

−

− 0.3 (0.3) 0.0 (0.0) 

 IN1 0.0 (0.0) 0.3 (0.3) 0.0 - 0.3 

−

−

−

−

−

 MI 0.3 (0.3) 0.3 (0.3) 0.0 - 0.0 

−

−

−

−

−

 MO1 7.7 (0.9) 7.7 (0.9) 4.3 - 7.7 

−

−

−

−

−

4 IL2 0.3 (0.3) 0.7 (0.7) 0.3 - 1.3 

−

−

−

−

−

 IN1 0.3 (0.3) 0.0 (0.0) 0.0 - 1.0 

−

−

−

−

−

 MI 0.0 (0.0)* 3.0 (0.6) 0.7 - 1.0 

−

−

−

−

−

 MO1 0.3 (0.3) 0.3 (0.3) 1.0 - 2.0 

−

−

−

−

−
Monsanto Company 10-SY-210U 

Table G-13 (continued). Abundance of Beneficial Arthropods in Beat Sheet Samples Collected from UnConventional Control, and the Commercial Reference Varieties from 2008 Field Trials  Beneficial Arthropod2 Lacewings Ladybird beetles Micro-parasitic wColl. Site1 MON 87708 (S.E.) Control (S.E.) Reference range MON 87708 (S.E.) Control (S.E.) Reference range MON 87708 (S.E.) Control (S.E.) 1 IL2 

−

−

−

−

−

−

−

−

 IN1 

−

−

− 0.0 (0.0) 0.0 (0.0) 0.0 - 0.3 

−

−

 MI 0.7 (0.7) 0.0 (0.0) 0.0 - 0.7 0.0 (0.0) 0.7 (0.3) 0.3 - 1.0 

−

−

 MO1 

−

−

− 1.0 (0.6) 0.7 (0.7) 0.3 - 1.7 

−

−

2 IL2 

−

−

−

−

−

−

−

−

 IN1 

−

−

−

−

−

−

−

−

 MI 4.7 (1.7) 5.7 (1.2) 2.3 - 8.0 0.7 (0.7) 0.7 (0.7) 1.7 - 5.0 

−

−

 MO1 1.0 (0.6) 0.7 (0.3) 0.0 - 1.7 1.7 (0.3) 2.3 (0.9) 0.0 - 1.0 

−

−

3 IL2 0.0 (0.0) 0.0 (0.0) 0.0 - 0.3 0.0 (0.0) † 0.0 (0.0) 0.0 - 0.0 

−

−

 IN1 0.0 (0.0) 0.0 (0.0) 0.0 - 0.3 

−

−

−

−

−

 MI 8.7 (1.2) 6.0 (3.1) 5.3 - 6.0 13.7 (6.7) 5.7 (0.7) 5.3 - 14.0 

−

−

 MO1 6.3 (5.4) 2.0 (0.6) 2.0 - 6.0 8.3 (0.9) 9.7 (4.3) 4.7 - 12.7 0.7 (0.7) 1.7 (0.9) 4 IL2 0.7 (0.3) 0.7 (0.3) 0.0 - 2.0 0.0 (0.0) 0.7 (0.3) 0.0 - 0.7 0.0 (0.0) 0.0 (0.0)  IN1 

−

−

−

−

−

−

−

−

 MI 4.0 (0.6) 2.0 (1.2) 1.3 - 6.0 3.7 (0.9) 4.0 (1.5) 2.7 - 7.3 1.0 (0.6) 1.3 (0.7)  MO1 1.7 (1.7) 1.3 (0.9) 0.7 - 3.0 7.3 (0.3) 4.7 (1.2) 2.0 - 8.3 

−

−
Monsanto Company 10-SY-210U 

Table G-13 (continued). Abundance of Beneficial Arthropods in Beat Sheet Samples Collected from UnConventional Control, and the Commercial Reference Varieties from 2008 Field Trials  Beneficial Arthropod2

Nabis spp. Opiliones Orius spp. 

Coll. Site1 MON 87708 (S.E.) Control (S.E.) Reference range MON 87708 (S.E.) Control (S.E.) Reference range MON 87708 (S.E.) Control (S.E.) Refer

ran

1 IL2 1.7 (0.7) 1.0 (0.6) 0.7 - 2.7 0.7 (0.3) 0.3 (0.3) 0.0 - 0.3 0.7 (0.7) 2.3 (0.9) 0.7 -  IN1 0.0 (0.0) † 0.0 (0.0) 0.0 - 0.0 0.3 (0.3) 0.3 (0.3) 0.0 - 0.7 0.0 (0.0) † 0.0 (0.0) 0.0 -  MI 0.0 (0.0) 0.3 (0.3) 0.0 - 0.7 

−

−

− 0.0 (0.0) † 0.0 (0.0) 0.0 - 

 MO1 0.0 (0.0) 0.0 (0.0) 0.0 - 1.0 

−

−

− 0.3 (0.3) 0.7 (0.7) 0.0 - 

2 IL2 1.3 (0.3) 1.3 (0.9) 1.0 - 2.0 

−

−

− 0.0 (0.0) 0.7 (0.7) 0.0 - 

 IN1 0.0 (0.0) 0.3 (0.3) 0.0 - 1.3 0.3 (0.3) 0.7 (0.3) 0.0 - 1.0 0.0 (0.0) 0.3 (0.3) 0.0 -  MI 2.3 (1.2) 4.0 (1.5) 1.7 - 2.3 

−

−

− 7.7 (1.3) 7.0 (1.5) 4.7 - 

 MO1 0.3 (0.3) 0.0 (0.0) 0.0 - 1.0 

−

−

− 1.7 (0.9) 2.0 (1.2) 1.3 - 

3 IL2 0.3 (0.3) 0.7 (0.3) 0.3 - 1.3 

−

−

− 0.0 (0.0) † 0.0 (0.0) 0.0 - 

 IN1 0.3 (0.3) 0.0 (0.0) 0.0 - 0.3 0.0 (0.0) 0.0 (0.0) 0.0 - 0.7 0.0 (0.0) 0.0 (0.0) 0.0 -  MI 3.3 (0.9) 1.3 (0.7) 2.0 - 4.0 

−

−

− 4.7 (1.2) 4.7 (1.8) 2.3 - 

 MO1 1.3 (1.3) 2.0 (1.5) 0.3 - 3.3 

−

−

− 4.7 (0.9) 2.3 (1.3) 3.7 - 

4 IL2 0.0 (0.0) 1.0 (0.6) 1.0 - 2.7 

−

−

− 3.3 (0.3) 1.3 (0.9) 0.3 - 

 IN1 0.0 (0.0) 0.7 (0.7) 0.0 - 0.7 1.0 (0.0) 1.3 (0.3) 0.3 - 1.0 0.7 (0.7) 0.0 (0.0) 0.0 -  MI 4.7 (0.7)* 1.7 (1.7) 2.0 - 6.3 

−

−

− 1.0 (0.6) 2.7 (0.9) 0.7 - 

 MO1 0.7 (0.7) 0.3 (0.3) 0.3 - 3.0 

−

−

− 0.0 (0.0) 3.0 (1.5) 0.7 - 
Monsanto Company 10-SY-210U 

Table G-13 (continued). Abundance of Beneficial Arthropods in Beat Sheet Samples Collected from UnConventional Control, and the Commercial Reference Varieties from 2008 Field Trials  Beneficial Arthropod2 Syrphid larvae Coll. Site1 MON 87708 (S.E.) Control (S.E.) Reference range 1 IL2 

−

−

−

 IN1 

−

−

−

 MI 

−

−

−

 MO1 

−

−

−

2 IL2 

−

−

−

 IN1 

−

−

−

 MI 1.7 (1.2) 3.0 (1.0) 1.0 - 4.3  MO1 

−

−

−

3 IL2 

−

−

−

 IN1 

−

−

−

 MI 1.3 (0.7) 0.7 (0.7) 0.0 - 2.3  MO1 

−

−

−

4 IL2 

−

−

−

 IN1 

−

−

−

 MI 

−

−

−

 MO1 

−

−

−

Note: The experimental design was a randomized complete block with three replications. Arthropod collection (Coll.) 1 was conducte

stage, and the three subsequent collections were conducted at approximately two week intervals thereafter. * Indicates a statistically significant difference between MON 87708 and the conventional control (

α=0.05). 

† No statistical comparisons were made due to lack of variability in the data. 

1Site codes are as follows: IL2 = Stark County, IL; IN1 = Boone County, IN; MI = Ottawa County, MI; MO1 = Shelby County, MO. 2MON 87708 and the conventional control values represent mean number of arthropods collected from three replications. S.E. = Standa

ranges were determined from the minimum and maximum mean values from among the commercial reference varieties at the site. Dash (

−) indicates arthropod not evaluated. 
Monsanto Company 10-SY-210U 481 of 721 

Table G-14. Abiotic Stress Response Evaluations of Dicamba-Treated MON 87708 

and Conventional Control from 2009 Field Trials Using an Observational Severity 

Scale 

Abiotic Stressor Number of 

observations 

across all sites1 

Number of observations where no 

differences were observed between 

MON 87708 and control 

Total 89 89 

Cold 1 1 

Compaction 3 3 

Drought2 21 21 

Flood3 17 17 

Frost damage 4 4 

Hail damage 13 13 

Nutrient deficiency 10 10 

Wind damage 20 20 

Note: The experimental design was a randomized complete block with four replications. 

Observations were made at approximately the following four crop developmental stages: 

Observation 1 at V3 –V5; Observation 2 at R1 - R2; Observation 3 at R3 - R5; Observation 4 

at R6 - R8. 

Data were not subjected to statistical analysis. 

1 Site codes are as follows: ARNE = Jackson County, AR; IARL = Jefferson County, IA; 

ILCY = Clinton County, IL; ILWY = Stark County, IL; INRC = Parke County, IN; INSH = 

Boone County, IN; KSLA = Pawnee County, KS; NEYO = York County, NE

2

Includes heat and dry. 

3

Includes wet soil, excess moisture. 
Monsanto Company 10-SY-210U 482 of 721 

Table G-15. Disease Damage Evaluations of Dicamba-Treated MON 87708 and 

Conventional Control from 2009 Field Trials Using an Observational Severity Scale 

Disease stressor Number of 

observations 

across all sites1 

Number observations where no 

differences were observed between 

MON 87708 and control 

Total 93 92 

 

Anthracnose 1 1 

Bacterial blight2 7 7 

Bacterial leaf spot 2 2 

Brown stem rot 3 3 

Cercospora3 5 5 

Charcoal rot 1 1 

Downy mildew 13 13 

Frogeye leaf spot4 8 8 

Leaf spot5 21 21 

Phytophthora root rot 7 7 

Powdery mildew 5 5 

Pythium 2 2 

Rhizoctonia 3 3 

Soybean mosaic virus 1 1 

Soybean rust 1 1 

Stem canker 1 1 

Sudden death 9 9 

White mold6 3 2 

 

Note: The experimental design was a randomized complete block with four replications. 

Observations were made at approximately the following four crop developmental stages: 

Observation 1 at V3 –V5; Observation 2 at R1 - R2; Observation 3 at R3 - R5; Observation 4 at 

R6 - R8. 

1

 Site codes are as follows: ARNE = Jackson County, AR; IARL = Jefferson County, IA; ILCY 

= Clinton County, IL; ILWY = Stark County, IL; INRC = Parke County, IN; INSH = Boone 

County, IN; KSLA = Pawnee County, KS; NEYO = York County, NE 

2 Includes Pseudomonas. 3 Includes Cercospora leaf blight and Cercospora leaf disease. 

4 Includes eye spot. 

5

Includes Septoria and Alternaria. 6

Indicates a difference observed between MON 87708 and the control for white mold at the 

ILWY site (Observation 2; slight vs. none). The damage rating for MON 87708 was outside of 

the reference range (no white mold was observed in the references). Data were not subjected to 

statistical analysis. 
Monsanto Company 10-SY-210U 

Table G-16. Arthropod–Related Damage Evaluations of Dicamba-Treated MON 87708 and Conventiona

Field Trials Using an Observational Severity Scale 

Arthropod 

Number of 

observations 

across sites1

Number of observations 

where no differences were 

detected between 

MON 87708(N) and the 

control2 

Statistically Significant Differen

Site3 Observation Number Arthropod Damage 

MON 87708(N) 

(S.E.) 

Co

(S

Aphids5 10 10 

−

−

−

Bean leaf beetle 23 22 KSLA 3 0.00(0.00) 0.08Blister beetle 1 1 

−

−

−

Cabbage looper 1 1 

−

−

−

Corn earworm 1 1 

−

−

−

Fall armyworm 1 0 

−

−

−

Grasshopper 18 16 INRC 3 0.45(0.09) 0.10 

 KSLA 3 0.20(0.00) 0.03

Green cloverworm 7 7 

−

−

−

Japanese beetle 10 10 

−

−

−
Monsanto Company 10-SY-210U 

Table G-16 (continued). Arthropod–Related Damage Evaluations of Dicamba-Treated MON 87708 and Cfrom 2009 Field Trials Using an Observational Severity Scale Arthropod 

Number of 

observations 

across sites1 

Number of observations 

where no differences were 

detected between 

MON 87708 and the control2

Statistically Significant Differen

Site3 Observation Number Arthropod Damage 

MON 87708 

(S.E.) 

Co

(S

Potato leaf hopper 3 3 

−

−

−

Soybean looper 2 2 

−

−

−

Stink bugs6 9 9 

Three cornered alfalfa hopper 3 3 

−

−

−

Velvet bean caterpillar 4 4 

−

−

−

Note: The experimental design was a randomized complete block design with four replications. Observations were conducted at apfour crop developmental stages: Observation 1 = V3-V5, Observation 2 = R1-R2, Observation 3 = R3-R5, and Observation 4 = R6-R8. 1 A total of 93 arthropod damage observations were made across sites. Lack of variability in the data precluded statistical comparisons 

Statistical comparisons could be made between MON 87708 and control for 59 of the observations. 

2 No statistically significant differences were detected (

α=0.05) or numerical differences between MON 87708 and the conventional 

values could not be generated due to lack of variability in the data. 

3 Site codes are as follows: ARNE = Jackson County, AR; IARL = Jefferson County, IA; ILCY = Clinton County, IL; ILWY = Stark 

County, IN; INSH = Boone County, IN; KSLA = Pawnee County, KS; NEYO = York County, NE

4 Means, standard errors (S.E.), and reference ranges are reported for a statistically significant difference that was detected (

α=0.05) bet

conventional soybean control. Reference range = minimum and maximum mean values among the commercial reference varieties. 

5 Aphids include soybean aphids. 6 Stink bugs include green stink bugs. 

− indicates that there were no statistically significant differences were detected (

α=0.05) between MON 87708 and the conventional soy
Monsanto Company 10-SY-210U 485 of 721 

References for Appendix G 

Heatherly, L.G. and R.W. Elmore. 2004. Managing inputs for peak production. Pages 

451-536 in Soybeans: Improvement, Production, and Uses. H.R. Boerma and J.E. Specht 

(eds.). American Society of Agronomy, Inc., Crop Science Society of America, Inc., Soil 

Science Society of America, Inc., Madison, Wisconsin. 

Kogan, M. and H.N. Pitre. 1980. General sampling methods for above-ground 

populations of soybean arthropods. Pages 30-60 in Sampling Methods in Soybean 

Entomology. M. Kogan and D.C. Herzog (eds.). Springer-Verlag, New York, New York. 

SAS Software Release 9.2 (TS1M0). ©2002-2008 by SAS Institute Inc., Cary, North 

Carolina. 
Monsanto Company 10-SY-210U 486 of 721 

Appendix H: Materials and Methods for Pollen Morphology and Viability 

Assessment 

H.1. Plant Production 

MON 87708, the near isogenic conventional soybean control A3525, and four 

commercial reference varieties were grown under similar agronomic conditions in a field 

trial in Clinton County, IL (Table G-1; IL1 site). The trial was arranged in a randomized 

complete block design with three replications. Each plot consisted of four rows 

approximately 20 feet in length. 

H.2. Flower Collection 

When the soybean plants were flowering, twenty whole flowers were collected from the 

fourth row of each plot. Four flowers were taken from each of five representative plants 

per plot. All flowers from a plot were placed into a single, clean container that was 

labeled with the plot number from which the sample originated. The containers were 

kept on ice for ≤ 8 hours until the pollen was prepared and stained. 

H.3. Pollen Sample Preparation 

Pollen samples were prepared in a laboratory. Clean microscope slides were labeled with 

the plot number. A circle of approximately one centimeter in diameter was drawn in the 

center of the slide with a pap hydrophobic barrier pen. Tweezers and a dissecting needle 

were used to open each of the collected flowers from a plot and brush the pollen into the 

circle on the slide. The utensils were cleaned between extractions. Approximately 20 μl 

of Alexander’s stain (Alexander, 1980) was added to the center of the circle containing 

the pollen. The pollen was stained at ambient temperature for at least ten minutes prior to 

examination. Pollen samples from all plots within a replicate were stained and evaluated 

on the same day. 

H.4. Data Collection 

Pollen characteristics were assessed by viewing samples under an Olympus Provis AX70 

light/fluorescence microscope equipped with an Olympus DP70 digital color camera. 

The microscope and camera were connected to a computer running Microsoft Windows 

2000 Professional and installed with associated DP Controller v1.2.1.108 and DP 

Manager v1.2.1.107 camera software and Image-Pro Plus v6.2.1.491 imaging software. 

H.4.1. Pollen Viability 

When exposed to the staining solution, viable pollen grains stained red to purple due to 

the presence of living cytoplasmic content. Nonviable pollen grains stained blue to green 

and may have appeared round to collapsed in shape, depending on the degree of 

hydration. The number of viable and nonviable pollen grains in each pollen sample was 

counted from a random field of view under the microscope. A minimum of 75 pollen 

grains were counted in each sample. Dense clusters of pollen or pollen grains adhering to 
Monsanto Company 10-SY-210U 487 of 721 

flower parts were not counted because they may not have absorbed the staining solution 

uniformly. 

H.4.2. Pollen Diameter 

Micrographs of ten representative pollen grains from each plot were taken at 400× 

magnification and imported into the imaging software. The software was used to 

measure pollen grain diameter along two perpendicular axes for each selected pollen 

grain. Mean pollen diameter for each plot was calculated from the 20 total measurements. 

H.4.3. General Pollen Morphology 

General pollen morphology was observed from micrographs of MON 87708, the 

conventional control, and commercial reference varieties that were also used for pollen 

diameter measurements. 

H.5. Statistical Analysis 

An analysis of variance was conducted according to a randomized complete block design 

using SAS® (Version 9.2). The level of statistical significance was predetermined to be 

5% (α=0.05). MON 87708 was compared to the conventional control for percent viable 

pollen and pollen diameter. No statistical comparisons were made between MON 87708 

and the commercial reference varieties. Instead, a reference range for each measured 

characteristic was determined from the minimum and maximum mean values from 

among the four commercial reference varieties. General pollen morphology was 

qualitative; therefore, no statistical analysis was conducted on these observations. 
Monsanto Company 10-SY-210U 488 of 721 

References for Appendix H 

Alexander, M.P. 1980. A versatile stain for pollen fungi, yeast and bacteria. Stain 

Technology 55:13-18. 

SAS Software Release 9.2 (TS1M0). ©2002-2008 by SAS Institute Inc., Cary, North 

Carolina. 
Monsanto Company 10-SY-210U 489 of 721 

Appendix I: Materials and Methods for Symbiont Assessment 

I.1. Materials 

The soybean materials for the symbiont interaction assessment included MON 87708, the 

near isogenic conventional soybean control A3525, and six commercial reference 

varieties (Table I-1). Nodules, root tissue, and shoot tissue collected from MON 87708, 

the conventional control, and the commercial reference varieties were evaluated. 

Table I-1. Starting Seed of MON 87708, Conventional Control, and Commercial 

Reference Varieties Used in the Symbiont Assessment 

Material Name Material Type Phenotype Monsanto Seed Lot # 

MON 87708 Test Dicamba-tolerant 11225299 

A3525 Control Conventional 10001822 

A2553 Reference Conventional 10000961 

LG C3884N Reference Conventional 11226859 

Stewart SB3454 Reference Conventional 11242910 

Garst 3585N Reference Conventional 10001517 

Hartz H5218 Reference Conventional 10001410 

A5560 Reference Conventional 10001114 

I.2. Characterization of the Materials 

For the MON 87708 and conventional control starting seed lots, the presence or absence 

of the dmo expression cassette was confirmed by event-specific polymerase chain 

reaction analyses.

I.3. Greenhouse Phase and Experimental Design 

Eight replicate 6-inch pots were prepared for each soybean material (MON 87708, the 

conventional control, and commercial reference varieties). The pots contained nitrogen 

deficient potting medium (Sunshine® Mix #2 Basic/LB2) composed of primarily peat, 

vermiculite, and perlite. At planting, each seed was inoculated with approximately 

1 × 107

 cells of Bradyrhizobium japonicum (VAULT® NP, Becker Underwood) in 

phosphate-buffered saline. Three seeds were planted in each pot. The soybean plants 

were grown in a greenhouse where temperatures ranged from approximately 17 to 35°C. 

Once pots were thinned to a single plant per pot, the pots were arranged in eight 

replicated blocks using a randomized complete block design. 

Replicates 1, 2, and 3 were planted on September 8, 2009; replicates 4, 5, and 6 were 

planted on September 9, 2009; and replicates 7 and 8 were planted on September 10, 

2009. In all cases, replicate pots had a minimum of one plant emerge within one week. 

A nitrogen-free nutrient solution (approximately 250 ml) was added weekly after plant 

emergence. 
Monsanto Company 10-SY-210U 490 of 721 

I.4. Plant Harvesting/Data Collection 

Six weeks after emergence, plants were excised at the surface of the potting medium and 

shoot and root plus nodule material were removed from the pots. The shoot material was 

cut into smaller pieces and placed in labeled bags. The plant roots with nodules were 

separated from the potting medium by washing with water. Excess moisture was 

removed using absorbent paper towels, and the roots with nodules were placed in labeled 

bags. On the same day that plants were harvested, nodules were removed by hand from 

the roots of each plant, enumerated, and weighed to determine the fresh weight of the 

nodules. 

The remaining root and shoot mass were determined for each plant. Nodules as well as 

root and shoot material were placed in a drying oven on the same day as collected or 

stored for up to four days at approximately 4°C before being placed in the drying oven. 

The plant material was dried for at least 72 hours at approximately 65°C to determine dry 

weight. The shoot tissue was ground after drying with a Harbil 5G high-speed paint 

shaker prior to total nitrogen analysis. Shoot total nitrogen was determined by 

combustion using a nitrogen analyzer (rapid N cube, Elementar Americas, Inc.). 

I.5. Statistical Analysis 

An analysis of variance was conducted according to a randomized complete block design 

using SAS® (Version 9.2). The level of statistical significance was predetermined to be 

5% (α=0.05). MON 87708 was compared to the conventional control for nodule number, 

nodule dry weight (g), shoot dry weight (g), root dry weight (g), and shoot total nitrogen 

(% and g/plant). No statistical comparisons were made between MON 87708 and the 

commercial reference varieties. Instead, a reference range for each measured 

characteristic was determined from the minimum and maximum mean values from 

among the six commercial reference varieties. 
Monsanto Company 10-SY-210U 491 of 721 

References for Appendix I 

SAS Software Release 9.2 (TS1M0). ©2002-2008 by SAS Institute Inc., Cary, North 

Carolina. 
Monsanto Company 10-SY-210U 492 of 721 

Appendix J: Petitioner’s Environmental Report 

J.1. Background 

This appendix provides information regarding four key areas to be covered in an 

environmental assessment: Purpose and Need, Alternatives, the Affected Environment, 

and Potential Environmental Impacts. This environmental report has been prepared by 

Monsanto to facilitate APHIS’s compliance with the National Environmental Policy Act 

(NEPA), including compliance with the Council on Environmental Quality’s (CEQ) 

regulations that implement NEPA13. 

Monsanto has produced dicamba-tolerant soybean plants (hereafter referred to as 

MON 87708) by inserting a gene that was discovered in a naturally occurring 

microorganism. The gene produces a dicamba mono-oxygenase (DMO) protein (referred 

to as the MON 87708 DMO) in the plant that inactivates the herbicide dicamba, thus 

rendering the plant tolerant to applications of the herbicide. Other broadleaf plants, 

including weeds in soybean fields, do not naturally contain the gene or exhibit tolerance 

to dicamba and when treated with dicamba do not survive. Numerous field trials 

conducted in the U.S. under APHIS notifications since 2005 have included MON 87708. 

Information has been collected from field trials, laboratory and greenhouse studies, and 

the literature to assess whether the tolerance to dicamba through production of the 

MON 87708 DMO protein and/or the gene insertion process has altered MON 87708 in 

any way that would make these plants more of a plant pest compared to conventional 

soybean or cause significant environmental impacts, including cumulative impacts. The 

purpose of this report is to provide relevant information regarding the potential for 

reasonably foreseeable, significant environmental impacts of widespread cultivation of 

MON 87708. 

MON 87708 will enable an expanded application window of dicamba in soybean, 

allowing for preemergence application through crop emergence (cracking), and 

postemergence in-crop applications through the R1/R2 growth stage. The current 

maximum allowable dicamba use pattern allowed on soybean, the proposed maximum 

allowable dicamba use pattern on MON 87708, and the maximum annual allowable 

dicamba rates are summarized in Table J-1. The anticipated use rates for dicamba on 

MON 87708, however, are expected to be less than the proposed maximum allowabable 

use rates. Section VIII.H.2 of the petition describes in detail the anticipated use rates and 

patterns for dicamba on MON 87708. 

 

13 Title 40 of the Code of Federal Regulation (40 CFR) Parts 1500-1508. 
Monsanto Company 10-SY-210U 493 of 721 

Table J-1. Summary of Dicamba Uses on Soybean 

Current Approved Uses Proposed Uses on MON 87708

Application 

Timing 

Maximum 

Single 

Application 

Rate 

(lbs a.e./acre)

Maximum 

Annual 

Application 

Rate 

(lbs a.e./acre)

Maximum 

Single 

Application 

Rate 

(lbs 

a.e./acre)

Maximum 

Annual 

Application 

Rate 

(lbs a.e./acre)

Preemergence 0.50a

2.0 

1.0b

2.0 

Postemergence 

Not labeled 

0.50 (V3) 

+ 

0.50

(R1/R2)c

Pre-harvest 

(7 days prior to 

harvest) 

1.0 Not labeled 

a 14-28 day planting interval based on product application rate 

b No planting interval c In-crop application through V3 with a sequential application through R1/R2 growth stage as 

needed. Total of all in-crop applications from emergence up to R1/R2 is 1.0 lb a.e./acre. 

An analysis of the potential impact from deregulation of MON 87708 on current soybean 

agronomic production systems, and related activities such as soybean processing, food 

and feed uses as well as marketing of soybean and soybean products is presented in this 

Environmental Report. Factors evaluated as part of the assessment include potential 

impacts to: 

 land use patterns, climate, water, soil and air quality, non-agricultural lands, farming 

practices, commodity and specialty soybean production, 

 marketability of soybean seed for planting and seed for specialty and commodity 

markets, and 

 public health, non-target organisms, threatened or endangered species, and 

biodiversity. 

The analysis conducted considers current and reasonably foreseeable conditions, the 

potential for deregulation of MON 87708 to impact these conditions, and potential 

cumulative impacts. In most cases, there are no impacts to current conditions (e.g., no 

differences between deregulation of MON 87708 versus continuing to regulate). Where 

differences are noted, these differences are described and their significance evaluated. 
Monsanto Company 10-SY-210U 494 of 721 

J.2. Purpose and Need for the Action 

APHIS’ mission is to protect the health and value of American agriculture and natural 

resources. 14 Under the authority of the Plant Protection Act (PPA), APHIS’ 

Biotechnology Regulatory Services (BRS) regulates the introduction (importation, 

interstate movement, or release into the environment) of certain genetically engineered 

(GE) organisms and products that may pose a risk to plant health.15 An organism is no 

longer subject to these regulations when APHIS determines that it is unlikely to pose a 

plant pest risk. A GE organism is considered a regulated article if APHIS has reason to 

believe it could pose a plant pest risk. A person may petition the agency to evaluate 

submitted data and determine that a particular regulated article is unlikely to pose a plant 

pest risk, and, therefore, should no longer be regulated as a potential plant pest.16 The 

petitioner is required to provide information related to plant pest risk that the USDA may 

use to assist in determining whether the regulated article is unlikely to present a plant pest 

risk.17 If, based on this information, as well as other sources of information, the USDA 

determines that the article is unlikely to pose a plant pest risk, the article may be granted 

nonregulated status. 

Monsanto Company (Monsanto) has submitted a petition (designated as 10-188-01p) to 

APHIS seeking the determination of nonregulated status in whole for MON 87708 

soybean plants genetically engineered to produce the MON 87708 DMO which is not 

typically found in soybean. Monsanto researchers have found that when soybean plants 

produce the MON 87708 DMO, they are tolerant to the broadleaf herbicide dicamba. 

Due to the unique broad spectrum herbicide properties of glyphosate, growers may use 

only glyphosate for their total weed management. As a result, in certain cropping and 

cultural systems (e.g., common corn and-soybean rotations), an increase in the number of 

crop acres with glyphosate resistant weed populations has occurred over the last decade 

and, recently, the number of growers incorporating other weed management practices has 

also grown over time. A prominent strategy to delay the development of herbicideresistant weeds is to increase the diversity of weed management options in agriculture, 

including use of herbicides with different modes-of-action, in a grower’s weed 

management program (Duke and Powles, 2009). MON 87708 has been developed to 

provide an effective and efficient method to incorporate another herbicide mode-of-action 

into the Roundup Ready soybean system (dicamba tolerance will be integrated into the 

Roundup Ready soybean system). This combination of herbicide-tolerance traits will 

allow the preemergence (including pre-plant) and postemergence use of both dicamba 

and glyphosate herbicides in an integrated weed management program to control a broad 

spectrum of grass and broadleaf weed species (Johnson et al., 2010). Increasing 

postemergence herbicide options is important, especially in conservation tillage 

situations, where the performance and consistency of postemergence herbicides has 

generally been greater than that of soil active residual products. Dicamba will improve 

the control of broadleaf weeds that are hard-to-control with glyphosate (e.g., common 

 

14 Souce is website: http://www.aphis.usda.gov/index.shtml. 

15 Source is 7 U.S.C. § 7701–7772, 7 CFR Part 340. 

16 Source is 7 CFR § 340.6 “Petition for Determination of Nonregulated Status”. 17 Source is 7 CFR § 340.6(c)(4). 
Monsanto Company 10-SY-210U 495 of 721 

lambsquarters, hemp sesbania, morningglory species, nightshade, Pennsylvania 

smartweed, prickly sida ̧ and wild buckwheat) and also offer an effective control option 

for glyphosate-resistant broadleaf weed species, namely marestail, common ragweed, 

giant ragweed, palmer pigweed, and waterhemp (Johnson et al., 2010). Dicamba will 

also offer an effective control option for broadleaf species resistant to ALS and PPO 

chemistries. In the case of PPO herbicides which are currently being heavily relied upon 

for control of glyphosate resistant amaranthus species, a primary dicamba benefit will be 

to provide options for delaying the further evolution and spread of PPO resistant 

amaranthus species (University of Tennessee, 2010).

Upon integration of MON 87708 into the Roundup Ready soybean system, growers will 

have the ability to continue to use established soybean production practices including 

crop rotation, tillage systems, labeled herbicides, and other cultural practices such as row 

spacing, thereby using the same planting and harvesting machinery currently being used. 

Growers will also continue to have the flexibility and simplicity in weed control provided 

by glyphosate that will allow growers to continue to reap the environmental benefits 

associated with the use of conservation-tillage that is facilitated by the use of glyphosate 

for postemergence weed control in the Roundup Ready soybean system (CTIC, 2011; 

CTIC, 2004). 

Monsanto has requested that APHIS make a determination that these soybean plants and 

their progeny will no longer be considered regulated articles under 7 CFR Part 340. 

APHIS’ action in this case is the determination whether or not to grant nonregulated 

status to MON 87708. APHIS’ purpose and need in making this determination is to 

fulfill its responsibilities under the PPA as defined under 7 CFR Part 340. 

While APHIS must consider impacts associated with the use of dicamba on the quality of 

the human environment as part of its determination to grant nonregulated status of 

MON 87708, the use of the pesticide is regulated at the federal level by the U.S. 

Environmental Protection Agency (U.S. EPA), not APHIS. APHIS’s authority under the 

Plant Protection Act does not allow it to specify conditions for the use of herbicides. 

Instead, EPA specifically approves labeling for any herbicide use including uses on 

agricultural crops. Before any dicamba product could be used over MON 87708 it must 

first be approved by the EPA as required by the Federal Insecticide Fungicide and 

Rodenticide Act (FIFRA); EPA must approve the pesticide (herbicide) product labeling 

for that specific use. Nevertheless, this environmental report examines the potential 

impact of dicamba use on MON 87708 on the human environment. 

J.3. Affected Environment 

This section describes the affected environment as it relates to soybean, Glycine max (L.) 

Merr., its uses and where soybean is grown. Related agricultural practices such as tillage, 

crop rotation, pesticide use, weed management, irrigation practices, and non-agricultural 

lands are also considered part of the affected environment, as is specialty soybean 

production, including organic soybean production and seed production. The information 

in this section, with minor exceptions as noted, is based upon information from Sections 
Monsanto Company 10-SY-210U 496 of 721 

II through IX of the petition, with those sections more specifically referenced throughout 

this section. 

J.3.1. Commercial Soybean Production and Uses 

Commercial soybean production and uses are discussed in Section VIII.B of the petition 

and summarized here; refer to the petition for more detail. Soybean is grown as a 

commercial crop in over 35 countries and is one of the most valued agricultural 

commodities because of its high protein and oil content. In 2008, soybean represented 

56% of world oilseed production, and approximately 33% of those soybean were 

produced in the U.S. In 2008, the U.S. exported 1.16 billion bushels (31.6 million metric 

tons) of soybean, which accounted for 40% of the world's soybean exports and was 

valued at $15.5 billion (ASA, 2009). 

Approximately 94% of the world’s soybean seed supply was crushed to produce soybean 

meal and oil in 2008 (Soyatech, 2010), and the majority was used to supply the feed 

industry for livestock use or the food industry for edible vegetable oil and soybean 

protein isolates. 

The U.S. soybean acreage in the past 10 years has varied from approximately 64.7 to 75.7 

million acres. Average soybean yields have varied from 33.9 to 43.3 bushels per acre 

over this same time period. According to data from USDA-NASS (2009a), soybean was 

planted on approximately 75.7 million acres in the U.S. in 2008, producing 2.96 billion 

bushels of soybean with a value of $27.4 billion (USDA-NASS, 2009b,c). 

In the U.S., soybean production occurs in three major soybean growing regions 

accounting for 99.1% of the soybean acreage: Midwest region (IL, IN, IA, KS, KY, MI, 

MN, MO, NE, ND, OH, SD, and WI), Southeast region (AL, AR, GA, LA, MS, NC, SC, 

and TN) and the Eastern Coastal region (DE, MD, NJ, NY, PA, and VA). Table J-1 

shows the relative productivity of the three regions in 2008. 

Table J-2. 2008 Soybean Productivity by Region 

Region 

% 2008 U.S. 

Soybean Acreage 

2008 Average Yield 

(bushels per acre) 

Range of Average State 

Yields (bushels per acre) 

Midwest/Great Plains 82.1 38.6 28.0 – 47.0 

Southeast 14.3 34.4 30.0 – 40.0 

Eastern Coastal 2.7 34.1 27.5 – 46.0 

Source: USDA-NASS (2009a,b). 

J.3.2. Specialty Soybean Production 

Specialty soybean are grown on fewer acres than commercial or commodity soybean and 

are typically grown, harvested and handled differently than commodity soybean, and 

premiums and incentives are paid for delivering a product that meets purity and quality 
Monsanto Company 10-SY-210U 497 of 721 

standards for the soybean variety. This category includes certified seed, organic 

production, soybean with specialty oil profiles, food grade soybean, and others. 

J.3.2.1. Certified Seed Production 

Certified seed production is discussed in Section VIII.B.2 of petition #10-SY-210U and 

summarized here; refer to the petition for more detail. Standardized seed production 

practices are responsible for maintaining high-quality seed stocks, an essential basis for 

U.S. agriculture. The value of seed quality (including genetic purity, vigor, and 

minimizing presence of weed seed, seed-borne diseases, and inert materials, such as dirt) 

has been identified as a major factor in determining crop yields. 

Soybean seed has four classes: 1) breeder, 2) foundation, 3) registered, and 4) certified 

(Association of Official Seed Certifying Agencies (AOSCA), 2009). Breeder seed is 

seed directly controlled by the originating or sponsoring plant breeding organization or 

firm. Foundation seed is first-generation seed increased from breeder seed and is handled 

to maintain purity and identity of a specific variety. Registered seed is the progeny of 

foundation seed that is handled to maintain satisfactory variety purity and identity. 

Certified seed is the progeny of breeder, foundation or registered seed, and is typically 

two generations from foundation seed. Not all soybean seed sold is officially certified; 

however, commercial soybean seed sold and planted for commodity soybean production 

typically meets or exceeds certified seed standards. 

Seed certification programs were initiated in the early 1900s in the U.S. to preserve the 

genetic identity and variety purity of seed. There are special land requirements, seed 

stock eligibility requirements, field inspections and seed labeling standards for seed 

certification. Seed certification services are available through various state agencies 

affiliated with the AOSCA. 

Soybean seed is produced throughout most of the U.S. soybean-growing regions by 

companies that produce and sell seed, and by toll seed producers, or tollers, which are 

companies that produce certified seed for other companies pursuant to a contract. Seed 

companies and tollers in turn contract acreage with growers to produce the required 

amount of soybean seed. Production or processing plants at these seed companies clean, 

condition, and bag the harvested soybean seed as well as monitor and inspect all the 

processes at the plant. 

The entire seed production process at the majority of the seed companies and tollers 

operates under standards established by the International Organization for 

Standardization (ISO), and includes internal and external audits (ISO, 2009). Field 

inspections are conducted on seed production fields throughout the soybean growing 

season to evaluate variety purity and ensure soybean plants are developing properly. 

Management practices are designed with the intent to keep the fields free of weeds, 

insects, and diseases, and to prevent mechanical mixing with other soybean varieties. 

The fields are also mapped to ensure the seed field has the minimum isolation 

requirement to prevent potential outcrossing. Isolation distances are agreed to by the 
Monsanto Company 10-SY-210U 498 of 721 

seed companies based on topography, varieties, volunteers, weeds, insects and weather 

patterns (AOSCA, 2009; ASTA, 2011). 

The field operations and management practices for producing soybean seed are similar to 

normal soybean production. However, special attention is needed in certain production 

practices to produce seed with high quality, high germination rates, and high genetic 

purity (Helsel and Minor, 1993). 

J.3.2.2. Organic Soybean Production 

Organic soybean production occurred on 125,600 acres in 2008 across Midwest and 

Southeast regions. Iowa, Minnesota, Michigan and Arkansas combined for over half the 

U.S. organic soybean acres. Over the past decade, organic soybean production peaked at 

174,500 acres in 2001 and ranged from 100,000 to 125,600 acres since 2000 (USDAERS, 2008a). Organic farming operations as described by the National Organic Program, 

which is administered by USDA’s Agricultural Marketing Service (AMS), requires 

organic production operations to have distinct, defined boundaries and buffer zones to 

prevent unintended contact with prohibited substances or products of excluded methods 

from adjoining land that is not under an organic production management plan. Organic 

production operations must also develop and maintain an organic production system plan 

approved by an accredited certifying agent. This plan enables the production operation to 

achieve and document compliance with the National Organic Standards, including the 

prohibition of the use of excluded methods.18 Excluded methods include a variety of 

methods used to genetically engineer organisms or influence their growth and 

development by means that are not possible under natural conditions or processes. The 

use of biotechnology such as that used to produce MON 87708 is an excluded method 

under the National Organic Program.19

Organic certification involves oversight by an accredited certifying agent of the materials 

and practices used to produce or handle an organic agricultural product. This oversight 

includes an annual review of the certified operation’s organic system plan and on-site 

inspections of the certified operation and its records. Although the National Organic 

Standards prohibit the use of excluded methods, they do not require testing of inputs or 

products for the presence of excluded methods. The presence of a detectable residue of a 

product of excluded methods alone does not necessarily constitute a violation of the 

National Organic Standards (USDA-AMS, 2011). The unintentional presence of the 

products of excluded methods will not affect the status of an organic product or operation 

when the operation has not used excluded methods and has taken reasonable steps to 

avoid contact with the products of excluded methods as detailed in an approved organic 

system plan. Organic certification indicates that organic production and handling 

processes have been followed, not that the product itself is “free” from any particular 

substance. 

 

18 Title 7 of the Code of Federal Regulation (7 CFR) Part 205. 

19 Source is 7 CFR § 205.2. 
Monsanto Company 10-SY-210U 499 of 721 

Organic soybean producers use production practices designed to prevent commingling of 

their crop with neighboring crops treated with herbicides and other pesticides (spray 

drift), or that may be using plant varieties produced by excluded methods (pollen 

movement). These well established practices include isolation zones, use of buffer rows 

surrounding the organic crop, adjusted planting dates, and varietal selection. 20 The 

implementation of management practices to avoid pollen from a biotechnology-derived 

crop in organic or conventional soybean production operations is facilitated by the nature 

of soybean pollination. Soybean is a highly self-pollinated species and exhibits a very 

low level of outcrossing (see Section IX.D). Outcrossing is the genetic transmission of a 

defined heritable characteristic from one group of individuals (population, crop variety) 

to another. Outcrossing most commonly results from cross-pollination. Since soybean is 

highly self-pollinating, organic or conventional soybean producers can and have 

effectively implemented practices (e.g., isolation during the growing season, equipment 

cleaning during harvest, and post-harvest separation of harvested seed) that allow them to 

reasonably avoid biotechnology-derived soybean and maintain organic or conventional 

production status (Brookes and Barfoot, 2004). Information about the National Organic 

Program, organic standards, and practices can be viewed on line (USDA-AMS, 2011). 

J.3.3. Agricultural Practices for Soybean 

J.3.3.1. Tillage 

Tillage is performed to prepare the soil for seed bed preparation and also serves as a weed 

control method because it uproots established plants and smothers them. Tillage in 

soybean production is discussed in Sections VIII.C.1 and VIII.C.2 of the petition and 

summarized here; refer to the petition for more detail. The benefits of conservation 

tillage or no-till systems relative to conventional tillage are well documented and include 

reduced soil erosion, reduced fuel and labor costs, conservation of soil moisture, 

improvement of soil structure, reduction of soil compaction and improvement of soil 

organic matter content. In 2007, approximately 27.5 million acres (39.6%) of soybean 

were planted in a no-till system (CTIC, 2007). In 2011 over 65% of U.S. soybean acres 

used some form of conservation tillage (USB, 2011a). The decision to plant soybean in a 

conservation tillage or no-till system is made long before planting as it may require 

special equipment. In addition, this decision is usually a long-term commitment, 

provided the system is successful. 

Slow soybean emergence and growth, plus lower yields, have been some of the concerns 

associated with adoption of conservation tillage systems in soybean, especially no-till. 

Research in Wisconsin and Minnesota shows that soil temperatures can be four to five 

degrees colder in no-till systems than in conventional tillage systems, which can slow 

emergence, but have little effect on soybean yield (Pedersen, 2008). Improved planters 

for establishment of good soybean populations and planting Roundup Ready® soybean 

varieties to effectively control weeds in no-till fields have made no-till a more viable 

 

20 Source is at website: http://attra.ncat.org/attra-pub/PDF/cropsfarmplan.pdf [Accessed on May 26, 2010]. ® Roundup Ready is a trademark of Monsanto Technology LLC. All other trademarks are the property of 

their respective owners. 
Monsanto Company 10-SY-210U 500 of 721 

production system for soybean. In 1995, before the introduction of Roundup Ready 

soybean, approximately 27% of the U.S. soybean acres used no-till production. In 2004, 

nine years after the introduction of Roundup Ready soybean system, no-till acreage 

increased to 36% of the total soybean acres (Sankula, 2006). The most recent surveys 

indicate that 39% of the soybean acres are produced using no-till methods (CTIC, 2007). 

Researchers still recommend some spring tillage on fine-textured and poorly drained soils 

for proper seedbed preparation (Pedersen, 2008). 

J.3.3.2. Crop Rotation 

The use of crop rotation in soybean production is discussed in Section VIII.I of petition 

#10-SY-210U and summarized here; refer to the petition for more detail. The wellestablished farming practice of crop rotation is still a key management tool for soybean 

growers. The purposes of growing soybean in rotation include: 

 improving yield and profitability of one or both crops over time; 

 decreasing the need for nitrogen fertilizer on the crop following soybean; 

 mitigating or breaking disease, insect, and weed cycles; 

 improving soil tilth and soil physical properties; 

 increasing residue cover; 

 reducing soil erosion; 

 increasing soil organic matter; and 

 reducing runoff of nutrients, herbicides, and insecticides (Boerma and Specht, 2004; 

Al-Kaisi et al., 2003). 

According to USDA Economic Research Service, 95% of the soybean-planted acreage 

has been in some form of a crop rotation system since 1991 (USDA-ERS, 2001). Cornand wheat-planted acreage has been rotated at a slightly lower level of 75% and 70%, 

respectively. Although the benefits of crop rotations can be substantial, the grower must 

make cropping decisions by evaluating both the agronomic and economic returns of 

various cropping systems. Crop rotations also afford growers the opportunity to diversify 

farm production in order to minimize market risks. 

Agronomic practices such as rotation patterns for soybean vary from state to state. 

However, there are similarities among states within certain growing regions. The 

majority of the U.S. soybean acreage (68.6%) is rotated to corn with approximately 

14.5% of the soybean acreage rotated back to soybean the following year. Wheat follows 

soybean on approximately 11.2% of the U.S. soybean acreage (see Table VIII-22 in the 

petition). 
Monsanto Company 10-SY-210U 501 of 721 

Continuous soybean production is uncommon in the Midwest. Soybean extension 

specialists encourage growers to avoid the practice as a way to reduce the risk of damage 

from diseases and nematodes (Hoeft et al., 2000; Al-Kaisi et al., 2003). Corn and 

soybean occupy more than 80% of the farmland in many of the Midwestern states, and 

the two-year cropping sequence of soybean-corn is used most extensively in this region. 

However, a soybean crop sometimes is grown after soybean and then rotated to corn in a 

3-year rotation sequence (soybean-soybean-corn) in the Midwest. Compared to corn, 

soybean shows a greater yield response to being grown after a number of years without 

soybean. The yield of both corn and soybean is approximately 10% higher when grown 

in rotation than when either crop is grown continuously (Hoeft et al., 2000). 

A combination of conservation tillage practices and crop rotation has been shown to be 

very effective in improving soil physical properties. Long-term studies in the Midwest 

indicate that the corn-soybean rotation improves yield potential of no-till systems 

compared to continuous corn production (Al-Kaisi, 2001). 

J.3.3.3. Irrigation 

The use of irrigation in soybean production is discussed in Section VIII.B of the petition 

and summarized here; refer to the petition for more detail. The productivity of soybean is 

highly dependent upon soil and climatic conditions. In the U.S., the soil and climatic 

requirements for growing soybean are very similar to corn. The soils and climate in the 

Midwestern, Eastern and portions of the Great Plains regions of the U.S. provide 

sufficient water under normal climatic conditions to produce a soybean crop. The general 

water requirement for a high-yielding soybean crop is approximately 20 inches of water 

during the growing season (Hoeft et al., 2000). Soil texture and structure are key 

components determining water availability in soils, where medium-textured soils hold 

more available water, allowing soybean roots to penetrate deeper in medium-textured 

soils than in clay soils. Irrigation is used on approximately 9% of the soybean acreage in 

the U.S. to supplement the water supply during dry periods in the Western and Southern 

soybean growing regions (USDA-ERS, 2008b). 

J.3.3.4. Management of Insects 

The management of insects in soybean production is discussed in Section VIII.D of the 

petition and summarized here; refer to the petition for more detail. Although insects are 

rated as less problematic than weeds in U.S. soybean production, management of insect 

pests during the growth and development of soybean is important for protecting the yield 

of soybean (Aref and Pike, 1998). Understanding the impact of insects on soybean 

growth is essential for proper management (Higley and Boethel, 1994). It is important to 

understand the way that insects injure soybean as well as how the soybean plant responds 

to insect injury. Insect injury can impact yield, plant maturity, or seed quality. The 

ultimate impact of injury is damage, as a measurable reduction in plant growth 

development or reproduction. Insect injury in soybean seldom reaches levels to cause an 

economic loss in the primary soybean production areas, as indicated by the low 

percentage (16%) of soybean acreage that receives an insecticide treatment (USDANASS, 2007b). 
Monsanto Company 10-SY-210U 502 of 721 

Characterizing soybean responses to insect injury is essential in establishing economic 

injury levels (Higley and Boethel, 1994). Most often, soybean insects pests are 

categorized or defined by the plant parts they injure, namely root-feeding, stem-feeding, 

leaf-feeding, or pod-feeding insects. The root- and stem-feeding insect groups are often 

the hardest to scout and typically are not detected until after they have caused their 

damage. The leaf-feeding insects comprise the biggest group of soybean insect pests, but 

not necessarily the most economically damaging insects. Recent research on defoliation 

has determined that a major effect of leaf injury is to reduce light interception by the 

soybean canopy which in turn can have a significant effect on yield (Higley and Boethel, 

1994). Soybean has an extraordinary capacity to withstand considerable defoliation early 

in the season without significant yield loss. By contrast, defoliation during the flowering 

and pod filling stages poses a greater threat to yield, because the soybean plant has less 

time to compensate for injury compared to other growth stages. Research indicates that 

the soybean plant can sustain a 35% leaf loss prior to the pre-bloom period without 

lowering yield (NDSU, 2002). However, from pod-set to maturity, the plant can tolerate 

only a 20% defoliation level before yield is impacted. 

J.3.3.5. Management of Diseases and Other Pests 

The management of diseases and other pests in soybean production is discussed in 

Section VIII.E of the petition and summarized here; refer to the petition for more detail. 

More than 100 pathogens are known to affect soybean, of which 35 are considered to be 

of economic importance (Heatherly and Hodges, 1999). The estimated yield losses to 

soybean diseases in the U.S. were 12.5, 13.2, and 13.0 million metric tons in 2008, 2009, 

and 2010, respectively (Wrather and Koenning, 2011), which equates to 15.5%, 14.4% 

and 14.4% losses of total soybean production, respectively (ASA, 2011). Pathogens can 

affect all parts of the soybean plant, resulting in reduced quality and yield. The extent of 

losses depends upon the pathogen, the state of plant development and health when 

infection occurs, the severity of the disease on individual plants, and the number of plants 

affected (Heatherly and Hodges, 1999). 

According to field surveys conducted in fifteen soybean-producing states during 1996 to 

2010, soybean cyst nematode (Heterodera glycines) caused the greatest soybean yield 

losses (Wrather and Koenning, 2011). Phytophthora root and stem rot (Phytophthora 

sojae), brown spot (Septoria glycines), charcoal rot (Macrophomina phaseolina), 

sclerotinia stem rot (Sclerotinia sclerotiorum), seedling diseases, and sudden death 

syndrome (Fusarium solani f.sp. glycines) followed in economical importance. As 

expected, yield losses vary by region (Wrather et al., 2001). 

Selecting resistant varieties is the primary tool growers have for disease control 

(Heatherly and Hodges, 1999). Resistant varieties may have morphological or 

physiological characteristics that provide immunity, resistance, tolerance or avoidance to 

certain pathogens. Cultural practices can also play an important role in disease 

management by reducing initial inoculums or reducing the rate of disease development 

(Heatherly and Hodges, 1999). 
Monsanto Company 10-SY-210U 503 of 721 

Preplant tillage can bury crop residue, which encourages the decomposition of fungalresting structures. Crop rotation is often recommended as a disease-management 

strategy. Rotating crops interrupts the disease cycle and allows time for the 

decomposition of inoculums. One exception is Rhizoctonia, a soil-inhabitant pathogen 

that grows on a wide variety of crops and can survive sufficiently in the soil to make crop 

rotation an impractical means of controlling this pest. Row spacing, plant population, 

and planting date can also be changed to manage soybean diseases. 

Soybean cyst nematode (SCN), Heterodera glycines, is one of the most damaging 

pathogens of soybean throughout the soybean growing regions of the U.S. with losses 

estimated to be about $1.5 billion (Pedersen, 2008). The simplest, least expensive 

method to reduce populations of this pest is to rotate soybean with a non-host crop such 

as corn, small grains, or sorghum. However, planting resistant varieties is regarded as the 

best and most effective management practice to prevent losses from this pest (Wrather 

and Mitchum, 2010). 

High-quality seed is essential for controlling seedling diseases. The most important 

seedling diseases in soybean are Phytophthora, Pythium, Rhizoctonia, and Fusarium

(Pedersen, 2008). Many soybean varieties demonstrate resistance to specific taxonomic 

races of Phytophthora. Treating soybean seed with a fungicide (e.g., metalaxyl or 

mefenoxam) is effective against damping-off disease (seedling blight) caused by common 

soil fungi, such as Phytophthora and Pythium. Fungicide seed treatments are 

recommended where there is a history of these seedling diseases. 

Asian soybean rust is a foliar fungal disease that typically infests soybean during 

reproductive stages of development and can cause defoliation and reduce yields 

significantly in geographies such as Brazil (Dorrance et al., 2007). Soybean rust is 

caused by the fungus Phakopsora pachyrhizi. This disease in the U.S. was first detected 

in Louisiana in 2004 (LSU, 2009). At this time, foliar application of fungicides is the 

standard disease-management practice to limit yield losses due to soybean rust. 

J.3.3.6. Management of Weeds 

The management of weeds in soybean production is discussed in Section VIII.F of 

petition #10-SY-210U and summarized here; refer to the petition for more detail. Annual 

weeds are perceived to be the greatest pest problem in soybean production, followed by 

perennial weeds (Aref and Pike, 1998). Weed control in soybean is essential to 

optimizing yields because weeds compete with soybean for light, nutrients, and soil 

moisture. Weeds can also harbor insects and diseases, and also can interfere with 

harvest, causing extra wear on harvest equipment (Pedersen, 2008). 

Foxtail spp. (Setaria spp.), pigweed (Amaranthus spp.), velvetleaf (Abutilon theophrasti), 

lambsquarters (Chenopodium album), and cocklebur (Xanthium strumarium) are common 

weeds in Midwest corn and soybean fields. However, growers consider giant ragweed 

(Ambrosia artemisiifolia), lambsquarters, Canada thistle (Cirsium arvense), cocklebur, 

and velvetleaf to be the top five most problematic weeds in corn and soybean because of 

the difficulty to control these weeds (Nice and Johnson, 2005). 
Monsanto Company 10-SY-210U 504 of 721 

The primary weed competition factors that affect a potential yield loss in soybean from 

weed competition are the weed species, weed density, and the duration of the 

competition. When weeds are left to compete with soybean for the entire growing 

season, yield losses can exceed 75% (Dalley et al., 2001). Generally, the competition 

between crops and weeds increases with increasing weed density. The time period that 

weeds compete with the soybean crop influences the level of yield loss. In general, the 

later the weeds emerge, the less impact the weeds will have on yield. Soybean plants 

withstand early-season weed competition longer than corn, and the canopy generally 

closes earlier in soybean than corn (i.e., plants in adjacent rows grow to a sufficient size 

such that their foliage touches between the rows blocking the sunlight from reaching the 

ground). In addition, canopy closure is much sooner when soybean is planted in narrow 

rows. 

The most effective weed management programs in soybeans use a combination of cultural, 

mechanical, and/or herbicide control practices, hereafter called diversified weed management 

practices. Herbicide practices include the use of several herbicides with different 

modes-of-action, either within or across seasons, applying herbicides at the labeled rate at 

the correct timing, and proper application of the herbicide. Cultural and mechanical 

weed control practices can also be important components of an effective diversified weed 

management program (Loux et al., 2009). Cultural practices such as crop rotation, 

narrow row spacing and planting date are a few of the crop management practices that are 

implemented to provide the crop with a competitive edge over weeds. Mechanical 

methods of weed control including tillage have been used for centuries to control weeds 

in crop production. Spring or fall preplant tillage and in-crop shallow cultivation can 

effectively reduce the competitive ability of weeds by burying the plants, disturbing or 

weakening their root systems, or causing sufficient physical injury to kill the plants. A 

consequence of in-crop cultivation for weed control is that it can injure crop roots and 

cause moisture loss. The planting of winter cover crops is another cultural practice that 

can also be utilized. The planting of cover crops, such as grasses, legumes or small 

grains, can protect and improve soil quality, help reduce erosion, and can serve as surface 

mulch in no-till cropping practices (Mannering et al, 2007). However the planting of a 

cover crop incurs additional costs to the grower and therefore cover crops are not a major 

weed management practice in major soybean growing areas (Iowa State University, 

2006). 

The use of herbicides has become an important part of managing weeds in soybean. In 

2006, approximately 98% of the soybean acreage received an herbicide application 

(USDA-NASS, 2007b). Over 35 different herbicide active ingredients are registered and 

available for use by soybean growers to control weeds (Tables VIII-7 and VIII-8). 

Herbicide weed control programs in conventional soybean consist of preemergence 

herbicides used alone or in mixtures with other preemergence herbicides. Mixtures of 

two preemergence herbicides are used to broaden the spectrum of control to both grasses 

and broadleaf weed species. Preemergence herbicides are followed by postemergence 

applications to control weeds that emerge later in the crop. Total postemergence weed 

control programs were seldom used in conventional soybean prior to 1995. Prior to 

Roundup Ready soybean, soybean planted in a no-till system would receive a preplant 

burndown herbicide application for broad-spectrum control of existing weeds at time of 
Monsanto Company 10-SY-210U 505 of 721 

planting, followed by different multiple soil residual herbicides at planting and possibly 

still other herbicides applied postemergence to the crop and the weeds. In conventional 

soybeans the typical herbicide program consisted of multiple soil residual herbicides 

applied preemergence to the crop and weeds and, possibly, other herbicides applied 

postemergence to the crop and weeds. Therefore, multiple herbicides and/or multiple 

applications were generally made in conventional and no-till non-Roundup Ready 

soybean. The average number of herbicide applications per acre in soybean rose from 1.5 

in 1990 to 1.7 applications in 1995 reflecting the use of at-plant and postemergence 

applications or two postemergence applications (Gianessi et al., 2002). 

Selective herbicides are designed to kill specific types of plants, usually grasses or broad 

leaf weeds, and have proved effective to reduce in-crop tillage or cultivation to control 

weeds in soybean production. The development of selective herbicides has progressed 

since the introduction of the first herbicide (2,4-D) for weed control in corn in early 

1940s. Although the primary purpose of tillage is for seedbed preparation, tillage still is 

used to supplement weed control with selective herbicides in soybean production. 

 The availability of herbicide-tolerant soybean products are an important aspect of weed 

management in U.S. soybean production, as discussed in Section VIII of the petition and 

summarized here; refer to the petition for more detail. Herbicide-tolerant soybean was 

introduced to provide growers with additional options by improving crop safety (no 

herbicide damage to the crop) and improving weed control. In 2006, herbicide-tolerant 

soybean (glyphosate-tolerant) was planted on 89% (67.2 million acres) of the of soybean 

acres (USDA-NASS, 2007a). The percentage of herbicide-tolerant soybean subsequently 

increased to 91% in 2009. With the high percentage of glyphosate-tolerant soybean and 

the use of glyphosate for preplant burndown and postemergence in-crop applications, it is 

not surprising that glyphosate was used on 97% of the total soybean acres in 2006. 

Because of the unique broad spectrum herbicide activity of glyphosate, it is possible for 

growers to choose to use only glyphosate for their total weed management. Some 

growers made this choice, choosing not to utilize diversified weed management practices 

and instead relying only on glyphosate for total weed management. As a result of these 

practices, there has been an increase in the number of crop acres with glyphosate resistant 

weed populations over the last decade. However, recently the number of growers 

incorporating other weed management practices along with the use of glyphosate, such as 

other herbicide modes-of-action or cultivation practices, has grown over time in part in 

response to the evolution of glyphosate resistant weeds and based upon recommendations 

from Monsanto, academics, and the pesticide industry. Academics and university 

extension services recommend growers use diversified weed management practices, as 

described above, as the guiding principle for managing resistance and shifts in weed 

population. The specific practices to be recommended by area or by farm will be 

communicated by local experts versed in the best methods for both proactive and reactive 

management of resistance. Since many soybean farmers practice conservation tillage and 

some may chose to plant soybeans repeatedly on the same land, the use of multiple 

herbicide modes-of-action with overlapping effectiveness on the targeted weed spectrum 

will be the primary method employed. Studies have demonstrated that using the same 

combination of herbicides with multiple modes-of-action and overlapping effectiveness 
Monsanto Company 10-SY-210U 506 of 721 

over multiple seasons is an effective way to proactively manage resistance (Beckie and 

Reboud, 2009). 

The incorporation of additional weed management practices has been most pronounced in 

Roundup Ready corn and Roundup Ready cotton but a growing trend in Roundup Ready 

soybeans has also occurred. In a 2005 grower survey, 15 to 21% of growers applied nonglyphosate herbicides in addition to glyphosate for weed control in glyphosate-tolerant 

soybean (Givens et al., 2009). These non-glyphosate herbicides were applied prior to 

planting, at planting and/or postemergence in soybean (see Section VIII.F.1 of the 

petition). In another grower survey conducted at the end of the 2009 growing season, the 

percent of growers applying non-glyphosate herbicides rose to 33% for those growing 

continuous Roundup Ready soybeans, and to 33% and 52% for growers growing 

Roundup Ready soybeans in rotation with Roundup Ready corn and Roundup Ready 

cotton, respectively (personnel communication from Dr. David Shaw, December 2011). 

These data indicate a trend towards increased diversification of weed management 

practices in glyphosate-tolerant crops. 

Further evidence of increased adoption of diversified weed management practices, 

including incorporation of multiple herbicide modes-of-action, across Roundup Ready 

corn, cotton and soybeans is presented by Prince et. al, (2011). This study reported that 

between 46% and 54% of surveyed Roundup Ready growers (corn, cotton and soybeans) 

who responded that they did not have glyphosate-resistant weeds on their farm used 

either a non-glyphosate residual and/or postemergence herbicide in the 2009 growing 

season. For growers indicating they have on-farm herbicide-resistant weed populations 

to other herbicides, the percentage of growers was higher at 72% to 75%. Furthermore, 

researchers report that approximately 40 to 50% of the growers utilizing glyphosatetolerant crops indicate that rotating herbicides or tank mixing glyphosate with other 

herbicides is an effective management practice to minimize the development of 

glyphosate resistance (Powles et al., 1996; Diggle et al., 2003; Beckie, 2006; Beckie and 

Reboud, 2009). Indeed, as described in detail below, a prominent strategy to delay the 

development of herbicide-resistant weeds is to increase the diversity of weed 

management options in agriculture, including use of herbicides with different modes-ofaction in a grower’s weed management program (Duke and Powles, 2009). 

Herbicide-resistant Weeds 

The use of any herbicide results in the potential for the selection of weeds resistant to that 

herbicide particularly when the herbicide is not used as part of a diversified weed 

management program. Within a weed species, individuals may possess an inherent 

ability to withstand the effects of a particular herbicide. Repeated use of that herbicide 

will expose the weed population to a "selection pressure," which may lead to an increase 

in the number of surviving resistant individuals in the population (HRAC, 2010). The 

increased and repeated use of glyphosate over glyphosate-tolerant crops without 

incorporation of diversified weed management practices, such as crop rotation, 

cultivation or use of multiple herbicide modes-of-action, has resulted in the selection of 

glyphosate-resistant weeds. Glyphosate-resistant weed biotypes found in soybean fields 

include Palmer pigweed (Amaranthus palmeri), tall waterhemp (Amaranthus 
Monsanto Company 10-SY-210U 507 of 721 

tuberculatus), common ragweed (Ambrosia artemisiifolia), giant ragweed (Ambrosia 

trifida), hairy fleabane (Conyza bonariensis), horseweed (Conyza canadensis), kochia 

(Kochia scoparia), goosegrass (Eleusine indica), Italian ryegrass (Lolium multifloru), 

rigid ryegrass (Lolium rigidum), Johnsongrass (Sorghum halepe) (Heap, 2011). As with 

other herbicide resistant weeds that have developed, the emergence of glyphosate 

resistant weed biotypes over the past decade represents a need for growers to adapt and 

implement improved weed management strategies. 

Weed resistance is common in the major non-glyphosate soybean herbicide groups. 

Table J-2 and Table J-3 summarize known resistance among the major weed species 

present in soybean within each of the key soybean herbicide groups and herbicide classes 

active on broadleaf weeds (Heap, 2011). Resistance to the ALS group of herbicides is 

present in most of the major broadleaf weed species commonly found in soybeans. For 

common ragweed and waterhemp there is known resistance to at least one member for 

several of the major soybean herbicide chemistry classes. While there are still effective 

options for managing common ragweed, waterhemp, palmer pigweed and other key 

broadleaf weeds, there is a need for additional herbicide modes-of-action to combat 

future resistance in soybeans and continued management of existing herbicide-resistant 

weed populations. Similarly, there has been an increase in the detection of weed 

populations with multiple resistance (i.e., resistance to multiple herbicide modes-ofaction) in some weed species, for example, Amaranthus spp. (Tranel et al, 2010). The 

emergence of these resistant biotypes and continued need to utilize diverse weed 

management practices supports the need for additional herbicide modes-of-action in 

major crops such as soybeans. 

Monsanto and academics recommend the use of multiple herbicide modes-of-action in 

the Roundup Ready soybean system regardless of whether glyphosate-resistant or 

hard-to-control broadleaf weeds are present. Monsanto specifically recommends the use 

of a soil residual as part of the weed management system. Growers may also choose to 

switch to other weed management systems in their soybean. APHIS has approved other 

herbicide-tolerant soybean including phosphinothricin-tolerant and ALS-tolerant soybean 

events (Table J-4). For growers who choose to use the Roundup Ready soybean system, 

Monsanto and university extension agents provide recommended control options for 

glyphosate-resistant weeds. These options include the use of residual and postemergent 

herbicides such as synthetic auxins (2,4-D), ACCase inhibitors (clethodim, sethoxydim), 

PPO inhibitors (lactofen, fomesafen), and ALS inhibitors (cloransulam). 22 These 

herbicides alone or combinations of these herbicides as well as traditional tillage methods 

are and will continue to be used to control glyphosate-resistant or hard-to-control 

broadleaf weeds. 

 

 

22Monsanto Technology Use Guide; www.weedresistancemanagement.com. 
Monsanto Company 10-SY-210U 

Table J-3. Known Weed Resistance in the Southern U.S. 1 Most Common Broadleaf Weeds (# states where listed as a top weed) 

Resistance 

Group 2 

ALS 

(Group 2) 

PPO 

(Group 14) 

PS II 

(Group 5) 

Glycine 

(Group 9) 

Chemistry 

Class 2

Sulfonyl 

Urea

Imidazol

inones

Triazoles Diphenyl 

ether

N-phenyl thalimide

Triazinones - 

Example chlorimuron imazapyr chloransulam lactofen

fomesafen

flumioxazin metribuzin glyphosate 

Morning glory (5) Sida (prickly sida (5) X Sicklepod (4) Hemp sesbania (3) Pigweed spp. 3 (3) X X X X Palmer pigweed (2) X X X X Cocklebur (1) X X X Horseweed (marestail) (1) 

X X X 

1Source: www.weedscience.org 2 Cross resistance is possible within a resistance group and/or chemistry class 3 Includes redroot pigweed and smooth pigweed 
Monsanto Company 10-SY-210U 

Table J-4. Known Weed Resistance in the Midwest U.S. 1 Most Common Broadleaf Weeds (# states where listed as a top weed) 

Resistance 

Group 2 

ALS 

(Group 2) 

PPO 

(Group 14) 

PS II 

(Group 5) 

Glycine 

(Group 9) 

Chemistry 

Class 2

Sulfonyl 

Urea

Imidazol

inones

Triazoles Diphenyl 

ether

N-phenyl thalimide

Triazinones - 

Example chlorimuron imazapyr chloransulam lactofen

fomesafen

flumioxazin metribuzin glyphosate 

Pigweed spp. 3 (12) X X X X Velvetleaf (11) Lambsquarters (10) X X X Cocklebur (9) X X X Common ragweed (7) X X X X X X Smartweed spp. (6) Morning glory (5) Waterhemp (5) X X X X X Horesweed (marestail) (3) 

X X X 

Giant ragweed (3) X X X X Kochia (2) X X X 
Monsanto Company 10-SY-210U 

1Source: www.weedscience.org 2 Cross resistance is possible within a resistance group and/or chemistry class 3 Includes redroot pigweed and smooth pigweed 
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J.3.3.7. Dicamba Herbicide Use in the U.S. 

Dicamba use in the U.S. is discussed in Section VIII.G of the petition and summarized 

here; refer to the petition for more detail. Dicamba is a broadleaf selective herbicide that 

was approved by the Environmental Protection Agency (U.S. EPA) for agricultural 

application uses in 1967 (U.S. EPA, 2009b). Dicamba herbicide is currently labeled for 

weed control in corn, soybean, cotton, sorghum, wheat, barley, oats, millet, pasture, 

rangeland, asparagus, sugarcane, and turf, grass grown for seed, conservation reserve 

programs, and fallow croplands. Dicamba treated acreage has ranged from 17.4 to 36.3 

million acres between 1990 and 2008. Usage of dicamba peaked during the period of 

1994 through 1997, where 1994 was the peak year when 36 million acres were treated 

with 9.4 million pounds of dicamba. Since 1994, the use of dicamba has steadily 

declined to 20.2 million treated acres with 2.67 million pounds applied in 2008. The 

decline is due to the competitive market introductions of sulfonylurea herbicides 

(chlorsulfuron, metsulfuron-methyl, and thifensulfuron-methyl) in wheat, new broadleaf 

herbicide active ingredients in corn, and glyphosate-tolerant corn. Usage in cotton is one 

exception, where dicamba-treated acres (preplant applications) have increased from 

140,000 to 590,000 acres from 2004 to 2008. Dicamba is formulated as a standalone 

herbicide product and marketed by several companies under various trade names such as 

Banvel®, Clarity®, Diablo®, Rifle®, Sterling®, and Vision. These dicamba products can 

also be tank mixed with one or more active ingredients depending on the treated crop. 

For example, Clarity can be tank mixed with over 75 herbicide products in labeled crops. 

Additionally, dicamba is formulated as a premix product with one or more other 

herbicide active ingredients such as glyphosate, 2,4-D, diflufenzopyr, atrazine, 

nicosulfuron, metsulfuron, primsulfuron, triazulfuron, rimsulfuron and halosulfuron. 

Based on USDA-NASS (2004, 2006b, 2007b, 2008) statistics, dicamba application rates 

ranged from 0.03 to 0.25 pounds per acre with the average number of applications 

ranging from 1 to 1.2 applications per cropping season. Dicamba rates are the lowest in 

barley, wheat, and oats, where typically more than one application is made in these crops 

per cropping season. The average application rate in corn is 0.19 pounds of dicamba per 

acre with slightly over one application per season. 

Dicamba is currently labeled for use in conventional or Roundup Ready soybean, 

although dicamba use is extremely limited because applications are restricted to preplant 

and/or preharvest applications due to crop tolerance concerns. The dicamba-treated 

acreage in 2008 soybean production was approximately 530,000 acres representing 0.7% 

of the total soybean acreage. 

J.3.4. Human Health and Worker Safety 

Soybean is a highly versatile crop which can be processed into a wide variety of food 

products. Soybean protein is used to enhance nutrition in a wide variety of food products, 

such as breakfast cereals and pasta. Soybean protein is also an important component in 

baked goods, alternative meat products, soups, energy bars, nutritional beverages infant 

formula and dairy replacement products (USB, 2011b). Soybean oil constitutes the 

majority (68%) of consumed edible fats and oils in the U.S. (ASA, 2011). It is present in 
Monsanto Company 10-SY-210U 512 of 721 

numerous food products including cooking oils, shortening, margarine, mayonnaise, 

salad dressings and a wide variety of fat or oil-based products (USB, 2011b). 

Humans consume soybean and have done so for thousands of years. Soybean improved 

with new traits produced by biotechnology pose no unique risks relative to other soybean 

developed using traditional breeding methods. Biotechnology-derived soybean is 

evaluated extensively prior to commercial introduction. All biotechnology-derived 

soybean products on the market today have satisfactorily completed the FDA 

consultation process established to review the safety of foods and feeds derived from 

biotechnology-derived crops for human and animal consumption. Biotechnology-derived 

soybean crop varieties that have been deregulated or are under consideration for 

deregulation are shown in Table J-5. 

 

Pesticides have been used extensively in the production of soybean (see Section J.3.3.6). 

The use of these pesticides is regulated by the Federal Insecticide, Fungicide, and 

Rodenticide Act (FIFRA). FIFRA was amended in 1988 to accelerate the reregistration 

of products with active ingredients registered prior to November 1, 1984. The amended 

Act calls for the development and submission of data to support the reregistration of the 

active ingredient, as well as a review of all data submitted to the EPA. During the 

reregistration process, EPA thoroughly reviews the scientific database underlying a 

pesticide's registration. The purpose of the Agency's review is to reassess the potential 

risks arising from the currently registered uses of a pesticide, to determine the need for 

additional data on health and environmental effects, and to determine whether or not the 

pesticide continues to meet the "no unreasonable adverse effects" criteria of FIFRA. 

In order to comply with FIFRA, EPA evaluates potential risks to humans and the 

environment, and may require applicants to submit more than 100 different scientific 

studies conducted according to EPA guidelines. The data required by EPA are used to 

evaluate whether a pesticide has the potential to cause adverse effects on humans 

(including chronic, reproductive and cancer risk), wildlife, fish, and plants (including 

endangered species and other non-target organisms, i.e., organisms that the pesticide is 

not intended to act against). If the pesticide may be used on food or feed crops, EPA also 

sets tolerances (maximum pesticide residue levels) for the amount of the pesticide residue 

that can legally remain in or on foods. EPA undertakes this analysis under the authority 

of the Federal Food, Drug, and Cosmetic Act (FFDCA), and must conclude that such 

tolerances will be safe, meaning that there is a reasonable certainty that no harm will 

result from aggregate (food, water and non-occupational residential/recreational) 

exposure to the pesticide residues. 

EPA has evaluated dicamba and concluded that is has a complete and comprehensive 

regulatory database (toxicity, environmental fate, and ecological toxicity) that has been 

evaluated by the EPA. EPA completed the reregistration process for dicamba and a 

Registration Eligibility Decision (RED) was issued in 2006 and subsequently amended in 

2008 and 2009 (U.S. EPA, 2009b). EPA concluded the available data submitted for 

dicamba are complete and adequate to support the continued registration of dicamba 

products and uses. EPA also considered toxicity data and available information 

concerning the variability of sensitive subpopulations, including infants and children. 
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The EPA concluded there is reasonable certainty that no harm will result to the general 

population, or to infants and children, as a result of aggregate exposure to dicamba 

residues. Thus, all then-current dicamba uses were eligible for reregistration (U.S. EPA, 

2009b). See Appendix M, Section M.2 and M.3 for additional details about the EPA 

registration process and the reregistraiton of dicamba. 

In the agricultural production of soybean, growers and workers may be exposed to 

pesticides applied to soybean by mixing, loading, or applying chemicals, or by entering a 

previously treated site. EPA conducts a comprehensive occupational worker safety 

evaluation and risk assessment of pesticides to assess the risk to agricultural workers 

during mixing, loading, and applying. EPA evaluated occupational risk to workers as a part 

of the dicamba RED and concluded that worker exposure to dicamba for all registered 

agricultural uses, including exposures associated with the current preemergence and 

postemergence pre-harvest soybean uses, meet the "no unreasonable adverse effects" 

criteria of FIFRA (U.S. EPA, 2009b). The use of dicamba on MON 87708 does not pose 

any new exposure considerations for workers. Therefore the use of dicamba on 

MON 87708 will not pose a risk to agricultural workers. See Appenidx M, Section M.2.2 

for additional details on applicator exposure. 

In addition, the Worker Protection Standard (WPS) provides additional protections to 

agricultural workers and pesticide applicators. The WPS contains requirements for 

pesticide safety training, notification of pesticide applications, use of personal protective 

equipment (PPE), restricted-entry intervals (REI) after pesticide application, 

decontamination supplies, and emergency medical assistance. Under the WPS, EPA 

requires the pesticide label to specify PPE and REI, based on the properties of the 

pesticide product, that will provide an appropriate level of protection23. 

 

23 www.epa.gov/pesticides/health/worker.htm 
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Table J-5. Deregulated or Submitted Biotechnology-derived Soybean Products 

Phenotype Event Institution Date Deregulated 

High Oleic Acid, Low 

Saturated Fat 

MON 87705 Monsanto December 16, 

2011 

Omega 3 Fatty Acid MON 87769 Monsanto Submitted 

Lepidopteran Resistant MON 87701 Monsanto June 28, 2011 

Herbicide-tolerant 

(2,4-D) 

DAS-68416-4 Dow 

AgroSciences 

Submitted 

Herbicide-tolerant 

(Glyphosate/Isoxaflutole) 

FG72 Bayer 

Crop Sciences 

Submitted 

Herbicide-tolerant 

(Imidazolinone)

BPS-CV127-9 BASF 

Plant Science 

Submitted 

High Oleic Acid DP-3Ø5423-1 Pioneer June, 2010 

Glyphosate- and ALStolerant DP-356043-5 Pioneer July, 2008 

Glyphosate-tolerant MON 89788 Monsanto February, 2007 

Phosphinothricin-tolerant GU262 AgrEvo October, 1998 

Phosphinothricin-tolerant A5547-127 AgrEvo May, 1998 

Altered Oil Profile G94-1, G94-19, G168DuPont May, 1997 

Phosphinothricin-tolerant W62, W98, A2704-

12, A2704-21, 

A5547-35 

AgrEvo August, 1996 

Glyphosate-tolerant 40-3-2 Monsanto May, 1994 

Source is website: http://www.aphis.usda.gov/brs/not_reg.html
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J.3.5. Animal and Plant Communities, Soil Microorganisms and Biodiversity 

J.3.5.1. Animal Communities 

Soybean production systems in agriculture are host to many animal species including 

deer, groundhogs, rabbits, raccoons, geese and small rodents (USDA-APHIS, 2011b). 

Mammals and birds, including migratory mammals and birds, may seasonally consume 

grain (Galle et al., 2009), and invertebrates can feed on the plant during the entire 

growing season. 

Animals that feed primarily on soybean are seed-feeding insects and rodents found in 

agricultural fields. Crop pest insects are considered less problematic than weeds in U.S. 

soybean production as indicated by the low percentage (14%) of soybean acreage that 

receives insecticide treatment (USDA-NASS, 2006). Management of insects in soybean 

production fields is discussed in Section VIII.D of the petition. Some rodents, such as 

mice or squirrels, may seasonally feed exclusively on soybean seeds. Thus, these animals 

may have a diet containing significant amounts of soybean seeds. Deer may also browse 

in soybean fields on the forage and on seed left after harvest. 

Animals that feed outside soybean fields are also considered in this section. The 

environment surrounding a soybean field, which may vary in plant composition 

depending on the region, may serve as a food source and habitat for mammals, birds, fish 

and insects. In certain areas, soybean fields may be bordered by other soybean, corn, or 

other crops; soybean fields may also be surrounded by woods and/or pasture/grassland 

areas, as well as aquatic environments. Therefore, the types of vegetation, including 

weeds, around a soybean field depend on the area where the soybean is planted. 

Fertilizers and/or water containing pesticides may run off into adjacent lands, pesticides 

may also move outside of the agroecosystem from drift and offsite movement. 

Regardless of the agricultural operation, animals and insects outside the field may be 

impacted directly from the use of fertilizers, pesticides, and erosion caused from 

agricultural operations, or indirectly, both positively and negatively, from effects on the 

plant community outside the soybean field. 

J.3.5.2. Plant Communities 

The affected environment for growing soybean plants can generally be considered the 

agroecosystem (managed agricultural fields) plus some area extending beyond the 

intended plantings that might be affected by agricultural operations. Plants, extraneous to 

the crop, which grow in planted fields can be considered weeds and are dealt with in 

section J.3.3.6 of this document. 

Plants growing outside soybean fields are considered in this section. The environment 

surrounding a soybean field varies in plant composition depending on the region. In 

certain areas, soybean fields may be bordered by other soybean, corn, or other crops; 

fields may also be surrounded by woods and/or pasture/grassland areas, as well as aquatic 

environments. Therefore, the types of vegetation, including weeds, around a soybean 

field depend on the area where the soybean is planted. A variety of weeds dwell in and 
Monsanto Company 10-SY-210U 516 of 721 

around soybean fields; those species may also vary depending on the region where the 

soybean is planted. These plants may be found in ditches, hedge rows, fence rows, wind 

breaks, yards, and other uncultivated areas, and may be annuals, biennials or perennials. 

Regardless of the agricultural operation, these plants may be impacted, both positively 

and negatively, by agricultural operations. Fertilizers and/or water may run off into 

adjacent lands, resulting in increased plant growth outside the agroecosystem. Negative 

impacts on plants adjacent to production fields can occur from herbicide runoff and drift. 

Finally, soybeans infrequently occur as a volunteer when soybean seeds remain in a field 

following harvesting and may be considered a weed in the subsequent crop. Volunteer 

soybean in rotational crops is not a concern in the Midwest region because the soybean 

seed is typically not viable after the winter period (Carpenter et al., 2002; OECD, 2000). 

In southern soybean growing areas of the U.S. where the winter temperatures are milder, 

it is possible for soybean seed to remain viable over the winter and germinate the 

following spring. If volunteer soybean should emerge after planting, shallow cultivation 

and/or use of another herbicide will control volunteers and effectively reduce competition 

with the crop. Several postemergence herbicides are also available to control volunteer 

soybean (conventional or glyphosate-tolerant soybean, and by extension dicamba-tolerant 

soybean) in each of the major soybean rotational crops (Section VIII.J. of petition #10-

SY-210U). Therefore, volunteer soybean normally is not a concern in rotational crops, 

such as corn, cotton, rice and small grains (e.g., wheat, barley, sorghum and oats), which 

are the primary rotational crops following soybean due to the availability of adequate 

control measures for volunteer soybean (Carpenter et al, 2002; OECD, 2000). 

J.3.5.3. Soil Microorganisms 

Soil microbial communities are highly complex and are often characterized by high 

microbial diversity (Tiedje et al., 1999). The occurrence and abundance of soil 

microorganisms are affected by 1) soil characteristics like tilth, organic matter, nutrient 

content, and moisture capacity, 2) typical physico-chemical factors such as temperature, 

pH, and redox potential, and 3) soil management practices. Agricultural practices such as 

fertilization and cultivation may also have profound effects on soil microbial populations, 

species composition, colonization, and associated biochemical processes (Buckley and 

Schmidt, 2001; 2003). Consequently, significant variation in microbial populations is 

expected in agricultural fields. 

Members of the bacterial family Rhizobiaceae and Bradyrhizobiaceae form a highly 

complex and specific symbiotic relationship with leguminous plants, including soybean 

(Gage, 2004). The nitrogen-fixing plant-microbe symbiosis results in the formation of 

root nodules, which provide an environment in which differentiated bacteria called 

bacteroids are capable of reducing or “fixing” atmospheric nitrogen. The product of 

nitrogen fixation, ammonia, can then be utilized by the plant. As a result of this 

relationship, nitrogen inputs are typically not necessary for agricultural production of 

soybeans. 
Monsanto Company 10-SY-210U 517 of 721 

J.3.6. Biodiversity 

Biodiversity refers to all plants, animals, and microorganisms interacting in an ecosystem. 

Among other benefits, biodiversity provides valuable genetic resources for crop 

improvement (Harlan, 1975) and also provides other functions beyond food, fiber, fuel, 

and income. These include pollination, genetic introgression, biological control, nutrient 

recycling, competition against natural enemies, soil structure, soil and water conservation, 

disease suppression, control of local microclimate, control of local hydrological processes, 

and detoxification of noxious chemicals (Altieri, 1999). The loss of biodiversity results 

in a need for costly external inputs in order to provide these functions to the crop (Altieri, 

1999). Agricultural land subject to intensive farming practices, such as that used in crop 

production, generally has low levels of biodiversity compared with adjacent natural areas. 

The use of broad spectrum insecticides and herbicides is one of the most severe 

constraints for biological diversity in crops. Tillage, seed bed preparation, planting of a 

monoculture crop, pesticide use, fertilizer use, and harvest may all limit the diversity of 

plants and animals (Lovett, Price, & Lovett, 2003). Herbicide use in agricultural fields is 

likely to indirectly impact biodiversity by decreasing weed species present in the field 

and those insects, birds and mammals that utilize these weeds. 

Conservation tillage practices can have a positive impact on wildlife, including beneficial 

arthropods (Altieri, 1999; Landis et al., 2005; Towery and Werblow, 2010). 

Conservation tillage practices benefit biodiversity due to decreased soil erosion improves 

surface water quality, retention of vegetative cover, crop residues serve as a food source,; 

and increased populations invertebrates which can serve as food sources to other 

organisms (Landis et al. 2005; Sharpe, 2010) 

J.3.7. Physical Environment 

J.3.7.1. Land Use 

In 2008, soybean was grown as a commercial crop on over 75 million acres in at least 27 

states in the U.S. (USDA-NASS, 2009a). Soybean acreage in the past five years has been 

relatively stable varying from 64.7 million to 75.7 million acres with a 10-year average of 

73.3 million acres (Table VIII-1). Soybean fields are typically highly managed 

agricultural areas that can be expected to be dedicated to crop production for many years. 

Fluctuations in soybean acreage are due to environmental, agronomic and economic 

factors, as well as government programs such as the conservation reserve program (CRP). 

Currently, biotechnology-derived herbicide tolerant soybean is planted on 91% of the 

soybean acreage (USDA-NASS, 2009a) and the Roundup Ready soybean system has 

become the standard weed control program in U.S. soybean production. There is no 

indication that the introduction and widespread adoption of biotechnology-derived crops 

in general has resulted in a significant change to the total U.S. acreage devoted to 

agricultural production. The cumulative land area in the U.S. planted to principal crops, 

which include corn, sorghum, oats, barley, winter wheat, rye, durum, spring wheat, rice, 

soybean, peanuts, sunflower, cotton, dry edible beans, potatoes, canola, proso millet, and 
Monsanto Company 10-SY-210U 518 of 721 

sugar beets, has remained relatively constant over the past 27 years. From 1982 to 1995, 

the average yearly acreage of principal crops was 323 million. This average is essentially 

unchanged at 326 million acres since the introduction of biotechnology-derived crops in 

1996 (USDA-NASS, 1984, 1988, 1990, 1992, 1995, 1998, 2000, 2003, 2006a, 2009a). 

J.3.7.2. Water Resources 

The soils and climate in the Midwestern, Eastern and portions of the Great Plains regions 

of the U.S. provide sufficient water under normal climatic conditions to produce a 

soybean crop. Irrigation is used on approximately 9% of the soybean acreage in the U.S. 

to supplement the water supply during dry periods in the Western and Southern soybean 

growing regions (USDA-ERS, 2008b). More information on water management in 

soybean production can be found in Sections VIII.B and J.3.3.3 of the petition. 

Groundwater may be impacted from soybean production by the movement of pesticides 

and fertilizers vertically through soil. Surface water may also be impacted from soybean 

production by runoff from soybean fields that carries soil particles and herbicides or other 

pesticides to streams, rivers, lakes, wetlands and other water bodies. As discussed below, 

based on existing data, the soil component of runoff is a much more important 

contributor to surface water impacts than is the pesticide component. 

Tillage causes widespread soil disturbance. Thus, erosion, topsoil loss and the resulting 

sedimentation and turbidity in streams are likely to increase with increased tillage. In 

2009, based on the states’ water quality reports, EPA identified sedimentation and 

turbidity as two of the top 10 causes of impairment to surface water in the U.S. in 

general; in 2007, EPA identified sedimentation/siltation as a leading cause of impairment 

to rivers and streams in particular (U.S. EPA, 2009a; EPA, 2007a). EPA has projected 

conservation tillage to be “the major soil protection method and candidate best 

management practice for improving surface water quality” (U.S. EPA, 2002). EPA 

identifies conservation tillage as the first of its CORE4 agricultural management practices 

for water quality protection (U.S. EPA, 2008a). 

Based on the states’ water quality reports to EPA, which EPA makes available through its 

National Assessment Database, pesticides in general and herbicides in particular are a 

relatively minor contributor to impairment of surface water in the U.S., compared to 

sedimentation/siltation and turbidity (U.S. EPA, 2008b). Pesticides accounted for less 

than one percent of reported causes of surface water impairment in all but four of the 17 

leading U.S. soybean-producing states. In those four states, pesticides accounted for two 

to eight percent of reported causes of impairment. Of the pesticides that were reported as 

contributing to impairment among the 17 leading soybean-producing states, almost all are 

highly persistent chemicals that are no longer registered for use in the U.S. (U.S. EPA, 

2008b). Dicamba is not included on this list. 

Water resources (Ground and surface water) may also be impacted from the use 

herbicides used to control weeds. Dicamba has been widely used in agriculture over the 

last four decades with dicamba’s peak use occurring in 1994 (see Section VIII.G of the 

petition). In the dicamba Reregistration Eligibility Decision (RED) document, EPA 
Monsanto Company 10-SY-210U 519 of 721 

considered potential risks associated with dicamba use, and its degradate DCSA when 

appropriate, due to surface or ground water using screening level (high-end exposure) 

models to estimate environmental concentrations. The EPA then compared these 

exposure estimates to appropriate endpoints from mammalian and aquatic animal and 

plant ecotoxicity studies to determine potential impacts on human health and the 

environment. The EPA used the models PRZM/EXAMS and SCIGROW to estimate 

levels of dicamba in surface and ground water, respectively, using the physical, chemical, 

and environmental fate properties, and approved high-end use patterns of dicamba (see 

Appendix M, Section M.3.2 of the petition). 

For drinking water resources, estimated surface water concentrations using the simulated 

sugarcane crop scenario for both ground and aerial applications of 2.8 lbs a.e./acre ranged 

from 9.7 to 13 μg/L dicamba a.e. and 0.66 to 0.81 μg/L for DCSA (U.S. EPA, 2005a). 

For environmental-ecological water quality, estimated surface water concentrations for a 

simulated soybean crop scenario for ground and aerial applications of 2.0 lbs a.e./acre 

ranged from 33.3 to 36.1 μg/L dicamba a.e. The U.S. Geological Survey National Water 

Quality Assessment (NAWQA) monitoring program also analyzed surface water in a 

1993-2003 survey of surface waters of the United States, which included geographical 

areas where dicamba use has historically been most intense. Dicamba had a low 

incidence of detections (approximately 3% of samples) and the highest levels detected 

were approximately 2 μg/L (U.S. EPA, 2005a). 

For groundwater, estimated concentrations using the simulated sugarcane crop scenario 

for both ground and aerial applications of 2.8 lbs dicamba a.e. per acre were 0.016 μg/L 

dicamba and 0.008 μg/L for DCSA, dicamba’s major environmental degradate (U.S. 

EPA, 2005a). The EPA also analyzed groundwater in a 1971-1991 national monitoring 

study. The frequency of dicamba detection was less than 2.5% based on 3172 samples 

analyzed, and dicamba was detected at levels from less than 0.01 to 44μg/L (U.S. EPA, 

1992), and peak dicamba detections occurred between 1985 and 1990. Dicamba also had 

a low incidence of detections (less 3% of samples) in the 1993-2003 NAWQA 

monitoring program where the highest detections were approximately 2.5 μg/L (U.S. 

EPA, 2005a). 

J.3.7.3. Soil Quality 

Soybean is cultivated across a wide variety of soils in the U.S. Cultivation and tillage 

practices can directly impact the attributes of soil, including its physical and biological 

properties. Microbial populations and associated biochemical processes are critical to 

maintaining soil health and quality. Additionally, maintaining soil pH in the range of 6.0 

to 7.0 will enhance the availability of inherent and fertilizer nutrients, reduce the 

availability of toxic elements, particularly aluminum and manganese, and enhance 

microbial activity (Hoeft, 2000). The increased microbial activity that is associated with 

the optimum pH level results in oxidation of organic matter and increased release of 

nutrients from the organic matter. 

Conservation tillage and no-till systems enhance soil quality relative to conventional 

tillage. Benefits of conservation tillage are well documented and include reduced soil 
Monsanto Company 10-SY-210U 520 of 721 

erosion, conservation of soil moisture, improvement of soil structure, reduction of soil 

compaction and improvement of soil organic matter content. In 2010, approximately 

27.5 million acres (39.6%) of soybean were planted in a no-till system (CTIC, 2010). In 

2011 over 65% of U.S. soybean acres used some form of conservation tillage (USB, 

2011a). 

J.3.7.4. Air Quality and Climate 

Many agricultural activities affect air quality including tillage, farm equipment, and 

nitrous oxide emissions from the use of nitrogen fertilizer. These agricultural activities 

individually have potential adverse environmental impacts on air quality and climate and 

may be impacted, positively or negatively, by changes in agricultural practices. Issues of 

concern include, but are not necessarily limited to, atmospheric emission of carbon 

dioxide, nitrogen oxide, sulfur oxide, and particulate matter. Agricultural practices have 

the potential to directly and indirectly impact air quality and contribute emissions which 

could lead to climate change. 

Tillage contributes to the release of greenhouse gases (GHG) because of the loss of 

carbon dioxide to the atmosphere and the exposure and oxidation of soil organic matter 

(Baker, et al., 2005). Emissions released from agricultural equipment (e.g., irrigation 

pumps and tractors) include carbon monoxide, nitrogen oxides, particulate matter, and 

sulfur oxides. Nitrous oxide may also be released following the use of nitrogen fertilizer. 

Agriculture, including land-use changes for farming, is responsible for an estimated 17 to 

32% of all human-induced GHG emissions (USDA-APHIS, 2011b). Generation of 

GHGs may have long term impacts on climate change as they function as retainers of 

solar radiation. The U.S. agricultural sector has been identified as the second largest 

contributor to GHG emissions (USDA-APHIS, 2011b). Herro (2008) proposes that if 

agriculture practices were modified, significant reductions in the release of GHGs could 

be achieved. 

J.3.8. Adjacent-Agricultural Crop and Non-Agricultural Plants 

Soybean is widely grown throughout the U.S. on land devoted to agricultural use. The 

biology of soybean is discussed in Section II of the petition and summarized here; refer to 

the petition for more detail. Soybean does not have any related wild relatives in the U.S. 

with which it can hybridize. Soybean does not survive outside of cultivation and is not 

invasive or weedy. Soybean is grown adjacent to large acre crops such as corn, wheat, 

and alfalfa, near vegetables, orchards, pastures, and adjacent to non-agricultural lands, 

such as forests, grasslands, streams, lakes, rivers and occasionally near urban lands. 

Herbicides are commonly used in the production of soybean and a detailed discussion of 

their use can be found in Section VIII.F.1 of the petition; refer to the petition for more 

detail. Impacts on adjacent agricultural crops and non-agricultural plants can occur from 

offsite movement of any herbicide, and these impacts are actively managed by farmers 

and applicators specially trained to use such products consistent with product labels and 

other state or local restrictions. Depending upon the herbicide, factors for managing the 

potential for drift and offsite movement include the selectivity and sensitivity of the 
Monsanto Company 10-SY-210U 521 of 721 

herbicide, local weather conditions at the time of application (wind, temperature, 

humidity, inversion potential), droplet size distribution, application volume, boom height 

(height of the application equipment above the crop canopy), sprayer speed, and distance 

from the edge of the application area (SDTF, 1997; Felsot et al., 2010). A variety of 

measures can be employed to control the potential for spray drift and offsite movement, 

including nozzle selection and application techniques and restrictions. 

The potential for offsite movement of an herbicide is regulated at a federal level by EPA. 

EPA considers possible effects from offsite movement as part of the pesticide registration 

process required under FIFRA. Specifically, in order to approve the use of a pesticide 

(herbicide), EPA must conclude that no unreasonable adverse effects on non-target 

vegetation will result from potential offsite movement when the pesticide is used 

according to the product label. When pesticides are applied in accordance with label 

instructions, offsite impacts can be avoided. EPA reassessed the potential risks to nontarget plants in its analysis in the dicamba RED, concluding that no specific additional 

drift mitigations were needed to support the continued registration of all dicamba uses, 

including use in conventional soybean with applications at early preemergence prior to 

planting and late postemergence prior to harvest (U.S. EPA, 2009b). A detailed 

discussion of the use of dicamba herbicide in the U.S. can be found in Section VIII.G of 

the petition. 

J.3.9. Animal Feed 

Soybean meal is the most valuable component obtained from processing soybean, 

accounting for roughly 50-75% of its overall value (USDA-ERS, 2005). The majority of 

soybean meal is used by the animal feed industry as a cost-effective protein and amino 

acid source to animal diets. Soybean meal can serve as a excellent protein source that 

can complement the limited amino acid profile of feeds derived from corn (Kerley and 

Allee, 2003). In 2009, approximately 36 million tons of soybean meal was produced, 27 

million tons of which was marketed for animal feed with the largest volumes consumed 

by poultry (48%), swine (26%), and beef (12%) (ASA, 2010). 

Dairy and livestock producers use soybean forage as feed. Soybean forage is an 

inexpensive, readily available, on-farm source of high-quality, high-protein forage 

adapted to growth during the summer months when other forage legume species typically 

are restricted in growth (USDA-ARS, 2006). Soybean forage can be used as hay or to 

produce silage (MAFRI, 2004). An additional use of soybean for feed can be full-fat 

(whole) soybean for dairy cattle and swine, but for swine it is limited due to the high oil 

content to a maximum of 20% of the total diet (Yacentiuk, 2008). 

Monsanto has completed the biotechnology consultation process with FDA for the safety 

and nutritional assessment of food and feed derived from MON 87708 soybean on 

October 11, 2011 (BNF No. 00125, Monsanto, 2011). As a part of its evaluation, FDA 

reviewed information on the identity, function, and characterization of the genes, 

including expression of the gene products in MON 87708 soybean, as well as information 

on the safety of the MON 87708 DMO and MON 87708 including a dietary risk 

assessment. 
Monsanto Company 10-SY-210U 522 of 721 

EPA has responsibility to regulate the use of pesticides (herbicides) that may be used on 

feed crops, and must establish pesticide tolerances (maximum pesticide residue levels) 

for the amount of pesticide residue that can legally remain in or on the feed crop. EPA 

undertakes this analysis under the authority of the Federal Food, Drug, and Cosmetic Act 

(FFDCA), and must conclude that such tolerances will be safe, meaning that there is a 

reasonable certainty that no harm to human health will result from the use of the 

pesticide. EPA reassessed all dicamba pesticide tolerances (food and feed tolerances) as 

part of the dicamba RED, including the 10 ppm soybean seed tolerance supporting the 

existing use in conventional soybean for early postemergence applications up to 0.5 

pound a.e. per acre and late postemergence prior to harvest up to 1.0 pound a.e. per acre 

(U.S. EPA, 2009b). A complete listing of dicamba feed tolerances can be found at 40 

CFR § 180.227. Monsanto has also petitioned (Pesticide Petition # 0F7725) the EPA to 

establish new feed tolerances on soybean forage (45 ppm) and soybean hay (70 ppm). 

Tolerances for soybean forage and hay for current dicamba uses in conventional soybean 

were not previously established because the current preharvest application is made past 

the stage where the crop would be useful as forage or hay. 

J.4. Alternatives 

The action of deregulation is governed by 7 CFR § 340.6 (d)(3)(i) which states that 

APHIS may approve the petition in whole or in part, resulting in three possible Agency 

actions in response to the petition: 

 No action 

MON 87708 would remain a regulated article 

 Approval in part 

MON 87708 would be granted deregulated status with some restrictions or 

conditions (e.g., geographic) 

 Approval in whole 

MON 87708 would be granted full deregulated status 

J.4.1. No Action Alternative 

Under the “no action” alternative, MON 87708 would remain a regulated article under 7 

CFR Part 340. MON 87708 could be grown under USDA notification or permit and 

confined release conditions. MON 87708 would not have unrestricted availability to 

commercial soybean growers. Under this alternative, growers will likely continue to use 

biotechnology-derived soybean that are commercially available and herbicides to control 

weeds in soybean fields. In 2009, 91% of the U.S. soybean was produced with herbicidetolerant biotechnology-derived soybean. In 2006, 98% of soybean fields received an 

herbicide application for weed control. Over 35 different herbicide active ingredients are 

available for use by soybean growers (Tables VIII-7 and VIII-8) and it is reasonably 

foreseeable that new herbicide active ingredients may be discovered or existing 
Monsanto Company 10-SY-210U 523 of 721 

herbicides used in other crops may be adapted for use on soybean in the future under this 

alternative. Growers may continue to use agronomic practices they currently use for 

production of soybean, including the Roundup Ready soybean system. Residual 

herbicides will continue to be recommended for use in the Roundup Ready soybean 

system to provide a second herbicide mode-of-action for weed resistance management. 

In areas where glyphosate-resistant or hard-to-control broadleaf weeds are present, 

growers may continue to choose to use the Roundup Ready soybean system and 

incorporate herbicides with other modes-of-action, including residual herbicides. 

Recommended control options for glyphosate-resistant broadleaf weeds include the use 

of residual herbicides, preemergent herbicides such as synthetic auxins (2,4-D), and 

postemergent herbicides such ACCase inhibitors (clethodim, sethoxydim), PPO inhibitors 

(lactofen, fomesafen), and ALS inhibitors (cloransulam). 24 Under the no action 

alternative, these herbicides alone and combinations of these herbicides, as well as tillage, 

could be used to control glyphosate-resistant and hard-to-control broadleaf weeds. 

Because of existing resistance to currently available soybean herbicides and in the 

absence of new herbicide options, the number of weed populations and species with 

multiple resistance in soybean production areas may continue to increase as well as 

resistance to herbicide classes that are currently being relied on to manage current levels 

of resistance to glyphosate and other herbicides (e.g., use of PPO herbicides to manage 

glyphosate resistant palmer pigweed and waterhemp). In the event new resistant biotypes 

to key herbicides in the PPO group evolve and increase, there will be fewer options 

available for herbicide control of key weed species such as in the Amaranthus genus. 

Under the no action alternative, growers may choose to use other biotechnology-derived 

herbicide-tolerant soybean products that have been deregulated by APHIS, including 

glyphosate-tolerant, ALS-tolerant (DP356043-5), or phosphinothricin-tolerant (e.g., 

A2704-12 and other events) soybean. Three other herbicide-tolerant soybean products 

are currently under consideration for deregulation by APHIS including: 2,4-D-tolerant 

(DAS-68416-4), glyphosate-tolerant and isoxaflutole-tolerant (FG72), and 

imidazolinone-tolerant (BPS-CV127-9). Thus, it is reasonably foreseeable that growers 

may choose these options and use of their companion herbicides under the no action 

alternative. 

Currently, 91% of the U.S. soybean crop is produced with herbicide-tolerant (principally 

glyphosate-tolerant) soybean (USDA-NASS, 2009a), indicating growers are using nonglyphosate based herbicides and/or traditional tillage methods for weed control on about 

10% of soybean acreage. Growers may choose to use the non-glyphosate based weed 

control systems and/or traditional tillage methods under the no action alternative. 

J.4.2. Alternatives Considered but Eliminated from Detailed Evaluation 

Approval in part based on plant pest risk 

 

24 Monsanto Technology Use Guides can be found at: 

http://www.monsanto.com/SiteCollectionDocuments/Technology-Use-Guide.pdf

. 
Monsanto Company 10-SY-210U 524 of 721 

Based on APHIS’ regulatory authority, MON 87708 could be granted “approval in part” 

dependent upon a finding of a plant pest risk in certain geographies or under certain 

conditions. APHIS may impose restrictions upon the cultivation or use of MON 87708 in 

specific geographies or circumstances to mitigate an identified plant pest risk. 

MON 87708 has been thoroughly characterized, and extensive information presented in 

Sections I through IX of petition #10-SY-210U demonstrates that MON 87708 does not 

present a plant pest risk under any circumstance. Therefore, from a plant pest risk 

perspective, there is no basis for imposing geographic or other restrictions on 

MON 87708. 

The safe use of dicamba for agricultural purposes was first established in 1967, and EPA 

recently reaffirmed its human and environmental safety for agricultural uses, including 

soybean production, with the reregistration in 2006 (U.S. EPA, 2009b). The decisions 

regarding the safe use of dicamba belong solely with EPA. APHIS has no regulatory 

authority to restrict the use of pesticides or impose measures to mitigate their risk. 

Monsanto has applied for a label for use of dicamba on MON 87708, and requested the 

establishment of new feed tolerances for soybean forage and hay. EPA will review the 

proposed label amendment and assess if the requested use pattern and use instructions 

meet its statutory standard of no unreasonable adverse effects. 

On the basis of this analysis demonstrating that there is no plant pest risk consideration or 

other risks that would lead USDA to consider approval in part, and because EPA has 

authority to regulate the herbicide dicamba, these alternatives were not further considered 

in this report. 

J.4.3. Approval in Whole Alternative 

Under the “approval in whole” alternative, MON 87708 would no longer be a regulated 

article under 7 CFR Part 340 and would be widely available for planting. Growers would 

have the option of treating their soybean crop with dicamba using an expanded window 

of herbicide application. MON 87708 will allow for preemergent applications up to crop 

emergence (cracking) and in-crop postemergent applications up to R1/R2 growth stage, 

compared to existing uses which permit only early preemergence applications 14-28-days 

prior to planting and late postemergence applications prior to harvest. With the approval 

by the EPA for the use of dicamba on MON 87708 and the integration of MON 87708 

into the Roundup Ready soybean system through traditional breeding, the combined 

crop-tolerances to both glyphosate and dicamba would allow growers to utilize 

glyphosate and dicamba herbicides in their weed management systems, as well as other 

herbicides currently registered for use in soybean. Monsanto is requesting approval in 

whole or full deregulated status for MON 87708. Information and assessments presented 

throughout this environmental report demonstrate that MON 87708 does not present a 

significant environmental impact when approved in whole. 

J.5. Potential Environmental Impacts 

The Council on Environmental Quality (CEQ) regulations require that significance be 

evaluated in terms of context (affected environment) and intensity (the severity of the 
Monsanto Company 10-SY-210U 525 of 721 

impact).25 Analysis of these factors considered the “no action” and the “approval in 

whole” alternatives. The differences between the two alternatives address the relevant 

issues in considering whether deregulation of MON 87708 results in a significant impact 

to the quality of the human environment. In most cases, there are no differences between 

the two alternatives. Where differences are noted, these differences are described and 

their significance evaluated. Factors evaluated as part of the assessment of significance 

include: potential impacts to land use patterns, farming practices, specialty and organic 

soybean production; potential impacts to non-agricultural lands; potential impacts to the 

marketability of soybean seed for planting and harvested seed for commodity markets; 

potential impacts to public health; and potential impacts to non-target organisms and 

threatened or endangered species, and biodiversity. Finally, cumulative impacts are 

considered in light of this action combined with past and reasonably foreseeable future 

actions. 

J.5.1. Methodology and Assumptions 

MON 87708 is intended to be a trait that will have utility across all of the acreage upon 

which soybean is currently grown and widely available in soybean varieties sold to 

growers. It is impossible to determine the exact amount of acreage on which 

MON 87708 may be grown if deregulated. However, for the purposes of this assessment, 

it is assumed that MON 87708 will occupy 40% of the U.S. soybean acreage at peak 

penetration (see Section VIII.H.2 of the petition). Over the past decade, glyphosatetolerant soybean varieties have been widely adopted in the marketplace. In most cases, 

glyphosate was applied to control narrowleaf and broadleaf weeds in U.S. soybean fields, 

where glyphosate provided excellent control of most weeds. In recent years, the 

development of glyphosate-resistant weeds and shifts in broadleaf weed populations to 

species that are inherently more tolerant to glyphosate have increased the use of 

additional herbicides that work through a different mode-of-action to achieve an 

acceptable level of weed control. As a result of the ongoing need to control the majority 

of weed species present in soybean fields, additional herbicides are being used, and 

multiple herbicide-tolerance traits are being developed to provide growers with additional 

weed control options that will compete with MON 87708. These herbicides and traits 

will likely be available at the time MON 87708 is introduced to the marketplace; thus, 

MON 87708 will compete for market share with established products, like Roundup 

Ready soybean 40-3-2 first introduced in 1996, and new herbicide-tolerance traits that 

will be available in the foreseeable future. Growers will ultimately select weed control 

systems that fit the needs for their individual farming operation, such that some 

proportion of growers will choose to use MON 87708 integrated into the Roundup Ready 

soybean system. 

It is EPA’s regulatory authority under the Federal Insecticide, Fungicide and Rodenticide 

Act (FIFRA) to register pesticide products for their intended uses, see Section J.3.4 for 

additional detail. Potential impacts of dicamba use associated with MON 87708 are a 

connected activity under the deregulation in whole alternative, and while pesticide use is 

regulated by EPA, dicamba use will also be discussed in order to assess potential impacts 

 

25 CEQ regulations are available in 40 CFR §1508.27. 
Monsanto Company 10-SY-210U 526 of 721 

to the quality of the human environment. Monsanto has submitted to EPA an application 

to amend EPA Reg. No. 524-582 to register a new use pattern for dicamba. On the basis 

of Monsanto’s assessment of EPA registration decisions and approved labels for 

alternative herbicides, the use of dicamba on MON 87708 poses less risk potential to 

human health and the environment than some existing alternative non-glyphosate 

herbicides (see Appendix L). The primary basis for this assertion is that the introduction 

of MON 87708 and the associated use of dicamba when compared to other nonglyphosate herbicides registered for use on soybean will: 1) reduce human health risk 

potential for applicators, bystanders, and consumers; 2) reduce risk potential to aquatic 

organisms in the environment; 3) positively impact the sustainability of soybean 

production in the U.S. due to the addition of a second mode-of-action into the Roundup 

Ready soybean system, thereby potentially delaying further development of glyphosateresistant broadleaf weed populations, and to broadleaf weed herbicides in general; and 4) 

support the continued use of conservation tillage with its well known environmental 

benefits. 

J.5.2. Physical Impacts; Land Use, Water Quality, Climate, Soil Quality and Air 

Quality 

J.5.2.1. Land Use Impacts 

Approval in whole alternative 

The difference between the approval in whole and the no action alternative is expected to 

be integration of MON 87708 into the Roundup Ready soybean system using traditional 

breeding techniques. For those acres where glyphosate resistant weeds may already be 

present, where application of an herbicide with a different mode-of-action would aid in 

weed control, or for grower implementation of weed resistant management practices, the 

cultivation of soybean containing the MON 87708 trait would be an option for growers. 

Herbicide-tolerant soybean has been deregulated and grown in the U.S. since 1996. 

Roundup Ready soybean currently occupies greater than 90% of total soybean acres. 

Fluctuations in total soybean acreage before and after herbicide-tolerant soybean was 

commercialized (USDA-NASS, 2011) indicates that factors unrelated to the availability 

of the herbicide-tolerant trait play a role in total soybean acres planted. Agricultural land 

use, and consequently crop production is dictated by many factors, the most significant of 

which are commodity prices. Accordingly, growers may increase acres dedicated to 

soybean production to meet increased demand, but they do so in response to commodity 

prices and market demand, not in response to availability or adoption of biotechnologyderived traits. 

With the exception of tolerance to dicamba, MON 87708 is phenotypically and 

agronomically unchanged from conventional soybean. Phenotypic and agronomic 

information collected from field trials conducted in 2008 using the same agricultural 

inputs showed no meaningful changes between MON 87708 and the conventional control 

(see Section VII.D of the petition). Information presented in the petition demonstrates 

that compared to conventional soybean, MON 87708 does not display increased 
Monsanto Company 10-SY-210U 527 of 721 

susceptibility to pests or diseases, and is not changed regarding crop emergence, growth 

development or yield. Additional laboratory and greenhouse-based experiments reached 

the same conclusion; MON 87708 was unchanged compared to the conventional control 

for seed germination and symbiotic relationship parameters. Therefore, production 

management practices (e.g., planting and harvest timing, fertilizer inputs, and pesticide 

use other than dicamba) are not expected to change with the introduction of MON 87708. 

Similarly, because there are no changes in growth and development or yield, there is no 

expectation that the introduction of MON 87708 and its use in development of soybean 

varieties will significantly alter the geographical range of commercial soybean cultivation. 

Thus, the introduction of MON 87708 is not anticipated to facilitate production of 

soybean in areas where it is not currently grown or have significant impact on total 

soybean production acres. Therefore, the approval in whole and no action alternatives 

are the same regarding their potential impact on land use. 

No action alternative 

Under the no action alternative MON 87708 and its progeny would continue to be 

regulated articles and would not be widely grown. Under this alternative dicamba 

herbicide would not become integrated into the Roundup Ready soybean system and 

dicamba use would likely remain similar to today’s use pattern in soybean. As discussed 

above, full deregulation of MON 87708 is not expected to result in changes to land use. 

Similarly, land use changes would not be expected with the no action alternative. As 

discussed, relatively minor fluctuations in soybean acreage would be expected with either 

alternative, resulting from environmental, agronomic, economic and governmental 

influences. Therefore, the no action and approval in whole alternatives are the same 

regarding their potential impact on land use. 

J.5.2.2. Water Quality Impacts 

Approval in whole alternative 

The difference between the approval in whole and the no action alternative is expected to 

be integration of MON 87708 into the Roundup Ready soybean system using traditional 

breeding techniques. For those acres where glyphosate resistant weeds may already be 

present, where application of an herbicide with a different mode-of-action would aid in 

weed control, or for grower implementation of weed resistant management practices, the 

cultivation of soybean containing the MON 87708 trait would be an option for growers. 

Impacts associated with MON 87708

Water quality could be impacted either directly by MON 87708 via plant material 

impacts on water resources, or indirectly via impacts from the use of dicamba or tillage 

practices associated with the planting of MON 87708. Conservation tillage, a system that 

leaves 30% or more of the previous crop residue covering the soil when planting another 

crop has been increasingly employed in commercial soybean acres, and helps minimize 

any impacts of soybean production on water quality by reducing soil erosion. 
Monsanto Company 10-SY-210U 528 of 721 

In terms of potential direct impacts on water quality, MON 87708 has been shown to be 

compositionally, agronomically and phenotypically equivalent to conventional soybean 

and is therefore unlikely to have any significant impact on surface water quality. The 

DMO protein contained in MON 87708 is a member of the larger family of oxygenase 

proteins that are ubiquitous in plants and microbes in the environment. The mode of 

action of this family of proteins is well known, and the introduced DMO protein itself 

was derived from a common soil bacterium (Stenotrophomonas maltophilia). 

MON 87708 DMO has been shown to have a high level of substrate specificity, and 

characterization data provided in Section V of petition #10-SY-210U demonstrate the 

safety of the DMO protein. Therefore, it is unlikely that the presence of DMO protein in 

MON 87708 will have a significant impact on water quality. 

Under full deregulation of MON 87708, there will be a decreased need for farmers 

employing conventional tillage practices in order to manage certain weed situations. 

There is a potential impact to soil conservation in those situations where tillage has been 

employed to manage resistant weeds (CAST, 2011). Dicamba’s complementary and 

supplementary postemergence activity to glyphosate will provide improved 

postemergence weed management options and thus support more sustainable 

conservation tillage practices because postermergence herbicide options are generally 

preferred by growers (Fawcett and Towery, 2002). Tillage causes widespread soil 

disturbance causing erosion and topsoil loss, impacting the sedimentation and turbidity of 

streams. EPA identified sedimentation and turbidity as two of the top 10 causes of 

impairment to surface water in the U.S.; similarly in 2007, EPA identified 

sedimentation/siltation as a leading cause of impairment to rivers and streams in 

particular (U.S. EPA, 2007a; 2009a;). EPA has projected conservation tillage to be “the 

major soil protection method and candidate best management practice for improving 

surface water quality” (U.S. EPA, 2002). EPA identifies conservation tillage as the first 

of its CORE4 agricultural management practices for water quality protection (U.S. EPA, 

2008a). Therefore, the approval in whole and no action alternatives are not significantly 

different regarding the impact of the cultivation of MON 87708 on water quality. 

Impacts from use of dicamba on MON 87708

Under full deregulation, dicamba would be an additional weed management tool for 

managing hard-to-control and herbicide-resistant broadleaf weeds found in soybean 

fields. The use of dicamba on soybean would be expected to increase relative to current 

and historical levels of use, up to 2.6 times the maximum historical annual level in 1994. 

However, potential impacts associated with any increased use of dicamba from the 

cultivation of MON 87708 have been adequately assessed by EPA as part of the dicamba 

Reregistration Eligibility Decision (RED), therefore it is reasonably foreseeable that EPA 

will register this specific use of dicamba under FIFRA. EPA considered potential risks 

associated with dicamba use, including its degradate DCSA when appropriate, on surface 

or ground water using screening level (high-end exposure) models to estimate 

environmental concentrations. The EPA then compared these exposure estimates to 

appropriate endpoints from mammalian, aquatic animal and plant ecotoxicity studies, and 

concluded dicamba meets the FIFRA standard for no unreasonable adverse effects on 

human health and the environment (see Section J.3.7.2 and Appendix M.5.2 of the 
Monsanto Company 10-SY-210U 529 of 721 

petition for additional detail on the EPA analysis). The EPA analysis, based on use 

patterns that exceed the proposed single and annual maximum use rates for dicamba on 

MON 87708, does not take into account normal variation in environmental 

concentrations that can occur, and assumes that greater than 85% of the water shed is 

treated with the herbicide at the maximum labeled rate on the same day. In addition, the 

EPA examined and considered available monitoring data as part the dicamba RED, where 

concentrations of dicamba in ground and surface water were detected at levels up to 

44 μg/L and 1.76 μg/L, respectively. Furthermore, potential impacts on ground and 

surface water from dicamba use on MON 87708 will be considered by EPA as part of 

Monsanto’s pending application to register the use of dicamba on MON 87708, and must 

meet the FIFRA standard for no unreasonable adverse effects on human health or the 

environment prior to approval. 

It is foreseeable that the frequency of dicamba detections in ground and surface water 

could increase as a result of the cultivation of MON 87708, however levels of dicamba in 

water are not expected to increase above the levels already evaluated and considered by 

EPA. Existing monitoring data provides additional support that water resources will not 

be impacted from any potential increase in dicamba use. Monsanto has compiled 

publicly available surface and ground water monitoring data from across the United 

States from 1990 through 2010, including sampling sites in areas where soybean and corn 

are grown (Upper Midwest, see Figure VIII-1 of the petition) and where dicamba use has 

historically been most intense (See Figure VIII-2 of the petition). Maximum labeled use 

rates during most of this timeframe (2.8 lb a.e. per acre single maximum and 7.7 lb a.e. 

per acre annual maximum) were much higher than presently allowed rates (1.0 lb a.e. per 

acre single maximum and 2.0 lb a.e. per acre annual maximum) and the rates proposed on 

Monsanto’s dicamba label for use on MON 87708. Therefore, an examination of 

available surface and groundwater monitoring data in these areas during the mid-1990s 

would be indicative of the anticipated levels of dicamba that may occur from the use on 

MON 87708. 

An evaluation of the compiled surface water data from 1994 through 1998 for the major 

soybean areas during the primary dicamba application months of April through July 

indicates that detected levels of dicamba (90th percentile concentration for all samples 

where dicamba was detected26) were less than 1 μg/L. Monitoring data from April 

through July were evaluated because these are the months where the majority of dicamba 

applications are made to soybean (preemergence) and corn (pre- and postemergence), and 

when surface water concentrations from with these applications would be expected to 

peak. The maximum level of dicamba in surface water during this same timeframe was 

9.4 μg/L. Similarly, the evaluation of the groundwater data for major soybean growing 

areas from 1994 through 1998 indicates that detected levels of dicamba (90th percentile 

concentration of all samples where dicamba was detected) were 0.25 μg/L or less. The 

maximum level of dicamba in groundwater during this same timeframe was 2.2 μg/L. 

Considering the available monitoring data for ground and surface water during the period 

of dicamba’s most intensive use and when application rates were significantly higher than 

 

26 EPA uses the 90th percentile as the relevant high-end endpoint when analyzing water monitoring data 
Monsanto Company 10-SY-210U 530 of 721 

the rates proposed for us on MON 87708, it is reasonable to assume that levels in ground 

and surface water that may result from the use of dicamba on MON 87708 would be 

below the levels (high-end exposure modeling and monitoring data) considered by the 

EPA in the dicamba RED, and where EPA concluded would no unreasonable adverse 

effects on human health or the environment. Therefore, the approval in whole and no 

action alternatives are similar regarding their potential impact on surface and ground 

water quality from the use of dicamba on MON 87708. 

No action alternative

Surface water may be impacted from soybean production by runoff from soybean fields 

that carries soil particles and herbicides or other pesticides to streams, rivers, lakes, 

wetlands and other water bodies. As discussed above, based on existing data, the soil 

component of runoff is a much more important contributor to surface water impacts than 

is the pesticide component. Similarly, ground water may be impacted from soybean 

production due to the use of herbicides or other pesticides for weed management. 

Under full deregulation, growers’ use of dicamba for managing hard-to-control and 

herbicide-resistant broadleaf weeds in soybean fields would be expected to increase. 

Under the no action alternative, growers would need to use other practices for dealing 

with hard-to-control and herbicide-resistant broadleaf weeds. These practices would 

likely consist of some combination of herbicide use and traditional tillage method. 

However, the specific combination of herbicides used would likely be different than with 

full deregulation, as dicamba would not be able to be used late preemergence or 

postemergence with MON 87708. Growers would likely use some combination of 

herbicides currently in use for soybean (discussed in Section VIII of the petition). 

If the no action alternative resulted in increased use of conventional tillage practices for 

weed control, overall adverse surface water impacts may be greater with the no action 

alternative than under full deregulation alternative. Tillage causes widespread soil 

disturbance. Thus, erosion, topsoil loss and the resulting sedimentation and turbidity in 

streams are likely to increase with increased tillage. Based on the states’ water quality 

reports to EPA, which EPA makes available through its National Assessment Database, 

pesticides in general and herbicides in particular are a relatively minor contributor to 

impairment of surface water in the U.S., compared to sedimentation/siltation and 

turbidity (U.S. EPA, 2008b). Pesticides accounted for less than one percent of reported 

causes of surface water impairment in all but four of the 17 leading U.S. soybeanproducing states. In those four states, pesticides accounted for 2% to 8% of reported 

causes of impairment. Of the pesticides that were reported as contributing to impairment 

among the 17 leading soybean-producing states, almost all are highly persistent 

chemicals that are no longer registered for use in the U.S. Only one currently used 

herbicide, atrazine, was reported (U.S. EPA, 2008b). 

In summary, based on EPA data, herbicides in general are minor contributors to surface 

water impairment in the U.S., while sedimentation/siltation and turbidity are major 

contributors. The no action alternative, compared to full deregulation of MON 87708, 

would likely result in a different combination of alternative herbicides being used and 
Monsanto Company 10-SY-210U 531 of 721 

may result in increased tillage to obtain effective weed control. Weed management is a 

primary reason for tillage and reduced herbicide options due to existing herbicide 

resistance, in some cases, may increase the need for tillage (CAST, 2011). Increased 

tillage could contribute to adverse surface water impacts through increased runoff of soil 

particles to surface water bodies. Therefore, the no action and approval in whole 

alternatives are not significantly different regarding their impact on water quality from 

the cultivation of MON 87708 and associated use of dicamba. 

J.5.2.3. Air Quality and Climate Impacts 

Approval in whole alternative 

The difference between the approval in whole and the no action alternative is expected to 

be the integration of MON 87708 into the Roundup Ready soybean system using 

traditional breeding techniques. For those acres where glyphosate resistant weeds may 

already be present, where application of an herbicide with a different mode-of-action 

would aid in weed control, or for grower implementation of weed resistant management 

practices, the cultivation of soybean containing the MON 87708 trait would be an option 

for growers. 

Agricultural activities have the potential to impact air quality. These activities include 

emissions from farming equipment, burning, nitrous oxide associated with the use of 

nitrogen fertilizer, and pesticide applications (Aneja et al., 2009; USDA-NRCS, 2006). 

Mechanical tillage practices also have the potential to impact air quality through the 

suspension of soil particulates in the air (USDA-NRCS, 2005; CTIC, 2011; Baker et al, 

2005). 

Agricultural practices are not expected to change significantly with the introduction of 

MON 87708. A discussion of the agricultural practices associated with soybean 

production in the U.S. is provided in Section VIII of the petition, and includes discussion 

of cultural, mechanical and herbicide practices for weed management. Deregulation of 

MON 87708 is expected to facilitate the trend toward increased adoption of conservation 

tillage methods by soybean growers because conservation tillage (specifically no-till) 

relies on the use of herbicides to control weeds that emerge in a field prior to or after 

planting the soybean seed into the previous crop stubble, thus avoiding disturbance of the 

soil. MON 87708 would help to maintain existing conservation tillage practices and 

facilitate the adoption of conservation tillage practices by simplifying weed control 

options for growers utilizing a non-glyphosate herbicide or where there are glyphosateresistant or hard-to-control broadleaf weeds present. Soybean represents the greatest 

number of acres of the major field crops utilizing conservation tillage and the highest 

percentage of total crop acres devoted to conservation tillage practices (CTIC, 2007). 

Considerable benefits to the physical environment, including those related to air quality, 

are obtained from use of conservation tillage methods including (CTIC, 2011; USDANRCS, 2005): 

 Dramatic reduction in soil erosion from wind and water; 
Monsanto Company 10-SY-210U 532 of 721 

 Less herbicide, water, and soil runoff from soils improving the quality of streams and 

lakes; 

 Overall healthier soils; 

 Increased carbon sequestration leading to reduced greenhouse gases; 

 Decreased fuel emissions due to reduced use of tractors to plow fields; 

 Reduced nitrogen applications (much of which is made from fossil fuels); and 

 Less overall water usage for agricultural purposes. 

While approval in whole of MON 87708 may facilitate some trend towards increasing 

conservation tillage, it is not expected to significantly impact climate or air quality. 

Therefore approval in whole and no action alternatives are not significantly different 

regarding their impacts on climate and air quality. 

No action alternative 

As discussed above, compared with full deregulation of MON 87708, the no action 

alternative may result in increased tillage, and decreases in conservation tillage. EPA 

reports conservation tillage as an agricultural practice that “increases carbon storage 

through enhanced soil sequestration” and that “may reduce energy-related CO2 emissions 

from farm equipment” (U.S. EPA, 2010). When carbon is stored, it is not available to be 

emitted in the form of carbon dioxide (CO2), a greenhouse gas. Thus, the no action 

alternative may result in increased tillage, which could cause some adverse, but probably 

not significant climate and air quality impacts compared with full deregulation. 

Under the no action alternative growers would still likely practice conservation tillage, 

and in certain situations they would rely on tillage and/or other soybean herbicides. 

Other herbicide-tolerant soybean events have been deregulated by APHIS or have been 

submitted to APHIS for deregulation. These events and their companion herbicides may 

be used to promote conservation tillage practices under the no action alternative. 

Therefore, the no action and approval in whole alternatives are not significantly different 

regarding their potential impact on air quality. 

J.5.2.4. Soil Quality Impacts 

Approval in whole alternative 

The difference between the approval in whole and the no action alternative is expected to 

be integration of MON 87708 into the Roundup Ready soybean system using traditional 

breeding techniques. For those acres where glyphosate resistant weeds may already be 

present, where application of an herbicide with a different mode-of-action would aid in 

weed control, or for grower implementation of weed resistant management practices, the 

cultivation of soybean containing the MON 87708 trait would be an option for growers. 
Monsanto Company 10-SY-210U 533 of 721 

Other than changes associated with herbicide use, MON 87708 will not alter the 

agronomic practices typically utilized in the cultivation of soybean. MON 87708 has 

been found to be compositionally, agronomically and phenotypically equivalent to 

conventional soybean. Therefore microbial populations and associated biochemical 

processes in soil are not expected to change with the introduction of MON 87708. The 

MON 87708-produced protein DMO demonstrates a high level of substrate specificity 

and is not expected to persist in the environment (see Section V.E in the petition). 

Studies have shown no impact to the symbiont interactions of MON 87708 (see Section 

VII.C.4 of the petition), or to NTOs such as beneficial and pest arthropods when exposed 

to MON 87708 DMO in the field (see Section VII.C.2.4). Based on these data, the 

cultivation of MON 87708 is not expected to impact microbial populations and associated 

biochemical processes. 

Multiple herbicides are already used in soybean production. Agricultural fields are 

purposefully managed to be weed-free resulting in greater economic benefit to the 

grower. A discussion of weed management practices is provided in Section VIII.F.1 of 

the petition. In the U.S., 98% of soybean acreage was treated with an herbicide in 2006 

(USDA-NASS, 2007b). Therefore, introduction of MON 87708 and treatment with 

dicamba is unlikely to affect soil quality in commercial soybean production systems 

differently than those herbicides already used in soybean. Dicamba has been registered 

by the EPA for use on a wide range of agricultural uses since 1967 (see Appendix M, 

Section M.1 of the petition). The EPA evaluated the environmental safety of dicamba 

and its metabolites as part of the RED (U.S. EPA, 2005b), and concluded that dicamba 

may accumulate with frequent and intensive use (2.0 and 2.8 lb per acre a.e. single 

application and 7.7 lb per acre a.e. annually). The EPA mandated reductions in dicamba 

use rates as part of dicamba’s continued registration to effect these and other potential 

impacts (U.S. EPA, 2009b). Based on the reduced application rates (1.0 lb per acre a.e. 

with a maximum annual rate of 2.0 lb a.e. per acre) dicamba is unlikely accumulate or 

persist in the environment. In addition, results of standardized tests with dicamba and 

dicamba formulations indicate no long-term effects on functional processes of soil 

microorganisms (carbon respiration and nitrogen transformation) at rates proposed for 

dicamba on MON 87708 (European Commission, 2007a). Based on this analysis, the 

approval in whole and no action alternatives are not significantly different regarding their 

impact on soil quality. 

No action alternative

Under the no action alternative agronomic practices currently utilized in the cultivation of 

soybean would not be altered. However, some combination of herbicides already used in 

soybean production acres and possibly increased tillage may be used more frequently to 

control problematic weeds. Under the no-action alternative, an increase in tillage may 

occur, and would negate many of the benefits of conservation tillage to soil including 

improvement of soil structure, reduction of soil compaction, conservation of soil moisture, 

reduction of soil erosion and improvement of soil organic matter content. Overall, the no 

action and approval in whole alternatives are similar regarding the potential impacts on 

soil quality. 
Monsanto Company 10-SY-210U 534 of 721 

J.5.3. Potential Impacts to Agricultural Practices 

Approval in whole alternative 

The difference between the approval in whole and the no action alternative is expected to 

be integration of MON 87708 into the Roundup Ready soybean system using traditional 

breeding techniques. For those acres where glyphosate resistant weeds may already be 

present, where application of an herbicide with a different mode-of-action would aid in 

weed control, or for grower implementation of weed resistant management practices, the 

cultivation of soybean containing the MON 87708 trait would be an option for growers. 

MON 87708 has been shown to be no different from conventional soybean in its 

agronomic and ecological characteristics (see Sections VII, VIII, IX and X of the 

petition), and has the same levels of tolerance to insects and diseases as conventional 

soybean. A summary of agronomic practices currently used for soybean production is 

presented in Section VIII of the petition. Except for weed management relative to 

dicamba use, the majority of agricultural practices, including tillage, insect and disease 

management, crop rotation practices, irrigation and volunteer management, will not 

change under full deregulation. 

For weed management under full deregulation growers would have the option of a wider 

window for treating their soybean crop with dicamba and the number of acres upon 

which dicamba is used will likely increase under this alternative. An estimate of the 

increase in treated acres can be made using the following assumptions. If dicambatreated acres reach 40% of the 75 million U.S. soybean acres, approximately 30 million 

acres of soybean would be treated with dicamba. Currently dicamba is used on 20.2 

million acres in all crops including soybean (soybean accounts for 0.53 million acres) and 

has historically been used on up to 36.3 million acres across all uses at its peak (see Table 

VIII-11 of the petition). The potential use of dicamba on MON 87708 would result in a 

total of 50.2 million acres treated with dicamba. Similarly, the total amount of dicamba 

applied in overall agriculture would also increase. Based upon an upper-end estimation 

of the anticipated commercial use pattern for dicamba in MON 87708 as described in 

Section VIII.H, an additional 22 million pounds of dicamba is estimated (high-end) to be 

added to U.S. soybean fields each season. According to NASS statistics (USDA-NASS, 

2007b), approximately 103 million pounds of herbicides were used on soybean in 2006 

(Table VIII-8 of the petition), and recently the trend has been towards increasing 

herbicide use for the management of problematic weeds including resistant populations 

as well as the incorporation of diversified weed management practices in soybean 

growing areas (see Section J.3.3.6 for additional details regarding recent trends). As 

discussed previously, dicamba will displace in part the use of some existing soybean 

herbicides. 

Dicamba use presents a relative reduction of risk potential in comparison to some of the 

alternative non-glyphosate herbicides currently available to soybean growers. The 

rationale and supporting information for the comparative alternative analysis is provided 

in Appendix L and summarized below: 
Monsanto Company 10-SY-210U 535 of 721 

• Dicamba has a more favorable toxicity profile and poses a lower health risk potential 

to applicators and consumers compared to some alternative herbicides (Appendix L, 

Table L-25); 

• Dicamba has lower toxicity to aquatic animals and plants and poses lower risk 

potential to aquatic organisms compared to some alternative herbicides (Appendix L, 

Table L-25); 

• Dicamba when used in conjunction with MON 87708 integrated into the Roundup 

Ready soybean system provides growers with a more flexible and reliable weed 

management system (Peterson et al., 2011). 

Dicamba tolerance would allow growers to utilize an expanded application window of 

dicamba in their weed management systems, while still allowing alternative herbicides 

currently registered for use in soybean to be applied. Breeding MON 87708 with 

glyphosate-tolerant soybean, such as Roundup Ready 2 Yield soybean, would allow for 

in-season application of glyphosate and dicamba, thereby improving the sustainability of 

weed efficacy for glyphosate, dicamba and other soybean herbicides. 

A reasonably foreseeable impact under full deregulation is the delay in the evolution and 

development of glyphosate- and dicamba-resistant broadleaf weeds as well as weeds 

resistant to other soybean herbicide classes, such as PPO herbicides, in soybean 

producing areas. This is because growers will likely use dicamba together with 

glyphosate on the combined dicamba- and glyphosate-tolerant soybean product because 

of the excellent crop tolerance and compatibility of the two herbicides. In addition, other 

herbicides will be recommended and used by growers especially in cases where the 

grower is managing a weed population already resistant to glyphosate. This will further 

assist in delaying resistance to dicamba and other herbicides used in the MON87708 

system. 

Dicamba is an excellent option to delay resistance to other herbicides because of its broad 

activity on broadleaf weeds and low level of weed resistance, specifically on the summer 

spectrum of weeds known to infest soybean acres. A prominent strategy to delay the 

evolution and development of herbicide-resistant weeds is to increase the diversity of 

weed management practices used in a particular cropping system. Diversified weed 

management practices use a combination of cultural (e.g., crop rotation), mechanical (e.g., 

cultivation), and herbicide control practices, including use of herbicides with different 

modes-of-action (Duke and Powles, 2009). See Section J.3.3.6 of the petition for 

additional details on diversified weed management practices. Thus, MON 87708 

integrated into the Roundup Ready soybean system provides the opportunity to increase 

the diversity of in-crop herbicide control options for growers and, in turn, supports the 

long term sustainability of the Roundup Ready soybean system with its established 

benefits. 

In summary, no significant changes in common agricultural practices are anticipated from 

to the cultivation of MON 87708 and the associated use of dicamba. Shifts in herbicide 

use will occur, with dicamba displacing some of the alternative herbicides. A decrease in 
Monsanto Company 10-SY-210U 536 of 721 

tillage practices may occur in some circumstances, and a delay in the development of 

glyphosate- and dicamba-resistant broadleaf weeds may be possible. Overall, the 

approval in whole and no action alternatives are similar regarding their impact on 

soybean agricultural practices. 

No action alternative 

Under the no action alternative MON 87708 and its progeny would continue to be 

regulated articles and would not be widely grown. Under this alternative dicamba 

herbicide would not become integrated into the Roundup Ready soybean system and 

dicamba use would likely remain similar to today’s use pattern in soybean. Growers 

would continue to use the Roundup Ready soybean system for broad spectrum weed 

control, other registered alternative herbicides alone or in combination with other 

herbicide-tolerant soybean for targeted hard-to-control weeds, and/or incorporate tillage 

into their practices. 

Integration of MON 87708 into the Roundup Ready soybean system has the potential to 

delay or prevent development of dicamba- and glyphosate-resistant weeds (see Appendix 

K of petition for additional detail). For soybean, the use of dicamba in conjunction with 

glyphosate provides growers with an herbicide system with two different modes of action. 

Thus, it is foreseeable that under the no action alternative, the inability to integrate 

MON 87708 into the Roundup Ready soybean system could increase the potential for 

glyphosate resistant weed populations to evolve and spread in soybean producing areas. 

In addition, the potential for resistance to evolve and spread for other soybean herbicides 

could also increase in these areas where growers do not use multiple modes-of-action. 

Also under the no action alternative, increased use of other non-glyphosate alternative 

herbicides, some with higher risk potential for human health and environmental 

characteristics compared to dicamba, and reduced flexibility for the grower (e.g., 

restricted plant-back intervals, rotational crop restrictions) would be expected. A number 

of weeds commonly found in the Midwestern and southern portions of the U.S. already 

display resistance to many of these alternative herbicides (Tables J-3 and J-4), while only 

four broadleaf weed species have been confirmed to be resistant to dicamba in the U.S., 

even though dicamba has been widely in use for over 40 years. Increasing the number of 

weed management options available to soybean growers, including other herbicidetolerant traits pending deregulation (see Table J-5), is an important element to delay and 

prevent further development of resistant weed populations. Given these observations, the 

no action alternative and corresponding lack of effective alternative modes of action may 

lead to an increase in weed resistant populations for these alternative herbicides. 

Herbicides are a critical element of conservation tillage practices. Since weed 

management is a primary reason for tillage, herbicides are the primary tool to replace 

tillage and thus are critical to the sustainability of conservation tillage practices. Under 

the no action alternative increased use of traditional tillage methods for the control of 

problematic weeds may occur in some situations and result in the potential loss of many 

of the benefits of conservation tillage. 
Monsanto Company 10-SY-210U 537 of 721 

Overall herbicides are widely used in production of a soybean crop. Information 

presented in Section VIII.F.1 of the petition highlights the dynamics of herbicide use in 

soybean. Considerable shifts in use patterns of herbicides occur over time based upon 

many factors including availability of new herbicides and herbicide-tolerant crops, 

herbicide efficacy, convenience and economics. Yet, herbicides in general remain a 

critical element of soybean production. 

From this analysis it is concluded that overall use of herbicides in soybean production is 

unlikely to be significantly different between the approval in whole and the no action 

alternatives although shifts in use patterns of herbicides may occur based on the decision 

to deregulate MON 87708. Additionally, an increase in tillage may occur in some 

circumstances under the no action alternative. Therefore, the no action and approval in 

whole alternatives are similar regarding potential impacts on soybean agricultural 

practices. 

J.5.3.1. Potential Impacts on Weed Resistance 

Approval in whole alternative 

The difference between the approval in whole and the no action alternative is expected to 

be integration of MON 87708 into the Roundup Ready soybean system using traditional 

breeding techniques. For those acres where glyphosate resistant weeds may already be 

present, where application of an herbicide with a different mode-of-action would aid in 

weed control, or for grower implementation of weed resistant management practices, the 

cultivation of soybean containing the MON 87708 trait would be an option for growers. 

A potential impact with the increased use of any herbicide is the development of 

herbicide-resistant weeds. A comprehensive discussion concerning the potential 

development of dicamba-resistant weeds may be found in Appendix K of the petition. 

The potential for the development of weed resistance to an herbicide is a function of the 

duration and frequency of herbicide use in the absence of other methods of weed control. 

Initially, resistant weed populations are localized at the field level, however resistant 

populations can become more widespread and effect larger areas as a result of the 

development of additional resistant populations occurring from selection pressure, or 

from gene flow and/or seed movement from existing resistant populations combined with 

inadequate management practices. In either case, the use of diversified weed 

management practices are essential to effectively manage weed resistance. 

Dicamba has been used for over 40 years and some resistant weeds have developed due 

to use of the herbicide and the selection pressure applied with continued use of this 

herbicide. The introduction of MON 87708 will likely result in limited additional 

selection pressure for the development of dicamba-resistant weeds for several reasons: 1) 

dicamba, as a broadleaf herbicide, would primarily be used in combination with 

glyphosate on MON 87708 integrated into the Roundup Ready soybean system because 

dicamba does not control narrowleaf weeds and broad spectrum control of broadleaf and 

narrowleaf weeds is the objective of all weed control systems; 2) the use of glyphosate 

plus dicamba would provide multiple modes-of-action on key broadleaf weeds which 
Monsanto Company 10-SY-210U 538 of 721 

would diminish the chance for selection of dicamba-resistant broadleaf weeds; 3) in cases 

where glyphosate plus dicamba will be applied to soybean fields with a known presence 

of glyphosate-resistant broadleaf weeds, a third herbicide mode of action will be 

recommended to growers that also has activity on the glyphosate-resistant broadleaf weed 

thereby providing two effective modes-of-action to control the glyphosate-resistant weed; 

and 4) the proposed dicamba herbicide label for MON 87708, existing glyphosate 

herbicide labels and separate Monsanto weed management recommendations (e.g., 

Monsanto’s annual TUG27 and publically available websites28) will specify the effective 

rate and timing of dicamba and glyphosate applications for optimal weed control, thereby 

reducing selection pressure for dicamba as well as glyphosate. 

Furthermore, in the unlikely case that broadleaf weeds were to evolve or develop with 

resistance to dicamba, existing cultivation and alternative herbicide tools (see Section 

VIII.F.1 for description of alternative herbicides) would remain potential options to 

provide effective control. Additionally, as discussed above in Section J.5.3, it is 

reasonably foreseeable that under full deregulation a delay in development and spread of 

glyphosate- and dicamba-resistant broadleaf weeds, as well as weeds resistant to other 

soybean herbicide classes such as PPO herbicides, will occur due to the likely use of 

dicamba and glyphosate on fields planted with the combined dicamba and glyphosate 

tolerance product. 

In summary, there is a low probability that additional dicamba resistant weed species or 

populations will evolve or develop as a result of the cultivation of MON 87708 and the 

use of dicamba and glyphosate in the weed management systems, and a delay in the 

development of glyphosate- and dicamba-resistant broadleaf weeds may be possible. 

Overall, the approval in whole and no action alternatives are similar regarding their 

impact on herbicide weed resistance. 

No action alternative 

Under the no action alternative MON 87708 and its progeny would continue to be 

regulated articles and would not be widely grown. Under this alternative dicamba 

herbicide would not become integrated into the Roundup Ready soybean system and 

dicamba use would likely remain similar to today’s use pattern in soybean. Growers 

would continue to use the Roundup Ready soybean system for broad spectrum weed 

control, other registered alternative herbicides alone or in combination with other 

herbicide-tolerant soybean, and/or incorporate tillage into their practices. There is a 

potential for glyphosate-resistant weed populations to increase and spread in soybean 

cultivation areas, as well as the potential for the development and spread of resistant 

populations to other alternative soybean herbicides. 

Integration of MON 87708 into the Roundup Ready soybean system has the potential to 

delay or prevent development of dicamba- and glyphosate-resistant weeds in soybean 

cultivation areas (Appendix K of petition). For soybean, the use of dicamba in 

 

27 http://www.monsanto.com/SiteCollectionDocuments/Technology-Use-Guide.pdf 

28 http://www.monsanto.com/weedmanagement/Pages/default.aspx 
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conjunction with glyphosate provides growers with an efficient, effective and flexible 

herbicide system with two different herbicide modes-of-action with overlapping activity 

on many broadleaf weeds. Thus, it is foreseeable, under the no action alternative and the 

inability to integrate MON 87708 into the Roundup Ready soybean system, that there is 

an increased potential for glyphosate resistant weed populations in soybean growing 

areas to evolve and spread, as well as an increased potential for resistance to evolve and 

spread for other soybean herbicides where growers do not effectively use multiple 

modes-of-action. However, existing alternative soybean herbicides will continue to be 

available. Therefore, the no action and approval in whole alternatives are similar 

regarding potential impacts on weed resistance. 

J.5.4. Potential Impact to Commercial Soybean Production 

Approval in whole alternative 

Soybean is a globally traded commodity with the U.S. being the top global producer 

(Soyatech, 2010). An overview of U.S. soybean production is provided in Section 

VIII.B.1 of the petition. Biotechnology-derived crops are subject to regulation in many 

countries. In order to support free trade in soybean, Monsanto will seek regulatory 

approval for MON 87708 and stacked products (i.e., products combined using traditional 

breeding techniques) with other biotechnology-derived soybean, where required, in all 

key soybean import countries with a functioning regulatory system to support the flow of 

international trade. As described in Section VIII.L of the petition, Monsanto adheres to 

the BIO Product Launch Policy 30 including: 1) conducting a market and trade 

assessment, 2) securing regulatory approvals in key export countries prior to full 

commercial launch, 3) following generally accepted best seed management practices to 

prevent unintended low level presence of the event in seed, 4) providing reliable 

detection methods to growers, processors and buyers prior to commercialization, and 5) 

communicating to stakeholders the company’s product launch stewardship policies. 

These actions protect against adverse impacts to trade of soybean due to the introduction 

of a new biotechnology-derived soybean. 

The difference between the approval in whole and the no action alternative is expected to 

be integration of MON 87708 into the Roundup Ready soybean system using traditional 

breeding techniques. For those acres where glyphosate resistant weeds may already be 

present or where application of an herbicide with a different mode of action would aid in 

weed control or the implementation of weed resistant management practices, the 

cultivation of soybean containing the MON 87708 trait would be an option for growers. 

Soybean is primarily a self-pollinated crop with minimal gene movement (see Section IX 

of the petition). Due to the biology of soybean flowers which results in low cross 

pollination potential, pollen movement between soybean fields is minimal. Thus the 

introduction of MON 87708 is not expected to effect commodity soybean production, due 

to low potential gene movement to neighbouring soybean crops. In 2009, 91% of the 

 

30 BIO’s Product Launch guidelines can be found at: 

http://www.excellencethroughstewardship.org/LinkClick.aspx?fileticket=ppgyTABguQs%3d&tabid=84. 
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U.S. soybean acres were planted to biotechnology-derived herbicide-tolerant soybean 

(USDA-NASS, 2009a). Thus, growers are accustomed to the presence of biotechnologyderived soybean in proximity to production fields and have developed practices to allow 

for production of a crop to meet customer expectations. Therefore, the approval in whole 

and no action alternatives are similar regarding potential impacts on commercial soybean 

production. 

No action alternative 

Under the no action alternative, MON 87708 would remain a regulated article and would 

not be available to growers. Growers could continue to use the Roundup Ready soybean 

system for broad spectrum weed control. Therefore, the no action and approval in whole 

alternatives are not different in their impact to commercial soybean production, because 

the majority of soybean grown in the U.S. is already biotechnology-derived and due to 

company stewardship policies accompanying the introduction of new biotechnologyderived products, as described above. Technology providers maintain a dialogue with 

stakeholders to communicate global approval status for biotechnology-derived soybean to 

avoid potential market disruption. Commodity and specialty soybean growers and 

handlers are accustomed to a diversity of soybean types in the marketplace, including 

soybean with modified oil profiles, food grade, clear hilum, and others. They have 

demonstrated an ability to provide soybean that meet their customers’ expectations. 

MON 87708 represents another biotechnology-derived soybean in the marketplace 

adding to the diversity of soybean available to growers and the market. Therefore the no 

action and approval in whole alternatives are expected to be the same regarding potential 

impacts on commercial soybean production.

J.5.5. Potential Impact to Certified Seed Production 

Approval in whole alternative 

Certified seed production is a carefully managed process (see Section VIII.B.2 of petition 

for additional detail). MON 87708 is not expected to impact certified seed production 

practices or production of specialty soybean seed for reasons described in this section. 

If MON 87708 is deregulated, seed production would occur within production systems 

already developed by seed producers for certified soybean seed. MON 87708 has been 

thoroughly characterized and (with the exception of its tolerance to dicamba) is not 

agronomically or phenotypically different from commercial soybean. The difference 

between the approval in whole and the no action alternative is expected to be integration 

of MON 87708 into the Roundup Ready soybean system using traditional breeding 

techniques. For those acres where glyphosate resistant weeds may already be present or 

where application of an herbicide with a different mode of action would aid in weed 

control or the implementation of weed resistant management practices, the cultivation of 

soybean containing the MON 87708 trait would be an option for growers. However, the 

implementation of management practices to avoid pollen from a biotechnology-derived 

soybean in organic, specialty or conventional soybean seed or commodity seed 

production operations is dictated by the nature of soybean pollination (see Section 
Monsanto Company 10-SY-210U 541 of 721 

VIII.B.2), and MON 87708 has been shown in field testing to not differ from commercial 

soybean varieties regarding pollen characteristics. Soybean is a highly self-pollinated 

species that exhibits very low levels of outcrossing. Numerous evaluations on soybean 

cross-pollination have been conducted, and the published results are summarized in Table 

IX-1 of the petition. Under typical soybean planting and cultivation conditions, crosspollination among adjacent plants in a row or among plants in adjacent rows ranged from 

0 to 6.3%. In experiments where supplemental pollinators (usually bees) were added to 

the experimental area, cross-pollination ranged from 0.5 to 7.74% in adjacent soybean 

plants or adjacent rows (Abrams et al., 1978; Chiang and Kiang, 1987). However, crosspollination even at the low levels observed in these controlled trials does not occur over 

long distances. Cross-pollination rates decrease to less than 1.5% beyond one meter from 

the pollen source, and rapidly decrease with greater distances from the source. The 

following cross-pollination rates at extended distances have been reported: 0.02% at 8.2 

m of separation (Caviness, 1966), 0.05% at 5.4 m (Ray et al., 2003), and 0% at 6.5 m 

(Abud et al., 2003). 

The low potential for cross pollination in soybean is recognized in certified seed 

regulations for foundation seed in the U.S., which require no measured isolation between 

different soybean cultivars in the field as long as there is adequate separation between the 

fields to prevent mechanical mixing (USDA-APHIS, 2006). Hence, certified soybean 

seed producers can and have effectively implemented practices (e.g., isolation distances 

during the growing season, equipment cleaning during harvest, and post-harvest 

separation of harvested seed) that allow them to maintain commercially acceptable levels 

of varietal purity. Because MON 87708 has been shown to be no different from 

conventional soybean relative to pollen morphology and viability, the cultivation of 

MON 87708 will not impact the ability to implement production practices required for 

the production of certified seed. Therefore, the approval in whole and no action 

alternatives are the same regarding the potential impact on certified seed production 

practices. 

No action alternative 

Under the no action alternative, MON 87708 would not be propagated to any extent by 

seed producers because there would be no commercial demand for seed containing 

MON 87708. Seed production practices would be the same as those described above for 

the deregulation in whole alternative and in Section VIII.B.2 of the petition. Seed 

producers already produce numerous varieties of soybean, the vast majority of which 

include a biotechnology-derived trait. Therefore, the no action and approval in whole 

alternatives are the same regarding the potential impacts on certified seed production. 

J.5.6. Potential Impacts to Organic Soybean Production 

Approval in whole alternative 

Organic soybean production is a carefully managed and regulated process. MON 87708 

and biotechnology-derived soybean in general, including those currently grown 

commercially, are not allowed for use in organic production systems because they were 
Monsanto Company 10-SY-210U 542 of 721 

developed through the use of excluded methods as defined by the National Organic 

Production (NOP) program standards (7 CFR § 205.2). With the exception of its 

tolerance to dicamba, MON 87708 has been shown to be no different from conventional 

soybean in its agronomic and ecological characteristics including characteristics of the 

pollen produced by MON 87708. Thus, MON 87708 is expected to be no different from 

other soybean in its ability to cross pollinate with other soybean and, therefore, no 

additional means beyond those already used to produce biotechnology-derived and 

organic soybean will be needed if MON 87708 were grown commercially. 

Similar to the exclusion of biotechnology-derived products in organic production, the use 

of synthetic herbicides, such as dicamba, are also excluded under the NOP. Dicamba is 

presently registered for early preemergence (early pre-plant) and late postemergence 

(pre-harvest) applications in soybean, and in other crops (corn, sorghum and wheat) 

commonly grown in rotation with soybean. It is likely that dicamba herbicide will be 

applied to MON 87708. As mentioned previously, herbicides are used extensively for 

production of soybean and the potential increased use of dicamba, along with its 

concurrent replacement of some alternative herbicides used to control glyphosate’s hardto-control and resistant weeds, is minimal considering the amount of herbicides (103M 

lbs and trending higher, see Table VIII-8 and discussion in J.5.3 of the petition) currently 

applied in commodity soybean production each year. For these reasons, production 

systems and practices in place now (discussed below) are sufficient to mitigate any 

impacts from the introduction of MON 87708 to organic soybean production. 

Currently, biotechnology-derived herbicide-tolerant soybean is planted on over 90% of 

the soybean acreage (USDA-NASS, 2009a). Despite the high adoption rates of 

biotechnology-derived soybean by growers, organic and conventional soybean 

production remains an option for growers who choose to produce soybean using these 

production practices. The decision to grow organic, conventional, or biotechnologyderived soybean is typically an economic one based on market dynamics. Organic 

soybean producers and those growing conventional soybean for non-biotechnology 

markets typically receive a market premium offsetting the additional production and 

record-keeping costs. While the widespread demand for Roundup Ready soybean has 

reduced the number of conventional soybean varieties that seed companies choose to sell, 

conventional and organically produced soybean seed is currently available from 

numerous seed suppliers (Table J-6). Additional information on organic seed sources is 

provided though the U.S. Agricultural Marketing Service (AMS). Thus, growers have a 

choice in the soybean they plant, and this is not expected to change with the introduction 

of MON 87708. 

Production systems designed prior to the introduction of MON 87708 or even prior to the 

introduction of biotechnology-derived soybean have allowed for production of soybean to 

meet varied customer demands. In addition to the market segments that produce organic 

or conventional soybean, distinct identity-preserved specialty soybean with such traits as 

clear hilum or high protein have also been grown and successfully marketed for specific 

food uses in domestic and export markets for many years (Cui et al., 2004). The NOP 

requires organic soybean producers utilize production practices designed to specifically 

avoid the presence of soybean products using conventional herbicide or other pesticide 
Monsanto Company 10-SY-210U 543 of 721 

treatments, as well as avoiding the use of biotechnology-derived crops. These well 

established practices to avoid the use of “excluded methods” will continue with the 

commercial introduction of MON 87708 integrated into the Roundup Ready system. 

They include isolation zones, use of buffer rows surrounding the organic crop, adjusted 

planting dates and varietal selection.31 The implementation of management practices to 

avoid pollen from a biotechnology-derived crop in organic or conventional soybean 

production operations is facilitated by the nature of soybean pollination. As noted 

previously in the petition, soybean is a highly self-pollinated species and exhibits a very 

low level of outcrossing. Hence, organic or conventional soybean producers can and 

have effectively implemented practices (e.g., isolation during the growing season, 

equipment cleaning during harvest, and post-harvest separation of harvested seed) that 

allow them to avoid the presence of biotechnology-derived soybean and maintain organic 

or conventional production status.32 

The difference between the approval in whole and the no action alternative is expected to 

be integration of MON 87708 into the Roundup Ready soybean system using traditional 

breeding techniques. There is potential for cultivation of soybean containing the 

MON 87708 trait to provide weed control solutions for problematic weeds and 

proactively manage and prevent the development of herbicide resistant weeds. However, 

the presence of a biotechnology-derived product like MON 87708 is unlikely in instances 

where producers utilize production practices designed to avoid biotechnology-derived 

products. Although the USDA’s National Organic Standards prohibit the use of excluded 

methods, the presence of products of an excluded method alone does not necessarily 

constitute a violation of the Standard or a loss of organic certification (USDA-AMS, 

2011). While the NOP does not set specific thresholds for the allowable presence of 

products of excluded methods, there may be some specifications in buyer allowances 

that permit between 0.1 to 5% biotechnology-derived soybean in organic soybean.33 

Similarly, international regulatory authorities have set allowable tolerances for the 

presence of biotechnology-derived material in conventional products to support food 

labeling and traceability laws. These tolerances allow from 0.9% (European Union) up to 

5% (Japan) of the food to be biotechnology-derived in products considered 

“conventional.” Levels above the threshold may trigger special labeling. 

Given that allowances for minor amounts of biotechnology derived material are allowed 

in soybean exported to key markets and that systems are in place for production of 

soybean that meet buyer expectations and the prevalence of biotechnology-derived 

soybean already on the market, the introduction of a new biotechnology-derived soybean 

product is unlikely to have an effect on organic soybean production. This conclusion is 

supported by USDA-ERS (2010) data that show the organic soybean market share was 

0.17% in 1995, the year prior to the introduction of Roundup Ready soybean, and was at 

0.2% in 2008; indicating availability of biotechnology-derived soybean has not 

significantly impacted the organic soybean industry. Therefore, the approval in whole 

 

31 Information on isolation methods can be found at: www.attra.ncat.org [Accessed on June 2, 2010]. 32 Source is website: http://attra.ncat.org/attra-pub/PDF/cropsfarmplan.pdf [Accessed on June 2, 2010]. 33 Source is website: http://attra.ncat.org/attra-pub/PDF/marketingorganicgrains.pdf [Accessed on May 27, 

2010]. 
Monsanto Company 10-SY-210U 544 of 721 

and no action alternatives are the same regarding the potential impact on organic soybean 

production. 

No action alternative 

The majority of soybean is currently produced using biotechnology-derived soybean. 

Growers have come to rely on biotechnology-derived soybean and their benefits. Given 

that several biotechnology-derived soybean have previously been deregulated by USDA 

and other events are under consideration for deregulation, the use of biotechnologyderived soybean is expected to remain relatively constant for the foreseeable future. 

Similarly, herbicide use is also expected to remain a key agronomic practice in 

production of commodity soybean. Thus, organic growers will continue to manage their 

production fields to avoid excluded methods and drift of pesticides under the no action 

alternative, and growers would continue to utilize the same practices they use now to 

produce their organic crop. Thus, the no action and approval in whole alternatives are the 

same regarding the potential impact to organic soybean production. 

Table J-6. Organic and Conventional Soybean Seed Sources 

Organic Soybean Seed Sources1

: Conventional Soybean Seed Sources 

Albert Lea Seed House AgVenture Seeds (modified oil) 

Blue River Hybrids Campbell Seed (modified oil) 

Golden Grains Becks Hybrids (food grade) 

Great Harvest Organics Monsanto (Asgrow) 

Greis Seed Farm Schillinger Seed 

Lancaster Ag Products Pioneer 

Lawler Farm Center Soy Genetics 

Prairie Gold Seeds Stewart Seed (modified oil) 

Superior Organic Grains, Ltd Stine Seed 

Walter Seed and Honey Co Syngenta - multiple brands 

 Terral Seed 

Various State Crop Improvement 

Organizations

1 Source is: www.organicgrains.ncsu.edu. 

J.5.7. Potential for Adjacent Agricultural Crop and Non-agricultural Impacts 

Approval in whole alternative 

Soybean (Glycine max) does not grow in the wild in North America (Hymowitz, 1987). 

Soybean does not grow and persist in unmanaged habitats and would not be expected to 

invade and/or persist in the natural environment, including streams, lakes, oceans or other 

aquatic environments. With the exception of the production of MON 87708 DMO that 
Monsanto Company 10-SY-210U 545 of 721 

confers tolerance to dicamba, MON 87708 is similar to other commercial soybean 

currently grown in the U.S. It is expected that under the approval in whole alternative 

MON 87708 will be integrated into the Roundup Ready soybean system using traditional 

breeding techniques. For those acres where glyphosate resistant weeds may already be 

present or where application of an herbicide with a different mode of action would aid in 

weed control or the implementation of weed resistant management practices, the 

cultivation of soybean containing the MON 87708 trait would be an option for growers. 

MON 87708 displays no altered plant pest characteristics compared to conventional 

soybean and is no more susceptible to insects or diseases that commonly infest soybean. 

On the basis of the information presented in petition #10-SY-210U on phenotypic and 

agronomic characteristics of MON 87708, and based upon the biology of soybean, 

MON 87708 is unlikely to effect adjacent vegetation. Therefore, the approval in whole 

and no action alternatives are the same regarding potential impacts on adjacent 

agricultural crops and non-agricultural areas. 

Herbicides are extensively used in U.S. soybean production, and their historic and current 

uses are described in Section VIII.F.1 of the petition. Herbicide drift and offsite 

movement are regulated by the U.S. EPA and actively managed by farmers and 

applicators specially trained to use such products consistent with product labels and other 

state or local restrictions. Depending upon the herbicide employed, relevant factors in 

managing the potential for spray drift include the selectivity and sensitivity of the 

herbicide, local weather conditions at the time of application (wind, temperature, 

humidity, inversion potential), droplet size distribution, application volume, boom height 

(height of the application equipment above the crop canopy), sprayer speed, and distance 

from the edge of the application area (SDTF, 1997; Felsot et al., 2010). A variety of 

measures can be employed to control the potential for spray drift and offsite movement, 

including nozzle selection and application techniques and restrictions. 

The approval in whole alternative would result in an increase in dicamba use in soybean 

compared to its current use. The U.S. EPA considers possible effects from offsite 

movement as part of the pesticide registration process. APHIS’s authority under the 

Plant Protection Act, on the other hand, does not authorize it to specify conditions or in 

any way regulate the use of herbicides. As a result, before any dicamba formulation 

could be employed over MON 87708, EPA’s approval of a FIFRA label for that specific 

use would be required. Such a label would address not only application rates, but as 

appropriate, could also include other measures to address the potential for offsite 

movement. EPA considers possible effects from offsite movement as part of the 

pesticide registration process. Specifically, in order to approve the use of a pesticide 

(herbicide) under FIFRA, EPA must conclude that no unreasonable adverse effects on 

non-target vegetation will result from offsite movement when the pesticide is used 

according to the product label, and when herbicides are applied according to the label 

requirements, offsite impacts can be avoided. EPA employed this analysis of offsite 

impacts in the dicamba RED, and concluded that no specific additional drift mitigations 

were needed to support the continued registration of all dicamba uses (U.S. EPA, 2009b). 

Since the proposed use pattern for dicamba on MON 87708 is consistent with use 

patterns evaluated in the dicamba RED, it would be reasonable to conclude that dicamba 

use on MON 87708 meets existing FIFRA standards related to drift and offsite movement. 
Monsanto Company 10-SY-210U 546 of 721 

The use of dicamba on MON 87708 in accordance with the FIFRA approved label does 

not pose any greater risk to non-target vegetation over existing dicamba agricultural uses 

approved by EPA. Nevertheless, Monsanto has already taken additional steps to manage 

dicamba offsite movement even though such steps are not required by EPA as stated in 

the RED. In the pending application to EPA, Monsanto requested the use of dicamba on 

MON 87708 on the low volatility DGA salt formulation (U.S. EPA Reg. No. 524-582) 

and limited dicamba application to ground application equipment as additional 

stewardship measures. Monsanto plans to further address the use of dicamba on 

MON 87708 with US EPA to evaluate whether any additional measures may be 

appropriate to further address potential drift and offsite movement. 

Monsanto will also implement a robust stewardship program to reinforce the EPA label 

requirements, including a strong emphasis on grower and applicator training by working 

with American Association of Pesticide Safety Educators (AAPSE) and other 

stakeholders in applicator training to further facilitate proper use of dicamba. 

Furthermore, as part of Monsanto’s stewardship program, Monsanto will encourage 

growers and applicators to consult with available sensitive crop registries prior to making 

dicamba applications to MON 87708. Many state lead agriculture agencies (IA, IL, IN, 

KS, MI, MN, MO, NE, OK, WI) have developed tools and resources to assist the 

applicator in the location of sensitive areas, such as vegetable or organic production 

fields, in an effort to minimize commercial impacts associated with herbicide offsite 

movement. 

Herbicides are widely used now in soybean production, and growers and commercial 

herbicide applicators have over 40 years experience in making dicamba applications in 

numerous crops including preemergence applications on conventional soybean. 

Alternative herbicides will continue to be used, as needed, for weed management or to 

implement weed resistance management practices, however it is anticipated that dicamba 

will displace in part the use of some alternative herbicides. EPA regulates the use of 

herbicides and has concluded that dicamba offsite movement from uses consistent with 

those proposed for MON 87708 does not pose unreasonable adverse effects to non-target 

vegetation. Furthermore, as indicated above, when herbicides are applied in accordance 

with the FIFRA label application use instructions, potential impacts on adjacent 

agricultural crops and non-agricultural plants can be avoided. Thus, potential impacts to 

these adjacent areas due to deregulation of MON 87708 and use of dicamba are similar 

when compared to the no action alternative (discussed below). Therefore, the approval in 

whole and no action alternatives are similar regarding potential impacts on adjacent 

agricultural and non-agricultural areas. 

No action alternative

Under the no action alternative, MON 87708 would not be widely available and growers 

would continue to use other practices for control of weeds in soybean fields. These 

practices would likely consist of some combination of herbicide use and tillage. 

However, the combination of herbicides used would likely be different than with full 

deregulation, as dicamba would only have a limited use pattern on soybean. Growers 

would likely use some combination of the herbicides currently in use for soybean 
Monsanto Company 10-SY-210U 547 of 721 

(discussed in Section VIII.F.1 of the petition). It is reasonably foreseeable that new 

herbicides will be invented, or that existing herbicides will be adopted for use on soybean 

or herbicides associated with soybean events under consideration for deregulation may be 

used (e.g., other auxin type herbicides like 2,4-D) (Table J-5). Offsite impacts to 

adjacent plants from these alternative herbicides would vary, depending on active 

ingredients, weather conditions, formulations, application methods and other factors. 

Because herbicides are currently used extensively in soybean production and will be for 

the foreseeable future, it is likely that the effects to adjacent crops and non agricultural 

plants would be similar under the no action and approval in whole alternatives. 

J.5.8. Potential Impacts to Raw or Processed Agricultural Commodities 

Approval in whole alternative

It is expected that under the approval in whole alternative MON 87708 will be integrated 

into the Roundup Ready soybean system using traditional breeding techniques. For those 

acres where glyphosate resistant weeds may already be present or where application of an 

herbicide with a different mode of action would aid in weed control or the 

implementation of weed resistant management practices, the cultivation of soybean 

containing the MON 87708 trait would be an option for growers. 

Within petition #10-SY-210U, extensive data have been presented relating to plant 

growth parameters, disease susceptibility, insect susceptibility, and forage and seed 

composition of MON 87708 compared to conventional soybean. These data indicate that 

there are no biologically relevant differences between MON 87708 and conventional 

soybean, except for its tolerance to dicamba. Biotechnology-derived crops used for food 

or feed undergo a voluntary food and feed consultation process with the FDA prior to 

release on the market. Monsanto will complete this consultation process with FDA prior 

to commercial introduction of MON 87708. 

Soybean compositional data on MON 87708 and conventional soybean are presented in 

Section VI of the petition. Information from this evaluation shows that soybean produced 

from MON 87708 is of comparable quality to soybean commercially produced in the 

U.S. for commodity markets. Dicamba residue levels in soybean seed harvested from 

MON 87708 treated with dicamba (1.0 lb a.e. per acre preemergent followed by two, 

sequential 0.5 lb a.e. per acre postemergent applications) at more than twice the 

anticipated commercial in-crop application rate were low, less than 0.1 ppm, and well 

below the established 10 ppm pesticide residue tolerance34 supporting dicamba use on 

commercial soybean (see Appendix M, Section M.4.1.1 of the petition). Monsanto is 

also petitioning the agency for the establishment of new tolerances on forage (45 ppm) 

and hay (70 ppm). 

Due the established safety of dicamba and allowable residues in food and feed, there is no 

evidence that the deregulation of MON 87708 would cause any impacts on either raw or 

 

34 EPA established tolerances for dicamba can be found at 40 CFR § 180.227. Dicamba residue is defined 

as dicamba, DCSA and 5-hydroxy dicamba. Analysis of MON 87708 included quantification of these 

defined dicamba residues, in addition to the minor metabolite DCGA. 
Monsanto Company 10-SY-210U 548 of 721 

processed soybean commodities resulting from the deregulation of MON 87708. EPA 

will evaluate any potential risks associated with the pending application for the new use 

of dicamba on MON 87708 and the establishment of new feed tolerances on soybean 

forage and hay as a part their review, and will conclude a reasonable certainty of no harm 

to human health when it approves the pending new use application. 

No action alternative 

MON 87708 would not be widely available or grown commercially under this alternative. 

Under this alternative, other herbicides and other biotechnology-derived soybean 

products or traditional tillage practices would be used instead of MON 87708 with its 

associated dicamba treatment. Therefore, the no action and approval in whole 

alternatives are similar regarding their impacts to raw and processed agricultural 

commodities. 

J. 5.9. Potential Impacts to Human Health and Safety 

It is expected that under the approval in whole alternative MON 87708 will be integrated 

into the Roundup Ready soybean system using traditional breeding techniques. For those 

acres where glyphosate resistant weeds may already be present or where application of an 

herbicide with a different mode of action would aid in weed control or the 

implementation of weed resistant management practices, the cultivation of soybean 

containing the MON 87708 trait would be an option for growers. 

Prior to the introduction of a biotechnology-derived crop product into the marketplace, 

Monsanto conducts tests to assure that the products are safe for their intended use and are 

appropriately labeled. For MON 87708, impacts on human health are considered for 

exposure to the biotechnology-derived soybean (MON 87708) and the associated 

dicamba herbicide. As mentioned previously, biotechnology-derived crops for food and 

feed use undergo a voluntary consultation process with the FDA prior to release onto the 

market. Although a voluntary process, Monsanto routinely completes a consultation with 

the FDA prior to placing a new biotechnology-derived crop product on the market. The 

consultation process with FDA on MON 87708 (BNF No. 125) was completed on 

October 11, 2011 (see Section I.C.I of the petition). 

Herbicide use and resulting residues associated with herbicide-tolerant crops improved 

through biotechnology are regulated under the Federal Food, Drug, and Cosmetic Act 

(FFDCA) by the EPA. Under the FFDCA (21 USC 301 et seq.), pesticide residues in or 

on raw agricultural commodities or processed foods are considered to be safe after a 

tolerance or exemption from tolerance has been established. The FDA enforces the 

tolerances set by the EPA. Currently, a pesticide residue tolerance exists for residues of 

dicamba on soybean seed at 10 ppm (40 CFR § 180.227), which is based on the approved 

late season postemergence (prior to harvest) use of dicamba on commercial soybean. As 

previously mentioned, Monsanto has submitted an application to EPA to register a new 

use pattern for dicamba on MON 87708 that will allow for preemergence and up to two 

postemergence in-crop applications of dicamba. Residue data generated on MON 87708 

to support this new dicamba use pattern using maximum label rates, show low levels of 
Monsanto Company 10-SY-210U 549 of 721 

dicamba residues on soybean seed (< 0.1 ppm dicamba a.e.; see Appendix M, Section 

M.4.1.1 of the petition) and confirm the current soybean seed tolerance is sufficient to 

support the new use on MON 87708. Monsanto has also petitioned the EPA to establish 

new tolerances for dicamba on soybean forage (45 ppm) and hay (70 ppm) to allow 

soybean commodities to be fed to or grazed by livestock. Existing dicamba tolerances 

for animal foodstuffs (e.g., meat, milk) are sufficient to address potential incremental 

dietary exposure to livestock from dicamba-treated MON 87708 forage and hay, as 

discussed in Section J.5.9.2. 

J.5.9.1. Human Health (MON 87708) 

Approval in whole alternative 

Under full deregulation, MON 87708 could be grown broadly across the U.S. Soybean 

and forage produced from MON 87708 would enter the food and feed chain and would 

be consumed by humans and animals. Agricultural workers would be exposed to 

MON 87708 and its associated agricultural practices. The potential human health 

impacts associated with the introduction of MON 87708 and increased applications of 

dicamba are separately discussed below. 

MON 87708 was developed through Agrobacterium-mediated transformation of soybean 

meristem tissue using the binary transformation plasmid PV-GMHT4355 (Section III; 

Figure III-1 and Table III-1). MON 87708 contains one copy of the insert at a single 

integration locus. No additional genetic elements from the transformation vector were 

detected in the genome of MON 87708, including backbone sequence from plasmid PVGMHT4355. Additionally the data confirm the organization and sequence of the insert, 

and demonstrate the stability of the insert over several generations. On the basis of these 

data, it is concluded that only the MON 87708 DMO is produced from the inserted DNA. 

For MON 87708, the available data demonstrate that harvested seed is as safe as 

conventional soybean for food and feed uses; thus it is safe and wholesome for 

consumption. To assess the impact of MON 87708 DMO on food and feed safety, 

bioinformatic analyses were used to establish the lack of both structurally and 

immunologically-relevant similarities between MON 87708 and allergens or toxins, 

based on the amino acid sequence of MON 87708 DMO. Furthermore, digestive fate 

experiments conducted with MON 87708 DMO demonstrate rapid digestion in simulated 

gastric fluid (SGF), a characteristic shared among many proteins with a history of safe 

consumption. Rapid digestion of MON 87708 DMO in SGF indicates that it is highly 

unlikely that MON 87708 DMO will reach absorptive cells of the intestinal mucosa. This, 

combined with the history of safe consumption of mono-oxygenases (the class of 

enzymes to which MON 87708 DMO belongs) and the lack of homology of the amino 

acid sequence to known allergens and toxins, supports a conclusion that MON 87708 

DMO has low allergenic and toxic potential. Finally, a high dose of MON 87708 DMO 

in a mouse acute oral toxicity evaluation demonstrated that it is not acutely toxic, and 

does not cause any adverse effect. The safety assessment supports the conclusion that 

exposure to MON 87708 DMO poses no meaningful risk to human or animal health. 
Monsanto Company 10-SY-210U 550 of 721 

Extensive analysis of the composition of MON 87708 seed and forage demonstrated that 

no biologically relevant changes were detectable. A detailed compositional assessment 

of soybean harvested seed and forage is presented in Section VI of petition #10-SY-210U. 

The levels of key nutrients, anti-nutrients, and other components in MON 87708 were 

examined and compared to that of the near-isogenic conventional soybean control, A3525, 

a conventional soybean variety with background genetics representative of MON 87708, 

but without the genetic modification. Additionally, tolerance intervals representing 99% 

of the values of each analyte for a commercial soybean population were established. 

Results demonstrate that the levels of key nutrients, anti-nutrients, and other components 

of MON 87708 are compositionally equivalent to the conventional control and within the 

range of variability of commercial soybean that were grown concurrently in the same trial. 

Furthermore, FDA completed its consultation on the food, feed and nutritional safety 

assessment on MON 87708 on October 11, 2011, confirming Monsanto’s conclusion on 

the safety of MON87708-derived food and feed. 

On the basis of the characteristics of MON 87708 DMO and the extensive compositional 

characterization of MON 87708 harvested seed, no impacts to human health are expected 

from the approval in whole alternative. Thus, the approval in whole and no action 

alternatives are similar regarding the potential impacts of MON 87708 on human health. 

No Action Alternative 

Under the no action alternative, MON 87708 would continue to be a regulated article. 

Human exposure to existing conventional and GE soybean would remain unchanged. 

Growers and consumers exposed to MON 87708 would be limited to individuals 

involved in the cultivation of MON 87708 under the conditions of regulation. Therefore, 

the no action and approval in whole alternatives are similar regarding the potential 

impacts of MON 87708 to human health. 

J.5.9.2. Human Health (Dicamba) 

Approval in whole alternative 

The toxicology or safety profile of dicamba has been extensively reviewed (U.S. EPA, 

2009b). Dicamba does not pose any unusual toxicological concerns and is not 

carcinogenic (U.S. EPA, 2009b; Durkin and Bosch, 2004; PMRA, 2008; European 

Commission, 2007b). EPA completed the reregistration of dicamba in 2006. The 

Reregistration Eligibility Decision (RED) document for dicamba and its associated salts 

concluded a high level of confidence exists for the dicamba hazard data base and the 

reliability of these data necessary to support the required safety finding for continued 

registration, including the pre-harvest use on commercial soybean. The dicamba RED 

document, and the related Health Effects Division (HED) of the EPA chapter (U.S. EPA, 

2005a), provide a detailed overview of the toxicological properties of dicamba. A 

summary of dicamba’s toxicity profile is presented in Section L.3.1 of Appendix L of the 

petition. 
Monsanto Company 10-SY-210U 551 of 721 

EPA evaluated the potential risks to humans from the use of dicamba as a part of the 

dicamba RED, concluding that aggregate exposure to dicamba, defined as dietary (food 

and water) and non-occupational (residential and recreational) exposures, meet the 

FIFRA determination of no unreasonable adverse effects and the FFDCA determination 

for reasonable certainty of no harm to human health (see Section M.4.1 of the petition for 

more detail). EPA has conducted acute and chronic dietary (food and water) risk 

assessments for dicamba based on a theoretical worst case exposure estimate. For food, 

this estimate assumes that dicamba is used on 100 percent of all the crops on which the 

pesticide is currently approved for use. It further assumes that the resulting pesticide 

residues found on all harvested food and feed crops and derived animal food 

commodities (e.g., meat and milk) are at the level of the legally established tolerance 

(i.e., the maximum allowable pesticide residue level). Residues of dicamba are defined 

as dicamba and its metabolites 5-hydroxy dicamba and 3,6-dichlorosalicylic acid (DCSA) 

in soybean commodities, and as dicamba and DCSA in animal food commodities, as 

currently regulated in 40 CFR § 180.227. For water, EPA assumed that dicamba could 

potentially move offsite to adjacent surface water bodies as a result of drift or runoff, or 

move through soil to groundwater. Since the estimated concentrations in groundwater 

were significantly lower compared to surface water, surface water estimates were used in 

the worse case dietary assessments (see Section J.5.2.2 and Section M.5.2 of the petition 

for details on dicamba levels in water resources). Based on the worst-case assumptions 

outlined above, acute and chronic dietary exposure was well below the Agency’s level of 

concern to satisfy the FIFRA and FFDCA standards (U.S. EPA, 2009b). 

Characterization of the nature of dicamba residues in MON 87708 confirms no additional 

residues of concern are created in MON 87708, and the current soybean seed dicamba 

residue definition is applicable for MON 87708. Residue levels in soybean seed 

harvested from MON 87708 treated with dicamba at the proposed maximum allowable 

application use pattern (1.0 pound a.e. per acre preemergence and two 0.5 pound a.e. per 

acre postemergence applications) were less than 0.1 ppm, well below the established 10 

ppm tolerance supporting the current use of dicamba on conventional soybean. These 

dicamba residue data for MON 87708 were submitted to the U.S. EPA on April 28, 2010 

(OPP Decision Number: D-432753, Registration Number 524-582), along with a 

proposed label for the use of dicamba on MON 87708. 

Presently, dicamba is applied to less than 1% of soybean acres using pre-plant and preharvest burn down applications (see Table VIII-12 in petition #10-SY-210U). Under the 

approval in whole alternative, dicamba will be used on more soybean acres and a higher 

percentage of soybean and soybean-derived foods will contain dicamba residues; 

however, dicamba residue levels in MON 87708 harvested seed or processed foods will 

be significantly lower compared to levels originating from the current pre-harvest 

soybean use (approximately 100-fold lower, based on <0.1 ppm. residue in MON 87708 

seed compared to established 10 ppm tolerance (see Appendix M, Section M.4.1.1 for 

residue levels in seed). It is difficult to determine the exact impact on actual dietary 

exposure from the expanded use of dicamba on MON 87708; however, the EPA 

concluded that residues of dicamba up to 10 ppm in soybean seed are safe (reasonable 

certainty of no harm as defined by FFDCA) for human and animal consumption, based 

on the EPA’s Tier 1 dietary and aggregate (dietary plus other non-occupational) exposure 
Monsanto Company 10-SY-210U 552 of 721 

assessments which assume 100% of soybean foods contain dicamba residues at the 

10 ppm tolerance level. 

Additionally, Monsanto has petitioned the EPA to establish new feed tolerances for 

soybean forage and hay to allow MON 87708 forage and hay to be fed to livestock, a 

practice that is presently prohibited for dicamba-treated commercial soybean (see 

Appendix M, Section M.4.1.1 for residue levels in forage and hay). This practice is 

presently not allowed because the current preharvest application is made past the stage 

where the crop would be useful as forage or hay. Thus, there has been no reason for 

pursuing these tolerances until earlier in-crop applications of dicamba were possible, as 

with MON 87708. Dietary exposure to livestock from the feeding of MON 87708 

soybean forage and hay does not result in an exceedance of the livestock maximum 

theoretical dietary burden established by the EPA, which is used to establish animal byproduct commodity tolerances (e.g., meat and milk). Therefore, the approval of soybean 

forage and hay tolerances does not result in a change to the current animal by-product 

commodity (food) tolerances and as a result does not increase potential dietary exposure 

of dicamba. 

Since no changes to the current dicamba food tolerances (soybean seed and animal byproducts) are needed, the dietary and aggregate risk assessments conducted in the RED 

are inclusive of the incremental exposure resulting from the use of dicamba on 

MON 87708. Therefore, the deregulation of MON 87708 would not present a significant 

impact to human health, and the approval in whole and no action alternatives are the 

similar regarding potential impacts on human health. 

No Action Alternative 

Under the no action alternative, MON 87708 would remain a regulated article and would 

not be commercially available to growers. It is likely that growers will continue to use 

herbicides in soybean production, and use the Roundup Ready soybean system. Growers 

will continue to use additional herbicides where needed or recommended to control hardto-control or glyphosate-resistant weeds. Growers may also choose to use other 

herbicide-tolerant soybean events, use a combination of alternative herbicides registered 

for use in soybean, or use traditional tillage practices to control weeds. In addition, 

dicamba will continue to be used on a small number of soybean acres (refer to Section 

VIII.G in the petition). Consumers will be exposed to residues of dicamba and the 

alternative herbicides through consumption of soybean and soybean-derived foods, and 

from residues on other food crops such as corn and wheat. On the basis of the analysis 

above, the approval in whole and no action alternatives involve the continued use of 

herbicides for production of soybean. Thus, the no action and approval in whole 

alternatives are similar regarding potential impacts to human health. 

J.5.9.3. Worker Safety 

Approval in whole alternative 
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It is expected that under the approval in whole alternative MON 87708 will be integrated 

into the Roundup Ready soybean system using traditional breeding techniques. For those 

acres where glyphosate resistant weeds may already be present or where application of an 

herbicide with a different mode of action would aid in weed control or the 

implementation of weed resistant management practices, the cultivation of soybean 

containing the MON 87708 trait would be an option for growers. 

There is no notable worker safety issue related directly to exposure to MON 87708. As 

described elsewhere, MON 87708 is as safe as conventional soybean for use as food or 

feed. MON 87708 DMO has no safety concerns that would result in adverse effects to 

workers exposed to MON 87708 plant tissues. 

Agricultural workers can be exposed to dicamba during the herbicide application or upon 

re-entry into treated MON 87708 fields. Under the proposed label, dicamba can be 

applied as a preemergent treatment on MON 87708 at rates up to 1 lb a.e. per acre and 

then again in two sequential 0.5 lb a.e. per acre postemergent treatments up to the R1/R2 

growth stage using a ground application method. The EPA conducted a comprehensive 

occupational exposure assessment as part of the dicamba reregistration in 2006 and 

concluded no unreasonable risk to agricultural workers from ground and aerial dicamba 

applications to soybean at rates up to 2 lb a.e. per acre when required personal protective 

equipment is worn (U.S. EPA, 2009b). See Section M.4.2 of the petition for additional 

detail. The application scenario evaluated by EPA encompasses the application method 

and rates for dicamba applied to MON 87708. Therefore, the deregulation of 

MON 87708 does not present a significant impact to the health of agricultural workers. 

No Action Alternative 

Under the no action alternative, MON 87708 would remain a regulated article and would 

not be commercially available to growers. It is likely that growers will continue to use 

herbicides in soybean production, and use the Roundup Ready soybean system and utilize 

additional herbicides where needed or recommended to control hard-to-control or 

glyphosate-resistant weeds. Growers may also choose to use other herbicide-tolerant 

soybean events, use a combination of alternative herbicides registered for use in soybean, 

or use traditional tillage practices. The continued use of dicamba on a small number of 

soybean acres would also be expected. Therefore, agricultural workers will be exposed 

to residues of dicamba and the alternative herbicides. On the basis of the analysis above, 

the deregulation in whole and no action alternatives involve the continued use of 

herbicides for production of soybean. Thus the impacts to agricultural worker health for 

either alternative are not considered different. 

J.5.10. Potential Impacts to Plant and Animal Communities Including Threatened 

or Endangered Species, Soil Microorganisms and Biodiversity 

Under the approval in whole alternative it is expected that MON 87708 will be integrated 

into the Roundup Ready soybean system using traditional breeding techniques. For those 

acres where glyphosate resistant weeds may already be present or where application of an 

herbicide with a different mode of action would aid in weed control or the 
Monsanto Company 10-SY-210U 554 of 721 

implementation of weed resistant management practices, the cultivation of soybean 

containing the MON 87708 trait would be an option for growers. 

The potential for MON 87708 to harm plant, animal and microbial communities as well 

as threatened and endangered species (TES) was evaluated by considering the biology of 

soybean, biochemical information and experimental data. Soybean does not have any 

sexually-compatible relatives in the U.S. Therefore, any effect due to movement of the 

dicamba tolerance trait is confined to soybean (Glycine max). The biochemical 

information and experimental data included product characterization information, 

information from the MON 87708 DMO safety assessments, the history of environmental 

exposure to mono-oxygenases (the class of enzymes to which MON 87708 DMO 

belongs), results from the environmental assessment described in the petition, and the 

demonstration of compositional, agronomic and phenotypic equivalence to conventional 

soybean. An analysis of the effects of MON 87708 on plant, animal and microbial 

communities as well as threatened and endangered species is found below. 

J.5.10.1. Gene Movement 

Approval in whole alternative 

In assessing the potential impact to plant and animal communities, the potential for gene 

movement and introgression from MON 87708 was evaluated because movement and 

establishment of the gene and trait to related species could have indirect impacts to plant 

and animal communities that extend beyond the original recipient organism. Additional 

discussion of the potential environmental impact due to gene movement may be found in 

Section IX.D of the petition. Monsanto considered two primary issues: 1) the potential 

for gene flow and introgression, and 2) the potential impact of introgression. The genus 

Glycine has approximately nine species, with commercial soybean (G. max) being placed 

in the subgenus Soja along with one other species, G. soja. G. max is sexually 

compatible only with G. soja and no other Glycine species. G. max is the only Glycine

species located in the United States. Therefore, the probability of gene flow and 

introgression of MON 87708 into other species in the U.S. is essentially zero (Stewart et 

al., 2003); thus, the potential impact of MON 87708 introgression to sexually compatible 

relatives on plant and animal communities is nonexistent if MON 87708 were approved 

in whole. The approval in whole and no action alternatives are the same regarding 

potential impacts on gene movement. 

No action alternative 

As discussed above under the approval in whole alternative, soybean is not sexually 

compatible with any other plant species in the U.S. Therefore, the no action and approval 

in whole alternatives are the same regarding potential impacts on gene movement. 

J.5.10.2. Animal Communities 

Approval in whole alternative 
Monsanto Company 10-SY-210U 555 of 721 

Soybean production systems in agriculture are host to many animal species. Mammals 

and birds, including migratory mammals and birds, may seasonally consume grain in the 

field, and invertebrates can feed on the plant during the entire growing season. Animals 

that feed primarily on soybean are seed-feeding insects and rodents found in agricultural 

fields. Rodents, such as mice or squirrels, may seasonally feed exclusively on soybean 

seeds. Thus, these animals may have a diet containing significant amounts of soybean 

seeds. Deer may also forage in soybean fields on forage and seed left after harvest (see 

Section IX.B.3.5 of the petition). 

Under the approval in whole alternative, the cultivation of MON 87708 is not expected to 

impact soybean agronomic practices, with the exception of a change in dicamba use 

pattern. Cultivation of MON 87708 would not alter agronomic inputs or the number of 

soybean acres under cultivation, and may have a small positive effect on tillage 

cultivation practices. Potential impacts to animals from widespread cultivation of 

MON 87708 would be primarily based on the possible effects of the introduced 

MON 87708 DMO that provides tolerance to dicamba and a broadened application 

window of dicamba to MON 87708. For instance, if MON 87708 is deregulated, 

MON 87708 and MON 87708 DMO will be present in soybean consumed by animals. 

As discussed previously, there is no meaningful risk to animal or human health from 

dietary exposure to MON 87708 DMO. There are no known toxic properties associated 

with MON 87708 DMO. Furthermore, the composition of the seed and forage produced 

by MON 87708 is compositionally equivalent to conventional soybean. This information 

on the safety of MON 87708 DMO and composition of MON 87708 soybean seed, as 

detailed in petition, indicate that there would be no negative effects to mammals that 

consume MON 87708 seed and forage. Similarly, it is expected that there would be no 

impact to birds or other animals, including migratory birds and animals, that may 

consume soybean forage or soybean seed from MON 87708. During field trials with 

MON 87708, no biologically relevant changes in insect feeding damage were observed 

(see Section VII of the petition) indicating similar insect susceptibility for MON 87708 

compared to conventional soybean. As MON 87708 exhibits no toxic effects on animals 

or insects, it is concluded that they will not be affected. In addition, the cultivation of 

MON 87708 does not impact the nutritional quality, safety or availability of animal feed 

(see Section J.5.11 of the petition). 

Dicamba is currently registered for early season preemergence (early preplant) and late 

season postemergence (pre-harvest) soybean applications. Upon deregulation, dicamba 

could also be applied late preemergence (up to cracking) and in-season postemergence 

applications up to R1/R2 growth stage. Therefore, with the exception of late preemergent 

and in-season applications of dicamba, the agronomic practices used to produce 

MON 87708 will be the same as those used to produce commercially available soybean. 

Animals may be affected by dicamba runoff and/or herbicide spray drift. However, a 

comprehensive safety evaluation and risk assessment conducted by EPA concluded that 

dicamba has low toxicity to mammals, is not a carcinogen, does not adversely affect 

reproduction and development, and does not bioaccumulate in mammals (U.S. EPA, 

2009b). An ecotoxicological risk assessment concluded that the use of dicamba does not 

pose an unreasonable risk of adverse effects to non-target species, such as birds and fish, 

when used according to label directions, nor does it pose an unreasonable risk of adverse 
Monsanto Company 10-SY-210U 556 of 721 

effects to insects outside of the application area (U.S. EPA, 2009b). Furthermore, outside 

the cultivated soybean field, dicamba is unlikely to affect forbs and beneficial arthropods 

that are dependent on plants for survival. On the basis of this analysis, approval in whole 

of MON 87708 will not result in significant impacts on animals, including insects and 

beneficial arthropods that live in or near soybean fields containing MON 87708, and the 

approval in whole and no action alternatives are the same regarding potential impacts on 

animals and animal communities. 

No action alternative 

Under the no action alternative MON 87708 and its progeny would continue to be 

regulated articles and would not be widely grown. Under this alternative dicamba 

tolerance trait would not become integrated into the Roundup Ready soybean system and 

dicamba use would likely remain similar to today’s use pattern in soybean. Adding other 

herbicides with different modes-of-action into the Roundup Ready system to mitigate 

development of glyphosate resistant weeds and control glyphosate resistant weeds would 

continue to remain an option. Additionally, conventional tillage may increase in some 

instances as an additional means to control problematic weeds. 

In order to manage weeds, growers would continue to use available herbicides, some of 

which may pose greater potential risks to animals or insects than dicamba; and cultivation 

practices may increase. Therefore, the no action and approval in whole alternatives are 

similar regarding their effects to animals and animal communities. 

J.5.10.3. Plant Communities 

Approval in whole alternative 

Soybean production systems in agriculture are host to many plant species. Likewise, the 

environment surrounding a soybean field varies in plant composition depending on the 

region. In certain areas, soybean fields may be bordered by other soybean, corn or other 

crops; fields may also be surrounded by wooded and/or pasture/grassland areas, as well 

as aquatic environments. Therefore, the types of vegetation, including weeds, around a 

soybean field depend on the area where the soybean is planted. A variety of weeds dwell 

in and around soybean fields; those species will also vary depending on the region where 

the soybean is planted. 

If MON 87708 is approved in whole, it will not compete with plants found outside of 

agricultural production, because, like commercially available soybean, MON 87708 does 

not exhibit characteristics associated with weedy growth. Weeds within fields of 

MON 87708 will be managed using existing agricultural practices, including mechanical, 

cultural, and chemical control measures. This will include in-season (up to R1/R2 

growth stage) applications of dicamba. 

Plants on adjacent land have the potential to be affected by dicamba runoff and offsite 

movement (spray drift and volatility), but when herbicides are applied according to the 

FIFRA label application instructions, offsite impacts can be avoided. EPA concluded in 

the dicamba RED (U.S. EPA, 2009b) that existing label language to mitigate offsite 
Monsanto Company 10-SY-210U 557 of 721 

movement was sufficient to reduce the potential risk of damage to adjacent vegetation. 

Since the proposed use pattern for dicamba on MON 87708 is consistent with the uses 

evaluated by EPA as part of the RED and the proposed label contains the offsite 

movement mitigation language, it is reasonable to conclude that the use of dicamba on 

MON 87708 also meets the FIFRA no unreasonable effects standard for drift and offsite 

movement. Nevertheless, Monsanto has taken additional steps to manage dicamba offsite 

movement as stewardship measures. In the pending application to EPA, Monsanto 

requested the use of dicamba on MON 87708 on the low volatility DGA salt formulation 

(U.S. EPA Reg. No. 524-582) and limited dicamba applications to ground application 

equipment. Monsanto also plans to further address the specific use of dicamba on 

MON 87708 with US EPA to evaluate whether additional measures, may be appropriate 

to address potential drift and offsite movement. 

Therefore, the presence of the dicamba tolerance trait in MON 87708 and the use of 

dicamba is not expected to have a significant impact on surrounding plant communities, 

and the approval in whole and no action alternatives are similar regarding potential 

impacts on plant communities. 

No action alternative 

Under the no action alternative MON 87708 and its progeny would continue to be 

regulated articles and would not be widely grown. Under this alternative dicamba 

tolerance trait would not become integrated into the Roundup Ready soybean system and 

dicamba use would likely remain similar to today’s use pattern in soybean. Adding other 

herbicides with different modes-of-action into the Roundup Ready system to mitigate 

development of glyphosate resistant weeds and control glyphosate resistant weeds would 

continue to remain an option. Growers will continue to use existing soybean herbicides 

to control glyphosate-resistant or hard-to-control weeds in soybean production fields. In 

addition, approximately 60% of soybean acres in the U.S. do not use conservation tillage 

practices, and it is anticipated that conventional tillage may increase in some instances as 

an additional means to control hard-to-control or resistant weeds. Therefore, the no 

action and approval in whole alternatives are similar regarding potential impacts to plants 

and plant communities. 

J.5.10.4. Threatened and Endangered Species 

Congress passed the Endangered Species Act (ESA) of 1973, as amended, to prevent 

extinctions facing many species of fish, wildlife, and plants. The purpose of the ESA is to 

conserve endangered and threatened species and the ecosystems on which they depend as 

key components of America‘s heritage. To implement the ESA, the U.S. Fish and 

Wildlife Service (USFWS) works in cooperation with the National Marine Fisheries 

Service (NMFS); other Federal, State, and local agencies; Tribes; non-governmental 

organizations; and private citizens. Before a plant or animal species can receive the 

protection provided by the ESA, it must first be added to the Federal list of threatened 

and endangered wildlife and plants. 
Monsanto Company 10-SY-210U 558 of 721 

A species is added to the list when it is determined by the USFWS/NMFS to be 

endangered or threatened because of any of the following factors: 

 The present or threatened destruction, modification, or curtailment of its habitat or 

range; 

 Overutilization for commercial, recreational, scientific, or educational purposes; 

 Disease or predation; 

 The inadequacy of existing regulatory mechanisms; and 

 The natural or manmade factors affecting its survival. 

Once an animal or plant is added to the list, in accordance with the ESA, protective 

measures apply to the species and its habitat. These measures include protection from 

adverse effects of Federal activities. Section 7 (a)(2) of the ESA requires that Federal 

agencies, in consultation with USFWS and/or the NMFS, ensure that any action they 

authorize, fund, or carry out is not likely to jeopardize the continued existence of a listed 

species or result in the destruction or adverse modification of designated critical habitat. 

It is the responsibility of the Federal agency taking the action to assess the effects of their 

action and to consult with the USFWS and NMFS if it is determined that the action “may 

affect” listed species or critical habitat. To facilitate APHIS’ ESA consultation process, 

APHIS has met with the USFWS from 1999 to 2003 to discuss factors relevant to APHIS’ 

regulatory authority and effects analysis for petitions for nonregulated status, and 

developed a process for conducting an effects determination consistent with the Plant 

Protection Act of 2000 (Title IV of Public Law 106-224). This process is described in a 

decision tree document, which have been included in recent Environmental Assessments 

(USDA-APHIS, 2011a) . APHIS has used this process to help fulfill its obligations and 

responsibilities under Section 7 of the ESA for biotechnology regulatory actions. 

The potential impact of MON 87708 on federally listed Threatened or Endangered 

Species (TES) and species proposed for listing has been evaluated. In this analysis, the 

biology of MON 87708 and the agricultural practices associated with the cultivation of 

MON 87708 have been considered for potential adverse impact on TES and their critical 

habitats. Additionally, the potential indirect effects of dicamba applications due to either 

the introduction or non-introduction of MON 87708 on TES are discussed. While APHIS 

does not have statutory authority to authorize or regulate the use of herbicides, APHIS 

considers the evaluation of potential impacts on TES from the application of dicamba, as 

conducted by the EPA, as part of its environmental analysis under NEPA. APHIS’ 

regulatory authority over GE organisms under the PPA is limited to those GE organisms 

for which it has reason to believe it may be a plant pest or those for which APHIS does 

not have sufficient information to determine that the GE organism is unlikely to pose a 

plant pest risk (7 CFR § 340.1). 

Approval in whole alternative 

It is expected that under the approval in whole alternative MON 87708 will be integrated 

into the Roundup Ready soybean system using traditional breeding techniques. For those 
Monsanto Company 10-SY-210U 559 of 721 

acres where glyphosate resistant weeds may already be present or where application of an 

herbicide with a different mode-of-action would aid in weed control or the 

implementation of weed resistant management practices, the cultivation of soybean 

containing the MON 87708 trait would be an option for growers. Monsanto intends to 

make MON 87708 available for cultivation across all soybean growing regions in the 

U.S. The cultivation of MON 87708 is not expected to change the agronomic practices 

or expand soybean production acres beyond levels and geographies of current 

commercial soybean varieties. Several lines of evidence can be used to assess the 

potential for MON 87708 to have adverse effects on TES. The first line of evidence is 

based on the characteristics and evaluation of MON 87708 DMO and natural constituents 

present in MON 87708. The second line of evidence is the potential for TES species to 

interact with MON 87708. The third line of evidence is based on the biology and 

competitiveness of MON 87708 compared to conventional soybean. Finally, as the 

introduction of MON 87708 may result in increased use of dicamba in soybean (see 

Section VIII.H of the petition for additional details), potential indirect effects of dicamba 

on TES are also discussed, although it is the EPA’s statutory obligation to comply with 

the ESA in the registration of this use. 

J.5.10.4.1. Evaluation of MON 87708 and MON 87708 DMO 

As previously noted, (see Section V.F.), bioinformatics analysis determined that 

MON 87708 DMO does not share amino acid sequence similarities with known 

allergens, gliadins, glutenins, or protein toxins. MON 87708 DMO was rapidly digested 

in in vitro assays using simulated gastric and intestinal fluids and did not show any 

adverse effects when administered to mice via oral gavage at levels far in excess of that 

reasonably expected to be consumed by humans or animals. Compared to conventional 

soybean, MON 87708 does not express any additional proteins or natural toxicants that 

are known to directly or indirectly affect a listed TES or species proposed for listing by 

the U.S. Fish and Wildlife Service. Compositional analysis of MON 87708 for nutrients 

and anti-nutrients indicated that the harvested seed and forage from MON 87708 were 

compositionally equivalent to conventional soybean. Thus, MON 87708 would not be 

expected to have any impacts on TES that would differ from conventional soybean. 

J.5.10.4.2. Potential Interactions of TES with MON 87708 

The only TES animal listed that occupies habitat that is likely to include soybean fields 

and that might feed on soybean is the Federally Endangered Delmarva Peninsula Fox 

Squirrel, Sciurus niger cinereus, found in areas of the mid-Atlantic Eastern seaboard.35 It 

is known to utilize certain agricultural lands readily, and its diet includes acorns, 

nuts/seeds of hickory, beech, walnut, and loblolly pine, buds and flowers of trees, fungi, 

insects, fruit, and an occasional bird egg. Given its varied diet, the safety of MON 87708 

DMO, and the demonstrated compositional, agronomic and phenotypic equivalence of 

MON 87708 to conventional soybean, it is concluded that no biologically significant 

changes to the habitat or diet of the Delmarva Peninsula Fox Squirrel are 

 

35 Source is from website: http://ecos.fws.gov/tess_public/SpeciesReport.do; [Accessed May 14, 2009]. 
Monsanto Company 10-SY-210U 560 of 721 

expected. Consequently, the planting of MON 87708 is not expected to impact the 

Delmarva Peninsula Fox Squirrel. 

Additionally, MON 87708 has demonstrated no characteristics that would allow it to 

expand agricultural production into new natural areas where other TES animal species 

could be found. Consequently, the planting of MON 87708 is not expected to affect any 

TES animal species. 

J.5.10.4.3. Potential Hybridization or Competition of MON 87708 with TES 

Soybean is not native to the U.S., and MON 87708 is not sexually compatible with any 

federally listed TES or a species proposed for listing. Like other G. max, MON 87708 

will likely be a poor competitor with native vegetation and will not survive outside of 

cultivation (Baker, 1965). Thus, MON 87708 is not expected to interbreed with any plant 

species or displace natural vegetation in the U.S. 

J.5.10.5. Potential Impact of Dicamba on TES 

As a result of joint discussions between USFWS and APHIS officials on June 15, 2011, 

the Agencies have agreed that it is not necessary for APHIS to perform an ESA effects 

analysis for the herbicide use associated with a biotechnology-derived crop. This is 

because EPA has both regulatory authority over the labeling of pesticides and the 

necessary technical expertise to assess pesticide effects on the environment under FIFRA. 

Similarly for MON 87708, since APHIS has no regulatory jurisdiction to authorize or 

regulate the use of dicamba by growers and the EPA has an application pending for the 

use of dicamba on MON 87708, the EPA will also have the obligation to conduct an ESA 

effects analysis for the use of dicamba on MON 87708. However, APHIS does consider 

the evaluation of potential impacts on TES from the application of dicamba as part of its 

environmental analysis. 

In order to register a pesticide under FIFRA, the EPA must reach a conclusion that the 

pesticide will not cause an unreasonable adverse effect when used as intended. Thus, the 

current and future proposed uses of dicamba on soybean must meet this standard to be 

registered by the EPA. Monsanto has requested approval of the use of dicamba on 

MON 87708 by amending the label for EPA Registration Number 524-582 (as described 

in Appendix L, Section L.2.4). In the EPA’s review of this registration request, they will 

apply the same statutory-based regulatory requirements as they do to all pesticides. 

In the 2006 dicamba RED (U.S. EPA, 2009b), EPA discussed the changes in the 

registered use pattern of dicamba that were required to assure that dicamba meets the 

FIFRA regulatory standards of no unreasonable adverse effects for pesticides in the U.S. 

Accordingly, EPA reduced the maximum single application rate to 1.0 lb a.e. per acre 

with a maximum annual rate of 2.0 lb a.e. per acre (reduced from 2.8 lb a.e. per acre 

single and 7.7 lb a.e. per acre maximum annual rate, respectively). EPA went further in 

the RED to state that no specific additional mitigations for offsite movement were needed 

(U.S. EPA, 2009b). 
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The EPA’s new application rates, among other measures implement by EPA as part of 

the dicamba RED, provide protection to non-target organisms. Because the proposed use 

of dicamba on MON 87708 falls within the use rate limits established by EPA in the 2006 

RED, it can be concluded that risks to non-target organisms from the use of dicamba on 

MON 87708 have been assessed. 

Furthermore, Monsanto has conducted a species-specific analysis on threatened and 

endangered (TE) birds, mammals, amphibians, reptiles, terrestrial invertebrates, and nonmoncotyledonous terrestrial and semi-aquatic plants. This refined analysis utilized 

dicamba- and species-specific information, in accordance with guidance and procedures 

outlined in the Overview of the Ecological Risk Assessment Process in the Office of 

Pesticide Programs, U.S. Environmental Protection Agency, Endangered and Threatened 

Species Effects Determinations, published in 2004 (U.S. EPA, 2004), and methods 

utilized in more recent threatened and endangered species effects determinations 

conducted by the EPA (U.S. EPA, 2007b; 2008c). This analysis concluded that TE birds, 

mammals, amphibians, reptiles, terrestrial invertebrates, and monocotyledonous plants 

will not be adversely affected from environmental exposure to dicamba at levels 

anticipated from the use on MON 87708. A summary of the refined species analysis may 

be found in Appendix N of the petition. The summary and supporting detailed analyses 

have been provided to EPA (Frank and Kemman, 2011 and 2012; Honegger, 2012; 

Mueth and Foster, 2010; Schuler et al., 2012; Wright et al., 2012). 

For the remaining federally listed TE non-monocotyledonous terrestrial plants, less than 

one percent of potential soybean production acres (defined as all cultivated crop acres 

listed in the 2001 National Land Coverage database in states with reported soybean 

production 36 are conservatively estimated to be in proximity to a TE nonmonocotyledonous terrestrial plant species.37 For the limited areas where a federally 

listed TE non-moncotyledonous terrestrial plant is in proximity to a MON 87708 field 

where dicamba may be applied, Monsanto has already undertaken steps to manage the 

offsite movement of dicamba (described in Section J.5.7) to further reduce potential 

impacts on these plant species. These measures include registering the use on 

MON 87708 on the low volatility DGA salt formulation of dicamba and limiting dicamba 

use on MON 87708 to ground application only. Monsanto will also consult with U.S. 

EPA in coming months to evaluate whether additional measures may be appropriate to 

address potential drift and offsite movement. Finally, upon commercialization of 

MON 87708, Monsanto will implement a web-based endangered species mitigation 

system, similar to PreServe.org, to further mitigate potential impacts of dicamba on TE 

non-monocotyledonous terrestrial plants. Therefore, the approval in whole and no action 

alternatives are similar regarding potential impacts on threatened or endangered species 

from dicamba use on MON 87708. 

 

36 Potential soybean production acres include all land of Class 81 (cultivated crops) and Class 82 

(pasture/hay) from the 2001 National Land Cover Database, in counties reporting soybean production in the 

1997, 2002, and 2007 Ag Census. 

37 Based on species location information available from the FIFRA Endangeres Species Task Force 

(FESTF) Multi-Jurisdictional Database (MJD) and other state and federal information sources. 
Monsanto Company 10-SY-210U 562 of 721 

No action alternative 

Under the no action alternative MON 87708 and its progeny would continue to be 

regulated articles and would not be widely grown. Under this alternative MON 87708 

would not become integrated into the Roundup Ready soybean system and dicamba use 

would likely remain similar to today’s use in soybean. Adding other herbicides with 

different modes-of-action into the Roundup Ready system to mitigate development of 

glyphosate resistant weeds and to control glyphosate resistant weeds would continue to 

remain an option. Alternatively, tillage may increase in some instances as an additional 

means to control problematic weeds. 

Since other herbicides are currently available and used in soybean production, and would 

likely be used instead of dicamba, tillage practices would likely continue as a method of 

weed control. Therefore, the approval in whole and the no action alternatives are similar 

regarding their effects to TES.

J.5.10.6. Soil Microorganisms 

Approval in whole alternative 

Potential impacts to soil microorganisms can arise from the exposure to the introduced 

gene and expressed protein in the GE crop product. In addition, agricultural practices 

such as pesticide applications and tillage are known to impact soil microbial populations, 

species composition, colonization, and associated biochemical processes. 

Soil microorganisms in general and symbiotic microbes associated with soybean roots are 

discussed in Section VII.C.4 and Appendix J, Section J.3.5.3 of the petition, and 

summarized here. The difference between the approval in whole and the no action 

alternative is expected to be integration of MON 87708 into the Roundup Ready soybean 

system using traditional breeding techniques. MON 87708 is not expected to alter the 

current agronomic practices for soybean cultivation. No adverse effects on soil 

microorganisms are associated with MON 87708 or its cultivation, nor do the 

characteristics of the MON 87708 DMO pose any concern to soil microorganisms. The 

B. japonicum-soybean symbiosis of MON 87708 was not changed either as a result of the 

introduction of the dicamba tolerance trait or as a result of the MON 87708-produced 

DMO. 

Dicamba was registered as an agricultural herbicide in 1967 (U.S. EPA, 2009b) and has a 

long history of use. Impacts on soil microorganisms have not been raised as a significant 

concern, and results of standardized tests with dicamba and dicamba formulations did not 

indicate any long term effects on soil microorganisms (see Section M.5.6.3 of the 

petition). Results of standardized tests with dicamba and dicamba formulations indicate 

no long-term effects on functional processes of soil microorganisms (carbon respiration 

and nitrogen transformation) at rates proposed for dicamba on MON 87708 (European 

Commission, 2007a). 

On the basis of these observations and in conjunction with related phenotypic 

measurements for MON 87708, no impact on soil microorganisms is expected from the 
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cultivation of MON 87708. Therefore, the approval in whole and no action alternatives 

are the same regarding potential impacts on soil microorganisms. 

No action alternative 

Under the no action alternative MON 87708 and its progeny would continue to be 

regulated articles, would not be commercially grown and use of dicamba herbicide would 

likely remain similar to today’s use pattern in soybean. 

Adding other herbicides with different modes-of-action into the Roundup Ready system 

to mitigate development of glyphosate resistant weeds and control glyphosate resistant 

weeds would continue to remain an option. Additionally, conventional tillage may 

increase in some instances as an additional means to control problematic weeds. 

Agricultural practices such as pesticide applications and tillage are known to impact soil 

microbial populations, species composition, colonization, and associated biochemical 

processes. However, alternative herbicides are already available and are used in soybean, 

and would likely be used instead of dicamba under the no action alternative. Similarly, 

approximately 40% of the U.S. soybean acres use conservation tillage and would likely 

continue to do so. Therefore, the no action and approval in whole alternatives are similar 

regarding potential impacts to microorganisms. 

J.5.10.7. Biodiversity 

Approval in whole alternative 

The difference between the approval in whole and the no action alternative is expected to 

be integration of MON 87708 into the Roundup Ready soybean system with the potential 

for cultivation of soybean seed containing the MON 87708. 

As confirmed in trials to evaluate plant growth and phenotypic characteristics, 

MON 87708 exhibits no traits that would cause increased weediness of soybean. Since 

soybean is not sexually compatible with any other plant species in the U.S., its 

unconfined cultivation would not lead to increased weediness of other sexually 

compatible relatives. Therefore, it is unlikely to have effects on non-target organisms 

common to agricultural ecosystems or TES recognized by the U.S. Fish and Wildlife 

Service and National Marine Fisheries Service, or to colonize adjacent non-agricultural 

ecosystems thereby compromising their biodiversity. 

MON 87708 DMO originates from S. maltophilia, an organism that is ubiquitous in the 

environment. MON 87708 DMO shares sequence identity and many catalytic and 

domain structural similarities with a wide variety of oxygenases present in bacteria and 

plants currently widely consumed, establishing that animals and humans are extensively 

exposed to these types of enzymes. Bioinformatics analysis determined that MON 87708 

DMO does not share amino acid sequence similarities with known allergens, gliadins, 

glutenins, or protein toxins. MON 87708 DMO was rapidly digested in in vitro assays 

using simulated gastric and intestinal fluids and did not show any adverse effects when 

administered to mice via oral gavage at levels that resulted in large margins of exposure. 
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MON 87708 DMO demonstrates a high level of substrate specificity. Additionally, 

studies have shown no impact to NTOs such as beneficial and pest arthropods when 

exposed to MON 87708 DMO in the field. Taken together these data indicate low 

probability that MON 87708 DMO will have a significant impact on biodiversity. 

The use of herbicides in agricultural fields is likely to indirectly impact biodiversity by 

decreasing weed species present in the field. Agricultural fields are purposefully 

managed to be weed-free resulting in greater economic benefit to the grower. In the U.S., 

98% of soybean acreage was treated with an herbicide in 2006 (USDA-NASS, 2007b). 

Therefore, introduction of MON 87708 and treatment with dicamba is unlikely to affect 

the animal or plant communities found in commercial soybean production systems 

differently than those already occurring due to the use of herbicides in soybean fields. 

Dicamba has an established history of safe use in agriculture. For an overview of the 

environmental safety of dicamba, see Appendix M of the petition. Based on this analysis, 

it is concluded that the potential effect of approval in whole of MON 87708 on 

biodiversity would not differ from impacts associated with current agricultural practices 

used for production of soybean. Therefore, the approval in whole and no action 

alternatives are similar regarding potential impacts on biodiversity. 

No action alternative 

Under the no action alternative MON 87708 and its progeny would continue to be 

regulated articles and would not be commercially grown. Use of dicamba herbicide 

would likely remain similar to today’s use pattern in soybean, and growers would likely 

continue to use alternative herbicides or incorporate more tillage in soybean production to 

manage hard to control weeds. Thus, the approval in whole and the no action alternatives 

are similar regarding potential impacts on biodiversity. 

J.5.11. Animal Feed 

The majority of the soybean cultivated in the U.S. is grown for animal feed and primarily 

fed as soybean meal. Soybean forage may also be used as feed for dairy cattle and 

livestock. Biotechnology-derived products may undergo a voluntary consultation process 

with the FDA prior to commercialization. Monsanto has completed the biotechnology 

consultation process with FDA for the safety and nutritional assessment of food and feed 

derived from MON 87708 soybean on October 11, 2011 (BNF No. 00125, Monsanto, 

2011). As a part of its evaluation, FDA reviewed information on the identity, function, 

and characterization of the genes, including expression of the gene products in 

MON 87708 soybean, as well as information on the safety of the MON 87708 DMO and 

MON 87708 soybean. 

Approval in whole alternative 

The difference between the approval in whole and the no action alternative is expected to 

be integration of MON 87708 into the Roundup Ready soybean system using traditional 

breeding techniques. Under the approval in whole alternative, there is potential for 

cultivation of soybean seed containing the MON 87708 trait to proactively manage and 
Monsanto Company 10-SY-210U 565 of 721 

prevent the development of herbicide resistant weeds. For those acres where glyphosate 

resistant weeds may already be present or where application of an herbicide with a 

different mode-of-action would aid in weed control or the implementation of weed 

resistant management practices, the cultivation of soybean containing the MON 87708 

trait would be an option for growers. 

Potential impacts to the safety of animal feed from widespread cultivation of 

MON 87708 would be primarily based on the possible effects of the introduced 

MON 87708 DMO that provides tolerance to dicamba and a broadened application 

window of dicamba to MON 87708. Upon deregulation, MON 87708 containing 

MON 87708 DMO and dicamba residues from the application of dicamba will be present 

in animal feed. As discussed previously, there is no meaningful risk to animal health 

from dietary exposure to MON 87708 DMO. There are no toxic properties associated 

with MON 87708 DMO. Furthermore, the composition of the seed and forage produced 

by MON 87708 is equivalent to conventional soybean. This information on the safety of 

MON 87708 DMO and composition of MON 87708 soybean seed as detailed in petition 

#10-SY-210U indicate that there would be no negative impact on the safety or nutritional 

quality of animal feed from the cultivation of MON 87708. The cultivation of 

MON 87708 will not change the number of soybean acres cultivated in the U.S., the 

extent in which soybean acres are cultivated will continue to be based on the same 

market-based drivers that exist today 

As discussed previously, dicamba is presently applied to less than 1% of soybean acres 

using pre-plant and pre-harvest burn down applications. Under the approval in whole 

alternative, dicamba will be used on more soybean acres and a higher percentage of 

soybean-derived animal feeds will contain dicamba residues. However, dicamba residue 

levels in MON 87708 harvested seed or processed meal will be significantly lower 

compared to levels originating from the current pre-harvest soybean use (approximately 

100-fold lower, based on <0.1 ppm residue in MON 87708 seed compared to established 

10 ppm tolerance - see Appendix M, Section M.4.1.1 for residue levels in seed). As part 

of the pesticide tolerance setting process for dicamba (40 CFR 180.227), the EPA 

concluded that residues of dicamba up to 10 ppm in soybean seed are safe (reasonable 

certainty of no harm as defined by FFDCA) for animal consumption. Additionally, as 

discussed in detail in Section J.5.9.2, Monsanto has petitioned the EPA to establish new 

feed tolerances for soybean forage and hay to allow MON 87708 forage and hay to be fed 

to livestock. Dietary exposure to livestock from the feeding of MON 87708 soybean 

forage and hay does not result in an increase in the maximum dietary (feed) exposure of 

dicamba evaluated and deemed safe by EPA. 

MON 87708, MON 87708 DMO and dicamba residues in MON 87708-derived feed 

components do not effect on the safety or nutritional quality of animal feed, therefore the 

deregulation of MON 87708 will not result in a significant impact on animal feed, and 

consequently to animal health. Therefore, the approval in whole and the no action 

alternatives are the same regarding their effects an animal feed. 

No action alternative 
Monsanto Company 10-SY-210U 566 of 721 

The no action alternative would not allow the widespread planting of MON 87708, and 

the use of dicamba herbicide would remain unchanged including the pre-harvest 

treatments which result dicamba residues in soybean seed up to the 10 ppm tolerance. 

Other herbicides are available and would be used in soybean as need to control weeds in 

conventional and herbicide-tolerant soybean. The availability of safe and nutritional 

animal feed from existing soybean crops, including crops containing dicamba or other 

herbicide residues from currently established uses, will continue. Therefore, the no 

action and approval in whole alternatives are similar regarding their potential impacts on 

animal feed. 

J.6. Socioeconomic Effects 

Approval in whole alternative

The decision to deregulate MON 87708 would allow for breeding of this product with 

conventional and previously deregulated biotechnology-derived soybean products of 

diverse genetic backgrounds. These varieties will include soybean with glyphosate 

tolerance (MON 89788; Roundup Ready 2 Yield soybean), to deliver products that 

enhance growers ability to manage weeds. It is expected that breeders and certified seed 

producers would use MON 87708 to develop varieties and to supply seed for planned 

commercial markets in the U.S. Monsanto anticipates that commercial use of 

MON 87708 will include export of soybean and soybean products, and has described 

import approval submission plans in Section I of petition #10-SY-210U. Monsanto’s 

pre-launch stewardship plan for this product is described in Section VIII-L of the petition. 

An extensive analysis of the socioeconomic impacts of herbicide-tolerant crops can be 

found in a recent series of articles published in the on-line journal AgBioForum.38 Much 

of the information below has been summarized from this series of articles. 

The adoption of herbicide-tolerant (Roundup Ready) soybean has been unprecedented 

since its commercialization in 1996. The technology was rapidly adopted by growers. 

Four years after being introduced, approximately 60% of U.S. soybean acreage was 

planted to Roundup Ready soybean; by 2005 over 80% of the soybean acres were planted 

to Roundup Ready soybean, and most recently in 2009 approximately 91% of soybean 

acres were planted to Roundup Ready varieties. Herbicide-tolerant crops have had a 

profound impact on agricultural production globally and similar adoption profiles have 

occurred in other geographies where herbicide-tolerant crops have been introduced (e.g., 

Argentina, Brazil, Canada). Early efforts to understand the reason Roundup Ready 

soybean and other herbicide-tolerant crops achieved such rapid adoption focused on 

profitability, yield and costs (Carpenter and Gianessi, 2001). Information from recent 

grower surveys cite other advantages such as simplicity, convenience, flexibility and 

safety as some of the primary reasons for using Roundup Ready crops (Hurley et al., 

2009). One of the most significant advantages of Roundup Ready cropping systems has 

 

38The source of information can be found at: http://www.agbioforum.org/v12n34/v12n34a00-frisvold.htm

[Accessed May 19, 2010]. 
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been the reduction in labor. The reduction in labor allowed growers more free time to 

pursue other activities and freed up farm management time for non-farm income. 

As with all herbicides, the intensive use of glyphosate in the absence of diverse weed 

management practices (i.e., other herbicide modes-of-action, mechanical cultivation, crop 

rotation or other cultural practices) has led to increased selection pressure and has 

contributed to weed shifts for the hard-to-control broadleaf weeds and/or the 

development of glyphosate-resistant weeds. Diversified weed management is recognized 

by Monsanto, academics and other weed science experts as the guiding principle for 

managing resistance and shifts in weed populations. Diversified weed management is the 

cornerstone of our stewardship messaging and weed management recommendations for 

Monsanto’s product portfolio, including MON 87708 soybean. It is well recognized that 

in situations where there is diversity in weed control methods, few to no resistant weeds 

develop (Dukes and Powles, 2009; Beckie and Reboud, 2009). One strategy 

recommended by experts to prevent or delay the development of resistant weed 

populations is to diversify herbicide weed control methods. 

MON 87708 was developed to provide soybean growers with a simple and flexible 

option to manage existing glyphosate-resistant weeds and better control of other hard-tocontrol weeds as well as delay or prevent further development of glyphosate-resistant 

broadleaf weeds and preserve the use of glyphosate in soybean production. Through 

traditional breeding methods, integration of MON 87708 into the Roundup Ready 

soybean system will allow for effective weed control through the application of 

glyphosate and dicamba. While either herbicide can be applied independently, it is 

envisioned that both glyphosate and dicamba will be tank mixed to simplify the 

application. Growers may also choose to treat or not treat with either herbicide 

depending on the unique situation for each farm and their overall diversified weed 

management plan. 

Monsanto considered the properties of herbicides available for use in soybean in 

developing a second herbicide-tolerance trait to complement the glyphosate tolerance 

trait. Dicamba was selected because it is very complementary to glyphosate in terms of 

the spectrum of broadleaf weeds controlled, its postemergence activity and because very 

few weeds have developed resistance to dicamba over many decades of use. Dicamba 

adds another postemergence herbicide mode-of-action for growers to use in their weed 

control system, thus diversifying the spectrum of weed control options and providing 

better overall weed control superior to current practices. 

The socio-economic benefits of MON 87708 are expected to complement and help 

maintain the benefits growers have realized using the Roundup Ready system. These 

include time and labor savings to growers through the simplicity and flexibility of the 

dicamba/glyphosate weed control system over alternative herbicides that are authorized 

for use in soybean production. In addition, the ability to use dicamba in combination 

with glyphosate will further preserve the benefits the Roundup Ready system has 

provided in the form of increasing adoption of conservation tillage acres. Monsanto and 

academics recommend the use of a third herbicide mode-of-action with soil residual 

activity as part of a comprehensive weed resistance management program to assure that 
Monsanto Company 10-SY-210U 568 of 721 

at least two effective modes-of-action are always used in the cultivated soybean field. 

The cultivation of MON 87708 provides an efficient and effective method to delay or 

prevent the development of glyphosate- or dicamba-resistant weeds, as well as weeds 

resistant to other herbicide class of chemistries, and is expected to outweigh additional 

costs of controlling resistant weeds through the dicamba/glyphosate weed control system. 

No action alternative 

Under the no action alternative, MON 87708 would not be available to commercial 

growers. Alternative herbicides would remain available for control of broadleaf weeds in 

soybean fields, and these herbicides will likely be used instead of dicamba. Monsanto, 

university extension services and other weed science organizations such as HRAC 

promote the use of integrated, diversified weed management systems with soybean 

growers; these recommendations include the rotation of herbicides with different 

modes-of-action or herbicide mixtures containing multiple modes-of-action in their weed 

management program. Growers have increasingly adopted these diversified weed 

management practices, specifically use of multiple modes-of-action, in major corn and 

soybean geographies of the U.S. (see Section J.3.3.6). Thus it is anticipated that the cost 

of the weed management to the soybean grower will not significantly change as a result 

of the cultivation of MON 87708. Technology providers are developing other herbicidetolerant crops that will likely be combined with glyphosate-tolerant crops to simplify 

weed control which may provide similar socioeconomic benefits to growers as those 

described above for MON 87708. On the basis of the above analysis, the approval in 

whole and no action alternatives are similar regarding potential increased costs and 

complexity in weed control for soybean growers. However, the no action alternative 

would not provide as many options to prevent or delay the development of herbicideresistant weeds, and not take advantage of MON 87708 to help sustain the long-term 

agronomic, environmental and socioeconomic value and benefits of the Roundup Ready 

soybean system. 

J.7. Cumulative Effects 

A cumulative impact may be an effect on the environment which results from the 

incremental impact of the proposed action when added to other past, present, and 

reasonably foreseeable future actions. 

J.7.1. Conventional Breeding with Other Biotechnology-derived or Conventional 

Soybean 

The potential effects associated with a determination of nonregulated status of a 

biotechnology-derived crop in combination with the future production of crop varieties 

with different GE traits that are no longer subject to the regulatory requirements of 7 CFR 

part 340 (i.e., “stacked” traits) would be considered a cumulative impact. 

Approval in whole alternative

As previously mentioned, several biotechnology-derived soybean products have been 

deregulated or are under consideration for deregulation, and a list of the events 
Monsanto Company 10-SY-210U 569 of 721 

deregulated by USDA is presented in Table J-4. Once deregulated, MON 87708 may be 

bred with these deregulated biotechnology-derived soybean products as well as with 

conventional soybean, creating new improved varieties. APHIS has determined that none 

of the individual biotechnology-derived soybean products it has previously deregulated 

displays increased plant pest characteristics or creates potentially significant impacts on 

the human environment. APHIS has also concluded that any progeny derived from 

crosses of these deregulated biotechnology-derived soybean products with conventional 

or previously deregulated biotechnology-derived soybean are unlikely to exhibit new 

plant pest properties. This presumption that combined trait biotechnology products are 

unlikely to exhibit new characteristics that would pose new plant pest risks or potential 

environmental impacts not observed in the single event biotech product is based upon 

several facts. Namely 1) stability of the genetic inserts is confirmed in each single event 

product across multiple generations; 2) stability of each of the introduced traits is 

continually and repeatedly assessed as new combined trait varieties are created by plant 

breeders and tested over multiple seasons prior to commercial release; 3) combined trait 

products are developed using the well established process of conventional breeding that 

has been safely used for thousands of years to generate new varieties; and 4) in both 

principle and practical application in the field, it has been shown that two unrelated 

biotechnology traits combined together by conventional breeding, do not display new 

characteristics or properties distinct from those present in the single event biotech 

products. 

Demonstrated Genetic and Phenotypic Stability

An assessment of the stability of the genetic insert in MON 87708 is discussed in Section 

IV of petition #10-SY-210U and summarized here; refer to the petition for more detail. 

Generational stability analysis was assessed by Southern blot analysis and demonstrated 

that the genetic insert present in MON 87708 was maintained across the five breeding 

generations evaluated. These data demonstrate that MON 87708 is stable in its progeny. 

Having established that the genetic material is stable and inherited in a Mendelian 

fashion, and based on experience with MON 87708 in Monsanto’s plant breeding 

program over the course of many generations and many field seasons39, it is concluded 

that the phenotype of MON 87708 is likewise stable. There are numerous examples in 

the literature that confirm that GE events and their associated phenotypes and overall 

characteristics are stably inherited across generations and across genetic backgrounds, 

including when they are combined by conventional breeding to produce combined trait 

products (McCann et al., 2005, Ridley et al., 2011, Zhou et al., 2011). 

Combined Trait Product Performance in Principle and in Practice 

Conventional plant breeding is routinely used to improve crop performance and is 

specifically employed to develop plant varieties that fit particular environments and 

production practices (Powell et al., 2003). The same biological and selective principles 

used for conventional and single GE event variety development are used to combine 

previously approved GE events. When conventional breeding is used to generate 

varieties with combined GE events, these varieties are screened over multiple generations 

 

39 Field trials have been conducted with MON 87708 since 2005. 
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and across diverse growing environments. Typically, product performance and 

agronomic features are evaluated and crop characteristics such as yield, field 

performance, and disease resistance are measured and tested to ensure that the traits are 

stable, heritable, and express the desired phenotype under a wide range of environmental 

conditions. The phenotypic characteristics evaluated during the screening of new 

combined GE event candidate varieties are the result of the plant’s genotype and are the 

culmination of the complex metabolic pathways that are activated in response to 

environmental conditions. The evaluation of phenotypic characteristics allows breeders 

to assess and screen for potential unintended effects produced as a result of the various 

traits (both GE traits and other inherent traits) combined in the variety. Selection during 

the conventional breeding process is valuable in removing undesirable characteristics and 

thereby helps to maintain the safety and quality of the food and/or feed product (Cellini et 

al., 2004; NRC, 2004; WHO, 1995). 

When assessing the safety of combined GE events, it is important to consider 

international guidance regarding conventional breeding and assessments of the 

substantial equivalence of GE events combined by conventional breeding. For example, 

the World Health Organization concludes that a substantial equivalence conclusion can 

be maintained in a combined GE trait variety if substantial equivalence had been 

demonstrated for each of the single event parents. Specifically, they argue that ‘‘...if 

substantial equivalence has been demonstrated both for a [genetically engineered] tomato 

with a gene producing a delayed ripening phenotype and for a [genetically engineered] 

tomato with a gene for herbicide resistance, then crossing these two varieties would result 

in a new variety that would be expected to be substantially equivalent to the parents’’ 

(WHO, 1995). Additional international groups, including the Food and Agriculture 

Organization/World Health Organization (FAO, 1996), International Seed Federation 

(ISF, 2005), and CropLife International (CLI, 2005) similarly advocate basing the safety 

of combined GE events on the safety of the parental GE events. The FDA has stated that 

‘‘narrow crosses are unlikely to result in unintended changes to foods that raise safety or 

other regulatory questions’’ – ‘‘including narrow crosses between different rDNAmodified [GE] lines’’ (U.S. FDA, 2001). 

Thus, the data packages that are developed for single GE events are useful in establishing 

the safety of the combined GE event product. Accordingly, single GE events previously 

assessed to be as safe as their conventional counterparts should continue to be safe when 

combined through conventional plant breeding. As a result, the rigorous safety 

assessments and plant pest risk assessments conducted on single GE events, which were 

deemed to pose no more risk than their conventional counterparts, also can be used to 

predict the safety and potential risk of the combined GE event product. As described 

below, this assertion is supported by direct experience gained through the commercial 

planting and utilization of combined GE event products as well as a body of information 

collected to support authorizations where combined GE event products require additional 

assessment. 

There are many examples of combined trait biotech products that have been produced 

commercially on millions of acres without incident over the past decade in both the US 
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and in other countries40. In the US, there are over 30 combined trait products that have 

been made commercially available. In 2011, 49% of the total corn acres planted (92 M 

acres) and 58% of total cotton planted (13M acres) contained multiple (2 or more) GE 

events (USDA-ERS, 2011) 41 . Combined trait biotech products were planted on 40 

million hectares or 25% of the global biotech area of 160 million hectares in 201142. 

The safety of commercially available combined trait biotech products has been 

well-demonstrated in multiple independent reports that document the continually 

increasing acceptance and use of these products by farmers globally (Brookes and 

Barfoot, 2009; James, 2009; Lemaux, 2008; Sankula, 2006). 

In addition, regulatory agencies in some countries request additional characterization of 

combined GE events and comparisons to single event parental controls and conventional 

comparators. These additional studies may include analysis of phenotype, molecular 

characteristics, protein characteristics, morphology and compositional evaluation. 

Analyses of combined GE events compared to parental controls have consistently 

revealed the following: no phenotypic differences from parental events; molecular 

characteristics that are the same as parental events with all events inherited stably; levels 

of introduced proteins comparable to the single event parents; no morphological 

differences compared to parental events; compositional equivalence based on nutrient and 

anti-nutrient evaluations, and no pleiotropic or toxic effects compared to the conventional 

non-GE crop (Pilacinski et al., 2011). 

Summary

Traditional soybean breeding has an established history of safe use, and use of 

MON 87708 in breeding programs is expected to behave in a manner similar to other 

conventional traits and biotechnology-derived traits. Given that there have been no plant 

pest characteristics associated with MON 87708, or with any of the previously 

deregulated events listed in Table J-5, no significant impacts are expected to other 

soybean through the use of MON 87708 in breeding programs and in combination with 

any of the previously deregulated biotechnology-derived soybean products. 

All biotechnology-derived soybean products on the market today have satisfactorily 

completed the FDA consultation process established to review the safety of foods and 

feeds derived from biotechnology-derived crops for human and animal consumption (see 

Table J-5). As mentioned previously, MON 87708 is intended to be combined with 

MON 89788 (Roundup Ready 2 Yield soybean) through traditional breeding. 

MON 87708 is expected to be utilized broadly in future additional combined trait 

soybean products, and thus will likely be bred with other biotechnology-derived soybean 

products that have been deregulated or have deregulation petitions pending before APHIS 

(e.g., MON 87701, MON 87705, MON 87769; see Table J-4 for current status). No 

impacts to public health (e.g., food or feed safety) are expected due to combination of 

these events through conventional breeding because the deregulated events have or will 

 

40 See http://www.biotradestatus.com for list of combined trait products that have previously been 

commercialized. 

41 See http://www.ers.usda.gov/data/biotechcrops/adoption.htm 42 http://www.isaaa.org/resources/publications/briefs/43/executivesummary/default.asp
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have completed a safety consultation with FDA and on the basis of knowledge of the type 

of modifications made to each of the deregulated events, and to the events under review, 

the biochemical pathways are unrelated to the trait introduced in MON 87708 and 

therefore not expected to interact or result in the production of novel properties or 

constituents. 

The decision to approve in whole MON 87708 would also allow for breeding of this 

product with conventional soybean of diverse genetic background. No impacts to public 

health (e.g., food or feed safety) or environmental safety are expected due to the breeding 

of MON 87708 with these other soybean because these varieties have an established 

history of safe use. 

Based on the above analysis, the approval in whole and no action alternatives are similar 

regarding potential cumulative impacts associated with the conventional breeding of 

MON 87708 with other conventional or biotechnology-derived soybean products. 

No action alternative 

Under the no action alternative, MON 87708 would not be used to produce new soybean 

varieties or in the commercial production of soybean because the trait would remain 

regulated. There are no effects that have been identified from combining MON 87708 

with other Monsanto biotechnology-derived soybean that have been deregulated by 

USDA; therefore, the no action and approval in whole alternatives are similar regarding 

their potential cumulative impacts associated with conventional breeding with other 

soybean products. 

J.7.2. Conservation Tillage 

Approval in whole alternative 

The single most damaging effect on land due to agriculture is loss of soil caused by 

tillage. Tillage is primarily performed for seed bed preparation and has the added benefit 

as a weed control measure. Roundup Ready crops, with Roundup Ready soybean being 

the most widely adopted, have enabled significant implementation/adoption of no-till or 

reduced tillage methods for weed control (Duke and Powles, 2009). 

In 1995, before the introduction of Roundup Ready soybean, approximately 27% of the 

U.S. soybean acres used no-till production. By 2004, no-till acreage increased to 36% of 

the total soybean acres (Sankula, 2006). The most recent surveys indicate that 39% of 

the soybean acres are produced using no-till methods (CTIC, 2007). A few states provide 

statistics on the adoption of no-till acres in their state. In 2006 in Illinois, no-till farming 

was used on 51% of the soybean acres. The University of Illinois Extension Service 

attributed that figure to the fact that 90% of the state’s soybean acres were planted to 

Roundup Ready soybean varieties, along with other factors, such as high fuel prices, 

improved equipment, higher yields using no-till practices, and better grower awareness of 

the advantages to soil and water quality from no-till farming. Approval in whole would 

allow for continued adoption and preservation of conservation tillage methods resulting 
Monsanto Company 10-SY-210U 573 of 721 

in positive cumulative environmental impacts. Therefore, the approval in whole and no 

action alternative are similar regarding the potential cumulative impacts on conservation 

tillage practices 

No action alternative 

Under this alternative, MON 87708 would not be grown commercially and growers 

would not be able to utilize dicamba to the same extent as they would if APHIS granted 

approval in whole to MON 87708. This alternative would limit grower’s options 

regarding the selection of herbicides available for use in conservation tillage. This option 

could potentially result in an increased use of tillage over time and potential loss of no-till 

acres in some soybean fields, thus decreasing the use of conservation tillage methods and 

environmental benefits associated with conservation tillage. Overall, the no-action and 

approval in whole alternatives are similar regarding the potential cumulative impacts on 

conservation tillage practices. 

J.7.3. Resistant Weeds 

Approval in whole alternative

Another cumulative effect includes the potential for development of herbicide-resistant 

weeds, including weed populations with multiple resistance. Integration of MON 87708 

into the Roundup Ready soybean system has the potential to delay or prevent 

development of dicamba- and glyphosate-resistant weeds, as well as resistance to other 

herbicide classes such as PPO herbicides. These issues have been discussed in Section 

J.5.3.1 and in Appendix K of petition #10-SY-210U, but warrant additional discussion 

under the assessment of cumulative effects. The development of resistant weed 

populations occurs over a period of time (Heap, 2009), and is an foreseeable consequence 

of herbicide use in the absence of diverse weed management practices as is discussed in 

additional detail in Section J.3.3.6. One of the strategies recommended by experts to 

prevent or delay the development of resistant weed populations is to diversify weed 

control methods. The use of herbicides such as dicamba in conjunction with glyphosate 

provides growers with an herbicide system with two different modes-of-action. Thus, it 

is foreseeable that integration of MON 87708 into the Roundup Ready soybean system 

with the use of dicamba and glyphosate together would have a low probability for 

increases in new weed populations developing resistance to either glyphosate or dicamba. 

In addition there would be the potential for fewer new species to evolve or develop 

resistance for either herbicide compared to a glyphosate or dicamba herbicide system 

alone. In the MON 87708 weed management system the use of glyphosate is not 

expected to increase. Even thought there will be an increase in the use of dicamba, it is 

clear that in situations where there is diversity in weed control methods, there will be a 

low probability for resistant weeds to develop (Dukes and Powles, 2009; Beckie and 

Reboud, 2009). In the unlikely case that broadleaf weeds were to develop with resistance 

to dicamba or with accumulated resistance to both glyphosate and dicamba, existing 

cultivation and alternative herbicide tools (see Section VIII.F.1 for description of 

alternative herbicides) would remain potential options to provide effective control. 
Monsanto Company 10-SY-210U 574 of 721 

Therefore, the approval in whole and no action alternatives are similar regarding potential 

cumulative impacts on weed resistance. 

No action alternative 

Other herbicide options for weed resistance management are available to growers. Under 

the no action alternative, grower’s options regarding the selection of herbicides available 

for use to control weeds in soybean and specifically herbicides with different modes-ofaction would be more limited than the approval in whole alternative as a result of the 

inability to use dicamba in the weed management system for soybean. Other soybean 

herbicides are available to growers to use either with or without the dicamba-glyphosate 

tolerant soybean system, and will remain available to growers in the future. However, 

herbicide weed management systems under the no action alternative are not as efficient 

and effective for managing resistant weeds and incorporating additional herbicide modesof-action. Thus, it is reasonable foreseeable that some increase in the development of 

additional herbicide-resistant weed populations or development of new herbicide resistant 

weed species compared to the approval in whole alternative will result. Therefore, the no 

action and approval in whole alternatives are similar regarding the potential cumulative 

effects on weed resistance. 

J.8. Summary 

MON 87708 has been thoroughly characterized and the extensive body of information 

presented in Sections I through IX of the petition demonstrate that MON 87708 does not 

present a plant pest risk, has no significant impact on threatened and endangered species 

or biodiversity, and will not impact the commercial interests of soybean growers or those 

involved in the marketing and sale of soybean and soybean products. The amount of land 

devoted to farming (specifically to soybean) has not changed with the introduction of 

biotechnology-derived crops. Similarly, no significant change in the use of agricultural 

land or amount of land devoted to farming would be expected to occur with the 

commercial introduction of MON 87708 integrated into the Roundup Ready soybean 

system. MON 87708 integrated into the Roundup Ready soybean system will offer 

growers the opportunity to expand the use of dicamba to late preemergence (up to 

cracking) and postemergence in-crop applications and, thereby, potentially delay or 

prevent the development of dicamba- and glyphosate-resistant weeds as well as resistance 

to other soybean herbicides in U.S. soybean fields. Other cultivation practices typically 

employed for production of commodity soybean, and management practices will remain 

unchanged. Growers are accustomed to the use of herbicides in their soybean production 

systems. For these reasons, the proposed action to grant nonregulated status to 

MON 87708 does not represent a significant impact to the environment. 

 
Monsanto Company 10-SY-210U 575 of 721 

References for Appendix J 

Abrams, R.I., C.R. Edwards, and T. Harris. 1978. Yields and cross-pollination of 

soybeans as affected by honey bees and alfalfa leafcutting bees. American Bee Journal 

118:555-560. 

Abud, S., P.I. Mello de Souza, C.T. Moreira, S.R.M. Andrade, A.V. Ulbrich, G.R. 

Vianna, E.L. Rech, and F.J. Lima Aragao. 2003. Pollen dispersal in transgenic soybean 

plants in the Verrado region. Pesquisa Agropecuária Brasileira 38:1229-1235. 

Al-Kaisi, M. 2001. Value of crop rotation in nitrogen management. Iowa State University. 

http://www.ipm.iastate.edu/ipm/icm/2001/4-23-2001/valuen.html. 

Al-Kaisi, M., M.H. Hanna, and M. Tidman. 2003. Crop rotation considerations for 2004 

management season rotation. Department of Entomology, Iowa State University. 

http://www.ent.iastate.edu/ipm/icm/2003/croprotation.html. 

Altieri, M.A. 1999. The ecological role of biodiversity in agroecosystems. Agriculture, 

Ecosystems and Environment 74: 19-31. 

Aneja, V. P., Schlesinger, W. H., & Erisman, J. W. (2009). Effects of agriculture upon 

the air quality and climate: Research, policy, and regulations. Environmental Science & 

Technology, 43, 4234–4240 

AOSCA. 2009. Seed certification handbook. Association of Official Seed Certifying 

Agencies, Moline, Illinois. 

Aref, S. and D.R. Pike. 1998. Midwest farmers’ perceptions of crop pest infestations. 

Agronomy Journal 90:819-825. 

ASA. 2009. Soy Stats. American Soybean Association, St. Louis, Missouri. 

ASA. 2010. Domestic utilization, U.S. soybean use by livestock 2009. American 

Soybean Association, St. Louis, Missouri. http://www.soystats.com/2010/Defaultframes.htm. Accessed July 26, 2010. 

ASA. 2011. Soy stats 2011. American Soybean Association, St. Louis, Missouri. 

http://www.soystats.com/2011/Default-frames.htm. Accessed February 10, 2012. 

ASTA. 2011. Existing U.S. industry production practices that address coexistence. 

American Seed Trade Association, Alexandria, Virginia. 

Baker, H.G. 1965. Characteristics and modes of origin of weeds. Pages 147-172 in The 

Genetics of Colonizing Species. H.G. Baker and G.L. Stebbins (eds.). Academic Press, 

New York, New York. 
Monsanto Company 10-SY-210U 576 of 721 

Baker, J., Southard, R., & Mitchell, J. (2005). Agricultural dust production in standard 

andconservation tillage systems in the San Joaquin Valley. Journal of Environmental 

Quality, 34, 1260-1269. doi: 10.2134/jeq2003.034 

Beckie, H.J. 2006. Herbicide-resistance: Management tactics and practices. Weed 

Technology 20:793-814. 

Beckie, H.J. and X. Reboud. 2009. Selecting for weed resistance: Herbicide rotation and 

mixture. Weed Technology 23:363-370. 

Boerma, H.R. and J.E. Specht. 2004. World distribution and trade of soybean and 

managing inputs for peak production; soybean production in the U.S.A. Pages 3, 502, 507 

and 522, In Soybeans: Improvement, production, and uses. Boerma H.R. and J.E. Specht 

(eds.). ASA, CSSA-SSSA, Madison, Wisconsin. 

Brookes, G. and P. Barfoot. 2004. Co-existence case study of North America: 

Widespread GM cropping with non GM and organic crops. PG Economics Ltd., 

Dorchester, United Kingdom. 

Brookes, G. and P. Barfoot. 2009. GM crops: Global socio-economic and environmental 

impacts 1996-2007. PG Economics, Ltd, Dorchester, United Kingdom. 

www.pgeconomics.co.uk/pdf/2009globalimpactstudy.pdf Accessed May 12, 2010. 

Buckley, D.H. and T.M. Schmidt. 2001. The structure of microbial communities in soil 

and the lasting impact of cultivation. Microbial Ecology 42: 11-21. 

Buckley, D.H. and T.M. Schmidt. 2003. Diversity and dynamics of microbial 

communities in soils from agro-ecosystems. Environmental Microbiology 5: 441-452. 

Carpenter, J.E. and L.P. Gianessi. 2001. Why U.S. farmers have adopted genetically 

modified crops and the impact on U.S. agriculture. AgBiotechNet 3:1-4. 

Carpenter, J., A. Felsot, T. Goode, M. Hammig, D. Onstad and S. Sankula. 2002. 

Comparative environmental impacts of biotechnology-derived and traditional soybean, 

corn, and cotton crops. Council for Agricultural Science and Technology, Ames, Iowa. 

CAST. 2011. Herbicide-resistant weeds threaten soil conservation gains: Finding a 

balance for soil and farm sustainability. Council for Agricultural Science and Technology, 

Ames, Iowa. 

Caviness, C.E. 1966. Estimates of natural cross-pollination in Jackson soybeans in 

Arkansas. Crop Science 6:211-212. 

Cellini, F., Chesson, A., Colquhoun, I., Constable, A., Davies, H.V., Engel, K.H., 

Gatehouse, A.M.R., Karenlampi, S., Kok, E.J., Leguay, J.J., Lehesranta, S., Noteborn, H., 

Pedersen, J., Smith, M., 2004. Unintended effects and their detection in genetically 

modified crops. Food and Chemical Toxicology 42, 1089–1125. 
Monsanto Company 10-SY-210U 577 of 721 

Chiang, Y.C., and Y.T. Kiang. 1987. Geometric position of genotypes, honeybee 

foraging patterns and outcrossing in soybean. Botanical Bulletin of Academia Sinica 

28:1-11. 

CLI. 2005. Position paper: Regulation of plant biotechnology products containing two or 

more traits combined by conventional plant breeding. Crop Life International, Brussels, 

Belgium. 

CTIC. 2004. National survey shows more farmers choose conservation. Conservation 

Technology Information Center, West Lafayette, Indiana. 

http://www.ctic.purdue.edu/ctic/ctic.html [Accessed November 9, 2004]. 

CTIC. 2007. Amendment to the national crop residue management survey summary. 

www.conservationinformation.org/pdf/National_Summary_2007_Amendment.pdf 

Accessed on June 14, 2010. 

CTIC. 2010. 2010 Amendment to the national crop residue management survey summary. 

www.conservationinformation.org/pdf/National_Summary_2007_Amendment.pdf 

Accessed June 14, 2010. 

CTIC. 2011. Top 10 conservation tillage benefits: Conservation for agriculture's future. 

Conservation Technology Information Center, West Lafayette, Indiana. 

http://www.ctic.purdue.edu/resourcedisplay/293/ Accessed September 21, 2011

Cui, Z., A.T. James, S. Miyazaki, R.K. Wilson and T.E. Carter. 2004. The breeding of 

specialty soybeans for traditional and new soyfoods. Pages 264-322 in Proceeding of the 

American Oil Chemists' Society, Urbana, Illinois. 

Dalley, C.D., K.A. Renner, and J.J. Kells. 2001. Weed competition in Roundup Ready 

soybean and corn. Michigan State University, Dept of Crop and Soil Science. 

http://www.ipm.msu.edu/CAT05_fld/pdf/5-26FactSheet.pdf Accessed on June 15, 2010. 

Diggle, A.J., P.B. Neve and F.P. Smith. 2003. Herbicides used in combination can reduce 

the probability of herbicide resistance in finite weed populations. Weed Research 

43:371–382. 

Dorrance, A.E., M.A. Draper, and D. Hershman. 2007. Using foliar fungicides to manage 

soybean rust. Ohio State University, Columbus, Ohio. 

Duke, S.O., and S.B. Powles. 2009. Glyphosate-resistant crops and weeds: Now and in 

the future. AgBioForum 12:346-357. 

Durkin, P. and S. Bosch. 2004. Dicamba - human health and ecological risk assessment - 

final report. Prepared for USDA, Forest Service. SERA TR 04-43-17-06d. 

http://www.fs.fed.us/foresthealth/pesticide/pdfs/112404_dicamba.pdf Accessed May 26, 

2010. 
Monsanto Company 10-SY-210U 578 of 721 

European Commission. 2007a. Annex B9. Ecotoxicology. Draft Assessment Report 

(DAR): Initial Risk Assessment Provided by the Rapporteur Member State Denmark for 

the Existing Active Substance Dicamba of the Third Stage (part B) of the Review 

Programme Referred to in Article 8(2) of the Council Directive 91/414/EEC. European 

Commission, Brussels, Belgium. 

European Commission. 2007b. Annex B6. Toxicology and metabolism. Draft 

Assessment Report (DAR): Initial Risk Assessment Provided by the Rapporteur Member 

State Denmark for the Existing Active Substance Dicamba of the Third Stage (part B) of 

the Review Programme Referred to in Article 8(2) of the Council Directive 91/414/EEC. 

European Commission, Brussels, Belgium. 

FAO. 1996. Joint FAO/WHO expert consultation on biotechnology and food safety. 

Paper 61. Food and Agriculture Organization, World Health Organization, Rome, Italy. 

Fawcett, R. and D. Towery. 2002. Conservation tillage and plant biotechnology: How 

new technologies can improve the environment by reducing the need to plow. 

Conservation Technology Information Center, Purdue, Indiana. 

Felsot, A.S., J.B. Unsworth, J.B.H.J. Linders, G. Roberts, D. Rautman, C. Harris and E. 

Carazo. 2010. Agrochemical spray drift; Assessment and mitigation—A review. Journal 

of Environmental Science and Health, Part B 46: 1-23. 

Frank AR, Kemman R. 2011. Information to Support an Endangered Species 

Assessment for Dicamba Use in Dicamba-Tolerant Soybeans: A County-Level Analysis 

for Animal Taxa. Monsanto Technical Report MSL-22547. EPA MRID 48644204. 

Frank AR, Kemman R. 2012. Information to Support an Endangered Species Assessment 

for Dicamba Use in Dicamba-Tolerant Soybeans: A County-Level Analysis for Plant 

Taxa. Monsanto Technical Report MSL-23442. EPA MRID 48644206

Gage, D.J. 2004. Infection and invasion of roots by symbiotic, nitrogen-fixing rhizobia 

during nodulation of temperate legumes. Microbiology and Molecular Biology Reviews

68:280-300. 

Galle, A.M., G.M. Linz, H.J. Homan and W.J. Bleier. 2009. Avian use of harvested crop 

fields in North Dakota during spring migration. University of Nebraska-Lincoln, USDA 

National Wildlife Research Center, Lincoln, Nebraska. 

Gianessi, L.P., C.S. Silvers, S. Sankula and J.E. Carpenter. 2002. Plant biotechnology: 

Current and potential impact for improving pest management in U.S. Agriculture, an 

analysis of 40 case studies. National Center for Food and Agricultural Policy. 

Washington, DC. 

Givens, W.A., D.R. Shaw, W.G. Johnson, S.C. Weller, B.G. Young, R.G. Wilson, M.D.K. 

Owen, and D. Jordan. 2009. A grower survey of herbicide use patterns in glyphosateresistant cropping systems. Weed Technology 23:156-161. 

Harlan, J.R. 1975. Our vanishing genetic resources. Science 188: 617-621. 
Monsanto Company 10-SY-210U 579 of 721 

Heap, I. 2009. International survey of herbicide resistant weeds. 

http://www.weedscience.org/in.asp Accessed November 11, 2009. 

Heap, I. 2011. International survey of herbicide resistant weeds. 

http://www.weedscience.org/in.asp Accessed December 11, 2011.Heatherly, L.G., and 

H.F. Hodges. 1999. Soybean production in the Midsouth. CRC Press, Boca Raton, 

Florida. 

Helsel, Z.R. and H.C. Minor. 1993. Soybean production in Missouri. University of 

Missouri, Department of Agronomy. Publication G4410. 

http://extension.missouri.edu/explore/agguides/crops/g04410.htm Accessed November 

11, 2009. 

Herro, A. 2008. Adjustments to agriculture may help mitigate global warming. 

Worldwatch Institute, Washington, D.C. http://www.worldwatch.org/node/5580 

[Accessed April 7, 2011]. 

Higley, L.G., and D.J. Boethel. 1994. Handbook of soybean insect pests. The 

Entomological Society of America, Lanham, Maryland. 

Hoeft, R.G., E.D. Nafziger, R.R. Johnson, and S.R. Aldrich. 2000. Soybean as a crop. 

Pages 31, 36, 38, 39, 41, 43, 47, 86, 89, 93, 96, 120, and 208, In Modern Corn and 

Soybean. Production MCSP Publications, Champaign, Illinois. 

Honegger, JL. 2012. Information to Support an Endangered Species Assessment for 

Dicamba Use in Dicamba-Tolerant Soybeans: Overview of an Evaluation of Potential 

Exposure and Biological Effects. Monsanto Technical Report MSL-22547. EPA MRID 

48644201 

HRAC. 2010. Herbicide Resistance Action Committee. Publications: Guideline to the 

management of herbicide resistance. 

http://www.hracglobal.com/Publications/ManagementofHerbicideResistance/tabid/225/D

efault.aspx Accessed June 29, 2010. 

Hurley, T.M., P.D. Mitchell, and G.B. Frisvold. 2009. Characteristics of herbicides and 

weed-management programs most important to corn, cotton and soybean growers. 

AgBioForum 12:269-280. 

Hymowitz, T. 1987. Introduction of the soybean to Illinois. Economic Botany 41:28-32. 

Iowa State University. 2006. Cover crops in the corn belt: survey finds underused 

potential as conservation tool. Leopold Center for Sustainable Agriculture. Winter 2006 

Letter. http://www.leopold.iastate.edu/news/leopold-letter/2006/winter/cover-crops-cornbelt-survey-finds-underused-potential-conservation Accessed February 12, 2012. 

ISF. 2005. Genetically modified crops and plant breeding. International Seed Federation, 

Nyon, Switzerland. 
Monsanto Company 10-SY-210U 580 of 721 

ISO. 2009. International Organization for Standardization. Selection and use of the ISO 

9000 family of standards. http://www.iso.org Accessed Jan. 9, 2009. 

James, C., 2009. Global status of commercialized biotech/GM crops: 2009. ISAAA Brief 

41. International Service for the Acquisition of Agri-Biotech Applications, Ithaca, New 

York. 

Johnson, B., B. Young, J. Matthews, P. Marquardt, C. Slack, K. Bradley, A. York, S. 

Culpepper, A. Hager, K. Al-Khatib, L. Steckel, M. Moechnig, M. Loux, M. Bernards and 

R. Smeda. 2010. Weed control in dicamba-resistant soybeans. Plant Management 

Network, St. Paul, Minnesota. 

http://www.plantmanagementnetwork.org/sub/cm/research/2010/dicamba/ Accessed 

September 20, 2010. 

Kerley, M.S. and G.L. Allee. 2003. Modifications in soybean seed composition to 

enhance animal feed use and value: Moving from a dietary ingredient to a functional 

dietary component. AgBioForum 6:14-17. 

Landis, D.A., F.D. Menalled, A.C. Costamagna and T.K. Wilkinson. 2005. Manipulating 

plant resources to enhance beneficial arthropods in agricultural landscapes. Weed Science

53:902-908. 

Lemaux, P.G., 2008. Genetically engineered plants and foods: A scientist’s analysis of 

the issues (part I). Annual Review of Plant Biology 59: 771–812. 

Loux, M.M., D. Doohan, A.F. Dobbels, W.G. Johnson, G.R.W. Nice, T.N. Jordan, and 

T.T. Bauman. 2009. Weed control guide for Ohio and Indiana. Purdue University and 

Ohio State. Publication WS16. http://www.btny.purdue.edu/Pubs/ws/ws-16/ Accessed 

on June 14, 2010. 

Lovett, S., P. Price and J. Lovett. 2003. Managing riparian lands in the cotton industry. 

Cotton Research and Development Corporation, Narrabri, Australia. 

LSU. 2009. Asian soybean rust: Background. Louisiana State University, Baton Rouge, 

Louisiana. 

http://www.agctr.lsu.edu/en/crops_livestock/crops/soybeans/Asian_Soybean_Rust/ 

Accessed June 17, 2010. 

MAFRI. 2004. Salvaging immature and frosted crops for silage. Manitoba Agriculture, 

Winnipeg, Manitoba. http://www.gov.mb.ca/agriculture/news/topics Accessed May 12, 

2010.

Mannering, J.V., D.R. Griffith and K.D. Johnson. 2007. Winter cover crops - Their value 

and management. AY-247. Purdue University Cooperative Extension Service, West 

Lafayette, Indiana. 
Monsanto Company 10-SY-210U 581 of 721 

McCann, M.C., K. Liu, W.A. Trujillo and R.C. Dobert. 2005 Glyphosate-Tolerant 

Soybeans Remain Compositionally Equivalent to Conventional Soybeans (Glycine max

L.) during Three Years of Field Testing. Journal of Agricultural and Food Chemistry 

53:5331-5335. 

Monsanto Company. 2011. Letter of completion of biotechnology consultaions on BNF 

00125. Food & Drug Administration, Department of Health & Human Services, Center 

for Food Safety and Applied Nutrition, Office of Food Additive Safety. October 11, 2011. 

http://www.fda.gov/Food/Biotechnology/Submissions/ucm282993.htm 

Mueth, MG and JE Foster. 2010. Determination of Dicamba Residue Decline in Forage 

after Application to Dicamba-Tolerant Soybean MON 87708 × MON 89788. Monsanto 

Technical Report MSL-22493. EPA MRID 48644205 

Nice, G. and B. Johnson. 2005. Indiana's top ten most problematic weeds. Purdue 

Extension Weed Science. http://www.btny.purdue.edu/WeedScience/2005/topten05.pdf 

Accessed November 11, 2009. 

NDSU. 2002. Soybean production guide for North Dakota and Northwestern Minnesota. 

North Dakota State University Extension Service and North Dakota Soybean Council, 

Fargo, North Dakota. http://www.ag.ndsu.edu/pubs/plantsci/rowcrops/a1172.pdf 

Accessed November 11, 2009. 

NRC. 2004. New approaches for identifying unintended changes in food composition. 

Pages 73-102 in Safety of Genetically Engineered Foods: Approaches to Assessing 

Unintended Health Effects. Institute of Medicine and National Research Council, 

National Academies Press, Washington, D.C. 

OECD. 2000. Consensus document on the biology of Glycine max (L.) merr. (soybean). 

ENV/JM/MONO(2000)9. Series on Harmonization of Regulatory Oversight in 

Biotechnology No.15. Organisation for Economic Co-operation and Development, Paris, 

France. 

Pedersen, P. 2008. Soybean production. Iowa State University Extension. 

http://extension.agron.iastate.edu/soybean Accessed November 11, 2009. 

Peterson, D., C. Mayo and S. Seifert-Higgins. 2011. Weed control in dicamba tolerent 

soybeans in Kansas. North Central Weed Science Society 66th Annual Meeting, 

Milwaukee, Wisconsin. 

Pilacinski, W., A. Crawford, R. Downey, B. Harvey, S. Huber, P. Hunst, L.K. Lahman, S. 

Macintosh, M. Pohl, C. Rickard, L. Tagliani and N. Weber. 2011. Plants with genetically 

modified events combined by conventional breeding: An assessment of the need for 

additional regulatory data. Food and Chemical Toxicology 49: 1-7. 

PMRA. 2008. Pest Management Regulatory Agency Re-evaluation – dicamba. Pest 

Management Regulatory Authority, Health Canada. Ottawa, Canada. 
Monsanto Company 10-SY-210U 582 of 721 

Powell, W.,R. Waugh, J. Bradshaw, J. Russell, L. Ramsay and B.P. Forster. 2003. 

Introduction to classical genetics and plant breeding. Pages 3-29 in Handbook of Plant 

Biotechnology. Volume 1. P. Christou and H. Klee (eds.). John Wiley and Sons, Inc., 

Hoboken, New Jersey. 

Powles, S.B., C. Preston, I.B. Bryan and A.R. Jutsum. 1996. Herbicide resistance: Impact 

and management. Pages 57-93 in Advances in Agronomy. Volume 58. D.L. Sparks (ed.). 

Academic Press, San Diego, California. 

Prince, J.M., D.R. Shaw, W.A. Givens, M.D.K. Owen, S.C. Weller, B.G. Young, R.G. 

Wilson and D.L. Jordan. 2011. Benchmark study: IV. Survey of grower practices for 

managing glyphosate-resistant weed populations. Weed Science Society of America. 

Accessed December 15, 2011. 

Ray, J.D., T.C. Kilen, C.A. Abel, and R.L. Paris. 2003. Soybean natural cross-pollination 

rates under field conditions. Environmental Biosafety Research 2:133-138. 

Ridley, W.P., G.G. Harrigan, M.L. Breeze, M.A. Nemeth, R.S. Sidhu and K.C. Glenn. 

2011. Evaluation of compositional equivalence for multitrait biotechnology crops. 

Journal of Agricultural and Food Chemistry 59:5865-5876. 

Sankula, S. 2006. Quantification of the impacts on U.S. agriculture of biotechnologyderived crops planted in 2005. National Center for Food and Agriculture Policy, 

Washington, D.C. 

Schuler LJ, Leopold VA, Mortensen SR. 2011. Information to Support an Endangered 

Species Assessment for Dicamba Use in Dicamba-Tolerant Soybeans: Refined Analysis 

for Terrestrial Animals. Monsanto Technical Report MSL-22546. EPA MRID 

48644203. 

SDTF. 1997. A summary of ground application studies. Spray Drift Task Force, St. Louis, 

Missouri. http://www.agdrift.com/PDF_FILES/Ground.pdf Accessed August 10, 2010. 

Sharpe, T. 2010. Tarheel wildlife - A guide for managing wildlife on private lands in 

North Carolilna. North Carolina Wildlife Resources Commission, Raleigh, North 

Carolina. 

Soyatech. 2010. Soya and oilseed bluebook: Statistics and reference. Southwest Habor, 

Maine. 

Stewart, C.N., M.D. Halfhill and S.I. Warwick. 2003. Transgene introgression from 

genetically modified crops to their wild relatives. Nature Reviews Genetics 4:806-817. 

Tranel, P.J., C.W. Riggins, M.S. Bell and A.G. Hager. 2010. Herbicide resistances in 

Amaranthus tuberculatus: A call for new options. Journal of Agricultural and Food 

Chemistry 59: 5808-5812. Tiedje, J.M., S. Asuming-Brempong, K. Nusslein, T.L. Marsh 

and S.J. Flynn. 1999. Opening the black box of soil microbial diversity. Applied Soil 

Ecology 13: 109-122. 
Monsanto Company 10-SY-210U 583 of 721 

Towery, D. and S. Werblow. 2010. Facilitating conservation farming practices and 

enhancing environmental sustainability with agricultural biotechnology: Exective 

summary. Conservation Technology Information Center, West Lafayette, Indiana. 

http://www.ctic.org/BiotechSustainability. Accessed January 27, 2012. 

USB. 2011a. U.S. soybean production: A comparison of sustainable production systems 

for conventional, biotech, and organic soybeans. United Soybean Board, Chesterfield, 

Missouri. 

USB. 2011b. Soyfoods Guide 2011. United Soybean Board, Chesterfield, Missouri. 

http://www.soyconnection.com/soyfoods/pdf/soyfoods_guide.pdf Accessed on January 

27, 2012U.S. EPA. 1992. Pesticides in groundwater database: A compilation of 

monitoring studies: 1971-1991. National Summary [EPA 734-12-92-001]. U.S. 

Environmental Protection Agency, Office of Prevention Pesticides and Toxic Substances, 

Washington, D.C. 

U.S. EPA. 2002. Environmental implications of conservation tillage: A systems 

approach. Office of Research and Development, National Exposure Research Lab. 

Document: EPA/600/D-85/147 (NTIS PB85237089). 

http://cfpub.epa.gov/si/si_public_record_report.cfm?dirEntryId=39100&fed_org_id=770

&SIType=PR&TIMSType=&showCriteria=0&address=nerl&view=citation&keyword=P

esticides&sortBy=pubDateYear&count=100&dateBeginPublishedPresented Accessed on 

May 19, 2010. 

U.S. EPA. 2004. Overview of the ecological risk assessment process in the Office of 

Pesticide Programs. U.S. Environmental Protection Agency, Endangered and Threatened 

Species Effects Determinations, Washington, D.C. 

U.S. EPA. 2005a. Dicamba: HED chapter of the reregistration eligibility decision 

document, phase 1. PC Code 029801; DP Barcode: D317720 (Memorandum, C. Olinger 

and Y. Yang, September 13). Health Effects Division, Office of Prevention, Pesticides 

and Toxic Substances, U.S. Environmental Protection Agency, Washington, D.C. 

U.S. EPA. 2005b. Environmental Fate and Ecological Risk Assessment for the 

Reregistration of Dicamba and Dicamba Sodium, Potassium, Diglycoamine, 

Dimethylamine and Isopropylamine Salts (Memorandum, W. Erickson, I. Abdel-Saheb 

and S. Borges, August 31). Environmental Fate Effects Division, Office of Prevention, 

Pesticides and Toxic Substances, U.S. Environmental Protection Agency, Washington, 

DC. 

U.S. EPA. 2007a. National water quality inventory: Report to Congress, 2002 reporting 

cycle: Findings. http://www.epa.gov/305b/2002report/ Accessed on May 19, 2010. 

U.S. EPA. 2007b. Risks of atrazine use to federally listed endangered Pallid Sturgeon 

Scaphirhyncus albus. 

http://www.epa.gov/oppfead1/endanger/liststatus/effects/pallid_sturgeon_eff_deter_08-

31-07.pdf. Accessed January 1, 2008.
Monsanto Company 10-SY-210U 584 of 721 

U.S. EPA 2008a. Agricultural management practices for water quality protection. 

http://www.epa.gov/watertrain/agmodule/ Accessed on May 19, 2010. 

U.S. EPA. 2008b. National assessment database. http://www.epa.gov/waters/ir/ 

Accessed on May 19, 2010. 

U.S. EPA. 2008c. Risks of glyphosate use to the Federally threatened California Reglegged frog. . http://www.epa.gov/owow/305b/2004report/ Accessed May 19, 2010. 

U.S. EPA. 2009a. National water quality inventory: Report to Congress, 2004 reporting 

cycle: Findings. http://www.epa.gov/owow/305b/2004report/ Accessed on May 19, 2010. 

U.S. EPA. 2009b. Reregistration eligibility decision for dicamba and associated salts. 

U.S. Environmental Protection Agency, Office of Prevention, Pesticides, and Toxic 

Substances, June 8, 2006, as amended June 17, 2009. 

U.S. EPA. 2010. Agricultural practices that sequester carbon and/or reduce emissions of 

other greenhouse gases. http://www.epa.gov/sequestration/ag.html Accessed on May 19, 

2010.U.S. FDA. 2001. Premarket notice concerning bioengineered foods. Federal 

Register 66: 4706-4738. 

University of Tennessee. 2010. 2010 weed control manual for Tennessee. University of 

Tennessee, Institute of Agriculture, Knoxville, Tennessee. 

USDA-AMS. 2011. Clarification of existing regulations regarding the use of genetically 

modified organisms in organic production and handling. Policy Memo 11-13. U.S. 

Department of Agriculture, Agricultural Marketing Service, Washington, D.C. 

USDA-APHIS. 2006. Soybean. U.S. Department of Agriculture, Washington, D.C. 

http://www.aphis.usda.gov/br/soybean.html Accessed January 20, 2006.USDA-APHIS. 

2011a. Determination of nonregulated status of insect resistant and glufosinate 

ammonium-tolerant (TwinLink) cotton, Gossypium hirsutum, events T304-40 x GHB119. 

U.S. Department of Agriculture, Animal & Plant Health Inspection Service, Washington, 

D.C. 

USDA-APHIS. 2011b. Monsanto petition (09-SY-2000U) for determination of nonregulated status of (MON 87769) stearidonic acid soybean: Draft environmental 

assessment. U.S. Department of Agriculture, Animal and Plant Health Inspection Service, 

Washington, D.C. 

USDA-ARS. 2006. Forage soybean cultivars: A source of high protein livestock feed. 

U.S. Department of Agriculture, Agricultural Research Service, Washington, D.C. 

http://www.ars.usda.gov/Research/docs.htm?docid=8813 Accessed April 24, 2006. 

USDA-ERS. 2001. How much U.S. cropland acreage is under crop residue management. 

Briefing Room. U.S. Department of Agriculture, Economic Research Service. 

http://www.ers.usda.gov/Briefing/AgChemicals/soilmangement.htm Accessed November 

14, 2009. 
Monsanto Company 10-SY-210U 585 of 721 

USDA-ERS. 2005. USDA soybean baseline 2005-14. United States Department of 

Agriculture, Economic Research Service, Washington, D.C. 

http://www.ers.usda.gov/briefing/soybeansoilcrops/2005baseline.htm Accessed June 10, 

2006 

USDA-ERS. 2008a. Data sets: Organic production table 7. U.S. certified organic beans, 

by state, 2008. U.S. Department of Agriculture, Economic Research Service. 

http://www.ers.usda.gov/Data/Organic/Data/Beans.xls Accessed January 17, 2012. 

USDA-ERS. 2008b. Data sets: U.S. and regional cost and return estimates in soybean for 

2008. U.S. Department of Agriculture, Economic Research Service. 

http://www.ers.usda.gov/data/CostsandReturns/testpick.htm Accessed January 18, 2010. 

USDA-ERS. 2010. Certified organic and total U.S. acreage, selected crops and livestock, 

1995-2008. U.S. Department of Agriculture, Economic Research Service, Washington, 

D.C.http://www.ers.usda.gov/Data/organic/Data/CertifiedandtotalUSacreageselectedcrop

slivestock.xls

USDA-ERS. 2011. Data sets - Adoption of genetically engineered crops in the U.S. U.S. 

Department of Agriculture, Economic Research Service, Washington, D.C. 

http://www.ers.usda.gov/Data/BiotechCrops/ 

USDA-NASS. 1984. Acreage - June 1984. U.S. Department of Agriculture, National 

Agricultural Statistics Service, Washington, D.C. 

USDA-NASS. 1988. Crop production – 1987 Summary. U.S. Department of Agriculture, 

National Agricultural Statistics Service, Washington, D.C. 

USDA-NASS. 1990. Acreage - June 1990. U.S. Department of Agriculture, National 

Agricultural Statistics Service, Washington, D.C. 

USDA-NASS. 1992. Acreage - June 1992. U.S. Department of Agriculture, National 

Agricultural Statistics Service, Washington, D.C. 

USDA-NASS. 1995. Acreage - June 1995. U.S. Department of Agriculture, National 

Agricultural Statistics Service, Washington, D.C. 

USDA-NASS. 1998. Acreage - June 1998. U.S. Department of Agriculture, National 

Agricultural Statistics Service, Washington, D.C. 

USDA-NASS. 2000. Acreage - June 2000. U.S. Department of Agriculture, National 

Agricultural Statistics Service, Washington, D.C. 

USDA-NASS. 2003. Acreage - June 2003. U.S. Department of Agriculture, National 

Agricultural Statistics Service, Washington, D.C. 

USDA-NASS. 2004. Agricultural chemical usage 2003 field crops summary. U.S. 

Department of Agriculture, National Agricultural Statistics Service, Washington, D.C. 
Monsanto Company 10-SY-210U 586 of 721 

USDA-NASS. 2006a. Acreage - June 2006. U.S. Department of Agriculture, National 

Agricultural Statistics Service, Washington, D.C. 

USDA-NASS. 2006b. Agricultural chemical usage 2005 field crops summary. U.S. 

Department of Agriculture, National Agricultural Statistics Service, Washington, D.C. 

USDA-NASS. 2007a. Acreage - June 2006. U.S. Department of Agriculture, National 

Agricultural Statistics Service, Washington, D.C. 

USDA-NASS. 2007b. Agricultural chemical usage 2006 field crops summary. U.S. 

Department of Agriculture, National Agricultural Statistics Service, Washington, D.C. 

USDA-NASS. 2008. Agricultural chemical usage 2007 field crops summary. U.S. 

Department of Agriculture, National Agricultural Statistics Service, Washington, D.C. 

USDA-NASS. 2009a. Acreage - June 2009. U.S. Department of Agriculture, National 

Agricultural Statistics Service, Washington, D.C. 

USDA-NASS. 2009b. Crop production 2008 summary. U.S. Department of Agriculture, 

National Agricultural Statistics Service, Washington, D.C. 

USDA-NASS. 2009c. Crop values 2008 summary. U.S. Department of Agriculture, 

National Agricultural Statistics Service, Washington, D.C. 

USDA-NASS. 2011. Quick stats: Soybeans - Acres planted. U.S. Department of 

Agriculture, National Agricultural Statistics Service, Washington, D.C. 

http://quickstats.nass.usda.gov/data/printable/4D84E77B-F692-32F3-B9AD8CDBDC688D03 Accessed December 20, 2011. 

USDA-NRCS. 2005. Residue and tillage management, no till/strip till/direct seed. U.S. 

Department of Agriculture, Natural Resources Conservation Service, Washington, D.C. 

USDA-NRCS. 2006. Conservation resource brief: air quality, number 0605 Retrieved 

November 9, 2010. 

http://www.nrcs.usda.gov/Internet/FSE_DOCUMENTS/nrcs143_023301.pdf 

WHO. 1995. Application of the principles of substantial equivalence to the safety 

evaluation of foods or food components from plants derived by modern biotechnology. 

World Health Organization, Rome, Italy. 

Wrather, J.A., W.C. Stienstra and S.R. Koenning. 2001. Soybean disease loss estimates 

for the United States from 1996 to 1998. Canadian Journal of Plant Pathology-Revue 

Canadienne De Phytopathologie 23:122-131. 

Wrather, J.A. and S. Koenning. 2011. Soybean disease loss estimates for the United 

States, 1996-2010. University of Missouri Delta Research Center, Agricultural 

Experiment Station, Portageville, Missouri. 

http://aes.missouri.edu/delta/research/soyloss.stm Accessed February 22, 2012
Monsanto Company 10-SY-210U 587 of 721 

Wrather, JA and M Mitchum. 2010. Soybean cyst nematode: Diagnosis and management 

(G-4450). University of Missouri Extension Service. Columbia, Missouri. 

http://extension.missouri.edu/publications/DisplayPub.aspx?P=G4450 Accessed 

February 10, 2012

Wright, JP, LJ Schuler, KH Carr, TB Orr, JL Honegger. 2012. Information to Support an 

Endangered Species Assessment for Dicamba Use in Dicamba-Tolerant Soybeans: An 

Evaluation of Potential Exposure and Biological Effects. Monsanto Technical Report 

MSL- MSL-22545. EPA MRID 48644202 

Yacentiuk, M. 2008. Full fat soybeans in swine rations. Manitoba Agriculture, Food and 

Rural Initiatives, Winnipeg, Manitoba. 

http://www.gov.mb.ca/agriculture/liestock/pork/bab02s57.html Accessed November 11, 

2008. 

Zhou et al. (2011) Stability in the Composition Equivalence of Grain from InsectProtected Maize and Seed from Glyphosate-Tolerant Soybean to Conventional 

Counterparts over Multiple Seasons, Locations, and Breeding Germplasms. J. Agric. 

Food Chem., 59:8822–8828.
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Appendix K: Herbicide Resistance 

K.1. Introduction 

Based upon theory of natural selection, plant populations can develop resistance to an 

herbicide due to the selection of individuals that carry specific genes that can render those 

individuals unaffected by the typical lethal effects of an herbicide. The application of an 

herbicide to the plant does not, itself, cause a mutation in subsequent generations. Rather, 

over time, those few plant biotypes containing resistant gene(s) become dominant in the 

population with repeated use of the herbicide in the absence of other control methods, 

such as use of other herbicides and/or use of cultural control methods. The development 

of resistant populations is common to all herbicides. The probability for resistance to 

develop is a function of: frequency of resistant allele(s)43, mechanism of resistance, 

dominance or recessive nature of the resistant allele(s), relative fitness of the resistant 

biotype, and frequency or duration of herbicide use in the absence of other control 

methods (Beckie, 2006; Jasieniuk, et al.,1996; Sammons et al., 2007). The probability of 

resistance is not the same for all herbicides, with some herbicides (e.g., ALS and ACCase 

classes) exhibiting resistance more quickly than other herbicides (e.g. auxin class , 

glyphosate, dinitroanilines class). 

Herbicide resistance can become a limiting factor in crop production if the resistant weed 

population cannot be controlled with other herbicides or cultural practices. In general, 

this has not been the case for any herbicide. In most crops, there are multiple herbicide 

options for growers to use. However, good management practices to delay the 

development of herbicide resistance have been identified and are being actively promoted 

by the public and private sectors (HRAC, 2010) and are being implemented by growers. 

Monsanto considers product stewardship to be a fundamental component of customer 

service and business practices. Stewardship of the dicamba herbicide to preserve its 

usefulness for growers is an important aspect of Monsanto’s stewardship commitment. 

Although herbicide resistance may eventually occur in weed species when an herbicide is 

widely used, resistance can be postponed, contained and managed through research, 

education and good management practices. These are the key elements of Monsanto’s 

approach to providing stewardship of dicamba used on MON 87708 integrated into the 

Roundup Ready soybean system. Monsanto will invest in research, and grower/retailer 

education and training programs to provide information on best practices to manage 

dicamba weed resistance in soybean production. This appendix provides an overview of 

Monsanto’s approach to the development of best management practices to mitigate 

dicamba weed resistance. Monsanto works closely with weed scientists in academia and 

with other companies to research and develop best management practices and to 

uniformly communicate such practices to growers. Evidence of this cooperative effort is 

the recent development and posting of herbicide resistant training modules on the WSSA 

website (www.wssa.net) and the publication of guidelines by the Herbicide Resistance 

Action Committee (HRAC) on their website (www.hracglobal.com). 

 

43 An allele is any of several forms of a gene, usually arising through mutation, that are responsible for 

hereditary variation.
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K.2. The Herbicide Dicamba 

Dicamba (3,6-dichloro-2-methoxybenzoic acid) is classified as a benzoic acid herbicide 

belonging to the synthetic auxin group of herbicides (HRAC, 2010). The herbicides in 

this group act as growth regulators similar to endogenous indole acetic acid (IAA) but are 

structurally diverse. The synthetic auxin group includes five chemical families (benzoic 

acid, pyridine-carboxylic acid, quinoline carboxylic acid, phenoxy-carboxylic acid and a 

separate class which includes one herbicide, benazolin ethyl). In addition to dicamba, 

specific herbicides in this group include 2,4-D, 2,4-DB, mecoprop, MCPA, clopyralid, 

picloram, quinclorac and several other active ingredients. Dicamba and other synthetic 

auxin herbicides are classified in Herbicide Group 4 by the Weed Science Society of 

America (HRAC, 2009). Most herbicides in this group are active on broadleaf weeds 

only, but a few have significant activity on grasses, e.g., quinclorac. The specific site of 

action among the different chemistry families may be different. Dicamba provides 

preemergence and postemergence control of over 95 annual and biennial broadleaf weed 

species and control or suppression of over 100 perennial broadleaf and woody species44. 

Dicamba is not active on grass weeds and is often used in combination with other 

herbicides to provide broad spectrum weed control. 

Dicamba herbicide was commercialized in the U.S. for agricultural use in 1967 and is 

currently labeled for preemergence and/or postemergence weed control in corn, soybean, 

cotton, sorghum, small grains (wheat, barley and oats), millet, pasture, rangeland, 

asparagus, sugarcane, turf, grass grown for seed, conservation reserve program land, and 

fallow cropland (U.S. EPA, 2009). Dicamba is sold as standalone formulation which can 

be tank mixed with one or more active ingredients depending upon the crop and the weed 

spectrum. Dicamba is also sold as a premix formulation with other herbicides. 

Dicamba kills plants by mimicking naturally-occurring plant growth hormones called 

auxins, thereby destroying tissue through uncontrolled cell division and growth (Ahrens, 

1994). Ahrens (1994) further states that dicamba has been found to affect cell wall 

integrity and nucleic acid metabolism whereas in other cases it has been found to increase 

cell wall permeability, leading to cell enlargement. At low concentrations, dicamba has 

been found to increase synthesis of DNA, RNA, and proteins, resulting in altered cell 

division and growth. At high concentrations, inhibition of cell division and growth occur. 

In general dicamba and other synthetic auxin herbicides have been found to affect 

multiple plant physiological systems. The molecular mechanism of auxin action is still 

not known in detail nor completely understood (Devine et al., 1993, Jugulam et al. 

2011). However, Grossmann (2010) in a review of auxin herbicides, outlined a proposed 

mechanism and mode of action for auxin herbicides and IAA at supraoptimal endogenous 

concentrations in dicot plant species. The proposal was based upon recent identification 

of receptors for auxins and hormone interaction in signaling between auxin, ethylene and 

the upregulations of abscisic acid biosynthesis which would account for a large part of 

the various auxin-herbicide-mediated responses that are seen in sensitive dicots. In 

addition research has indicated that there is a high level of redundancy in auxin receptors 

 

44 Clarity product label http://www.cdms.net/LDat/ld797002.pdf 
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which may account for the lack of development of widespread resistance to this herbicide 

group (Walsh et al., 2006). 

Dicamba is taken up by plants through the roots, stems, and foliage (Ahrens, 1994; NPIC, 

2002). Dicamba translocates to all plant tissues but accumulates in growing tissues. 

Translocation of dicamba is typically slower in tolerant plants such as grasses compared 

to broadleaf plants. Dicamba has a relatively low soil-binding coefficient. 

K.3. Herbicide-Resistant Weeds and Resistance Management Strategies 

The development of herbicide-resistant weeds is not a new phenomena and resistance is 

not limited to certain select herbicides. In 1957, the first U.S. herbicide-resistant weed, a 

spreading dayflower biotype resistant to 2,4-D, was identified in Hawaii (Heap, 2010). 

Currently, there are 73 individual weed species that have known herbicide-resistant 

biotypes to one or more herbicides in the U.S. For example, there are 42 weed species 

resistant to ALS herbicides, 15 to ACCase inhibitors, 24 to photosystem II inhibitors, and 

10 to glycine herbicides (Heap, 2010). Growers have been managing herbicide-resistant 

weeds for decades with the use of alternative herbicides and/or cultural methods such as 

tillage or crop rotation. 

The occurrence of an herbicide-resistant weed biotype does not end the useful lifespan or 

preclude the effective use of the herbicide in question as part of an overall diversified 

weed management system. The three herbicide classes with the highest number of 

resistant species, ALS, ACCase and triazine herbicides, are still effectively used by 

growers today. 

It is important to distinguish herbicide resistance from herbicide tolerance. A herbicidetolerant weed species is one that is naturally tolerant to a herbicide, for example a grass 

species is not killed by the application of a broadleaf herbicide. Furthermore, certain 

weed species, while neither resistant nor tolerant, are inherently difficult to control with a 

particular herbicide, requiring more careful herbicide use and weed management 

practices. 

Since the first confirmed cases of herbicide resistance, research has been directed at 

determining which practices are best for managing existing resistance situations and how 

best to reduce the development of herbicide resistance. Resistant management practices 

most often recommended by University/Cooperative Extension Service (CES) and 

industry are: 1) use of multiple herbicide modes-of-action in mixture, sequence, or in 

rotation, 2) crop rotation, 3) use of cultural control measures such as tillage and time of 

planting, and 4) use of the labeled herbicide rate at the recommended timing of 

application (Gressel and Segel, 1990; Beckie, 2006). Recent research by Beckie and 

Reboud (2009) indicates that in some cases herbicide mixtures offer a better management 

option than rotating herbicides. Simultaneously using two herbicides with different 

modes-of-action significantly reduce the probability of weeds developing resistance to 

either or both herbicides (Beckie and Reboud, 2009). Crop rotation is also an effective 

method for resistance management due to the fact that it fosters the use of additional 

herbicide modes- of-action and, potentially, use of additional cultural practices to manage 
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weeds over time. The use of multiple methods of weed control in a single location is the 

technical basis for developing management programs to delay the development of 

resistance. This general concept has been referred to as applying “diversity” within a 

crop or across a crop rotation (Beckie, 2006; Powles, 2008). 

It is generally accepted that conservation tillage practices (minimum-till and no-till) 

create environments where herbicide resistance is more likely to develop (Beckie, 2006). 

This is probably due to selection pressure put on weeds by herbicides in these nondiverse environments and the absence of tillage as a cultural weed management practice 

to supplement herbicide use. However, this is not always the case. Legere et al. (2000) 

found that an increase in the use of ACCase inhibitors in a conservation tillage system 

(e.g. aryloxyphenosy propionates and phenylpyrazolines herbicide families) did not result 

in an increased incidence of wild oat populations resistant to ACCase inhibitors. In 

conclusion, conservation tillage practices should not be considered a primary contributing 

factor to the development of resistance in all cases 

K.4. Characteristics of Herbicides and Herbicide Use Influencing Resistance 

While the incidence of weed resistance is often associated with repeated applications of 

an herbicide, the actual probability for the development of resistant populations is related, 

in part, to the specific herbicide active ingredient, chemical family and the herbicide 

group. Some herbicides are more prone to the development of resistance than others 

(Heap, 2010). The graph in Figure K-1 illustrates the global instances of weed resistance 

to various herbicide groups. The different slopes of observed resistance are largely due 

to the factors described above, which relate to the specific herbicide active ingredient as 

well as to the group and herbicide family and its function. 
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Figure K-1. Weed Resistance to Various Herbicide Families1

As can be seen in Figure K-1, weed resistance to the synthetic auxin group of herbicides has been 

slower to develop than for other herbicide groups even though these were the first synthetic 

herbicides discovered and used commercially. Possible reasons for this are discussed below.

 

1

Global number of resistant biotypes 

K.5. Mechanisms of Resistance and Inheritance of Resistance 

To date, the three known basic mechanisms by which weed species develop resistance to 

a herbicide have been identified: 1) target site alteration (target site), 2) enhanced 

metabolism of the herbicides (metabolism), and 3) reduced absorption and/or 

translocation of the herbicide such that the herbicide does not get to the site of action 

within the plant cell (exclusion) (Sammons et al., 2007). 

Herbicide resistance via target site alteration is the most common resistance mechanism 

among the various herbicide groups and chemical families. It has been found that a target 

site mechanism is the most common mechanism for ALS inhibitors, ACCase inhibitors, 

and triazines, but is less common for other herbicide groups, such as glyphosate (Powles 

and Yu, 2010). The most common type of target site alteration is one where amino acid 

substitution(s) in the protein that is the target of the herbicide occurs such that the 

alteration prevents the binding of the herbicide to the protein and as a result the activity 

of the targeted protein is not altered and the plant grows normally. 

In the case of synthetic auxin herbicides, resistance has been speculated to be due to 

mutation(s) in genes encoding an auxin-binding protein causing reduced herbicide 

binding (Zheng and Hall, 2001; Goss and Dyer, 2003). In several studies, differential 
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herbicide absorption, translocation, and metabolism were ruled out as possible 

mechanisms of resistance in kochia (Cranston et al., 2001) and in wild mustard (Zheng 

and Hall, 2001). However, current research has not presented convincing evidence for a 

single mechanism of resistance and this inability to elucidate the mechanism of resistance 

may be due to a lack of thorough understanding of auxin mechanism of activity 

(Jasieniuk et al., 1996). Walsh et al. (2006) identified seven alleles at two distinct genetic 

loci that conferred significant resistance to picolinate auxins (picloram) in Arabidopsis, 

yet had minimal cross-resistance to 2,4 D and IAA. 

Multiple mechanisms for inheritance of dicamba resistance have been reported in the 

literature. Jasieniuk et al. (1995) reported results indicating that inheritance of dicamba 

resistance in wild mustard is determined by a single, completely dominant nuclear allele. 

However, Cranston et al. (2001) reported results indicating that dicamba resistance in 

kochia is determined by a quantitative trait (two or more genes). 

In summary, the slow development of weed resistance to synthetic auxin herbicides may 

in part be due to their proposed multiple sites of physiological action in plants (Jasieniuk 

et al., 1996) and to the possibility that inheritance, at least in some species, is determined 

by a quantitative trait (Cranston et al., 2001). 

K.6. Weeds Resistant to Dicamba 

As noted earlier, like other herbicides, the use of dicamba may lead to the development of 

dicamba-resistant weed species. To date there are four species with known resistant 

biotypes to dicamba in the U.S./Canada after over 40 years of use; common hempnettle 

(Galeopsis tetrahit), kochia (Kochia scoparia), prickly lettuce (Lactula serriola) and wild 

mustard (Sinapis arvensis) (Heap, 2010). Additionally, a population of lambsquarters 

(Chenopodium album) has been confirmed to be resistant in New Zealand for a total of 

five species worldwide with confirmed resistant biotypes to dicamba. For the synthetic 

auxin group of herbicides there is a total of 29 species globally with biotypes having 

confirmed resistance to at least one member of this group but only eight species in the 

U.S. and four species in Canada (Heap, 2010). All of these populations are, but for two 

(wild carrot in OH and MI and waterhemp in Nebraska), found in western states or 

western Canadian provinces. In some species, cross-resistance between different 

herbicides within the auxin group has been confirmed (cross-resistance is a plant’s 

resistance to another herbicide as a result of exposure to a similarly acting herbicide). 

Therefore, consideration has to be given to the possibility that dicamba resistance could 

extend to some of the other broadleaf species listed as resistant to other synthetic auxin 

herbicides (Miller et al., 2001; Jasieniuk et al., 1995; Cranston et al., 2001). However, 

because of differences in sites of action among chemistries within this group (i.e., 

benzoic acids compared to pyridine-carboxylic acids) cross resistance between the 

herbicide groups is not a certainty (Monaco et al., 2002). 

With the introduction of MON 87708 into the Roundup Ready soybean system, where 

dicamba will be applied in combination with glyphosate, it is important to note that 

kochia is the only broadleaf species with resistant biotypes to either synthetic auxins or 

glyphosate. There are no known kochia biotypes resistant to both of these herbicides. 
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The development of a dicamba-glyphosate biotype is unlikely because both dicamba and 

glyphosate, each with two distinct modes-of-action, will likely be applied in the same 

season to MON 87708 in the Roundup Ready soybean system. If populations with 

resistance to multiple herbicides were to occur, there are other herbicide options for 

managing the weed in soybean (e.g., clomazone and flumioxazin) and in its rotational 

crops (e.g., atrazine and isoxaflutole in corn) (Table K-1). In addition, kochia is a weed 

found primarily in the western soybean growing regions where soybean would be rotated 

with corn and wheat for which there are multiple options for kochia control (Casey, 

2009). The glyphosate-resistant kochia biotype is also found in western soybean growing 

areas, but it is isolated to small areas where soybean is grown in limited acreage. 

K.7. Sustainable Use of Dicamba as a Weed Management Option in Soybean 

Dicamba is a broadleaf herbicide that does not provide control of grass weeds. For that 

reason, MON 87708 will be sold only in soybean varieties that also contain other 

herbicide-tolerant traits, such as with the Roundup Ready soybean system (e.g., 

MON 89788). Soybean varieties containing both MON 87708 and MON 89788 will 

enable dicamba to be applied with glyphosate or other soybean herbicides in an 

integrated weed management program, ideally as a mixture, to control a broad spectrum 

of grass and broadleaf weed species. Dicamba applications on MON 87708 will provide 

effective control of glyphosate-resistant broadleaf weeds and improve the control of 

annual and perennial broadleaf weed species, some of which are difficult to control with 

glyphosate. Dicamba will also help delay development and/or combat existing weed 

resistance issues that can limit the use of the PPO- and ALS-inhibiting herbicide groups 

by providing an additional mode of action for management of certain broadleaf species 

that are known to be prone to resistance to many of the current options for weed 

management (i.e. Amarathus spp.). MON 87708 will foster the adoption of Integrated 

Pest Management (IPM) practices in soybeans by allowing growers to continue to 

primarily focus on postemergence in-crop weed control, as they have practiced with the 

Roundup Ready soybean system. This will allow growers to delay some herbicide 

treatments until field scouting indicates a need for additional postemergence weed control 

which is consistent with the principles of IPM. Increasing postemergence herbicide 

options in soybeans is important, especially in conservation tillage situations, where 

consistency of postemergence herbicides has generally been greater than that of soil 

active residual products and thus a driving factor in the adoption of conservation tillage 

systems in the U.S. 

Upon the inclusion and integration of MON 87708 into the Roundup Ready soybean 

system and approval of the use of dicamba on MON 87708 (Monsanto has submitted to 

EPA an application to amend Registration Number 524-582 to register a new use pattern 

for dicamba on MON 87708), preplant/preemergence applications of dicamba can be 

made up to 1.0 lb a.e./A up through crop emergence (cracking) followed by two in-crop 

postemergence applications up to 0.5 lb a.e./A through the R1/R2 growth stage in 

soybean. However, the majority of weed control scenarios in MON 87708 will not 

require the use of the maximum labeled rate, and the anticipated commercial preplant/preemergence and in-crop use rates are between 0.25 to 0.375 lb a.e./A (based on 

established weed control rates for soybean weeds), with an average application rate of 
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0.35 lb a.e./A as described in Section VIII.H). Residual herbicides also will be 

recommended for use, in addition to glyphosate and dicamba, to provide early season 

weed control and to supplement dicamba activity on certain hard-to-control and 

glyphosate-resistant weed biotypes, such as glyphosate-resistant palmer amaranth where 

weed populations can be very substantial. See section K.8.1 for specific weed 

management recommendations. 

Dicamba, as a complementary herbicide to glyphosate, will provide new weed control 

options in soybean that strengthen the utility and sustainability of glyphosate as a weed 

control tool in the Roundup Ready soybean system. Likewise, glyphosate, as a 

complementary herbicide to dicamba, would strengthen the utility and sustainability of 

dicamba as a weed control tool for MON 87708 to be integrated into the Roundup Ready 

soybean system. 

In the event there is known or suspected presence of a dicamba-resistant weed, it will be 

possible to provide the affected grower(s) with options for managing the dicambaresistant biotype. There are multiple preemergence (including soil residuals) and 

postemergent herbicide options for managing broadleaf weed populations that are 

resistant or may potentially develop resistance to dicamba in soybean as well for crops 

grown in rotation with soybean. These options are noted in Table K-1. 
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Table K-1. Management Recommendations for Control of Dicamba- and Other Synthetic Auxin-ResistanWeed Species Primary Crop Soybean 

Rotational Crops / Other Uses 

Corn Wheat Pastures/Roadsid

Kochia Saflufenacila Atrazinea Saflufenacila(Kochia scoparia) Clomazonea Saflufenacila Glyphosatea

Flumioxazina Isoxaflutolea Bromoxynil/MCPAa

Glyphosatea Mesotrionea

Paraquata Glyphosatea

Prickly Lettuce Saflufenacila Saflufenacila Saflufenacila

(Lactuca serriola) Chlorimuron/metribuzina Atrazinea Triasulfurona

Glyphosate + imazethapyra Carfentrazone + 

atrazinea

Metsulfuron + 

thifensulfurona

Isoxaflutole + 

atrazinea

Wild Carrot Glyphosatec Glyphosatec

(Daucus carota) Chlorimuronc Atrazinec

Chlorimuron/metribuzinc Primisulfuronc

NicosulfurondHalosulfurond

Field Bindweed Glyphosatea Glyphosatea Glyphosatea(Convolvulus arvensis) 

 Glyphosate + 

imazethapyra

 Glyphosate + 

Imazamoxa 
Monsanto Company 10-SY-210U 

Table K-1 (continued). Management Recommendations for Control of Dicamba- and Other Synthetic AuWeed Primary Crop Soybean 

Rotational Crops / Other Uses 

Corn Wheat Pastures/Roadsid

Yellow Starthistle (Centaurea solstitialis) 

 Chlorsulfuron 

Aminopyralide 

Spreading Dayflower (Commelina diffusa) 

 

Lambsquarters Metribuzinb Isoxaflutolea Bromoxynila

(Chenopodium Cloransulamb Atrazinea Chlorsulfuron/Metsulfurona

album)

g Saflufenacila Saflufenacila Glyphosatea

Imazamoxb Mesotrionea Saflufenacila

Glyphosateb Bromoxynilb

aBernards et al., 2010. bLoux et al., 2010.cMichigan State University Extension, 2010. dKells and Stachler, 1997. ePNWE, 2010. fUniversity of Arkansas CES, 2010. gResistance to lambsquarters has only been confirmed in New Zealand.
Monsanto Company 10-SY-210U 598 of 721 

K.8. Stewardship of Dicamba Use on MON 87708 

In order to steward the use of agricultural herbicides and herbicide-tolerant cropping 

systems such as MON 87708 integrated into the Roundup Ready soybean, Monsanto has 

conducted investigations and worked extensively with academics and other herbicide 

manufacturers to understand and recommend best practices to manage herbicide 

resistance. These investigations have demonstrated that one of the major factors 

contributing to the development of resistant weed biotypes has been poor weed control 

management practices. The lack of adequate management includes: 1) application of 

herbicides at rates below those indicated on the product label for the weed species, and 2) 

sole reliance on a particular herbicide for weed control without the use of other herbicides 

or cultural control methods (Beckie, 2006; Peterson et al., 2007). 

K.8.1. Weed Control Recommendations 

The pending label for dicamba use on MON 87708 is based on the maximum allowable 

use rates and patterns. Prior to launch of MON 87708 in the Roundup Ready soybean 

system, Monsanto, in cooperation with academics, will conduct trials to confirm the 

optimum rate and timing for dicamba alone and in combination with glyphosate and other 

herbicides. Recommendations to growers will be developed from this information and 

will be provided in herbicide product labels, Monsanto’s Technology Use Guide (TUG), 

and in other education and training materials to be broadly distributed. Specifically, 

current research conducted by Monsanto to define the optimum weed management 

systems indicate the following: 1) in the absence of glyphosate resistant populations, the 

recommendation will be to apply a soil active residual herbicide followed by a 

postemergence application of glyphosate plus dicamba to control weed escapes, and 2) in 

the presence of glyphosate resistant populations, the same system will be recommended 

with a potential second application of glyphosate plus dicamba if needed. In this latter 

case, the preemergence herbicide to be recommended will be one with activity against the 

targeted glyphosate resistant species. This will ensure more than one mechanism of 

action against the targeted species, which is a fundamental component of a good weed 

resistance management program. These management systems will reduce the potential for 

further resistance development to glyphosate, dicamba as well as other critical soybean 

herbicides. In conservation tillage systems, a preplant application of glyphosate plus 

dicamba may be recommended in some situations in addition to the in-crop applications 

described above. This is not expected to increase selection pressure on either product 

since the preplant weed spectrum is generally different from the in-crop spectrum. 

K.8.2. Dispersal of Technical and Stewardship Information 

Monsanto will use multiple methods to distribute technical and stewardship information 

to growers, academics and grower advisors. Monsanto’s TUG will set forth the 

requirements and best practices for the cultivation of MON 87708 including 

recommendations on weed resistance management practices. Growers who purchase 

varieties containing MON 87708 will be required to enter into a limited use license with 
Monsanto Company 10-SY-210U 599 of 721 

Monsanto and must sign and comply with the Monsanto Technology Stewardship 

Agreement (MTSA), which requires the grower to follow the TUG. 

The weed resistance management practices that will be articulated in the TUG will also 

be broadly communicated to growers and retailers in order to minimize the potential for 

the development of resistant weeds. These practices will be communicated through a 

variety of means, including direct mailings to each grower purchasing a soybean variety 

containing MON 87708, a public website45, and reports in farm media publications. The 

overall weed resistance management program will be reinforced through collaborations 

with U.S. academics who will provide their recommendations for appropriate stewardship 

of dicamba in soybean production, as well as by collaboration with crop commodity 

groups who have launched web-based weed resistance educational modules. Finally, 

Monsanto will urge growers to report any incidence of repeated non-performance of 

dicamba on weeds in fields planted with MON 87708, and Monsanto will investigate 

cases of unsatisfactory weed control to determine the cause as defined in K.9. 

The EPA is the U.S. federal regulatory agency that administers the federal law governing 

pesticide sale and use under the Federal Insecticide, Fungicide and Rodenticide Act 

(FIFRA). EPA encourages pesticide manufacturers to provide growers with information 

regarding an herbicide’s mode-of-action to aid growers in planning herbicide use 

practices and to foster the adoption of effective weed resistance management practices as 

specified by EPA in Pesticide Registration (PR) Notice 2001-5 (U.S. EPA, 2001). In that 

document EPA states that “this approach to resistance management is sound and would 

be highly beneficial to pesticide manufacturers and pesticide users.” EPA approves all 

pesticide label use instructions based on its evaluation of supporting data supplied by the 

pesticide registrant or manufacturer. By approving a label, EPA has concluded that the 

product will not cause unreasonable adverse effects to the environment when used in 

accordance with the label’s directions. After EPA approves a pesticide label, it is a 

violation of federal law to use the pesticide for a use or in a manner not in accordance 

with the label directions. Monsanto incorporates EPA’s guidelines for pesticide 

resistance management labeling on its agricultural herbicide labels, and will continue to 

do so in the future. Monsanto will adopt a similar approach to pesticide resistance 

management guidance on its dicamba product labels. 

In summary, Monsanto will require weed resistance management practices through the 

MTSA and TUG for its biotechnology-derived herbicide-tolerant products, such as 

MON 87708 integrated into the Roundup Ready soybean system, and to promote these 

practices through product labeling and educational outreach efforts as an effective means 

to manage weed resistance development for both dicamba and glyphosate. 

K.8.3. Weed Resistance Management Practices 

Monsanto will provide information to growers and grower advisors on best management 

practices to delay the development of resistance to dicamba. The weed resistance 

management recommendations for the use of dicamba in conjunction with soybean 

 

45 http://www.monsanto.com/weedmanagement/Pages/default.aspx 
Monsanto Company 10-SY-210U 600 of 721 

varieties containing MON 87708 will be consistent with the Herbicide Resistance Action 

Committee’s guidelines for prevention and management of herbicide resistance (HRAC, 

2010) 46 . These guidelines recommend an integrated approach to weed resistance 

management including crop management (i.e., row spacing, etc), cultivation techniques, 

and the use of multiple herbicide modes-of-action to manage a weed population. 

In cases where resistance is confirmed for dicamba in soybean producing areas, 

Monsanto and University/Cooperative Extension Service (CES) personnel will provide 

recommendations for alternative herbicide control methods to growers. These 

recommendations would be made available through Monsanto supplemental labels, 

Monsanto and university publications, and internet sites to growers, consultants, retailers 

and distributors. For all existing cases of dicamba-resistant weeds in the U.S. and 

globally today, alternative herbicides and cultural methods are available to growers to 

effectively control these biotypes. Examples of recommended alternative herbicides 

from University/CES personnel that are applicable to weed species known to be resistant 

to dicamba and other synthetic auxin herbicides are found in Table K-1. It is important to 

note that there are many alternative options in each situation. 

K.9. Monsanto Weed Performance Evaluation and Weed Resistance Management 

Plan 

An important part of a weed resistance management plan is the timely acquisition of 

information regarding product performance. Monsanto has an extensive technical, sales 

and marketing presence in the soybean markets where MON 87708 will be grown. 

Through our relationships with farm advisors, key University/CES personnel, and 

growers using our seeds and traits products, Monsanto will acquire important and timely 

information regarding product performance. This will allow the timely recognition of 

performance issues that could arise related to weed resistance or other means. Field 

employees and hired consultants are trained and provided processes for responding to 

product performance inquiries. Individual performance issues that could be related to 

potential resistance are promptly handled. In addition performance inquiries are 

periodically reviewed by Monsanto for trends that could indicate the need for follow up 

action on a broad scale. 

If resistance is confirmed, the scientific and grower communities will be notified and a 

weed resistance mitigation plan will be implemented by Monsanto in cooperation with 

the University/CES. The mitigation plan will be designed to manage the resistant biotype 

through effective and economical weed management recommendations implemented by 

the grower. The scope and level of intensity of the mitigation plan may vary depending 

on a combination of the following factors: 1) biology and field characteristics of the weed 

(seed shed, seed dormancy, etc.), 2) importance of the weed in the agricultural system, 3) 

resistance status of the weed to other herbicides with alternate modes-of-action, and 4) 

availability of alternative control options. These factors are analyzed by Monsanto and 

 

46 The Herbicide Resistance Action Committee (HRAC) is an international body founded by the 

agrochemical industry for the purpose of supporting a cooperative approach to the management of 

herbicide resistance and the establishment of a worldwide herbicide resistance database. 
Monsanto Company 10-SY-210U 601 of 721 

University/CES personnel in combination with economic and practical management 

considerations to develop a tailored mitigation strategy. The plan considers what is 

technically appropriate for the particular weed and incorporates practical management 

strategies that can be implemented by the grower. 

After a mitigation plan is developed, Monsanto communicates the plan to the grower 

community through the use of supplemental labeling (labeling which includes newly 

approved use directions, or other instructions), informational fact sheets, retailer training 

programs, agriculture media and/or other means, as appropriate. 

In addition to the grower inquiry initiated process, Monsanto, alone and/or in cooperation 

with University/CES, will conduct field studies to understand the potential for weed 

resistance and weed shifts as the result of various weed management programs 

implemented for MON 87708 integrated into the Roundup Ready soybean system. These 

studies will allow researchers to better track specific factors that can influence the 

development of resistance to specific weeds. 

K.10. Summary 

Development of weed resistance is a complex process that can be difficult to accurately 

predict. Multiple methods for managing weed resistance are available and no single 

option is best. No single agronomic practice will mitigate resistance for all herbicides or 

all weeds. As a result, weed resistance needs to be managed on a case-by-case basis and 

tailored for the particular herbicide and weed in order to meet grower needs. Using good 

weed management principles, built upon achieving high levels of control through proper 

application rate, choice of cultural practices, and appropriate companion weed control 

products will allow dicamba herbicides to continue to be used effectively. In cases where 

weed populations have developed resistance to dicamba, effective management options 

are available and experience has shown that growers will continue to find value in using 

dicamba in their weed control programs. 

The key principles for effective stewardship of dicamba use, including the integration of 

MON 87708 in the Roundup Ready soybean system, include: 1) basing weed 

management and weed resistance management practices on local needs and using the 

tools necessary to optimize crop yield, 2) using proper rate and timing of application, 3) 

not relying solely on one herbicide weed control option across a cropping system, 4) 

responding rapidly to instances of unsatisfactory weed control, and 5) providing up-todate weed management and weed resistance management training. 

Overall, there is a low potential for dicamba-resistant broadleaf weed populations to arise 

from the use of dicamba applied to MON 87708 integrated into the Roundup Ready 

soybean system. The reasons are as follows: 

Dicamba will be used in combination with glyphosate in a majority of cropping situations, 

and weed recommendations will also include the concurrent use of residual herbicides for 

complementary weed control and different modes-of-action. These use patterns mean 
Monsanto Company 10-SY-210U 602 of 721 

that there will be multiple modes-of-action against the major broadleaf species present in 

soybean production. This is a primary way to delay the development of resistance. 

The development of resistance to auxin herbicides has been found to be relatively slow. 

This observation is hypothesized to be due to multiple sites of action within plants and 

evidence suggesting that resistance is determined by multiple genes (quantitative traits), 

at least in some species. 

Only four broadleaf weed species have been confirmed to be resistant to dicamba in the 

U.S., and relatively low numbers of broadleaf species have been confirmed to be resistant 

to synthetic auxin herbicides even though dicamba has been widely in use for over 40 

years. 

Known resistant broadleaf populations to dicamba and other auxin herbicides are 

primarily found in the western U.S. and, thus, are not present in the major soybean 

geographies. In addition, the known dicamba-resistant biotypes are not major weed 

species present in the U.S. soybean crop. 
Monsanto Company 10-SY-210U 603 of 721 

References for Appendix K 

Ahrens, W.H. 1994. Dicamba. 3,6-dichloro-2-methoxybenzoic acid. Pages 91-94 in 

Herbicide Handbook. Seventh Edition. Weed Science Society of America, Champaign, 

Illinois. 

Beckie, H.J., and X. Reboud. 2009. Selecting for weed resistance: Herbicide rotation and 

mixture. Weed Technology 23:363-370. 

Beckie, H.J. 2006. Herbicide-resistant weeds: Management tactics and practices. Weed 

Technology 20:793-814. 

Bernards, M.L., R. E.Gaussoin, R.N. Hliein, S.Z. Knezevic, D.J. Lyon, L.D. Sandell and 

R.G Wilson. 2010. Guide for weed management in Nebraska (EC130). University of 

Nebraska Extension, Lincoln, Nebraska 

Casey, P.A. 2009. Plant guide for kochia (Kochia scoparia). United States Department of 

Agriculture, Natural Resources Conservation Service, Manhattan, Kansas. http://plantmaterials.nrcs.usda.gov/technical/publications/newpublications/pg_kosc.pdf Accessed 

June 1, 2010. 

Cranston, H. J., A. J. Kern, J. L. Hackett, E. D. Maxwell, B. D. Maxwell, and W. E. Dyer. 

2001. Dicamba resistance in kochia. Weed Science 49:164-170. 

Devine, M., S.O. Duke, and C. Fedtke. 1993. Physiology of herbicide action. PTR 

Prentice Hall, Inc. Englewood Cliffs, NJ. pp. 295-309. 

Goss, G.A., and W. E. Dyer. 2003. Physiological characterization of auxinic herbicideresistant biotypes of Kochia (Kochia scoparia). Weed Science 51:839-844. 

Gressel, J. and L. A. Segel. 1990. Modelling the effectiveness of herbicide rotations and 

mixtures as strategies to delay or preclude resistance. Weed Technology 4:186-198. 

Grossmann, K. 2010. Auxin herbicides: current status of mechanism and mode of action. 

Pest Management Science 66: 113-120. 

Heap, I. 2010. International survey of herbicide resistant weeds. 

http://www.weedscience.org/in.asp Accessed July 1, 2010. 

HRAC. 2009. Classification of herbicides according to mode of action. Herbicide 

Resistance Action Committee, Corvallis, Oregon. 

http://www.hracglobal.com/Publications/ClassificationofHerbicideModeofAction/tabid/2

22/Default.aspx Accessed June 29, 2009. 

HRAC. 2010. Guideline to the management of herbicide resistance. Herbicide Resistance 

Action Committee, Calgary, Alberta. http://www.hracglobal.com Accessed April 1, 2010. 
Monsanto Company 10-SY-210U 604 of 721 

Jasieniuk, M., I.N. Morrison, and A.L. Brûlé-Babel. 1995. Inheritance of dicamba 

resistance in wild mustard (Brassica kaber). Weed Science 43:192-195. 

Jasieniuk, M., A.L. Brûlé-Babel, and I.N. Morrison. 1996. The evolution and genetics of 

herbicide resistance in weeds. Weed Science 44:176-193. 

Jugulam, M., C. Hall, W.G. Johnson, K.B. Kelley and D.E. Riechers. 2011. Evolution of 

resistance to auxinic herbicides: Historical perspectives, mechanisms of resistance, and 

implications for broadleaf weed management in agronomic crops. Weed Science 

Manuscript: 1-45 

Legere, A., H.J. Beckie, F.C. Stevenson, and A.G. Thomas. 2000. Survey of management 

practices affecting the occurrence of wild oat (Avena fatua) resistant to acetyl-coA 

carboxylase inhibitors. Weed Technology 14:366-376. 

Loux, M.M., D. Doohan, A.F. Dobbels, W.G. Johnson, G.R.W. Nice, T.N. Jordan and 

T.T. Bauman. 2010. Weed control guide for Ohio and Indiana. Bulletin 789. Purdue 

University and Ohio State University Extension, West Lafayette, Indiana. 

Kells, J.J. and J.M. Stachler. 1997. Controlling wild carrot, as published in Michigan 

State University Extension Soybean Facts Winter 1997 (FACT9707). Michigan State 

University Extension. http://web1.msue.msu.edu/imp/mods1/fact9707.html Accessed: 

May 19, 2010. 

Michigan State University Extension. Controlling Wild Carrot. 

http://fieldcrop.msu.edu/documents/controlling%20wild%20carrot.pdf Accessed: May 19, 

2010. 

Miller, T.W., S.L. Shinn, and D.C. Thill. 2001. Cross-resistance in and chemical control 

of auxinic herbicide-resistant yellow starthistle (Centaurea solstitialis). Weed 

Technology 15:293-299. 

Monaco, T.J., S.C. Weller and F.M. Ashton. 2002. Growth regulator herbicides. Pages 

291-310 in Weed Science: Principles and Practices. Fourth Edition. John Wiley & Sons, 

Inc., New York, New York. 

NPIC. 2002. Dicamba technical fact sheet. National Pesticide Information Center, 

Corvallis, Oregon. http://npic.orst.edu/factsheets/dicamba_tech.pdf Accessed April 30, 

2010. 

PNWE. 2010. Control of problem weeds - Starthistle. Pacific Northwest weed 

management handbook. Pacific Northwest Extension Service, Portland, Oregon. 

http://uspest.org/pnw/weeds Accessed May 25, 2010. 

Peterson, D., B. Olson, K. Al-Khatib, R. Currie, J.A. Dille, J. Falk, P. Geier, D. Regehr, P. 

Stahlman and C. Thompson. 2007. Glyphosate stewardship. MF-2767. Kansas State 

University Agricultural Experiment Station and Cooperative Extension Service, 

Manhattan, Kansas. 
Monsanto Company 10-SY-210U 605 of 721 

Powles, S.B. 2008. Evolved glyphosate-resistant weeds around the world: lessons to be 

learnt. Pest Management Science 64:360-365. 

Powles, S.B. and Qin Yu. 2010. Evolution in action: Plants resistant to herbicides. 

Annual Review of Plant Biology 61:317-347. 

Sammons, R.D., D.C. Heering, N. DiNicola, H. Glick, and G.A. Elmore. 2007. 

Sustainability and stewardship of glyphosate and glyphosate-resistant crops. Weed 

Technology 21:347-354. 

U.S. EPA. 2001. Pesticide Registration (PR) Notice 2001-5, Guidance for Pesticide 

Registrants on Pesticide Resistance Management Labeling. 

http://www.epa.gov/opppmsd1/PR_Notices/pr2001-5.pdf. 

U.S. EPA. 2009. Reregistration Eligibility Decision (RED) for Dicamba and Associated 

Salts. Case No. 0065. U.S. Environmental Protection Agency Office of Prevention, 

Pesticides and Toxic Substances, Special Review and Reregistration Division, 

Washington, D.C. 

University of Arkansas Cooperative Extension Service (CES). 2010. Recommended 

chemicals for weed and brush control (MP-44). Page 78. University of Arkansas, U.S. 

Department of Agriculture and County Governments Cooperating. 

Walsh, T.A., R. Neal, A.O. Merlo, M. Honma, G.R. Hicks, K. Wolff, W. Matsumura, and 

J.P. Davies. 2006. Mutations in an auxin receptor homolog AFB5 and in SGT1b confer 

resistance to synthetic picolinate auxins and not to 2,4-dichlorophenoxyacetic acid or 

indole-3-acetic acid in Arabidopsis. Plant Physiology 142:542-552. 

Zheng, H. and J.C. Hall. 2001. Understanding auxinic herbicide resistance in wild 

mustard: physiological, biochemical, and molecular genetic approaches. Weed Science 

49:276-281. 
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Appendix L: Comparative Analysis of Dicamba and Alternative Soybean 

Herbicides 

L.1. Introduction 

Dicamba is a selective broadleaf herbicide belonging to the auxin agonist class, the oldest 

known class of synthetic herbicides, and is a member of the benzoic acids sub-group. 

Dicamba mimics the action of the plant hormone indole acetic acid and causes rapid 

uncontrolled cell division, and growth leading to plant death. Dicamba has been 

registered for agricultural uses in the U.S. since 1967 and has been widely used in 

agricultural production for over forty years. Dicamba is presently approved for use on 

asparagus, corn, cotton, grass seed production, pasture and rangeland grasses, small 

cereals including barley, oats, rye, and wheat, sorghum, soybean, and sugarcane. 

Dicamba is also used for industrial vegetation management (e.g., forestry and roadsides), 

professional turf management (e.g., golf courses, sports complexes), and residential turf 

(U.S. EPA, 2009b). 

Monsanto Company has developed biotechnology-derived soybean MON 87708 that is 

tolerant to dicamba (3,6-dichloro-2-methoxybenzoic acid) herbicide. MON 87708 offers 

growers expanded use of dicamba in soybean production. The excellent crop tolerance of 

MON 87708 to dicamba facilitates a wider window of application in soybean, allowing a 

preemergence application up to emergence (cracking) and in-crop postemergence 

applications through the R1/R2 stage of growth. Dicamba provides effective control of 

over 95 annual and biennial weed species, and control or suppression of over 100 

perennial broadleaf and woody plant species. MON 87708 will be combined with 

MON 89788 (Roundup Ready 2 Yield soybean) utilizing traditional breeding 

techniques. The combination of herbicide-tolerance traits allows the use of dicamba and 

glyphosate herbicides in an integrated weed management program to control a broad 

spectrum of grass and broadleaf weed species, such that the two herbicides can be used in 

sequence or tank-mixed. Monsanto has submitted an application to the Environmental 

Protection Agency (U.S. EPA) to register this new use pattern for dicamba on 

MON 87708 OPP Decision Number D-432752). 

The availability of MON 87708 integrated into the Roundup Ready soybean system will 

result in a simple and effective dual mode-of-action herbicide system that will control 

hard-to-control broadleaf weeds, assist in the management of resistant broadleaf weeds, 

including glyphosate-resistant biotypes, and subsequently displace some soybean 

herbicides currently in use today (also referred to as alternative herbicides). 

The intent of this comparative analysis is to define current herbicide use in U.S. soybean 

production, and to compare dicamba’s human health and environmental properties to 

herbicides currently used by growers for weed control. In order for a pesticide 

(herbicide) to be registered by EPA it must meet the FIFRA and FFDCA standards for 

 

 Roundup Ready 2 Yield is a registered trademark of Monsanto Technology LLC. All other trademarks 

are the property of their respective owners. 
Monsanto Company 10-SY-210U 607 of 721 

safety to human health and the environment. The EPA must conclude that the herbicide 

when used according to the label does not pose an unreasonable risk to humans or the 

environment, and, in order to establish a tolerance for the use of a herbicide on a food or 

feed crop, find there is a reasonable certainty of no harm to human health from nonoccupational (food, water and residential/recreational) exposures to the herbicide. 

Consequently all alternative soybean herbicides can be used safely, and do not pose a risk 

to humans or the environment. 

Nonetheless, in some instances dicamba offers a reduction in risk potential compared to 

some alternative herbicides, in the same risk category (e.g., acute human risk, aquatic 

plant risks). In other instances dicamba presents a similar risk potential compared to the 

some alternatives. In some instances, dicamba presents a greater risk potential compared 

to some alternatives. 

Overall, the use of dicamba on MON 87708 incorporated into the Roundup Ready 

soybean system will offer a benefit compared to alternative soybean herbicides. For 

human health, aquatic plants and animals, and preemergence application flexibility. 

dicamba use on MON 87708 provides an overall reduction of potential risk (combination 

of “Yes” and “No” entries in Table L-17) compared to the alternatives except for 

flumiclorac-pentyl. Specifically,: 

 Formulations based on the diglycolamine salt of dicamba, such as Clarity® or M1691 

herbicide, have favorable acute toxicity profiles that reduce the risk of acute adverse 

effects for applicators, agricultural workers and bystanders compared to six 

alternative herbicide products (Section L.5.1.1). 

 The active ingredient (a.i.) dicamba has favorable chronic, reproduction, 

carcinogenicity and developmental toxicity characteristics, which reduce the potential 

for risks from exposure to applicators, agricultural workers and consumers, compared 

to exposure from eight alternative herbicide products (Section L.5.1.2). 

 Dicamba has very low toxicity to fish and aquatic invertebrates, and based on EPA 

high end exposure screening level assessment methodology will not affect listed and 

non-listed aquatic plants and animals, which reduces the potential risk to aquatic 

organisms as compared to seven alternative products(Section L.5.2.1 and L.5.2.2). 

 Dicamba use on MON 87708 will strengthen and extend the benefits of glyphosatebased weed control in soybean, which has many well-known and recognized 

environmental and human health benefits as acknowledged in previous EPA Office of 

Pesticide Programs (OPP) decisions to grant reduced risk status to multiple 

glyphosate-tolerant crop uses, i.e., corn, canola, and sugar beet.47 Additionally, the 

use of glyphosate-based weed control in soybean has provided growers with more 

profit opportunities than conventional soybean by reducing input costs (Gianessi, 

2005). By utilizing both active ingredients, which have different herbicidal modes-of- 

 

47 http://www.epa.gov/opprd001/workplan/completionsportrait.pdf
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action, the risk to soybean production posed by weeds that are hard-to-control with 

glyphosate alone, or have developed resistant to glyphosate, is reduced. Hard-tocontrol weeds generally require a higher rate and/or application at a smaller growth 

stage in order to consistently achieve commercially acceptable control, and includes 

copperleaf, hemp sesbania, morningglory, prickly sida, velvetleaf, waterhemp, and a 

number of other broadleaf and grassy weeds. Refer to the Roundup WeatherMax 

label (U.S. EPA Reg. No. 524-537) for a listing of these weeds. 

 Dicamba use on MON 87708 will also provide growers with the option to use an 

herbicide with a different mode-of-action to manage weed species (e.g., Pigweed) 

where certain biotypes have demonstrated resistance to herbicide classes other than 

glyphosate, such as protoporphyrinogen oxidase (PPO) or acetolactate synthase 

(ALS) inhibitors (Section L.5.3.3). Herbicide resistant weeds are those listed on the 

International Survey of Resistant Weeds website (www.weedscience.org). 

 Dicamba use on MON 87708 will reduce the risk to soybean production that ensues 

when growers utilize alternate herbicides that have long rotation restriction periods or 

pose potential for substantial crop injury and loss of yield (Section L.5.3.2.1). 

 Planting glyphosate tolerant soybean thereby allowing the use of glyphosate to 

effectively control weeds in no-till fields has made no-till a viable production system 

for soybean (Pedersen, 2008). The use of dicamba on MON 87708 will reduce the 

risk of growers reverting back to conventional tillage practices from conservation 

tillage practices due to concerns over weed control or resistance and thereby foster 

continued adoption of conservation tillage practices, which is an important goal for 

the agro-ecosystem and the long term sustainability of U.S. agriculture. The 

integration of MON 87708 into the Roundup Ready soybean system will allow the

flexibility to incorporate a second herbicide and mode-of-action in preemergent or 

postemergent applications and support the continued use of conservation tillage 

production (Section L.5.3.3). 

In conclusion, dicamba provides a similar and in some cases a more favorable 

comparative risk profile compared to other alternative herbicide products; the use of 

dicamba on MON 87708 will positively impact integrated pest management practices and 

sustainability of soybean production in the United States by providing a valuable weed 

management tool for this important agricultural crop that imparts greater flexibility and 

weed control options, and will help to slow the selection for herbicide-resistant weed 

biotypes for all herbicide modes-of-action (i.e., ALS, HPPD, PPO, glyphosate) currently 

registered for use in soybean; and will continue to support adoption of no-till and 

conservational tillage practices. 

 
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L.2. Background

L.2.1. Chemical Name and Structure 

Dicamba (3,6-dichloro-2-methoxybenzoic acid) is a carboxylic acid that can form salts in 

aqueous solution. The chemical structure is provided in Figure L.1. Dicamba products 

registered for agricultural uses are formulated with various dicamba salts. The 

formulated products Clarity and M1691 contain the diglycolamine salt of dicamba at a 

nominal level of 56.8% by weight, which is equivalent to 38.5% by weight dicamba acid 

(also referred to acid equivalents or a.e.). 

O

CH3

Cl

Cl

O OH

Figure L-1. Structure of Dicamba 

L.2.2. Herbicide Class and Herbicidal Mode-of-Action 

Dicamba belongs to the “benzoic acid class” of herbicides. The Weed Science Society of 

America (WSSA) places dicamba in herbicide group number 4.48 It is an auxin agonist; it 

is a plant hormone (indole acetic acid, IAA) mimic that causes rapid uncontrolled cell 

division and growth, leading to plant death. Dicamba is mainly used to control broadleaf 

weeds and woody plants. 

L.2.3. Current U.S. Uses and Established Tolerances 

Dicamba is a selective herbicide labeled for control of certain broadleaf weeds and 

woody plants. It was first registered for agricultural use in the U.S. in 1967 and is widely 

used today in agricultural, industrial, and residential settings. Dicamba salts have 

approved uses on asparagus, corn, cotton, grass seed production, pasture and rangeland 

grasses, small cereals including barley, oats, rye, and wheat, sorghum, soybean, and 

sugarcane. Dicamba is also used for industrial vegetation management (e.g., forestry and 

roadside right-of-ways), professional turf management (e.g., golf courses, sports 

complexes), and residential turf (U.S. EPA, 2009b; Durkin and Bosch, 2004). 

 

48 There are several systems of herbicide mode-of-action classification. Among the most widely used are 

those of the Herbicide Resistance Action Committee (HRAC) and the WSSA. The classifications are 

compared in a chart at: http://www.plantprotection.org/HRAC/Bindex.cfm?doc=MOA.html. Accessed 

May 27, 2010 
Monsanto Company 10-SY-210U 610 of 721 

Established food and feed tolerances are listed at 40 CFR 180.227, and in addition to the 

crop plants, they include residue limits for meat, milk, and meat by-products that may 

arise when livestock consume dicamba-treated commodities. 

L.2.4. Proposed use pattern for dicamba on MON 87708 

Monsanto has submitted to the EPA an application to amend EPA Reg. No. 524-582 to 

register a new use pattern for dicamba. The new use pattern allows for sequential 

applications of dicamba, formulated as the diglycolamine salt, to MON 87708: 

 Preplant / preemergent applications, totaling up to 1 pound per acre of dicamba a.e. 

 One or two postemergent in-crop applications (up to 0.5 pounds a.e./acre each), timed 

between soybean emergence (cracking) and early flowering (the R1/R2 growth stage) 

of the soybean. 

The proposed use pattern for dicamba on MON 87708 is within the maximum single and 

annual application rates that have previously been assessed by EPA. Once approved by 

EPA, this label will become a legally-binding constraint on the use of dicamba on 

MON 87708. 

Residue analyses conducted by Monsanto, in which MON 87708 was treated at the 

proposed maximum application rates, were found to produce total dicamba-derived 

residues in soybean seeds at a median level of <0.065 ppm (range of <0.041 to 0.471 

ppm). These levels are substantially less than the current established food tolerance; the 

highest measured residue from the MON 87708 residue study (0.471 ppm) was only 

4.7% of the established 10 ppm dicamba soybean tolerance. Therefore, the proposed 

maximum dicamba treatments to MON 87708 (allowing up to 1 lb a.e. of dicamba per 

acre applied at a preemergent timing plus up to two postemergent in-crop applications of 

up to 0.5 lb a.e./acre each), will not increase dietary exposure to dicamba residues in 

soybean seed beyond that which is already permitted within the existing legal food 

tolerance. 

Residues were also measured in soybean forage and hay of treated MON 87708. 

Monsanto is seeking the establishment of feed tolerances for soybean forage and hay to 

allow feeding of these dicamba-treated MON 87708 commodities to livestock. However, 

these dicamba residues will not increase the livestock dietary burden used to establish the 

current animal commodity food tolerances, so all dietary exposures from the proposed 

new dicamba use on MON 87708 have already been accounted for within the existing 

dicamba food tolerances. 

L.3. Properties of Dicamba Herbicide

L.3.1. Human Health 

MON 87708, treated with preemergent and over-the-top applications of dicamba 

herbicide has the opportunity to reduce risks to human health by: 
Monsanto Company 10-SY-210U 611 of 721 

 Replacing, in part, currently used or foreseeable future alternative herbicide products 

with less favorable risk profiles, and 

 Providing an effective tool to manage hard-to-control broadleaf weeds or broadleaf 

weeds that are resistant to glyphosate, PPO inhibitors, or ALS inhibitors, thereby 

preserving the ability of growers to manage broadleaf weed problems and maximize 

yields in a less risky manner, recognizing the human health benefits offered by the 

superior toxicity and reduced risk profile of glyphosate and the risk profile 

demonstrated by dicamba. 

Both dicamba, as the Clarity or the M1691 formulation, and glyphosate, as various 

Roundup-branded formulations, have “CAUTION!” signal words (CAUTION is the most 

favorable of three possible label signal words that can be required by EPA) and favorable 

chronic toxicity profiles. In a comparative analysis with other alternative soybean weed 

control products, dicamba products offer better, or at least equivalent, human health 

safety profiles, as discussed below. 

In 2006, EPA issued the Reregistration Eligibility Decision (RED) document for dicamba 

and its associated salts (U.S. EPA, 2009b). The RED document, and the related Health 

Effects Division (HED) chapter (U.S. EPA, 2005b), presented an overview of the 

toxicological properties of dicamba, which is summarized below. 

The measurement of human health is the result of conventional laboratory testing against 

standard indicator species (generally rats, mice, and dogs) and is required for the 

registration of a pesticide by the EPA. Results are presented using standard toxicity 

indices, such as the concentration or dose required for 50% lethality (LC50 or LD50), the 

highest dosing level that produced No Observable Adverse Effects (NOAEL), or the 

lowest dosing level that produced an Observable Adverse Effect (LOAEL). The results 

of the acute (single exposure) toxicity studies for dicamba are presented in Table L-1. 

Results for developmental, reproduction, mutagenic and neurotoxicological studies are 

presented in Table L-2. Subchronic, chronic and carcinogenicity study results are 

presented in Table L-3. 

Table L-1. Dicamba Acid Acute Toxicity Study Findings 

Study Endpoint EPA Category1

Acute oral (rat) LD50 2740 mg/kg III 

Acute dermal (rat) LD50 2000 mg/kg III 

Acute inhalation (rat) LC50 5.3 mg/L IV 

Primary eye irritation (rabbit) Irritant II 

Primary dermal irritation (rabbit) Irritant II 

Dermal sensitization (guinea pig)2

 Negative NA 

1

EPA acute toxicity categories range from I (worst) to IV (best).

2

Determination of the potential to cause or elicit skin sensitization reactions (allergic contact dermatitis) is 

an important element in evaluating a substance’s toxicity.
Monsanto Company 10-SY-210U 612 of 721 

Table L-2. Dicamba Acid Reproductive, Developmental, Mutagenic, and 

Neurotoxicologic Findings 

Study 

Systemic Toxicity Endpoint 

(mg/kg body wt/day) 

Offspring Toxicity 

Endpoint (if any) 

(mg/kg body wt/day) 

Developmental (rat) Maternal NOAEL 160; LOAEL 400. 

Clinical signs: decreased food 

consumption and weight gain, 

increased mortality. 

Developmental 

NOAEL 400 (HDT1). 

Developmental 

(rabbit) 

Maternal NOAEL 62.5; LOAEL 150. 

Clinical signs: decreased motor 

activity, ataxia, increased abortion. 

Developmental 

NOAEL 62.5; LOAEL 

150. Clinical signs: 

Increased abortion. 

Developmental 

Neurotoxicity 

Not Required 

Reproduction, 

multigeneration (rat) 

Parental NOAEL 122/136 (M/F2); 

LOAEL 419/450 (M/F). Clinical 

signs: reduced righting reflex. 

Reproductive NOAEL 122; LOAEL 

419. Delayed F1 male maturation. 

Offspring NOAEL 45; 

LOAEL 136. Clinical 

signs: Decreased pup 

weights, all 

generations. 

Acute Neurotoxicity 

(rat) 

NOAEL not established; LOAEL 

300. Clinical signs: Impaired gaits 

and righting reflex, impaired 

respiration, rigidity. 

Subchronic 

Neurotoxicity (rat) 

NOAEL 401/472 (M/F); LOAEL 

768/1029 (M/F). Clinical signs: 

Rigidity, slightly impaired gait and 

righting reflex. 

Gene Mutation – 

Salmonella 

Not mutagenic. 

Chromosome 

aberration (CHO3

) 

Aberrations not induced at any tested 

concentration with or without S9 

activation. 

Unscheduled DNA 

Synthesis (UDS) 

No Evidence of UDS up to 3000 

μg/mL 

1

HDT stands for the highest dose tested. 

2

M/F stands for males/females. 

3

Chinese hamster ovaries. 
Monsanto Company 10-SY-210U 613 of 721 

Table L-3. Dicamba Acid Subchronic, Chronic and Cancer Findings 

Study 

Toxicity Endpoints 

(mg/kg body weight/day) 

Subchronic Oral (rat) NOAEL 479/536 (M/F1); LOAEL 

1000/1065 (M/F). Clinical signs: decreased 

weight gains, liver effects. 

28-day dermal (rat) NOAEL 1000 (HDT2). 

Chronic / Carcinogenicity (rat) NOAEL 107/127 (M/F; HDT). Not 

carcinogenic. 

Chronic (dog) NOAEL 52 (HDT). 

Carcinogenicity (mouse) NOAEL 358/354 (M/F); (HDT). Not 

carcinogenic. 

1

M/F stands for males/females. 

2

HDT stands for the highest dose tested. 

The EPA has classified dicamba as “Not Likely to Cause Cancer in Humans” (the most 

favorable among EPA’s cancer categories), and concluded that dicamba is not mutagenic 

and is not a developmental toxin. There was no evidence of behavioral or neurological 

effects on offspring and, therefore, a developmental neurotoxicity study was not required 

by the EPA. 

In the human health risk assessments for dicamba, the EPA employed a chronic 

Population Adjusted Dose (cPAD) of 0.45 mg/kg/day, based on a 100-fold safety factor 

applied to the offspring NOAEL in the multigeneration rat study. For evaluating acute 

exposures to dicamba, the EPA employed an acute Population Adjusted Dose (aPAD) of 

1 mg/kg, based on a 300-fold safety factor (for use of the LOAEL rather than the 

NOAEL) applied to the LOAEL in the rat acute neurotoxicity study (U.S. EPA 2008a). 

Using a dietary exposure model to estimate combined exposures from all presently 

approved uses, including both food and water exposure routes, and assuming that 100% 

of all labeled crops are treated with dicamba and that the resulting foods have tolerancelevel residues, total dietary exposure reached only 4.4% of the aPAD level and 2.7% of 

the cPAD level for the general U.S. population; and 11% of the aPAD and 6.8% of the 

cPAD for the most highly exposed subpopulations of children 1-2 years old (U.S. EPA 

2008a). The proposed dicamba use on MON 87708 does not require an increase in the 

soybean seed food tolerance. Monsanto has requested new tolerances for soybean forage 

and hay; however, these do not increase the livestock dietary burden utilized in the RED 

assessment, so all dietary exposures from the proposed new dicamba use on MON 87708 

are already accounted for in these assessments. 

 
Monsanto Company 10-SY-210U 614 of 721 

L.3.2. Toxicology of Dicamba Plant and Animal Metabolites 

MON 87708 has been genetically enhanced to express a dicamba metabolizing enzyme 

(dicamba mono-oxygenase). The enzyme catalyzes a mono-oxygenation reaction 

resulting in an oxidative demethylation of dicamba, forming 3,6-dichloro-2-

hydroxybenzoic acid, also known as 3,6-dichlorosalicylic acid (DCSA). In the dicambatreated MON 87708, glucoside conjugates of DCSA were the major plant metabolites. 

Cl OH

Cl

O OH

Figure L-2. Structure of DCSA 

DCSA is a known metabolite of dicamba in soil, plants, and livestock. It is presently 

included in the residue expression specified in the food and feed tolerance for dicamba in 

40 CFR 180.227(a)(3). Therefore, the existing food tolerance for soybean seed includes 

DCSA residues. 

In the RED, EPA considered that DCSA has structural similarity to dicamba, and 

concluded that it would have similar toxicity to the parent dicamba. Monsanto has 

conducted and submitted additional toxicity studies involving direct dosing with DCSA 

(U.S. EPA OPP Decision Number D432752) to further substantiate the conclusion 

reached by the EPA in the RED. The results of these studies, summarized in Table L-4, 

can be compared to those of dicamba (Tables L-1 through L-3). Monsanto has submitted 

these studies to the EPA in support of our application to register the use of dicamba on 

MON 87708. 
Monsanto Company 10-SY-210U 615 of 721 

Table L-4. Summary of Toxicological Findings from Testing of DCSA 

Study 

Systemic Toxicity 

Endpoint (mg/kg body 

wt/day) 

Offspring Toxicity 

Endpoint (if any) 

(mg/kg body wt/day) 

Acute oral (rat) LD50 2641 (Category III) NA 

Developmental (rat) Maternal NOAEL 100 Developmental NOAEL 

100

Developmental (rabbit) Maternal NOAEL 25 Developmental NOAEL 65 

(HDT)

Reproduction, 

multigeneration (rat) 

Parental NOAEL 42 (M/F1

combined)

Offspring NOAEL 42 

Gene mutation – S.

typhimurium & E. coli 

Not mutagenic NA 

In vitro chromosome 

aberration (CHO) 

Aberrations not induced NA

Micronucleus (mouse) Negative NA

Subchronic (90-day) oral 

(Rat) 

NOAEL 362/222 (M/F) NA

Chronic (12-month)(rat) NOAEL 171/206 (M/F). NA

Subchronic (90-day) 

(dog) 

NOAEL 50 NA

In Vitro cytogentics 

(human lymphocytes) 

Weakly positive with S9 

activation 

NA

In Vivo cytogenetics (rat) Negative NA

Carcinogenicity Ongoing NA

NA denotes Not Applicable. 

1

M/F stands for males/females. 

Smaller amounts of a glucoside of another known metabolite, 5-dichloro-3,6-

dihydroxybenzoic acid (3,6-dichlorogentisic acid, DCGA) were also identified in the 

soybean metabolism study. Levels of the DCGA glucoside were less than 10% of total 

MON 87708 soybean-contained radioactivity. Monsanto conducted a limited set of 

toxicity studies on DCGA; and has provided these studies to EPA to support our 

application to register dicamba on MON 87708. 

The results of the DCSA and DCGA toxicity studies substantiate the EPA conclusion that 

dicamba metabolites will have similar toxicity to parent dicamba. 

L.3.3. Dicamba Ecological Effects – Non-target Species 

The following tables summarize the hazard potency of dicamba to non-target species. 

These data were taken from the Environmental Fate and Effects Division (EFED) 

Chapter of the dicamba RED (U.S. EPA, 2005a). These measures are the result of 

conventional laboratory testing against standard indicator species of fish, birds, 

mammals, and plants as required for EPA registration. Results are presented using 
Monsanto Company 10-SY-210U 616 of 721 

standard toxicity indices, such as the concentration or dose required for 50% lethality 

(LC50 or LD50) or the concentration required for a 25 or 50% reduction in growth or 

biomass (EC25 or EC50). In some cases the “No Observable Effect Concentration” 

(NOEC) or “No Observable Adverse Effect Level” (NOAEL) is also listed. A summary 

of the findings of the ecotoxicity studies conducted on dicamba acid is provided in Tables 

L-5 through L-7. Ecotoxicity studies have also been conducted on the diglycolamine 

(DGA) salt of dicamba and are summarized in Table L-8. 

Table L-5. Dicamba Acid Ecotoxicity Findings on Mammals, Birds and Fish (U.S. 

EPA. 2005a) 

Study Toxicity Endpoint Comment 

Mammals 

Dicamba acid acute oral (rat) LD50 2740 mg/kg body wt 

Multigeneration reproduction (rat) 

NOAEL/LOAEL 

45 /136mg/kg/day 

Birds 

Dicamba acid acute oral (quail) LD50 188 mg/kg body wt 

Dicamba acid acute oral (duck) LD50 1373 mg/kg body wt 

Dicamba acid sub-acute oral (quail) LC50 >10,000 mg/kg diet 

Dicamba acid sub-acute oral (duck) LC50 2009 mg/kg diet 

Dicamba acid sub-acute oral (duck) LC50 >10,000 mg/kg diet Replicate 

Dicamba acid reproduction (quail) 

NOEC/LOEC 

1390 (HDT1

) mg/kg 

diet 

Dicamba acid reproduction (duck) 

NOEC/LOEC 

695/1390mg/kg diet 

Fish 

Dicamba acid (trout) LC50 28 mg/L 

Dicamba acid (trout) LC50 135 mg/L Replicate 

Dicamba acid (trout) LC50 153 mg/L Replicate 

Dicamba acid (Bluegill) LC50 135 mg/L 

Dicamba acid (Bluegill) LC50 >50 mg/L Replicate 

Dicamba acid (Sheepshead) LC50 >180 mg/L 

Dicamba acid fish early life stage NA 

Dicamba acid fish life cycle NA 

NA denotes Not Applicable. 

NOEC stands for No Observable Effect Concentration. 

LOEC stands for Lowest Observable Effect Concentration. 

1

HDT stands for the Highest Dose Tested. 

 
Monsanto Company 10-SY-210U 617 of 721 

Table L-6. Dicamba Acid Ecotoxicity Findings on Aquatic and Terrestrial 

Invertebrates (U.S. EPA. 2005a) 

Study Toxicity Endpoint Comment 

Aquatic Invertebrates 

Dicamba acid (Daphnia) LC50 111 mg/L 

Dicamba acid (Daphnia) LC50 >100 mg/L Replicate 

Dicamba acid (Sowbug) LC50 >100 mg/L 

Dicamba acid (Scud) LC50 >100 mg/L 

Dicamba acid (Grass Shrimp) LC50 >132 mg/L 

Dicamba acid (Fiddler Crab) LC50 >173 mg/L 

Dicamba acid (Oyster) LC50 >1 mg/L 

Dicamba acid (Glass Shrimp) LC50 >56 mg/L 

Invertebrate life cycle NA 

Terrestrial Invertebrates 

Honeybee LD50 >90.6 μg/bee 

NA denotes Not Applicable. 

Table L-7. Dicamba Acid Ecotoxicity Results on Aquatic and Terrestrial Plants 

(U.S. EPA. 2005a) 

Study Toxicity Endpoint Comment 

Aquatic Plants 

Dicamba acid duckweed EC50 / NOEC >3.25 / 0.2 mg/L 

Dicamba acid green alga EC50 / NOEC >3.7 / 3.7 mg/L 

Dicamba acid marine diatom EC50 / NOEC 0.49 / 0.011 mg/L 

Dicamba acid blue-green alga EC50 / NOEC 0.061 / 0.005 mg/L 

Dicamba acid freshwater diatom EC50 / NOEC 2.3 / 0.5 mg/L 

Terrestrial Plants 

Dicamba acid seedling emergence monocot 

EC25 / NOEC 

0.004 / <0.032 lb/a Onion 

Dicamba acid seedling emergence dicot EC25 / 

NOEC 

0.0027 / < 0.0022 lb/a Soybean 

Dicamba acid vegetative vigor monocot EC25 / 

NOEC 

0.15/ 0.13 lb/a Onion 

Dicamba acid vegetative vigor dicot EC25 / 

NOEC 

0.0068 / <0.004 lb/a Soybean 
Monsanto Company 10-SY-210U 618 of 721 

Table L-8. Dicamba, Diglycolamine (DGA) Salt Ecotoxicity Findings (U.S. EPA. 

2005a) 

Study Toxicity Endpoint Comment 

Birds 

Dicamba DGA salt acute oral (quail) LD50 262 mg a.e./kg body 

weight 

Dicamba DGA salt sub-acute oral (quail) LC50 >1522 mg a.e./kg diet 

Dicamba DGA salt sub-acute oral (duck) LC50 >1522 mg a.e./kg diet 

Fish 

Dicamba DGA salt (trout) LC50 >270mg a.e./L 

Dicamba DGA salt (bluegill) LC50 >270mg a.e./L 

Aquatic Invertebrates 

Dicamba DGA salt (Daphnia) LC50 >270 mg a.e./L 

 As a part of the reregistration evaluation under FIFRA, EPA conducted an ecological 

screening assessment for dicamba. This assessment compared the results from 

toxicity tests with dicamba conducted with various plant and animal species to 

conservative (high end) estimates of the concentration of dicamba to which an 

organism might be exposed in the environment. These estimates, called the 

Estimated Environmental Concentrations (EECs), are point estimates for specific 

types of exposure (e.g., aquatic or dietary) that assume constant high concentrations 

throughout the lifespan of the organism. These estimates do not take into account 

normal variation in environmental concentrations, the dilution or dissipation of those 

concentrations, or the frequency of exposure of wildlife to the pesticide. Such 

assumptions provide a screening level of assessment where conclusions of no harm 

can be drawn with high confidence; however, it is also possible to reach a conclusion 

of no harm with more refined exposure assumptions. EPA derived exposure 

estimates for a number of use patterns (U.S. EPA, 2005a), including two that are 

reasonably close to the proposed use pattern of dicamba on MON 87708. These use 

patterns are: 1) for wheat, with two 1.0 lb a.e./acre applications; and 2) for sorghum, 

with two 0.5 lb a.e./acre applications. For both crops, the two applications were 

considered to occur on May 1st and June 1st. Based on these highly conservative 

screening level assessments, EPA’s Environmental Fate and Effects Division (EFED) 

identified the concerns for non-target species effects (terrestrial plants, Risks to 

terrestrial plants; acute risks to birds, and chronic risk to mammals in their risk 

assessment for the 2006 Dicamba RED (U.S. EPA, 2009b), however EPA concluded 

in the final reregistration decision document that no specific additional drift 

mitigations were needed to support the continued registration of all dicamba uses. 

 
Monsanto Company 10-SY-210U 619 of 721 

L.3.4. Environmental – Soil, Air, and Water Quality 

The following tables summarize the physical and chemical properties of dicamba that can 

influence its mobility and persistence in the environment. These measures are the result 

of conventional laboratory testing, as required for EPA registration. Results are 

presented using standard environmental fate indices, such as the time required for 50% 

degradation (half-life) of the initial concentration. 

Table L-9. Environmental Fate and Physical Properties of Dicamba Acid 

Environmental Physical Properties Results 

Dicamba acid vapor pressure 3.4 x 10-5 torr (25° C) 

Dicamba acid acidity (pKa) 1.87 

Dicamba acid water solubility 6100 mg/L 

Dicamba acid octanol / water partition coefficient 0.1 

Dicamba acid Koc (Freundlich soil-binding constant) 3.5 – 21.1 mL/g 

Dicamba acid field dissipation half-life (conducted 

with salt formulations) 

3 – 19.8 days 

Dicamba acid aerobic soil half-life, Laboratory 6 days 

Dicamba acid anaerobic soil half-life, Laboratory 141 days 

Dicamba acid aerobic aquatic half-life, Laboratory 20.2 – 24.3 days 

Dicamba acid aqueous photolysis half-life 38.1 days 

Dicamba acid soil photolysis degradation rate ~20% after 30 days 

Hydrolysis half-life Stable 

L.3.4.1. Risks of Leaching and Runoff 

The high water solubility, relatively low soil-binding coefficient, and a pKa that requires 

dicamba to exist primarily as an anion at normal soil pH suggest that dicamba may have 

risk of leaching or runoff. These indicators of low soil-binding are largely offset by the 

rapid soil microbial breakdown of both dicamba and its DCSA metabolite under aerobic 

conditions, leading overall to only a low to moderate risk of leaching or runoff. Using 

the Pesticide Root Zone Model/Exposure Analysis Modeling System (PRZM/EXAMS51) 

and Screening Concentration in Ground Water (SCI-GROW52) models, EPA estimated 

potential drinking water concentrations originating from ground and surface water for use 

in the dicamba dietary risk assessment. Model results indicated potential dicamba ground 

and surface water concentrations of 0.016 μg/L and 13.8 μg/L, respectively (U.S. EPA, 

 

51 PRZM-EXAMS Model Description. Environmental Fate and Effects Division, Office of Pesticide 

Programs, U.S. Environmental Protection Agency, Washington, D.C. 

http://www.epa.gov/oppefed1/models/water/models4.htm#przm. Accessed May 28, 2010

52 SCI-GROW Model Description. Environmental Fate and Effects Division, Office of Pesticide Programs, 

U.S. Environmental Protection Agency, Washington, D.C. 

http://www.epa.gov/oppefed1/models/water/scigrow_description.htm. Accessed May 28, 2010. 
Monsanto Company 10-SY-210U 620 of 721 

2008). Surface water modeling using PRZM-EXAMS described in the EFED RED 

Chapter (U.S. EPA, 2005a) predicted levels ranging from 4.9 to 18 μg/L for a one lb per 

acre dicamba a.e. application rate to soybean (the highest single proposed application rate 

for MON 87708). Dicamba has been in use for many years on a significant number of 

acres in the corn belt, with approximately 36 million treated-acres in 1994 alone (Table 

VIII-11), so that data from groundwater monitoring studies provide additional evidence 

pertaining to the potential for dicamba to leach. There has been a low incidence of 

dicamba detections (approximately 3% of sites) in the U.S. Geological Survey National 

Water Quality Assessment (NAWQA) monitoring program (U.S. EPA, 2005b). The 

highest detection levels were approximately 2 μg/L. Altogether, the modeling 

predictions and the NAWQA monitoring results show that dicamba concentrations that 

might occur in drinking water are very low, and it confirms that the potential risk of 

dicamba leaching or runoff does not threaten human health or acceptable water quality. 

The lifetime Health Advisory Level (HAL) 53for dicamba is 4,000 μg/L (U.S. EPA, 

2011). 

L.3.4.2. Other Hazards 

Dicamba does not include any fluorine atoms, which are characteristic of ozone depletion 

risk. Based on the vapor pressure presented in Table L-9, dicamba is not sufficiently 

volatile to be present in the stratosphere at ozone-threatening levels. Dicamba acid, nor 

its water soluble salts or derived formulation, does not have the properties of 

explosiveness, flammability, or corrosion. 

L.3.4.3. Efficacy and Weed Management Practices for Dicamba on MON 87708 

Since the introduction of the Roundup Ready® soybean crop product in 1996, glyphosate 

herbicides have become the predominant weed control tool in soybean production. That 

has been beneficial for growers, because glyphosate use in the Roundup Ready soybean 

system offers unmatched flexible weed control at a relatively low price; the high adoption 

rate of the Roundup Ready soybean system by growers is a testament to its ease of use 

and ability to provide full spectrum weed management. The adoption of the Roundup 

Ready soybean system has provided additional benefits from a risk perspective, because 

glyphosate is widely-recognized as an herbicide that has a favorable profile in terms of 

human health, non-target animal species, and ground and surface water. A significant 

step that can be taken toward minimizing future potential risk from herbicides is to 

strengthen and support the continued sustainability of glyphosate use in soybean 

production by providing a complementary tool to manage hard-to-control weeds and, as a 

result, optimize soybean yield. As stated earlier, EPA has acknowledged the value of the 

glyphosate-based weed control system and the need to have other herbicide modes of 

action available for IPM purposes, including weed resistance management, particularly in 

 

53 The concentration of a chemical in drinking water that is not expected to cause any adverse noncarcinogenetic effects for a lifetime of exposure. 

® Roundup Ready is a registered trademark of Monsanto Technology LLC. All other trademarks are the 

property of their respective owners. 
Monsanto Company 10-SY-210U 621 of 721 

its decision to continue registration of herbicides in cotton (U.S. EPA, 2009a). In this 

revised RED decision on MSMA (an organic arsenic herbicide), EPA reversed it decision 

on continued registration of this active ingredient in cotton based in part on the 

aforementioned IPM benefits. 

According to 2006 market data 75.59 million acres of soybeans were planted in 2006, and 

74.6 million acres were harvested (USDA-NASS, 2007a). Soybean-growing acres 

received approximately 2.0 herbicide treatments (AgroTrak, 2009), leading to 

approximately 149 million herbicide treated acres. Based on the most recently available 

USDA-NASS figures from 2006, 97% of the herbicide total treated acres were made with 

glyphosate herbicides, and the remaining 32% of treatments were made with more than 

25 other active ingredients (USDA-NASS 2007b). 

In 2008, dicamba-treated soybean acres only accounted for 0.53 million acres, or 0.7% of 

the soybean acres (Table VIII-12). This is primarily because current commercial soybean 

varieties are not tolerant to dicamba, and therefore dicamba is only labeled for application 

at timings that avoid contact with the growing plant or when some level of injury is 

acceptable: (1) preplant treatments 2 to 4 weeks prior to planting, depending on rate and 

rainfall, or (2) preharvest treatments after the seeds have matured, at least 14 days before 

harvest. 

As a consequence of the intensive use of glyphosate for weed control in Roundup Ready 

soybean over the past 13 seasons, certain problematic weeds (glyphosate hard-to-control 

and-resistant) have become more common, although glyphosate is still considered to 

provide very broad spectrum weed control. Since the registration and market 

introduction of glyphosate, it has been known that certain weeds are more difficult to 

control than others at any given herbicide rate and timing of application. Glyphosate, as 

with any herbicide, when used repeatedly season-after-season without other weed 

management practices, can select for those weeds that fall in this “difficult-to-control” 

category, including weeds such as morning glory, nightshade, and wild buckwheat, so 

that their populations can increase if the correct rate and application timing are not 

practiced. For some weed species biotypes, selection and resistance have occurred in 

some geographical locations. In addition, resistance has developed for some populations 

of weeds that were formerly well-controlled by glyphosate in soybean; these include 

glyphosate-resistant waterhemp, Palmer amaranth, horseweed, common ragweed, and 

giant ragweed. As a group, the glyphosate hard-to-control and glyphosate-resistant 

weeds are generally broadleaf dicotyledonous weeds. Accordingly, in Roundup Ready 

soybean fields where these problem weeds are widespread, affecting yield and 

complicating harvest, growers have begun to use a variety of broadleaf herbicides 

sequentially or as a mixture with glyphosate. 

Monsanto, and other companies, have recognized this weed control need, and now 

recommend the incorporation of multiple modes-of-action for weed control. Companies 

are also searching for a companion herbicide for use with glyphosate to manage these 

populations of problematic broadleaf weeds. A complementary herbicide would provide 

new weed control options that strengthen the utility and sustainability of glyphosate as a 

weed control tool in the Roundup Ready soybean system. In the absence of MON 87708 
Monsanto Company 10-SY-210U 622 of 721 

integrated into the Roundup Ready soybean system, growers will need to rely on multiple 

herbicide applications at different timings to gain similar weed control. Dicamba is 

considered an optimum herbicidal companion to glyphosate in control of broadleaf 

weeds, since soybean injury can be eliminated through the introduction of MON 87708, 

because it: 1) has a different mode-of-action from glyphosate; 2) dicamba has shown 

little propensity toward resistance development; 3) is very effective in controlling the 

problematic broadleaf weed spectrum described above; 4) has a short soil half-life, 

without the potential for carry-over problems or plant-back restrictions; 5) can offer 

superior crop safety relative to other postemergent herbicides available for use in 

soybean; 6) is available at a cost appropriate to its benefit; and 7) is compatible in 

mixtures with glyphosate and other herbicides. 

Monsanto has developed MON 87708 by introducing a gene that encodes an enzyme to 

catalyze the breakdown of dicamba. The dicamba mono-oxygenase enzyme cleaves a 

specific carbon-oxygen bond and converts the aromatic methoxyl moiety into a hydroxyl 

group, to form the non-herbicidal metabolite 3,6-dichlorosalicylic acid (DCSA). The rate 

of dicamba breakdown in MON 87708 is sufficiently rapid to provide complete 

protection from herbicidal symptoms, thereby allowing dicamba applications anytime up 

to crop emergence (cracking) as well as two postemergent in-crop applications between 

cracking and the R1/R2 growth stage. Monsanto seeks to offer growers soybean varieties 

that include both the glyphosate- and dicamba tolerance traits (MON 89788 × 

MON 87708), allowing use of either herbicide alone or together in sequence or as a 

mixture, to provide wide flexibility to meet diverse and specific weed control needs in 

individual fields. 

L.4. Properties of Alternative Herbicide Products 

As discussed in L.3.4, ninety-seven percent (97%) of soybean treated acres receive an 

application of glyphosate; the remaining acres are treated with more than 25 other active 

ingredients. In some of the soybean acres, these other active ingredients are applied on 

acres that also receive a glyphosate application. The ten most widely used herbicides are 

shown in Table L-10. Integration of MON 87708 into the Roundup Ready soybean 

system and the subsequent use of dicamba will result in the displacement of some 

currently used, or foreseeable future use herbicides, and therefore the properties of these 

alternative herbicides are summarized in this section to provide a baseline for comparison 

to dicamba use on MON 87708. 
Monsanto Company 10-SY-210U 623 of 721 

Table L-10. Ten Most Widely Used Alternative Herbicides in U.S. Soybean 

Production 

Herbicide Treated acres (millions)1

2,4-D (acid, salts, and esters, combined) 6.32 

flumioxazin 3.45 

imazethapyr 3.35 

cloransulam-methyl 2.90 

chlorimuron-ethyl 2.66 

fomesafen 2.26 

clethodim 2.16 

pendimethalin 2.15 

tribenuron 2.05 

flumiclorac-pentyl 1.32 

1

AgroTrak, 2009 

Table L-11 also summarizes key information from alternate herbicide product labels that 

are included in the comparative analysis in Section L.5. Table L-12 lists the eighteen 

active ingredients that make up the products in Table L-11. 2,4-D, being used primarily 

as a pre-plant application, is the most widely-used herbicide in this alternate herbicide 

list, representing about 10% of treated acres; whereas acifluorfen, carfentrazone-ethyl, 

and flufenacet are the least used among these, representing <0.5% of treated acres. 

Mesotrione has not been used in soybean production previously; the use on soybean was 

only recently registered by the EPA (2009d). Table L-12 also lists general regulatory 

information about each herbicide. Note that only paraquat is classified as a Restricted 

Use pesticide among this group, on the basis of acute toxicological concern. 
Monsanto Company 10-SY-210U 

Table L-11. Alternative Registered Soybean Herbicides 

Brand (U.S. EPA Reg. No.) 

Active Ingredient 

Signal Word1

Active Ingredient Content 

Re-entry Interval (REI)2

Max. Soybean lb/a (single treatment)3

Max. Soybean lb/a (season) Label Warnings or Sp

Aim® (279-3241) Carfentrazoneethyl Caution 2 lb/gal 12 hr 0.008 5 0.023 "toxic to fish";"toxic to 

do not feed foliage; som

Authority® First 

DF (279-3246) 

Sulfentrazone Caution 0.62 lb/lb 12 hr 0.31 0.31 "known to leach"; "tox

estuarine invertebrates.rotation restrictions, up O.M. limits (sands <1% 

Cloransulammethyl 0.08 lb/lb 0.04 0.04 

Authority MTZ (279-3340) 

Sulfentrazone Caution 0.18 lb/lb 12 hr 0.028 0.046 "known to leach"; "tox

estuarine invertebrates. Over The Top); sensitivpossible 

Metribuzin 0.27 lb/lb 0.042 0.07 

Basagran®

(7969-45) 

Bentazon Caution 4 lb/gal 48 hr 1 2 "known to leach";30-da

treated forage and hay; 

Butoxone® 7500 

(71368-49) 

2,4-DB Caution 0.75 lb/lb 48 hr 0.375 soil type limits 

Butyrac® 200 

(42750-38) 

2,4-DB DMA salt Danger 2 lb/gal 48 hr 0.4 0.4 "toxic to fish";60-day P

occur, especially with t

Cadet® (279-

3338) 

Fluthiacet-methyl Warning 0.91 lb/gal 12 hr 0.0065 0.009 do not feed foliage; min

Callisto® (100-

1131) 

Mesotrione Caution 4 lb/gal 12 hr 0.1875 0.1875 “high potential for runo

restrictions up to 18 mtbleaching" may occur; only, no in crop use 
Monsanto Company 10-SY-210U 

Table L-11 (continued). Alternative Registered Soybean Herbicides 

Brand (U.S. EPA Reg. No.) 

Active Ingredient 

Signal Word1

Active Ingredient Content 

Re-entry Interval (REI)2

Max. Soybean lb/a (single treatment)3

Max. Soybean lb/a (season) Label Warnings or Spe

Classic® (352-

436) 

Chlorimuronethyl Caution 0.75 lb/lb 12 hr 0.14 0.14 6 60-day PHI; crop rotatio

to 30 mts and complica3 different intervals specand soil pH; do not feed limits; "temporary leaf y

Cobra® (59369-

34) 

Lactofen Danger 2 lb/gal 12 hr 0.2 0.4 6 "toxic to fish"; Do not a

growth stage R6 / 45-dainjury 

Extreme ®

(241-405) 

Imazethapyr Warning 0.17 lb/gal 48 hr 0.064 6 0.064 6 "properties & characteri

with chemicals detected crop rotation limits 

 Glyphosate-IPA 2 lb/gal 0.75 0.75 FirstRate (62719-275) 

Cloransulammethyl Caution 0.84 lb/lb 12 hr 0.04 0.055 65-day PHI; crop rotatio

to 30 mts; soil types; 14hay feeding restriction 

Flexstar ®

(100-1101) 

Fomesafen Warning 1.88 lb/gal 24 hr 0.35 0.375 6 "cause tumors"; "known 

PHI; do not feed foliagelimits 

Gangster ® Copack (59639-

131) 

Flumioxazin Caution 51% 12 hr 0.096 0.096 "toxic to aquatic inverte

"Preemergent only. “procharacteristics Associatedetected in ground wateinvertebrates." 

Cloransulammethyl 84% 0.032 0.032 

Gramaoxone Inteon ® (100-

1217) 

Paraquat dichloride 

Danger 2 lb/gal 

(cation basis) 

12 hr 1.5 2.9 "toxic to wildlife"; Restr

Over-the-Top use 
Monsanto Company 10-SY-210U 

Table L-11 (continued). Alternative Registered Soybean Herbicides 

Brand (U.S. EPA Reg. No.) 

Active Ingredient 

Signal Word1

Active Ingredient Content 

Re-entry Interval (REI)2

Max. Soybean lb/a (single treatment)3

Max. Soybean lb/a (season) Label Warnings or Spec

Ignite® (264-829) Glufosinateammonium Warning 2.34 lb/gal 12 hr 0.66 0.8 "runoff potential"; "toxic t

plants"; 70-day PHI; some limits up to 180 days; only Liberty Link soybean 

Liberty® (264-660) Glufosinateammonium Warning 1.67 lb/gal 12 hr 0.44 0.8 "runoff potential"; "toxic t

plants"; 70-day PHI; do nocrop rotation limits up to 

Phoenix®

(59639-118) 

Lactofen Caution 2 lb/gal 12 hr 0.3 0.4 6 "toxic to fish"; Do not app

growth stage R6 / 45-day 

Pursuit® (241-310) Imazethapyr Caution 2 lb/gal 4 hr 0.063 0.063 "properties & characterist

chemicals detected in grouday PHI; do not feed forag

Pursuit® Plus 

(241-331) 

Pendimethalin Caution 2.7 lb/gal 24 hr 0.84 0.84 "properties & characterist

chemicals detected in grouto fish"; 85-day PHI; crop to 40 months 

Imazethapyr 0.2 lb/gal 0.063 0.063 

Raptor ® (241-379) Imazamoxammonium Caution 1 lb/gal 4 hr 0.04 0.04 "phytotoxic to all plants"; 

rotation limits up to 26 mowith complicated warning
Monsanto Company 10-SY-210U 

Table L-11 (continued). Alternative Registered Soybean Herbicides 

Brand (U.S. EPA Reg. No.) Active Ingredient 

Signal Word 1

Active Ingredient Content 

Reentry Interval (REI)2

Max. Soybean lb/a (single treatment)3

Max. Soybean lb/a (season) Label Warnings or Specia

Reflex® (100-993) Fomesafen Danger 2 lb/gal 24 hr 0.375 0.375 6 "known to leach"; 45-day P

limits up to 18 mts; minor igeographical restrictions (5 with different rate structure

Resource®

(59639-82) 

Flumicloracpentyl Warning 0.86 lb/gal 12 hr 0.081 0.11 "toxic to shrimp"; 60-day P

forage or hay to livestock; tspotting or burn to soybean 

Scepter ®70 DG 

(241-306) 

Imazaquin Caution 0.7 lb/lb 12 hr 0.123 0.123 6 "properties & characteristic

chemicals detected in grounPHI; do not feed forage or hcrop rotation limits up to 40 limitations (3 regions) 

Sencor® (DF 75%) 

(264-738) 

Metribuzin Caution 0.75lb/lb 12 hr 0.66 6 1.3 6 "can seep or leach"; 70-day 

day PHI on feeding forage Over-the-Top application, dOK; injury in high pH or loon certain crop varieties, crup to 18 mts 

Synchrony® XP 

(352-648) 

Thifensulfuron Caution 0.069 lb/lb 12 hr 0.013 0.013 45-day planting restriction 

soybean planting / emergencomplicated crop rotation rregions, 4 intervals) with limts; do not feed forage or hsoil types; injury if adjuvan

UltraBlazer (70506-60) 

Acifluorfen sodium 

Danger 2 lb/gal 48 hr 0.374 0.5 50-day PHI; minor injury 

 Chlorimuron-ethyl 0.215 lb/lb 0.04 0.04 
Monsanto Company 10-SY-210U 

Table L-11 (continued). Alternative Registered Soybean Herbicides 

Brand (U.S. EPA Reg. No.) 

Active Ingredient 

Signal Word 1

Active Ingredient Content 

Reentry Interval (REI)2

Max. Soybean lb/a (single treatment)3

Max. Soybean lb/a (season) Label Warnings or Specia

Valor® SX 

(59639-99) 

Flumioxazin Caution 0.51 lb/lb 12 hr 0.096 0.096 "runoff potential"; "toxic to 

invertebrates."; preemergenin crop use; do not feed foralivestock; crop rotation limisoil type limits; injury undeconditions or poorly drained on use with flufenacet, alacmetolachlor, or dimenthena

Valor® XLT 

(59639-117) 

Flumioxazin Caution 0.3 lb/lb 12 hr 0.094 0.094 "toxic to aquatic invertebrat

preemergence only, no in crfeed forage or hay to livestolimits up to 30 mts; injury uconditions or poorly drained 

 Chlorimuronethyl 0.103 lb/lb 0.032 0.032 Weedone® (650, 

638, LV4, LV6) and other 2,4-D brands (71368-3, -6, -10, -11, -14, -19) 

2,4-D; 2,4-D salts; 2,4-D esters 

 Varies Varies 0.93 0.93 Weedone 650 as an exampl

aquatic invertebrates."; do nsoils (<1% O. M.); preplant weeds only , no in crop useforage or hay to livestock. 

1The EPA-required statement to convey to applicators the overall acute toxicity hazard posed by the product. Caution is more favorab

more favorable than Danger. 

2The period of time following application during which worker reentry into the treated area is restricted, according to EPA’s Worker Prote3The highest single-treatment and seasonal rates that can be applied to soybean according to the product Directions for Use label. 4Lists specific statements extracted from the product label that represent specific hazards or limitations that may reduce the utility of the 

control 

5Higher rates with directed / hooded sprayers. 6Regional or soil type limitations may lower this rate. 7Soybean label not yet publically available. Corn label comments are cited PHI – preharvest interval, O. M. – organic matter, mts - months. 
Monsanto Company 10-SY-210U 

Table L-12. Active Ingredients Contained in Alternative Herbicides 

Active Ingredient 

First 

Registered1

2006 Treated 

Soybean 

Acreage 

(%)2

Registration 

Review 

Status3 RED Date4

Max. 

Soybean lb/a 

(single 

treatment)5

Max. 

Soybean 

lb/a 

(season) 

Tolera

(40 C

180

glyphosate salts 3-May-76 97 open 2009 Sep-93 1.5 6 36dicamba-diglycolamine salt 2-Feb-56 <0.5 NA Jun-09, corrected 19 29 222,4-D acid, salts, and esters 

3-Jun-52 10 2013 Jun-05 0.93 0.93 14

flumioxazin 12-Apr-2001 3 unscheduled NA 0.096 0.096 56imazethapyr 30-Jan-87 3 2014 Jun-06 0.0648 0.0648 44cloransulam-methyl 29-Oct-1997 1 2011 NA 0.04 0.055 51chlorimuron-ethyl 4-Apr-86 4 2011 Sep-04 TRED 0.14 0.148 42fomesafen 10-Apr-87 2 open 2007 TRED Aug-07 0.375 0.3758 43flumiclorac-pentyl 23-Mar-94 1 open 2009 Aug-05 TRED 0.081 0.11 47sulfentrazone 22-Nov-93 1 open 2009 NA 0.31 0.31 49thifensulfuron 25-Apr-86 1 2011 NA 0.013 0.013 43imazaquin 20-Mar-86 1 2014 TRED Dec-05 0.123 0.1238 42imazamox-ammonium 17-Apr-95 <0.5 2014 NA 0.04 0.04 122paraquat dichloride 8-Jan-80 1 2012 Aug-97 1.0 2.9 20lactofen 1-Apr-87 <0.5 open 2007 TRED Sep-03 0.3 0.4 43glufosinate-ammonium 29-May-91 <0.5 open 2008 NA 0.66 0.8 472,4-DB 30-Jun-66 <0.5 2014 Jan-05 0.4 0.4 33fluthiacet-methyl 14-Apr-99 <0.5 unknown NA 0.0065 0.009 55acifluorfen sodium 29-May-81 <0.5 unscheduled Sep-09 0.374 0.5 38Mesotrione 4-Jun-01 0.0 unscheduled NA 0.1875 0.1875 57
Monsanto Company 10-SY-210U 

TRED denotes Tolerance Reregistration Eligibility Decision 1The date the herbicide was first approved for any use (e.g., industrial) by U.S. EPA. 2The percentage of the herbicide-treated soybean acres that were treated with each herbicide in AR, IA, IL, IN, KS, KY, LA, MI, MN, M

SD, TN, VA, and WI in 2006 (USDA-NASS, 2007b) . 

3The herbicide’s progress in the ongoing EPA program named as Registration Review. Year indicates when the official docket was o

required by law to re-evaluate pesticides periodically, generally every 10-15 years. 

4The date when EPA issued a Reregistration Eligibility Decision document. Reregistration was an earlier re-evaluation program designe

data are up-to-date for a.i.s first registered before 1984. TRED means Tolerance Reassessment Eligibility Decision, which refers to an a

some post-1984 a.i.s followed. 

5The maximum amount of the herbicide that can be applied to soybean in a single treatment or during the entire season, according to produ6The number of the paragraph in the Code of Federal Regulations where that herbicide’s food and feed tolerances are listed. 7An EPA pesticide classification that restricts a product, or its uses, to use by a certificated pesticide applicator or under the direct 

applicator. See 40 CFR 152.160.

8Regional or soil type limitations may lower this rate. 

9Maximum treatment rates for the proposed dicamba label. 
Monsanto Company 10-SY-210U 631 of 721 

L.4.1. Human Health Effects of Alternative Herbicide Products 

Table L-13 provides information concerning human health parameters for each 

alternative herbicide compared to dicamba. The listed parameters include: 

 Acute Toxicity Categories for the herbicide. 

 Cancer Classification of the herbicide. 

 FQPA (Food Quality Protection Act) safety factor employed in EPA’s risk 

assessment process for the herbicide. 

 Level of Exposure representing the acceptable safe range for acute (acute 

population adjusted dose, aPAD) and chronic (chronic population adjusted dose, 

cPAD) exposures. 

 Extent to which all presently approved uses exhaust the acceptable safe exposure 

range (% aPAD and % cPAD utilized for the most highly exposed population 

subgroups), according to recent Federal Register Rules or other public risk 

assessment documents. 

A variety of chemical-specific public data sources were used to compile this comparison. 

Columns 9 to 11 in Table L-13 (to the right of the vertical gray bar) pertain specifically to 

the use of the herbicide in soybean: 

 The established soybean seed food tolerance in 40 CFR 180 that supports the uses 

in soybean. 

 The Theoretical Maximum Residue Concentration (TMRC) arising from this 

soybean tolerance using the DEEM dietary exposure software, assuming that 

residues are at tolerance levels and that 100% of the crop has been treated. 

 The percentage of the acceptable chronic exposure (cPAD) that is contributed by 

the soybean TMRC dietary exposure. 
Monsanto Company 10-SY-210U 

Table L-13. Human Health Risk Parameters for Alternative Herbicides 

Active Ingredient 

Acute (Oral, Dermal, Inhalation, Eye Irr., Skin Irr., Sens.)1

Cancer 

Classification2

cPAD 

mg/kg/day3

% cPAD 

Utilized4,5

aPAD 

mg/kg/day6

% aPAD 

Utilized

6,5

FQPA 

SF7

Soy See

Tol. 

(ppm) 8

glyphosate acid / potassium salt 

IV / IV / NA / II / IV / N; IV / IV / III / III / IV / N E 1.75 7 NA NA 1X 20 

dicamba acid diglycolamine salt12

 III / III / IV / II / II / N; III / III / IV / III / III / N not likely 0.45 6.6 1 11 1x 10 

2,4-D acid /salts / esters 

III / III / III/ I / III-IV / 

N; III / III / IV / III / IV / N 

D 0.005 38 0.067 58 1X 0.02 

flumioxazin IV/III/IV/III/IV/N not likely 0.02 18 0.03 8 1X 0.02 imazethapyr IV / III / IV / IV / III / N not likely 2.5 < 1 NA NA 1X 0.1 cloransulam-methyl IV / III / III / III / IV / N not likely 0.1 <1 NA NA 1X 0.02 chlorimuron-ethyl IV / III / IV / III / IV / N not likely 0.09 0 NA NA 1X 0.05 fomesafen III / II /III / I / II-III / Y not likely 0.0025 31 NA NA 1X 0.05 flumiclorac-pentyl IV / III/ IV/ IV/ II/ Y no evidence 1 < 0.01 NA NA 1X 0.01 sulfentrazone III/ III / IV / I / IV / Y not likely 0.14 <1 0.25 1 1X 0.05 thifensulfuron IV / III / IV / IV / III / N not likely 0.07 <1 1.59 <0.1 1X 0.1 imazaquin IV / III / III / IV / IV / N no evidence 0.25 <1 NA NA 1X 0.05 imazamoxammonium IV / III / IV / III / IV / N not likely NA NA NA NA 1X ex '03 paraquat dichloride II / III / I /II / IV / N E 0.00045 26 0.0042 66 1X 0.7 
Monsanto Company 10-SY-210U 

Table L-13 (continued). Human Health Risk Parameters for Alternative Herbicides 

Active Ingredient 

Acute (Oral, Dermal, Inhalation, Eye Irr., Skin Irr., Sens.)1

Cancer 

Classification2

cPAD 

mg/kg/day3

% cPAD 

Utilized4,5

aPAD 

mg/kg/day6

% aPAD 

Utilized

6,5

FQPA 

SF7

Soy See

Tol. 

(ppm) 8

lactofen IV / III / IV / III / IV / N likely/unlikely 0.008 <0.1 0.17 <0.1 3X (A) 0.01 glufosinateammonium III / III / III / III / IV / N no evid. of 0.006 27 0.0063 48 1X 2 2,4-DB III / III / IV / III / IV / not likely 0.03 2 0.6 <1 1X 0.5 fluthiacet-methyl IV / III / IV / IV / IV /N likely (7.5x10-7) 0.001 <1 NA NA 1X 0.01 acifluorfen sodium III / III / IV / I / II / N likely/unlikely 0.013 <1 0.02 <1 1X/3X /10X 0.1 mesotrione IV / III / IV / III / IV / N not likely 0.007 5.8 NA NA 3X 0.01 

Sources of the information summarized in this table are listed in the Alternative Herbicide Specific References Section. N denotes negative for dermal sensitization NA stands for not applicable. 1EPA categories for the standard six acute toxicity tests of the active ingredient. Categories I and IV denote the least and most favora

Category I findings are highlighted in bold font. 2The conclusion reached by the EPA Office of Pesticide Programs Cancer Assessment Review Committee. The system of classification hresulting in a combination of different terminology. Generally, Group E, “not likely” or “no evid.” are the most favorable conclusions

uncertainty, and “likely/not likely” or “likely” indicate that a potential to induce cancer exists. More informat

http://www.epa.gov/opp00001/health/cancerfs.htm. |Accessed March 26, 2010| “Likely” findings are highlighted in bold font. 3The EPA-determined chronic Population Adjusted Dose, against which chronic exposure, primarily from combined food and water r

human health risk assessment. This key risk assessment parameter is derived from consideration of all the chronic toxicity studies, and in

factors. Higher values indicate herbicides with less severe chronic toxicity effects. 4The percentage of the cPAD that is represented by all presently approved uses of that herbicide for the most highly-exposed population 

by summing estimated chronic exposures from dietary and water, and dividing by the level of exposure that is considered safe (i.e., cP

indicate that current estimated exposure is a smaller proportion of the safe level, and therefore implies a greater margin of safety. EPA when new risk assessments are conducted, such as when a new food tolerance is petitioned. 

5Most highly exposed population subgroup. 6The acute risk assessment parameters correspond to those described above for chronic exposure. In some cases, EPA’s review of the acu

result in an acute effect of concern, and no aPAD is needed, indicated in the table as NA. 7The Safety Factor EPA has utilized according to the requirements of the Food Quality Protection Act. FQPA that requires EPA utilize a

safety factor to protect infants and children, unless the scientific results indicate that a different level is protective. If the database is com
Monsanto Company 10-SY-210U 

and developmental toxicity studies do not indicate that pre- and postnatal exposure results in increased sensitivity, EPA often reduces the F

indication of increased sensitivity, or the necessary data are lacking, SFs of 3X or 10X are sometimes used. These higher safety fac

analysis to denote higher risk to infants and children, and such cases are highlighted in bold font. 

8The 3 columns to the right of the gray bar pertain to the use of each herbicide on soybean only, other current uses are excluded from this a9The soybean seed food tolerance established in the relevant numbered paragraph in 40 CFR 180. 10The Monsanto-calculated theoretical maximum dietary exposure to the most highly-exposed U.S. population subgroup if 100% of all 

that herbicide, and residues were at tolerance levels. This theoretical exposure does not occur in real life, but allows a consistent coherbicides. The calculation is made using the same DEEM dietary exposure model that EPA routinely uses http://www.durango-software

|Accessed March 26, 2010|. 

11The ratio of the prior column divided by the cPAD. It denotes the percentage of the safe exposure level that is attributed to soybean r

index. High numbers are less favorable. Any value above 100% requires further risk refinement to be deemed acceptable, such as that fo

than 10-times that of dicamba are highlighted in bold font. 

12Dicamba diglycolamine data are for Clarity (U.S. EPA Reg. No. 7969-137) and M1691 formulations (U.S. EPA Reg. No. 524-582). 
Monsanto Company 10-SY-210U 635 of 721 

L.4.2. Ecological Effects of Alternative Herbicide Products – Non-target Species 

L.4.2.1 Fish and Aquatic Invertebrates.

Table L-14 provides information about the hazards, exposures, and risks of dicamba and 

each of the eighteen alternative herbicides to fish and aquatic invertebrates (considering 

2,4-D acid, salts, and esters as one alternative herbicide). The listed parameters include: 

 LC50 endpoints from acute fish toxicity studies, as reported in EPA’s one-liner 

database55, published in a RED or Registration Review risk assessments. The 

highest and lowest available LC50 values for any fish study are listed, regardless 

of species, including both fresh and marine species together. The purpose is to 

define a range of concentrations that spans the expected fish-toxic levels. 

 EC50 endpoints from acute aquatic invertebrate studies, as reported from the same 

sources cited above. The highest and lowest available EC50 values for any 

invertebrate study are listed, regardless of species, including both fresh and 

marine species together. The purpose is to define a range of concentrations that 

spans the expected aquatic invertebrate-toxic levels. 

 Estimated environmental exposure concentrations (EECs) in surface water for 

each of the eighteen alternative herbicides. The third column in Table L-14, 

identified as “Calculated EEC”, provides a simple standard estimate based on the 

maximum single application rate in soybean, using EPA’s standard field-farm 

pond scenario. This scenario examines a 1-acre pond in a 10-acre field in which 

(1) 5% of the application drifts into the 6-foot-deep pond and (2) 5% of the 

application onto the 10 acres runs off into the same pond. The fourth column in 

Table L-14 lists other model estimates of surface water concentrations as 

provided by one or more modeling programs, as cited by EPA in public 

documents, such as Federal Register final tolerance rule drinking water 

assessments. The purpose is to define a range of concentrations that spans 

available estimates of potential aquatic exposure levels. 

 Calculated Risk Quotients (RQs) for aquatic animals, comprised of fish and 

aquatic invertebrates combined together. Rather than calculate a single RQ for 

each species, Monsanto has calculated a range of potential RQs for each 

herbicide, bracketed by the best- and worst-case values. The “best” RQ is derived 

from the ratio of the lowest reported EEC concentration divided by the highest 

LC50 or EC50 for any aquatic animal. Conversely, the “worst” RQ is derived from 

the ratio of the highest EEC concentration divided by the lowest LC50 or EC50 for 

any aquatic animal. (Note: the RQ figures are rounded to two decimal places, so 

that entries that appear as “0.00” mean that the specific RQ is less than 0.005.) 

The purpose is to define a range of RQs that span and describe the risk posed by 

 

55 National Information System – Regional IPM Centers. OPP Pesticide Ecotoxicity Database. 

http://www.ipmcenters.org/Ecotox/index.cfm [Accessed May 27, 2010]. 
Monsanto Company 10-SY-210U 636 of 721 

the alternative herbicide to aquatic animals. The RQs that exceed the EPA’s 

Level of Concern (LOC) of 0.5 are marked in bold font. 

Using the worst case risk quotient, ten of 18 alternative herbicides have risk quotients 

greater than or equal to 0.01, while the worst case risk quotient for dicamba and seven 

other herbicide is <0.01. Only three of these 10 herbicides have risk quotients greater 

than 0.05 or 0.1, the levels of concern for threatened or endangered species and acute 

restricted use, respectively. Two of these 10 herbicides have RQ values greater than 0.5, 

the highest acceptable level of concern. Monsanto believes that based on risk quotients, 

dicamba offers a lower risk to aquatic animals relative to three of the 18 alternative 

herbicides: 2,4-D esters, flumioxazin, and lactofen. This conclusion is tabulated in Table 

L-17. 
Monsanto Company 10-SY-210U 

Table L-14. Aquatic Toxicity Parameters for Fish and Aquatic Invertebrates for Alternative Herbicides 

Active Ingredient 

Maximum 

Soybean 

lb/acre 

(single 

treatment)1

Calculated 

EEC (ppm)

2,3

FIRST, GENEEC 

or PRZX/EXAMS 

Surface Water 

ppm (RED or 

Tolerance Rule)4

Fish LC50 Range 

(ppm)5

Aquatic 

Invertebrate LC50 

or EC50 Range 

(ppm)f

Risk Quotie

Aquatic An

Range

 low high low high best 

glyphosate salts 1.5 0.050 0.0008 - 0.021 

45 >1000 55 780 0.00 

dicamba /DGA salt 1 0.034 0.01 - 0.036 

28 > 270 >100 >270 0.00 

2,4-D acid + salts 0.93 0.031 0.064 - 0.118 >80 2244 25 820 0.00 2,4-D esters 0.93 0.031 0.064 - 0.118 >0.15 14.5 2.2 12 0.00 < flumioxazin 0.096 0.003 0.018 - 0.034 2.3 21 0.23 5.5 0.00 imazethapyr 0.064 0.002 0.006 > 112 423 > 109 > 1000 0.00 cloransulam-methyl 0.04 0.001 0.002 > 86 > 154 > 111 > 121 0.00 chlorimuron-ethyl 0.14 0.005 0.003 - 0.005 > 2 8.4 >10a > 10 0.00 fomesafen 0.375 0.013 0.006 - 0.012 126 > 163 25 376 0.00 flumiclorac-pentyl 0.081 0.003 0.00024 >0.189 > 24 >0.189 >38 0.00 sulfentrazone 0.31 0.010 0.004 - 0.016 94 > 120 1 60.4 0.00 thifensulfuron 0.013 0.000 0.0003 - 0.004 >100 > 100 >1000 > 1000 0.00 
Monsanto Company 10-SY-210U 

Table L-14 (continued). Aquatic Toxicity Parameters for Fish and Aquatic Invertebrates for Alternative H

Active Ingredient 

Maximum 

Soybean 

lb/acre 

(single 

treatment)1

Calculated 

EEC (ppm)

2,3

FIRST, GENEEC 

or PRZX/EXAMS 

Surface Water 

ppm (RED or 

Tolerance Rule)4

Fish LC50 Range 

(ppm)5

Aquatic 

Invertebrate LC50 

or EC50 Range 

(ppm)5

Risk Quotie

Aquatic An

Range

 low high low high best 

imazaquin 0.123 0.004 0.004 - 0.008 280 420 280a 280 0.00 imazamox-ammonium 0.04 0.001 0.002 > 94.2 > 122 >94.3 > 122 0.00 paraquat dichloride 1.0 0.033 0.0015 > 1 156 1.2 1.2 0.00 < lactofen 0.3 0.011 0.000008 - 0.4 0.46 0.46 0.02 4.85 0.00 glufosinate-ammonium 0.66 0.022 0.043 - 0.094 > 320 > 1000 8 668 0.00 2,4-DB 0.4 0.013 0.013 - 0.015 2 18 25a 25 0.00 fluthiacet-methyl 0.0065 0.000 0.0005 - 0.0008 0.043 0.16 0.3 >2.3 0.00 acifluorfen sodium 0.374 0.013 0.0024 - 0.010 31 204 28.1 > 1000 0.00 mesotrione 0.1875 0.011 0.004 - 0.02 > 114 520 3.3 840 0.00 

1The highest single-treatment rate permitted by the herbicide’s product labels. This rate is used to calculate potential acute exposure to 

via spray drift or runoff. 

2Based on the maximum single treatment rate, with 5% spray drift and 5% runoff from 10 treated acres into a 1-acre 6-foot-deep pond. 3The Estimated Environmental Concentration in surface water. It was calculated by Monsanto using the EPA’s “standard f

http://www.epa.gov/oppefed1/models/water/geneec2_description.htm |Accessed May 28, 2010|. In this scenario, it is assumed that th

10 acre farm field containing a 1 acre pond that is six feet deep. The pond experiences 5% of the application rate by spray drift and 5% 

soil enters the pond via runoff. This concentration estimation is a simple, conservative Tier 1 procedure that utilizes only the applicati
Monsanto Company 10-SY-210U 

exposures, and allows quick comparison of many different herbicides. Other more increasingly-detailed computer models can be uEEC estimates, but these require the user to input various physical chemical parameters and weather data for each product to be m

methods are listed in the next column. 

4Modeling estimates of potential aquatic exposure levels. When EPA conducts risk assessments, it uses computer models to obtain est

water concentrations. These are published in the RED for each herbicide, or in tolerance rules in the Federal Register. The range o

can be compared to the single number in the prior column. Sources of these estimates are listed in the “ALTERNATIVE 

REFERENCES“ section. 

5“Fish LC50 Range (ppm)” and “Aquatic Invertebrate LC50 or EC50 Range (ppm)”. These four columns describe the range of hazard 

sources representing the toxicity of each herbicide versus freshwater and marine animals. The LC50 or EC50 means the water conc

immobilize half of the test species, which is a standard potency descriptor. The highest and lowest values found for any fish species (etc.) were tabulated. Likewise, the highest and lowest values found for any aquatic invertebrate species (Daphnia, shrimp, crab, etc.) these data are listed in the “ALTERNATIVE HERBICIDE-SPECIFIC REFERENCES“ section and in available public databases (Nati

Regional IPM Centers. OPP Pesticide Ecotoxicity Database. http://www.ipmcenters.org/Ecotox/index.cfm) |Accessed May 28, 2010|. 6EPA’s EFED uses a Risk Quotient (RQ) method for ecological risk assessment. The RQ equals the potential exposure level divided b

exposures or more potent hazard findings lead to higher RQs. EFED has established Levels of Concern (LOCs) for various non-target the RQ exceeds the LOC, further refinement is needed to determine whether risk mitigation might be needed. For non-listed aquatic a

risk is 0.5, for acute risk restricted use is 0.1, and for threatened or endangered species it is 0.05. In this analysis, Monsanto calculated the lowest EEC estimate by the highest hazard (LC50 or EC50) value, and calculated the worst-case RQ from the highest EEC and 

purpose was to bracket a range that typified the aquatic animal risk presented by each herbicide. When neither the worst-case nor th

LOC of 0.05, Monsanto concluded that risk to aquatic animals is minimal. Instances where the LOC threshold of 0.5 is exceeded are h

7Lists instances where the product label includes warning statements about aquatic animal exposure. 
Monsanto Company 10-SY-210U 640 of 721 

L.4.2.2. Aquatic Plants 

Table L-15 provides information about the hazards, exposures, and risks of dicamba and 

each of the eighteen (18) alternative herbicides to aquatic plant species, specifically 

duckweed and aquatic algae species (considering 2,4-D acid, salts, and esters as one 

alternative herbicide). The data format, sources, and methods of Estimated 

Environmental Exposure Concentration (EEC) calculation are identical to those described 

above for the aquatic animals (Table L-14). A Level of Concern (LOC) value of 1.0 has 

been used for judging RQ exceedances in the case of aquatic plants, consistent with EPA 

EFED’s normal practices. 

The assessment and comparison summarized in Table L-15 establishes that dicamba 

poses little acute risk to aquatic plants at use rates of 0.05 – 1.0 lb dicamba a.e./acre, 

which is consistent with EFED’s assessment published in the RED EFED Chapter. 

Monsanto was unable to identify aquatic plant hazard data for three of the alternative 

herbicides (imazaquin, chlorimuron-ethyl, and flumiclorac-pentyl). For nine (9) of the 

eighteen (18) alternative herbicides, the range of RQs is < 0.005 to 0.75; that is, none of 

these nine a.i.s present an aquatic plant risk, which even in the worst-case calculation, 

reach EFED’s Level of Concern (LOC) for aquatic plants. However, for seven of the 

alternative herbicides, the worst-case RQs did exceed EFED’s LOC of 1.0. It is not 

surprising that some herbicides are quite toxic to aquatic plants, and the worst-case RQs 

for three of the alternate herbicides (flumioxazin, lactofen, and paraquat dichloride) 

exceeded the LOC by a factor of more than 50-fold. 

Monsanto believes that dicamba offers a lower risk to aquatic plants relative to seven of 

the 18 alternative herbicides (2,4-D, flumioxazin, sulfentrazone, thifenslufuron, paraquat 

dichloride, lactofen, and mesotrione). This conclusion is tabulated in Table L-17.
Monsanto Company 10-SY-210U 

Table L-15. Aquatic Toxicity Parameters for Aquatic Plants for Alternate Herbicide Active Ingredients 

Active Ingredient 

Maximum 

Soybean 

lb/acre 

(single 

treatment)1

Calculated 

EEC 

(ppm) 2,3

FIRST, 

GENEEC or 

PRZX/EXAMS 

Surface Water 

ppm (RED or 

Tolerance Rule)4

Duckweed and 

Algae EC50 Range 

(ppm)5

Risk Quotient for 

Aquatic Plants 

Range6

 low high best worst 

glyphosate salts 1.5 0.050 0.0008 - 0.021 0.77 38.6 0.00 0.06 dicamba /DGA salt 1 0.034 0.01 - 0.036 0.06 > 3.7 0.00 0.60 2,4-D acid + salts 0.93 0.031 0.064 - 0.118 0.29 156 0.00 0.41 2,4-D esters 0.93 0.031 0.064 - 0.118 0.066 >19.8 0.00 1.79 flumioxazin 0.096 0.003 0.018 - 0.034 0.0005 0.019 0.16 68.00 imazethapyr 0.064 * 0.002 0.006 0.008 59.2 0.00 0.75cloransulam-methyl 0.04 0.001 0.002 0.003 135 0.00 0.67 chlorimuron-ethyl 0.14 0.005 0.003 - 0.005 NA NA NA NA fomesafen 0.375 0.013 0.006 - 0.012 0.09 71 0.00 0.14 flumiclorac-pentyl 0.081 0.003 0.00024 NA NA NA NA sulfentrazone 0.31 0.010 0.004 - 0.016 0.002 0.033 0.12 8.0 thifensulfuron 0.013 0.000 0.0003 - 0.004 0.0016 > 0.026 0.02 2.50 imazaquin 0.123 0.004 0.004 - 0.008 NA NA NA NA imazamox-ammonium 0.04 0.001 0.002 0.011 > 0.038 0.03 0.18 paraquat dichloride 1.0 0.033 0.0015 0.00055 2.84 0.00 90.9 lactofen 0.3 0.011 0.000008 - 0.4 0.001 0.001 0.00 400 glufosinate-ammonium 0.66 0.022 0.043 - 0.094 1.5 7.8 0.00 0.06 2,4-DB 0.4 0.013 0.013 - 0.015 >0.932 >0.932 <0.01 <0.02 fluthiacet-methyl 0.0065 0.000 0.0005 - 0.0008 0.0022 >0.018 <0.01 0.36 acifluorfen sodium 0.374 0.013 0.0024 - 0.010 > 0.26 0.38 0.01 <0.05 mesotrione 0.1875 0.011 0.004 - 0.02 0.018 132 0.00 1.11 
Monsanto Company 10-SY-210U 

The first three columns in this table are identical to- those in Table L-14. 1The highest single-treatment rate permitted by the herbicide’s product labels. This rate is used to calculate potential acute exposure to aqu

spray drift or runoff. 

2Based on the maximum single treatment rate, with 5% spray drift and 5% runoff from 10 treated acres into a 1-acre 6-foot-deep pond. 3The Estimated Environmental Concentration in surface water. It was calculated by Monsanto using the EPA’s “standard f

http://www.epa.gov/oppefed1/models/water/geneec2_description.htm |Accessed May 28, 2010|. In this scenario, it is assumed that the h

acre farm field containing a 1 acre pond that is six feet deep. The pond experiences 5% of the application rate by spray drift and 5% of 

enters the pond via runoff. This concentration estimation is a simple, conservative Tier 1 procedure that utilizes only the applicatio

exposures, and allows quick comparison of many different herbicides. Other more increasingly-detailed computer models can be used to estimates, but these require the user to input various physical chemical parameters and weather data for each product to be modeled. Exam

listed in the next column. 

4Modeling estimates of potential aquatic exposure levels. When EPA conducts risk assessments, it uses a computer models to obtain est

water concentrations. These are published in the RED for each herbicide, or in tolerance rules in the Federal Register. The range of estim

compared to the single number in the prior column. Sources of these estimates are listed in the “ALTERNATIVE HERBICIDE-SPECIFIC 

5These two columns describe the range of hazard data found in public data sources representing the toxicity of each A.I. versus freshwat

EC50 means the water concentration needed to kill or prevent growth of half of the test species, which is a standard potency descriptor

values found for any aquatic plant species (diatom, duckweed, alga, etc.) were tabulated. Sources of these data are listed in the “ALTE

SPECIFIC REFERENCES” section of this document and in available public databases cited in footnote 37 (National Information System OPP Pesticide Ecotoxicity Database. http://www.ipmcenters.org/Ecotox/index.cfm |Accessed May 28, 2010|) 6As described above for Table L-15, EPA’s EFED uses a Risk Quotient (RQ) method for ecological risk assessment. For non-listed aqua

or endangered species the LOC is 1.0. In this analysis, Monsanto calculated best-case RQs by dividing the lowest EEC estimate by the hi

and calculated the worst-case RQ from the highest EEC and lowest hazard value. The purpose was to bracket a range that typified the aby each herbicide. When neither the worst-case nor the best-case RQs exceed a LOC of 1.0, Monsanto concluded that risk to aquatic pl

where the LOC threshold of 1.0 is exceeded are highlighted in bold font. 
Monsanto Company 10-SY-210U 643 of 721 

L.4.3. Looking Toward the Foreseeable Future 

As noted, weeds that are difficult to control with glyphosate and weeds that are 

glyphosate-resistant, PPO-resistant, or ALS-resistant represent an opportunity for 

improved weed control in soybean. To address this need, Monsanto is seeking to 

commercialize MON 87708, which will allow dicamba to be used as a weed control tool 

in this important U.S. crop. Monsanto is also aware that other companies are developing 

biotechnology-derived soybean enhanced with other herbicide-tolerant traits. 

Biotechnology-derived soybean developed to be tolerant to applications of 2,4-D has 

been submitted to USDA-APHIS for deregulation. 56 Monsanto proposes that 2,4-D 

presents more relative risk than dicamba in the area of acute human health and chronic 

exposure risk, and for 2,4-DB relatively more ecological risk for aquatic animals. 

Applications of 2,4-D will also require management of offsite movement. 

In addition, biotechnology-derived soybean enhanced to be tolerant to herbicides that 

inhibit 4-hydroxyphenylpyruvate dioxygenase (HPPD) is one such trait that Monsanto 

believes is in development, based on public announcements from companies other than 

Monsanto; evidence of these developments is apparent in the recent EPA approval of 

mesotrione for use on mesotrione-tolerant soybean (U.S. EPA, 2009e). HPPD-tolerance 

allows use of these broadleaf herbicides in soybean production, which is a good technical 

fit as highlighted by Table L-16 which shows many of the troublesome soybean weeds 

are effectively controlled by HPPD herbicide products such as Balance Pro (active 

ingredient: isoxaflutole), Laudis (active ingredient: tembotrione) or Callisto Herbicide 

(active ingredient: mesotrione). Since some of these products are not yet registered for 

soybean use, and the label for Callisto defining application parameters in soybean is not 

yet commercially available, it is not possible to undertake a rigorous application-ratebased risk comparison with dicamba, but it is possible to make a hazard comparison, 

using isoxaflutole and tembotrione as typical examples (mesotrione is included above in 

the discussion of alternate herbicides, since has been approved by EPA). 

Inhibition of HPPD in plants results in a disruption of carotenoid biosynthesis, which 

leaves the plant’s chlorophyll pigments unprotected from rapid degradation via 

photooxidation, and results in a very characteristic white bleached symptomology of 

plant parts that are normally green. HPPD is also an animal liver enzyme involved in the 

catabolic breakdown of tyrosine, and its inhibition in laboratory animals results in 

elevated tyrosine plasma concentrations (tyrosinemia), which can cause adverse ocular, 

developmental, liver, and kidney effects. Stated simply, the mechanism of herbicidal 

efficacy based on HPPD inhibition is inherently linked to a potential for negative human 

health effects. 

Publicly available study results show that isoxaflutole and tembotrione both caused 

ocular and liver effects in test animals, and both caused developmental toxicity at non-

 56 Dow Agrosciences press release, December 15, 2009. 

http://www.dowagro.com/newsroom/corporatenews/2009/20091215a.htm. [Accessed March, 26, 

2010]. 
Monsanto Company 10-SY-210U 644 of 721 

maternally toxic levels. Because of the potent toxic effects, EPA has calculated cPADs 

for isoxaflutole and tembotrione that are lower than that of dicamba by factors of 225 and 

1125, respectively. EPA is also considering the need for a cumulative risk assessment 

approach for these two chemicals, along with mesotrione and other HPPD-inhibiting 

herbicides, because they share a common toxic mechanism (U.S. EPA, 2009d, e). In 

addition, both isoxaflutole and tembotrione had carcinogenic effects in long-term testing 

summarized by EPA. For tembotrione, carcinogenicity was limited to rats only, but 

isoxaflutole was found to have carcinogenic effects in two species, and was categorized 

by EPA as a B2 carcinogen in 1998 when first registered. Isoxaflutole products, such as 

Balance Pro, are Restricted Use pesticides because of a very high level of concern about 

damage to non-target plants caused by spray drift, and isoxaflutole labels also bear 

warning statements about the likelihood of persistence and leaching. There have also 

been concerns about isoxaflutole use leading to levels of herbicidally-active isoxaflutole 

metabolites in surface water utilized for irrigation purposes. Because of these properties 

of HPPD chemicals, Monsanto believes that dicamba offers a lower risk relative to these 

potential future soybean herbicides. 

L.4.4. Efficacy and Weed Management Practices of Alternative Herbicide Products 

Table L-16 provides weed control effectiveness of formulated products containing 

dicamba, glyphosate and alternate herbicides. Weed control less than 70% is considered 

insufficient (white), control between 70 and 85% is considered as marginal effectiveness 

(black-white), and control of more than 85% is considered commercially acceptable 

(black). The data presented in Table L-16 are derived by combining state and dealer57

herbicide guidance for soybean production across major soybean-producing states (Ohio 

and Indiana 58 , Iowa 59 , Tennessee 60 , North Dakota 61 ). Weed control herbicide 

recommendations provided by University Extension scientists to control specific weeds 

were converted to a common scale and combined to reflect an average herbicide weed 

control rating across geographies. Weed control ratings specific to resistant weeds were 

based mainly on recommendations from Ohio and Indiana. Monsanto weed scientists 

applied their own expert scientific judgment during the conversion of University 

Extension recommendations into a common scale. The weeds chosen for inclusion in 

Table L-16 represent current problem weeds in soybean, either because they exhibit 

herbicide resistance or because they are generally hard-to-control species. 

As Table L-16 shows, the alternative herbicides can be used in preemergent applications, 

postemergent applications, or, like dicamba and glyphosate, as both pre- and 

 

57 2009 Crop Protection Guide - Information for Dealers http://www.agrisolutionsinfo.com/ Accessed 

May 28, 2010

58 2010 Ohio and Indiana Weed Control Guide (Bulletin 789; Pub. WS16) 

www.btny.purdue.edu/Pubs/WS/WS-16/ Accessed May 28, 2010 59 2009 Herbicide Guide for Corn and Soybean Production - http://www.weeds.iastate.edu [Accessed May 

28, 2010] 60 2010 Weed Control Manual for Tennessee 

http://www.utextension.utk.edu/publications/pbfiles/pb1580.pdf Accessed may 28, 2010 61 2010(a) NDSU Weed Control Guide - http://www.ndsu.edu/weeds Accessed May 28, 2010
Monsanto Company 10-SY-210U 645 of 721 

postemergent applications. Several herbicides, such as 2,4-D, 2,4-DB, sulfentrazone and 

paraquat, do not have sufficient soybean safety for application in-crop, so their use is 

limited to control of existing weeds and preemergent control of later emerging weeds via 

soil residual activity, if any. Others, such as the protoporphyrinogen oxidase (PPO) 

inhibitors acifluorofen, lactofen, and fomesafen, are most effective as postemergent 

treatments, even though they may cause some soybean leaf injury. Acetolactate synthase 

(ALS) inhibitors like chlorimuron-ethyl and cloransulam-methyl can be used at either 

timing. Glufosinate, like glyphosate and dicamba, does not have intrinsic soybean 

selectivity and can only be used as a postemergent application over soybean that is 

genetically enhanced to provide glufosinate tolerance (i.e., Liberty Link). 

Table L-16 also highlights expected weed efficacy trends. Glyphosate does not provide 

commercial control of glyphosate-resistant weed biotypes such as common ragweed or 

horseweed, but does control the wild type plants of these species. Similarly, ALSinhibiting herbicides provide poor control of ALS-resistant weeds, and PPO inhibitors do 

not adequately control PPO-resistant weeds. Table L-16 focuses on problem weeds 

found in soybean and does not include any weeds with auxin resistance, although 2,4-D 

provides good control of many herbicide-resistant broadleaf weed biotypes in this group. 

Extreme, which contains two modes-of-action (glyphosate and imazethapyr), provides 

better control of this target weed spectrum than most of the herbicide products having 

only a single mode-of-action. 

Further discussion of the comparative effectiveness of dicamba versus alternative 

herbicide products against herbicide-resistant and hard-to-control weeds can be found in 

L.5.3. 
Monsanto Company 10-SY-210U 

Table L-16. Herbicide Efficacy Comparison: Herbicide Resistant Weeds and Hard-to-Control Weeds in Soy

2 >85% control1 70% - 85% control0 < 70% control

Herbicide Herbicide Brand

Class

GR Waterhemp

GR Palmer amaranth

GR Horseweed

GR C. ragweed

GR G. ragweed

GR Johnsongrass

ALS C. ragweed

ALS+PPO C. ragweed

ALS+GR C. ragweed

ALS G. ragweed

ALS+GR G. ragweed

ALS+GR Horseweed

ALS Kochia

TRI Lambsquarters

A. Morningglory

B. Nightshade

Cockelbur

C. Ragweed

G. Ragweed

Glyphosate Roundup EPSPS

Dicamba Clarity auxin

2,4-D 2,4-D auxin

2,4 DB Butyrac 200 auxin

Chlorimuron + Metribuzin Canopy DF ALS/PSIIChlorimuron + Tribenuron Canopy EX ALS

Imazaquin Scepter ALS

Sulfentrazone + Imazethapyr Authority Assist PPO/ALS

Sulfentrazone + Metribuzin Authority MTZ PPO/PSII

Sulfentrazone + Cloransulam Authority First / Sonic PPO/ALS

Flumioxazin Valor PPO

Flumioxazin + Chlorimuron Valor XLT PPO/ALS

Flumioxazin + Cloransulam Gangster PPO/ALS

Cloransulam Firstrate ALS

Paraquat Gramoxone Inteon PSI

Isoxaflutole Balance PRO HPPD

Mesotrione Callisto HPPDMesotrione Callisto HPPD

Tembotrione Laudis HPPD

Tropramezone Impact HPPD

Ha

HPPD POSTEMERGENCE5

Resistant Weeds1

PREEMERGENCE/BURNDOWN3

HPPD PREEMERGENCE4
Monsanto Company 10-SY-210U 

Table L-16 (continued). Herbicide Efficacy Comparison: Herbicide Resistant Weeds and Hard-to-Control W

2 >85% control1 75% - 85% control0 < 75% control

Herbicide Herbicide Brand

Class

GR Waterhemp

GR Palmer amaranth

GR Horseweed

GR C. ragweed

GR G. ragweed

GR Johnsongrass

ALS C. ragweed

ALS+PPO C. ragweed

ALS+GR C. ragweed

ALS G. ragweed

ALS+GR G. ragweed

ALS+GR Horseweed

ALS Kochia

TRI Lambsquarters

A. Morningglory

B. Nightshade

Cockelbur

C. Ragweed

G. Ragweed

Hemp sesbania

Glyphosate Roundup EPSPS

Dicamba Clarity auxin

Imazethapyr + Glyphosate Extreme ALS/EPSPS

Fomesafen Flexstar PPO

Imazethapyr Pursuit ALSCloransulam Firstrate ALS

Flumiclorac Resource PPO

Chlorimuron Classic ALS

Imazamox Raptor ALS

Lactofen Cobra / Phoenix PPO

Fomesafen Reflex PPO

Acifluorfen Ultra Blazer PPO

Glufosinate Ignite Glu

Chlorimuron + Thifensulfuron Synchrony XP ALS

Fluthiacet Cadet PPO

Hard-t

Resistant Weeds1

POSTEMERGENCE5

Horseweed is synonymous to marestail. GR = Glyphosate resistant ALS = Acetolactate synthase Tri = TriazinePPO = protoporphyrinogen oxidase
Monsanto Company 10-SY-210U 

Note: This table was based primarily on State University Extension weed control recommendations for soybean growing areas, Table L

which each product controls the targeted weeds in soybean. The legend describes the meaning of the symbols. See L.5.3 for additional de

1The section of the table describes control of populations (biotypes) of weeds that are known to have genetic resistance to specific herbicid2This section describes control of weeds that are difficult to control in soybean with existing herbicide treatments. Generally these a

removal would require herbicide rates that would severely damage the crop. 

3This section describes weed control by herbicides that are applied prior to soybean emergence. The weeds may or may not be emerged. 4This section describes foreseen use of herbicides that are not yet approved for use in soybean that have inhibition of 4-hydroxyphenylpyru5The section describes weed control by herbicides that are applied after the soybean emergence, generally over-the-top of the crop.
Monsanto Company 10-SY-210U 649 of 721 

L.5. Comparison of Dicamba Use in the Dicamba-Tolerant Soybean System 

(MON 87708) to Alternative Herbicides 

The intent of the comparative analysis is to define current herbicide use in U.S. soybean 

production, and to compare dicamba’s human health and environmental properties to 

herbicides currently used by growers for weed control. In order for a pesticide 

(herbicide) to be registered by EPA it must meet the FIFRA and FFDCA standards for 

safety to human health and the environment. The EPA must conclude that the herbicide 

when used according to the label does not pose an unreasonable risk to humans or the 

environment; and in order to establish a tolerance for the use of an herbicide on a food or 

feed crop, find there is a reasonable certainty of no harm to human health from nonoccupational (food, water and residential/recreational) exposures to the herbicide. 

Consequently all alternative soybean herbicides can be used safely, and do not pose a risk 

to humans or the environment. 

L.5.1. Comparisons for Human Health Risks 

Table L-13 provides information concerning human health parameters for dicamba and 

the alternative herbicide products. These data allow a comparison of the relative human 

health safety among the optional weed control products on the basis of: 

 Acute toxicity. 

 Chronic toxicity. 

 Cancer risks and classification. 

 Risk to infants and children. 

 Magnitude of potential exposure that is considered to be within the acceptable 

safe range. 

 Extent to which all presently approved uses exhaust the acceptable safe acute and 

chronic dietary exposure ranges for the most highly exposed population 

subgroups. 

 The proportion of present dietary exposure that arises through use on soybean. 

A compilation of Monsanto’s comparative determinations of the risks posed by the 18 

alternate herbicides compared to dicamba in the four basic human health risk categories 

(acute, cancer, chronic and infants/children) is tabulated in Table L-17. A “Yes” entry 

indicates a benefit for dicamba compared to that alternative herbicide. Entries not 

indicated with a “Yes” mean that dicamba is either comparable or less favorable than the 

alternative herbicide. A “Neutral” entry means that dicamba and the alternative herbicide 

have similar risks, and a “No” entry denotes alternative herbicides that have a benefit 

compared to dicamba. Table L-17 is intended as a 1-page scorecard summary of these 
Monsanto Company 10-SY-210U 650 of 721 

benefit comparisons. Monsanto has concluded that dicamba use on MON 87708 will 

provide benefits compared to the alternate herbicides used for soybean weed control. 

L.5.1.1. Acute Health Risks 

Dicamba acid has a Category II classification for eye and skin irritation, but once the acid 

is neutralized to form the DGA or other salt forms used in herbicide product 

formulations, all acute Categories are III or IV. These classifications are the most 

favorable acute toxicity categories in EPA’s acute hazard paradigm. Several alternative 

herbicides (acifluorfen, sulfentrazone, and some forms of 2,4-D) have high risk of eye 

irritation (Category I). Principal formulations of fomesafen, paraquat, lactofen, 2,4-DB 

(DMA salt), and acifluorfen have a “DANGER!” signal word (Table L-11). The 18 

alternate herbicide actives, as a group, also have relatively low acute (oral, dermal or 

inhalation) toxicity. For eight of the eighteen, EPA determined that an acute dietary risk 

assessment is not needed because there are no relevant acute toxicological effects. One 

notable exception is paraquat, which has high risk (Category I) of acute toxicity by the 

inhalation route, and present uses occupy 66% of the aPAD (the safe exposure level). 

2,4-D and glufosinate have relatively low aPADs compared to their dietary residues, and 

present uses occupy 58% and 48% of their respective aPADs. Overall, Monsanto 

concludes that dicamba has a lower acute toxicity risk compared to six of the eighteen 

alternate herbicides and their formulated products (paraquat, 2,4-D, glufosinate, 

acifluorfen, sulfentrazone, and fomesafen). These six are indicated by a “Yes” in the 

“Acute Toxicity Risk” column of Table L-25. Flumioxazin and 2,4-DB were judged to 

have similar acute toxicity risk as dicamba, and are marked as “Neutral” in Table L-17. 

Ten herbicides, which either did not have relevant acute toxicological effects or which 

utilized 1% or less of the allowable acute exposure (aPAD), were judged to have less 

acute toxicity risk than dicamba, and are marked as “No” in Table L-17. 

L.5.1.2. Chronic Risk 

Chronic dietary risk can be evaluated by consideration and comparison of the percentage 

of the safe exposure level (chronic population adjusted dose: cPAD) that is used up by all 

currently registered uses of an active ingredient. Table L-17summarizes the % cPAD 

utilized for each alternative herbicide, according to recently published Federal Register 

Final Rule information. A number of the alternative herbicides have very low use rates, 

and result in very low residues in treated food or feed and, accordingly, utilize only a 

small percentage of the cPAD. For dicamba and twelve of the alternate herbicides, the 

percentage of the cPAD utilized for all approved uses is <10%, so that this group has at 

least an added 10-fold margin of safety beyond that which EPA has determined is 

protective of human health. For imazamox, EPA has determined that residues in food or 

feed are not likely to approach levels of concern, and exempted it from the requirement of 

food and feed tolerances. The remaining five herbicides (2,4-D, flumioxazin, fomesafen, 

paraquat, and glufosinate) utilize a somewhat higher portion of the cPAD, ranging from 

18% to 38%. Although the 10% cutoff is an arbitrary one, and the increased risk of even 

the worst-case alternative herbicide for chronic risk is only 6-fold greater than that of 

dicamba, Monsanto concludes that dicamba offers a lower chronic toxicity risk compared 

to 2,4-D, flumioxazin, fomesafen, paraquat and glufosinate. 
Monsanto Company 10-SY-210U 651 of 721 

Another way to compare chronic dietary risk would be to focus directly on the soybean 

seed residues and their contribution to cPAD utilization. In Table L-13, columns 9 to 11 

(those to the right of the gray bar) present the established soybean seed tolerance for each 

herbicide. Using DEEM (a computer dietary exposure model used commonly by EPA,) 

and a theoretical worst-case approach (assume 100% of soybean is treated with a given 

herbicide that results in residues at the tolerance level), Monsanto calculated the 

theoretical maximum residue contribution (TMRC, a standard Tier 1 dietary risk method) 

to dietary exposure of each herbicide via soybean seed in μg/kg/day and as a percentage 

of the cPAD, as shown in Table L-13. This analysis highlighted paraquat and glufosinate 

as alternative herbicides with notably higher cPAD utilization by the soybean residue 

component, both herbicides were considered to present more relative risk than dicamba in 

the previous comparative method as well. The TMRC soybean calculation for paraquat 

yields a number that is 2.6-fold higher than the cPAD, because the EPA’s risk assessment 

methodology for paraquat assumes a market penetration in soybean of <5%. If this 

market share were increased, the risk attributed to soybean residue could become a 

substantial portion of the cPAD. 

Based on this reasoning, Monsanto concluded that dicamba offers lower chronic toxicity 

risk than five alternative herbicides (2,4-D, flumioxazin, fomesafen, paraquat and 

glufosinate), and these a.i.s are marked with a “Yes” in the chronic toxicity column of 

Table L-17. Imazamox-ammonium was considered by EPA to have such low toxicity that 

no food or feed tolerances were required by EPA, and therefore imazamox-ammonium 

was judged by Monsanto to have lower chronic toxicity risk than dicamba, indicated by a 

“No” in Table L-17. The alternate herbicide other a.i.s under consideration were judged 

to have similar chronic toxicity risk as dicamba, and are marked “Neutral” in Table L-17. 

L.5.1.3. Cancer Risk 

EPA classified dicamba as “not likely” for human carcinogenicity. Fourteen of the 

alternative herbicides are classified similarly – “not likely”, “no evidence”, or “Group E 

(Evidence of Non-Carcinogenicity for Humans)”. 2,4-D is classified as “D”, meaning 

that although the studies are acceptable, the evidence is unclear and some uncertainty 

remains. Two of the alternative herbicides (lactofen and acifluorfen) are classified as 

“not likely” at low doses but “likely” at high doses, due to liver and other effects. A 

peroxisome proliferation mechanism is established for acifluorfen. Since acifluorfen is a 

metabolite of lactofen, the similarity in toxicology and cancer classification is 

appropriate. One alternative herbicide, fluthiacet-methyl, is categorized as a “likely” 

human carcinogen, due the occurrence of dose-related tumors in both rats and mice; a Q* 

of 7.5 X 10-7 has been calculated, which is a measure of carcinogenic potency that EPA 

utilizes in risk assessment. Overall, Monsanto concludes that dicamba has a cancer risk 

benefit compared to four of the alternative herbicides (2,4-D, lactofen, acifluorfen, and 

fluthiacet-methyl), and this judgment is indicated by a “Yes” in the Cancer Risk column 

of Table L-17. Four alternative herbicide a.i.s (flumiclorac-pentyl, glufosinate, 

imazaquin, and paraquat) have been categorized by EPA as Group E or “no evidence”, 

and Monsanto judges that these four have lower cancer risk than dicamba, and are 

marked as “No” in Table L-17. The other alternate herbicide a.i.s that are in the “not 
Monsanto Company 10-SY-210U 652 of 721 

likely” category have similar cancer risk to dicamba, and are marked as “Neutral” in table 

L-17. 

L.5.1.4. Risks to Infants and Children 

The Food Quality Protection Act (FQPA) requires that EPA take special care in its risk 

assessments to establish that infants and children do not have increased sensitivity to 

pesticides and thereby experience greater risks from residues in food than the general 

U.S. population. EPA’s implementation of this requirement is embodied in the additional 

FQPA safety factor, which is established by the statute at 10-fold, unless there is 

evidence that another level is protective. When making determinations about the 

magnitude of the FQPA safety factor, the EPA considers the completeness of the 

database and specifically the findings from the developmental and reproductive toxicity 

studies to determine if there is evidence that infants and children are more sensitive than 

adults. Therefore, the magnitude of the FQPA safety factor (usually 1X, 3X, or 10X) is a 

comparative parameter that may be used as an indication of the potential risks to infants 

and children and for overall developmental and reproductive toxicity findings. 

Dicamba risk assessments for both acute and chronic effects utilize an FQPA safety 

factor of 1X. This is also the case for 15 of the 18 alternate herbicides, although in some 

cases, an acute dietary risk assessment was not necessary due to a lack of acute toxicity 

effects. For lactofen, acifluorfen, and mesotrione a 3X or 10X FQPA safety factor was 

used for the acute and/or chronic assessments, indicating a higher level of risk or 

uncertainty. Therefore, Monsanto concludes that dicamba offers an improved risk profile 

for infants and children compared to these three alternative herbicides, which are marked 

with a “Yes” in the Infants & Children Risk column in Table L-17. All other alternate 

herbicide a.i.s which have an FQPA safety factor of 1X are judged to have similar 

potential risk as dicamba, and are marked with “Neutral” in Table L-17. 
Monsanto Company 10-SY-210U 

Table L-17. Summary of Comparative Analysis of Dicamba and Alternative Herbicides 

Active Ingredient 

Mode-ofAction 

(WSSA 

Group1) 

Human Health Risk Measures2

Aquatic Non-Target 

Species Risk 

Measures3 Known 

Resistant 

Weed 

Species4 

Herbicidal 

Efficacy5

(< 50% of 

dicamba) 

Long 

Rotational 

Crop 

Restriction6

S

I

Po

Acute 

Toxicity 

Risk 

Cancer 

Risk 

Chronic 

Risk 

Infants & 

Children 

Risk 

Aquatic 

Animal 

Risk 

Aquatic 

Plant 

Risk 

2,4-D acid / esters Aux (4) 

Yes Yes Yes Neutral Yes Yes 

28 

Neutral Neutral N

2,4-DB Neutral Neutral Neutral Neutral Neutral Neutral Yes Neutral Nimazethapyr ALS (2) No Neutral Neutral Neutral Neutral Neutral

107 (Yes) 

Yes Yes N

cloransulammethyl ALS (2) No Neutral Neutral Neutral Neutral Neutral Yes Yes Nchlorimuronethyl ALS (2) No Neutral Neutral Neutral Neutral NA Yes Yes Nthifensulfuron ALS (2) No Neutral Neutral Neutral Neutral Yes Yes Yes Nimazaquin ALS (2) No No Neutral Neutral Neutral NA Yes Yes Nimazamoxammonium ALS (2) No Neutral No Neutral Neutral Neutral Yes Yes Nflumioxazin PPO (14) Neutral Neutral Yes Neutral Yes Yes Yes Yes Nfomesafen PPO (14) Yes Neutral Yes Neutral Neutral Neutral Neutral Yes flumicloracpentyl PPO (14) No No Neutral Neutral Neutral NA Yes Neutral Nsulfentrazone PPO (14) Yes Neutral Neutral Neutral Neutral Yes 5 Yes Neutral Nlactofen PPO (14) No Yes Neutral Yes Yes Yes Neutral Neutral fluthiacetmethyl PPO (14) No Yes Neutral Neutral Neutral Neutral Yes Neutral acifluorfen sodium PPO (14) Yes Yes Neutral Yes Neutral Neutral Yes Neutral paraquat dichloride BiPyr (22) Yes No Yes Neutral Neutral Yes 24 Yes Neutral Nglufosinateammonium Glu (10) Yes No Yes Neutral Neutral Neutral No reports Neutral Neutral Nmesotrione HPPD (28) No Neutral Neutral Yes Neutral Yes No reports Yes Yes N
Monsanto Company 10-SY-210U 

Table L-17 is intended to be a 1-page scorecard to track the benefits of dicamba use on MON 87708 according to the different listed csignifies that Monsanto has concluded that dicamba represents a benefit versus the relevant alternative herbicide on the basis of the ri

heading. The basis for entering a “Yes” under each risk factor is further explained in the relevant portions of this document. ”Neutral” en

exist for dicamba and the alternative herbicide. “No” means the alternative herbicide offers a risk benefit compared to dicamba

1

The herbicidal biochemical mechanism of weed-killing activity, according to the Weed Science Society of America. 

www.plantprotection.org/HRAC/Bindex.cfm?doc=MOA.html |Accessed May 27, 2010| 2A tally of the benefits dicamba use on MON 87708 offers over the alternative herbicide in the four categories of human health risk

described in L.5.1. 

3A tally of the benefits dicamba use on MON 87708 offers over the alternative herbicides in the two categories of aquatic non-target spe

criteria described in L.5.2. 

4A listing of the worldwide numbers of known resistant weeds for each herbicide based on its mode-of-action group. Dicamba has 5 known 

“Yes” indicates that the number of resistant weeds in this herbicide class is many more than the known five dicamba resistant species bioty

each individual herbicide in the class is not provided. See L.5.3.3. www.weedscience.org/summary/MOASummary.asp |Accessed May 28, 201

5Alternative herbicides that provide commercial control of fewer than 50% of targeted problem weeds in soybean compared to dicamb

Table L-16, where commercial control is considered to be >85%, as indicated by a fully-black circle symbol. See L.5.3.1. 6Alternative herbicides that require long waiting periods between application and subsequent planting of a crop other than soybean. This c

to growers. See Table L-18 and L.5.3.2. 

7“Alternative herbicides that can substantially injure the soybean crop when applied for weed control, potentially reducing soybean yield. 8Tabulation of the number of “Yes” entries in each row, indicating a total score for improved risk profile for dicamba use on MON 87708 

herbicide. This summation is not presented as a net value (i.e., subtracting where an alternative herbicide has a benefit over dicamba). 9Tabulation of the number of “No” entries in each row, indicating a total score for worse risk profile for dicamba use on MON 87708 o

herbicide. This summation is not presented as a net value (i.e., subtracting where an alternative herbicide has a benefit over dicamba). 

NA – not available.
Monsanto Company 10-SY-210U 655 of 721 

L.5.2. Comparisons for Ecological Effects

MON 87708 can be treated with preemergence and postemergence in-crop applications 

of dicamba. Such a weed control treatment regime has the opportunity to reduce risks to 

aquatic fish and invertebrates by: 

 Replacing currently-used or foreseeable future alternative herbicide products that 

have higher aquatic toxicity risk profiles, and 

 Addressing hard-to-control broadleaf weeds or broadleaf weeds that are resistant 

to glyphosate, PPO inhibitors, or ALS inhibitors, thereby preserving the ability of 

growers to manage weed problems and maximize soybean yield in the least risky 

manner, recognizing the ecological benefits offered by the superior hazard and 

reduced risk profile of glyphosate and the favorable profile of dicamba compared 

to some of the alternative herbicides.. 

L.5.2.1. Aquatic Animals

As mentioned, Table L-14 provides information concerning hazards, potential exposures, 

and risks to fish and aquatic invertebrates for each alternate herbicide and for dicamba. 

These data allow a comparison of the relative aquatic animal safety among the optional 

weed control products on the basis of: 

 Potency against the indicator species. 

 Estimates of potential exposure to aquatic animals. 

 Calculated RQs for aquatic animals, by combining the hazard and exposure 

parameters. The RQs that exceed the EPA’s LOC for non-listed aquatic animal 

species of 0.5 are marked in bold font. 

The assessment and comparisons summarized in Table L-17 establish that dicamba poses 

little acute risk to aquatic animals, which is consistent with EFED’s assessment published 

in the RED EFED Chapter and described in more detail in L.3.3. Furthermore, for 16 of 

the 18 alternate herbicides, aquatic animal RQs range from < 0.005 to 0.05, and do not 

present a risk to aquatic animals even using a worst-case upper bound exposure 

estimation. Only 2,4-D (the esters form) and lactofen exceed the LOC of 0.5 using 

conservative “worst-case” exposure estimates. These are highlighted in bold font. 

Monsanto notes that EPA considers the LOC for aquatic animals that are listed as 

endangered or threatened species to be 0.05 and the LOC for acute restricted use to be 

0.1. If the 10-fold to 5-fold lower level of concern were applied, flumioxazin would also 

exceed the LOC based on this “worst-case” exposure estimate. 

In addition to the calculated RQs, product labels for five of the alternative herbicides bear 

EPA-required warning statements for toxicity to fish or invertebrates, based on the hazard 

values (low LC50 or EC50) of those herbicides. These are flumioxazin, cloransulammethyl, flumiclorac-pentyl, lactofen, and 2,4-DB. 
Monsanto Company 10-SY-210U 656 of 721 

Therefore, Monsanto concludes that dicamba presents a risk benefit for aquatic animals 

compared to three of the 18 alternate herbicides: 2,4-D, flumioxazin, and lactofen. This 

conclusion is tabulated in the Table L-17 scorecard by a “Yes” in the relevant column. 

The alternative herbicide a.i.s that have a “worst-case” RQ < 0.05 are similar to dicamba 

in regards to aquatic animal risk, and are marked “Neutral” in Table L-17. 

L.5.2.2. Aquatic Plants 

As mentioned, Table L-15 provides information concerning hazards, potential exposures, 

and risks to aquatic plants for each alternate herbicide and for dicamba. These data allow 

a comparison of the relative aquatic plant safety among available weed control products 

on the basis of: 

 Potency against the indicator species. 

 Estimates of potential exposure to aquatic plants. 

 Calculated RQs for aquatic plants, by combining hazard and exposure parameters. 

The RQs that exceed the EPA’s LOC for non-listed aquatic plant species of 1.0 

are marked in bold font. 

The data format, sources, and methods are identical to those described above for the 

aquatic animals (Table L-14). A Level of Concern (LOC) value of 1.0 has been used for 

judging RQ exceedances in the case of aquatic plants, consistent with EPA EFED’s 

normal practices. 

The assessment and comparison summarized in Table L-17 establishes that dicamba 

poses little acute risk to aquatic plants, which is consistent with EFED’s assessment 

published in the RED EFED Chapter (U.S. EPA, 2005a) and discussed above. Monsanto 

was unable to locate public aquatic plant hazard data for three of the alternate herbicides 

(imazaquin, chlorimuron-ethyl, and flumiclorac-pentyl). For 9 of the 18 alternatives, the 

worst case RQs ranged between < 0.02 and 0.75; and do not present a risk to aquatic 

plants even using a worst-case upper bound exposure estimation. However, for seven 

alternative herbicides, the worst-case RQs did exceed EFED’s LOC of 1.0. It is not 

surprising that some herbicides are quite toxic to aquatic plants. The worst-case RQs for 

three of the alternate herbicides (flumioxazin, paraquat dichloride, and lactofen) exceeded 

the LOC by a factor of more than 50-fold. 

Therefore, Monsanto concludes that dicamba presents less risk to aquatic plants 

compared to seven of the 18 alternative herbicides: 2,4-D (ester form), flumioxazin, 

sulfentrazone, thifensulfuron, paraquat dichloride, lactofen, and mesotrione. This 

conclusion is tabulated in the Table L-17 scorecard by a “Yes” in the relevant column. 

The other alternative herbicide a.i.s, for which RQs are less than EPA’s LOC have 

similar non-target aquatic plant risk as dicamba, and are marked in Table L-17 with 

“Neutral”. The three herbicides for which no relevant data were available are marked as 

“NA”.
Monsanto Company 10-SY-210U 657 of 721 

L.5.3. Comparison for Efficacy and Weed Management, Including Weed Resistance 

L.5.3.1. Comparison of Weed Management Efficacy 

As mentioned, Table L-16 compares weed control effectiveness of formulated products 

containing dicamba and the alternative herbicides. The various products’ overall 

effectiveness can be compared using the simple method of counting the fully-black circle 

symbols, which identify instances of commercial level weed control. Dicamba provides 

commercial control for 23 of the listed weeds (primarily broadleaf species), which is the 

most of any of the herbicides in the table; dicamba offers commercial control for 13 of 

the herbicide-resistant biotypes and 10 of the hard-to-control species. After dicamba, 

preemergent treatments with 2,4-D, or postemergent treatments with glufosinate, 

tembotrione, and imazethapyr plus glyphosate are the next most effective herbicides 

against this target group of weeds. The least effective herbicides in this analysis against 

these targeted weeds and resistant biotypes are 2,4-DB, paraquat, imazethapyr alone, 

flumiclorac-pentyl, chlorimuron-methyl and fluthiacet. To summarize the comparative 

herbicidal effectiveness in this analysis, the scorecard in Table L-16 contains a column in 

which those herbicides that provide commercial control of 50% or fewer weeds compared 

to the number of weeds controlled by dicamba (i.e., eleven or fewer fully-black circles in 

Table L-16) are marked with a “Yes”, to denote a clear dicamba advantage. Alternative 

herbicide a.i.s that provide commercial control of greater than 50% of the number of 

weeds controlled by dicamba (i.e., 12 or more fully-black circles in Table L-16) are 

marked in Table L-17 with “Neutral”, meaning are judged similar to dicamba. The 50% 

threshold criterion is Monsanto’s arbitrary choice that is intended to identify herbicide 

options that are expected to provide substantially poorer weed control compared to 

dicamba. 

L.5.3.2. Comparison for Weed Management Practices 

Beyond direct efficacy against the key weeds, other advantages of MON 87708 treated 

with dicamba exist.

L.5.3.2.1. Increased Weed Control Flexibility 

MON 87708 will allow more flexibility for control of weeds just prior to or at planting of 

the crop, due to elimination of preplant intervals or plant back restrictions on present 

dicamba labels. These restrictions were in place due to concern over potential soybean 

injury, which is not a concern for MON 87708. Current common practice is to use 2,4-D 

for preemergent burndown treatment. When applied too close to soybean planting, 2,4-D 

can potentially reduce crop stands and cause injury to new seedlings (Thompson et al., 

2007). Restrictions have been implemented for most products and are as follows: rates 

up to 0.5 lb a.i. per acre must be applied at least seven days prior to planting; rates 

between 0.5 and 1 lb a.i. per acre must be applied at least 30 days prior to planting. 

Additionally, several sulfonylurea herbicides can also be utilized for preemergent 

burndown weed control in soybean. For example, Canopy herbicide can be applied prior 

to planting with the following restrictions: rates of 2.2 oz per acre or less should be 
Monsanto Company 10-SY-210U 658 of 721 

applied at least seven days prior to planting; rates of 2.2 to 3.3 oz per acre should be 

applied at least 14 days before planting. 

Current recommendations to control glyphosate-resistant biotypes of waterhemp and 

Palmer amaranth include the use of a residual herbicide treatment and postemergent 

applications of PPO-inhibiting herbicides such as Cobra (lactofen), Ultra Blazer 

(acifluorfen), or Cadet (fluthiacet). It is commonly known that these postemergent 

herbicides can cause excessive injury to soybean, especially under hot and sunny 

conditions. Soybean injury, caused by acifluorfen from early (V2 to V3 growth stage) or 

late postemergence (V5 to V6) applications, was seen as increased chlorosis and stunting 

that translated into yield reduction (Young et al., 2003). Furthermore, Legleiter et al.

(2009) illustrated the effectiveness of preemergent herbicides for the control of resistant 

waterhemp populations and demonstrated inconsistent control with PPO-inhibiting 

herbicides or herbicide combinations for the control of waterhemp populations with 

multiple resistance to glyphosate and PPO-inhibiting herbicides. The introduction of 

MON 87708 will provide an effective management option for herbicide-resistant biotypes 

of waterhemp and Palmer amaranth that is not expected to cause either crop injury or 

yield loss. 

Residual herbicides containing herbicides including sulfentrazone, dimethenamid, 

pendimethalin, metribuzin, or metolachlor have been shown to control waterhemp and 

Palmer amaranth; however, such products can present other challenges. Limitations 

include the need of adequate soil moisture for activation, potential crop injury, crop 

rotation restrictions, and use restrictions based on soil type. As an example, metribuzin 

should not be used on sands, loamy sands, and sandy loams with less than 1% organic 

matter62, and pendimethalin applied after crop-emergence can result in soybean injury63. 

For some residual herbicides there are extensive rotational crop restrictions, ranging from 

four to 40 months after application, to avoid subsequent crop injury to the rotated crop 

caused by herbicide remaining in the soil. These limitations reduce the choice of crops 

that can be planted in case the soybean crop is destroyed by weather or even the 

following growing season. Examples of planting limitations among the alternate 

herbicides are shown in Table L-18. 

 

62 Product label for Metribuzin 75 DF (http://www.cdms.net/LabelsMsds/LMDefault.aspx?pd=7614&t=). 

[Accessed 5/14/2010] 

63 Product label for Pendimax 3.3 

(http://aesop.rutgers.edu/~plantbiopath/links/bbcpestweb/GrapeLabels/pendimax33.pdf). [Accessed 

5/14/2010] 
Monsanto Company 10-SY-210U 659 of 721 

Table L-18. Planting Restrictions (months) for Alternative Herbicide Products 

CROP 

Herbicide Product 

(Active Ingredient) 

Authority MTZ 

(sulfentrazone 

+ metribuzin) 

Pursuit 

(imazethapyr) 

CANOPY EX 

(chlorimuron + 

tribenuron) 

Raptor 

(imazamox) 

Field corn 4 8.5 8 8.5 

Wheat 4 4 4 9 

Cotton 12 18 8 9 

Peanut 12 0 15 9 

Sorghum 12 18 9 3 

Onions 18 40 30 9 

Dicamba does not have rotational restrictions for such extended time periods, and 

provides a substantial advantage in flexibility. For the majority of crops, no rotational 

restrictions apply after 120 days following dicamba applications. Specifically, there are 

no rotational restrictions for planting corn following a dicamba application. For cotton, a 

rotational restriction of 21 days is recommended. To summarize this advantage, Table L17 includes a column in which a “Yes” is marked for those alternative herbicides that 

include substantial rotational or replanting restrictions. Entries for alternative herbicide 

a.i.s that do not require long rotation intervals are judged similar to dicamba, and are 

marked in Table L-17 with a “Neutral” in the relevant column. 

Furthermore, in situations where sequential herbicide applications were employed to 

control common waterhemp populations, reduced soybean yields have been reported. 

Yield reductions up to 19% compared to a non-treated control were reported when 

acifluorfen or fomesafen was the postemergent herbicide (Soltani et al., 2009). These 

reduced yields were associated with crop injury (chlorosis and stunting) following 

postemergent applications of certain herbicides, especially PPO inhibitors (Baumann et 

al., 2010; Loux et al., 2009). 

Dicamba treatments in MON 87708 provide excellent crop safety. Table L-17 includes a 

column in which the crop injury potential versus dicamba is summarized; a “Yes” entry 

indicates a substantial potential for visible soybean injury. Entries for alternative 

herbicide a.i.s with good soybean crop safety are judged to be similar to dicamba, and are 

marked with a “Neutral” in the relevant column of Table L-17. In conclusion, the 

introduction of MON 87708 will provide an additional mode-of-action for postemergent 

weed control with excellent crop safety. 

L.5.3.2.2. Weed Spectrum Benefits

Dicamba provides control of over 95 annual and biennial weed species, and control or 

suppression of over 100 perennial broadleaf and woody species. Dicamba provides more 

effective preemergent weed control than 2,4-D on cutleaf evening primrose, clover, and 

chickweed (Loux et al., 2009). Furthermore, dicamba provides excellent control 

compared to 2,4-D on summer annuals including those with a prostrate growth habit such 
Monsanto Company 10-SY-210U 660 of 721 

as knotweed and purslane. With regard to perennial weeds, research conducted at North 

Dakota State University indicates that dicamba is more effective in controlling Canada 

thistle compared to 2,4-D and effectively controls field bindweed (Zollinger, 2000; 

NDSU, 2010b). 

Dicamba also provides excellent control of wild buckwheat while 2,4-D has only limited 

activity and provides inadequate control (Zollinger et. al, 2006). Other preemergent or 

postemergent herbicides often provide incomplete control of wild buckwheat including 

dinitroanilines or PPO inhibitors. The most effective herbicides for buckwheat are 

dicamba, and some sulfonylurea products; however, some of the sulfonylurea herbicides 

may persist and carry over for more than one growing season, especially in high pH soils. 

Dicamba has been valued as more efficacious on lambsquarters than fomesafen or 

acifluorfen based on university weed control guidelines (Moechnig et al., 2010; 

University of Illinois, 2008; Legleiter et al., 2009; Loux et al., 2009). In addition, 

dicamba exhibits improved control of sicklepod (Loux et al., 2009), kochia and common 

ragweed (Legleiter et al., 2009), and waterhemp (Soltani et al., 2009) compared to 

fomesafen and acifluorfen. 

L.5.3.3. Comparison for Herbicide-Resistant Weeds 

The development of weed resistance reduces the effectiveness of all major herbicide 

classes used in soybean production today, including glyphosate, thereby jeopardizing 

soybean yields and requiring the introduction of new tools to control populations of 

resistant weeds. It is widely recognized that utilizing herbicides with different modes-ofaction in conjunction with established products is especially effective combating further 

weed resistance development and to provide control of existing populations (Beckie, 

2006). Preplant / preemergent or early-postemergent in-crop applications of dicamba in 

MON 87708 will introduce a new mode-of-action; thus the introduction of dicamba use 

on MON 87708 holds great promise for addressing current and future weed management 

needs in soybean. The primarily basis for this promise is the wide spectrum of activity of 

dicamba on broadleaf weed species, which are the most common hard-to-control species 

and resistant weed biotypes in soybean production today. 

Dicamba belongs to the auxin agonist class of herbicides, which is the oldest class of 

known synthetic herbicides. This class includes 2,4-D, 2,4-DB, mecoprop, MCPA, 

clopyralid, and several other active ingredients, and is WSSA Herbicide Group Number 

4.64 On the basis of their structural and chemical properties, auxinic herbicides have been 

classified into several sub-groups, i.e., phenoxyalkanoic acids (e.g., 2,4-D, MCPA), 

benzoic acids (e.g., dicamba, chloramben), pyridines (e.g., picloram, clopyralid), and 

quinolinecarboxylic acids (e.g., quinclorac, quinmerac). Generally, auxinic herbicides 

are effective against broadleaf (dicotyledonous) plant species, allowing them to be used 

in production of narrow leaf (monocotyledonous) crops such as corn and wheat. The 

 

64 There are several systems of herbicide mode-of action classification. Among the most widely used are 

those of the Herbicide Resistance Action Committee (HRAC) and the Weed Science Society of America. 

The classifications are compared in a chart at 

http://www.plantprotection.org/HRAC/Bindex.cfm?doc=MOA.html [Access May 28, 2010] 
Monsanto Company 10-SY-210U 661 of 721 

relative occurrence of herbicide-resistant weeds differs between the different sub-groups 

of auxinic herbicides. The largest number of resistant weed biotypes has been found for 

2,4-D. Considering that auxinic herbicides have been widely used in agriculture for more 

than 60 years, the occurrence of weed resistance to this class is relatively low (28 species 

worldwide, to date) and its development has been slow especially when compared to the 

speed of appearance of resistance to ALS inhibitors (107 species) or triazine-resistant 

populations (68 species).65 The relatively low incidence of auxinic herbicide resistance is 

believed to be attributable to the fact that there are multiple mechanisms of action for 

these herbicides (Gressel et al., 1982; Morrison and Devine, 1994). 

Only five weed species have been reported to date to be resistant to dicamba worldwide: 

kochia (Kochia scoparia) 66 , lambsquarters (Chenopodium album) 67 , prickly lettuce 

(Lactuca serriola), common hempnettle (Galeopsis tetrahit) and wild mustard (Brassica 

caber)

68. Of these five species, resistant populations of lambsquarters have only been 

reported in New Zealand. Regarding the two species with resistant populations in the 

U.S. (kochia, prickly lettuce) and Canada (common hempnettle and wild mustard), all 

were common to cereal production areas in the Western U.S. and Canada. No dicambaresistant populations have been reported in the main soybean production areas, including 

the Midwest, the South and the East coast of the U.S. Table L-17 shows the number of 

U.S. weed species known to have resistance to each of the major herbicide groups (or 

sub-groups within groups, as appropriate) to which the alternative herbicides belong. It 

also contains a “Yes” marker for those herbicides that have many more known resistant 

weed biotypes than the five known for dicamba, indicating a potential lower risk for 

weed resistance development for dicamba compared to alternative herbicides. Entries not 

indicated with a “Yes” mean that dicamba is either comparable or less favorable than the 

alternative herbicide. A comparison of each individual herbicide in the class is not 

provided. 

Although dicamba resistance exists, weed populations that are resistant to dicamba in 

U.S. soybean cropping areas have not been overly problematic to date, possibly because 

selection pressure on summer annual weeds has been low. Dicamba has seen very 

limited use in soybean (0.7% of treated soybean acres as a preplant and pre-harvest 

applications) and currently has relatively low usage in the crops that are commonly 

rotated with soybean (9.4% of corn acres as preplant and in-crop applications, 6.3% of 

cotton acres as a preplant application, and 8.4% of wheat acres, respectively) 

(Table VIII-12), although historically dicamba was used more extensively in corn. In 

addition, there are over 20 commercially-available pre-mixed multiple-herbicide 

formulations that contain dicamba, so dicamba is often used in combination with other 

 

65 Weed Science Society of America. http://www.weedscience.org/summary/MOASummary.asp

[Accessed May 28, 210] 

66 “Dicamba Resistance in Kochia”. H. J. Cranston, A. J. Kern, J. L. Hackett, E. K. Miller, B. D. Maxwell 

and W. E. Dyer, Weed Science 49:164-170, 2001. 

67 “Chemical Control Options for the Dicamba Resistant Biotype of Fathen (Chenopodium album)”. A. 

Rahman, T. K. James, and M. R. Trolove, New Zealand Plant Protection 61: 287-291, 2008. 68 “Inheritance of Dicamba Resistance in Wild Mustard (Brassica Kaber)”. M. Jasieniuk, I. N. Morrison 

and A. L. Brûlé-Babel, Weed Science 43:192-195, 1995. 
Monsanto Company 10-SY-210U 662 of 721 

herbicide modes-of-action. It is expected that selection pressure favoring the 

development of dicamba-resistant weeds will continue to be low even after in-crop use of 

dicamba on MON 87708 is approved. Monsanto believes this is because dicamba will 

predominantly still be used in combination with other herbicides exhibiting different 

modes-of-action, principally glyphosate, but also with other soil-active herbicides. The 

presence of multiple herbicides in the weed management system greatly diminishes the 

likelihood of weed resistance to dicamba developing to a level of predominance in weed 

populations. Dicamba is an excellent complement to the weed control spectrum of 

glyphosate, and it has a relatively low cost and low potential for crop injury to 

MON 87708 relative to other broadleaf weed options. Taken together, these factors will 

help to ensure that more fields receive weed-control treatments using multiple herbicide 

modes-of-action, in a proactive way, aimed at slowing the evolution of resistance. In 

general, this will serve to further reduce the development of weed resistance problems to 

all herbicides targeting broadleaf weeds in soybean production, and in crops rotated with 

soybean. 

As part of the projected role of dicamba as a companion herbicide for glyphosate, 

dicamba will provide growers with a new mode-of-action for use in-crop against summer 

annual broadleaf weeds. Dicamba will help prevent and/or combat existing weed 

resistance issues that can limit effectiveness of the PPO- and ALS-inhibiting herbicide 

classes. Herbicides from these two herbicide classes have historically dominated the 

non-glyphosate broadleaf weed control tools available in soybean (13 of the 18 

alternative herbicides considered here are PPO- or ALS-inhibiting herbicides (Table L17)), and were the predominant modes-of-action used for weed control in soybean 

production prior to the introduction of the Roundup Ready soybean system; and remain 

the primary options recommended for management of glyphosate-resistant or hard-tocontrol broadleaf weeds in soybean. For example, fomesafen and flumioxazin, both PPO 

Inhibitors, are the primary herbicides recommended for control of glyphosate-resistant 

Palmer amaranth (pigweed) in soybean. 

Dicamba will foster the adoption of IPM practices in soybean by allowing growers to 

continue to primarily focus on postemergent in-crop weed control, as they have practiced 

with Roundup Ready soybean, and will make current practices more effective by 

providing an additional mode-of-action herbicide. This will allow growers to delay some 

herbicide treatments until field scouting indicates a need for additional weed control 

which is consistent with the principles of Integrated Pest Management. Dicamba, as a 

companion product to glyphosate, will also continue to foster adoption of and maintain 

the use of conservation tillage practices, because of grower preference to use 

postemergent products, such as dicamba on MON 87708, compared to reliance on soilactive preemergent products 

In summary, the ability to use dicamba as part of a weed management program in U.S. 

soybean production will provide significant benefits for managing broadleaf weed 

resistance not only relative to glyphosate but also to other herbicides such as those 

included in PPO- and ALS-inhibitor classes. There is evidence in the scientific literature 

from data generated in field studies and from model simulations that the application of 

multiple herbicides, each effective in controlling a weed spectrum, with more than one 
Monsanto Company 10-SY-210U 663 of 721 

mode-of-action can significantly delay the evolution of resistant populations within a 

field (Powles et al., 1996; Beckie, 2006). In addition, there is evidence that resistance 

would be delayed longer by use of herbicide mixtures than by using an herbicide rotation 

strategy (Diggle et al., 2003; Beckie and Reboud, 2009). Based on this general 

information on resistance, Monsanto believes that application of dicamba on MON 87708 

integrated into the Roundup Ready soybean system will reduce the development of 

herbicide-resistant broadleaf populations to glyphosate and other herbicides. This 

conclusion is based on the following: 1) the efficacy and broad spectrum of glyphosate 

and weed control spectrum of dicamba, 2) the low level of dicamba-resistant broadleaf 

weeds in the major soybean production areas, 3) the low number of species resistant to 

glyphosate, 4) the expected use of dicamba applied to MON 87708, and 5) the fact that 

dicamba will be used in combination with glyphosate, and other alternative herbicides as 

necessary to control problematic weeds. Using available information, Monsanto 

conservatively estimates that dicamba use on MON 87708 could reduce the growth of 

resistant weed populations on five to ten percent of the expected 75 million U.S. planted 

soybean acres (Table VIII-1), which is equal to approximately 3.6 to 7.2 million acres. 

Furthermore, Monsanto believes the opportunity for dicamba use on MON 87708 will 

foster continued adoption of conservation tillage practices, an important goal for the 

agro-ecosystem and the long term sustainability of U.S. agriculture. Presently, 41.5% of 

an estimated 74 million acres of U.S. soybean acres, or 30.6 million acres, employ no-till 

or conservation tillage production systems (CTIC, 2007). The introduction of 

MON 87708 into the Roundup Ready soybean system would allow the flexibility to 

incorporate an additional herbicide mode-of-action in both pre- or postemergent 

applications, and support the continued use of conservation tillage production systems. 

The benefits of conservation tillage are well known and demonstrated including soil and 

water conservation, improved environmental (e.g., water) quality, and a reduced carbon 

footprint (Arriaga and Balkcom, 2005; Reicosky, 2008). These conservation tillage acres 

will represent a significant portion of the 30 million total acres projected for MON 87708 

planting, in turn representing a significant number of acres where a delay in the 

development of glyphosate-resistant biotypes could be expected to occur. 

L.6. Statement of Benefits for Dicamba-Tolerant Soybean MON 87708 

Monsanto intends to commercialize MON 87708 in combination with the Roundup 

Ready 2 Yield soybean platform (glyphosate-tolerant soybean), so that both herbicide 

tolerance traits occur in the same soybean plant. The addition of MON 87708 to the 

Roundup Ready 2 Yield soybean platform enables the use of dicamba withglyphosate or 

other herbicides as a mixture or sequential applications for pre-plant burndown and inseason weed control, resulting in a highly effective weed management system for 

soybean growers. Combined use of dicamba plus glyphosate will provide excellent grass 

and broadleaf weed control, including those broadleaf weeds that are hard–to-control or 

glyphosate-resistant. The use of dicamba and glyphosate should deter the spread of 

existing glyphosate-resistant broadleaf weeds, and possibly delay the development of 

new resistant populations through use of multiple modes-of-action. The decision by 

growers to adopt MON 87708 x Roundup Ready 2 Yield soybean system and to include a 

dicamba herbicide treatment, either preemergent or in-crop, will be dependent up on three 
Monsanto Company 10-SY-210U 664 of 721 

factors: weed spectrum in a grower’s field, efficacy of dicamba against broadleaf weeds, 

and costs and flexibility versus alternative herbicide programs. 

The use of dicamba in combination with glyphosate for burndown treatments prior to 

planting on no-till acres will provide growers with a more flexible weed management tool 

than is presently available. MON 87708 (in combination with application of dicamba) 

will provide growers with greater flexibility in timing of application of burndown 

treatments, eliminating the need for any interval between dicamba application and 

planting, and improving overall efficacy on hard-to-control and larger broadleaf weeds. 

In addition, dicamba is an effective complement to glyphosate in cropping systems where 

glyphosate-resistant weeds such horseweed (Conyza canadensis) have developed. 

Approximately 40% of the total soybean acres in the United States are produced in a notill system (CTIC, 2007). In these cropping systems, it is anticipated that dicamba will 

displace other herbicides, such as 2,4-D, because a preplant interval is not needed with 

MON 87708 to avoid potential crop injury, and because dicamba has a better weed 

spectrum profile, as discussed previously. However, the potential future introduction of 

2,4-D-tolerant soybean will likely allow for a decreased preplant interval for soybean 

containing this trait. 

In addition to soybean production under no-till cropping systems, MON 87708 offers an 

effective solution for improving weed control in conventional tillage cropping systems. 

Herbicides with soil residual and postemergent efficacy are frequently used in 

combination with glyphosate to control weeds that are hard-to-control, resistant to 

glyphosate, or are capable of producing multiple flushes. Examples include, but are not 

limited to, the following species: Amaranthus tuberculatus (tall waterhemp), Palmer 

amaranth (Amaranthus palmeri), giant ragweed (Ambrosia trifida) and lambsquarters 

(Chenopodium album). In these cropping situations, it could be assumed that dicamba 

would replace some of the less effective PPO inhibitors (such as acifluorfen and lactofen) 

that have greater potential for crop injury. The opportunity to use dicamba as yet a third 

mode-of-action in these situations would also reduce the potential for selection pressure 

for resistance to any single herbicidal mode-of-action. 

Assuming that the U.S. soybean crop is grown on approximately 75 million acres (Table 

VIII-1), it is estimated that dicamba herbicides could be used for weed control on a total 

of approximately 30 million acres. There are currently 11 active ingredients that account 

for 90% of the herbicide usage (based on expenditure) in soybean production today 

(AgroTrak, 2009). The estimated percentage of treated soybean acreage by active 

ingredient is presented in Table L-12 based on the latest USDA-NASS use data from 

2008. The use of dicamba as a preplant burndown or in-crop application has the potential 

to reduce the rates, acreage, and/or the number of applications of these alternative 

herbicides in soybean production. 

There are widely varying estimates of the current number of soybean acres that have 

populations of weeds that are resistant to one or more of the classes of herbicides 

currently registered for use in soybean. It is difficult to determine the validity and 

comparability of these estimates, as different regions and weed scientists have a different 

approach or method for deriving these estimates. Regardless of the accuracy of these 
Monsanto Company 10-SY-210U 665 of 721 

estimates of the current state of herbicide resistance in soybean production, the 

application of dicamba on MON 87708 will provide a benefit in broadleaf weed 

resistance management for all the herbicide modes-of-action currently used in soybean 

for the following reasons: 1) the broad spectrum of broadleaf weed control provided by 

dicamba, 2) the limited number of dicamba-resistant biotypes present after decades of 

dicamba use, 3) the projected soybean acreage (40%) adopting MON 87708 into the 

Roundup Ready soybean system and the combination with other herbicide modes-ofaction on these acres, and 4) the compatibility of dicamba end-use products with 

herbicides having other modes-of-action. 
Monsanto Company 10-SY-210U 666 of 721 

References for Appendix L 

AgroTrak. 2009. Unpublished 2008 agricultural survey datra. Gfk Kynetec Marketing 

Research, Inc. http://www.gfk.com/gfk-kynetec/. 

Arriaga, F.J. and K.S. Balkcom. 2005. Benefits of conservation tillage on rainfall and 

water management. 2005 Georgia Water Resources Conference, University of Georgia, 

Athens, Georgia. 

Baumann, T.T., A.F. Dobbels, W.G. Johnson, M.M. Loux, G.R.M. Nice, and S. J.M. 

2010. Weed Control Guide for Ohio and Indiana. Purdue University and Ohio State 

GUniversity Extension Bulletin 789. 

Beckie, H.J. 2006. Herbicide-resistant weeds: Management tactics and practices. Weed 

Technology 20:793-814. 

Beckie, H.J. and Xavier Reboud. 2009. Selecting for weed resistance: Herbicide rotation 

and mixture. Weed Technology 23:363-370. 

CTIC. 2007. 2007 amendment to the national crop residue management survey summary. 

Conservation Technology Information Center, West Lafayette, Indiana. 

http://www.conservationinformation.org/pdf/National_Summary_2007_Amendment.pdf 

Accessed June 14, 2010. 

Crop Protection Guide - Information for Dealers. 2009. 

http://www.agrisolutionsinfo.com/ [Accessesd March 26, 2010]. 

Diggle A.J., P. Neve, and F.P. Smith. 2003. Herbicides used in combination can reduce 

the probability of herbicide resistance. Weed Research 4: 371-382. 

Durkin, P. and S. Bosch. 2004. Dicamba - Human health and ecological risk assessment - 

Final report. SERA TR 04-43-17-06d. U.S. Department of Agriculture, Forest Service, 

Washington, D.C. http://www.fs.fed.us/foresthealth/pesticide/pdfs/112404_dicamba.pdf 

Accessed May 26, 2010. 

Gianessi, L.P. 2005. Economic and herbicide use impacts of glyphosate-resistant crops. 

Pest Management Science 61: 241-245. 

Gressel, J., and Segel, L. A. 1982. Interrelating factors controlling the rate of appearance 

of resistance: the outlook for the future. Pages 325–347in Herbicide Resistance in Plants. 

H.M. LeBaron and J. Gressel, eds. John Wiley & Sons, Inc., New York, New York. 

Legleiter, T.R., Bradley, K.W., and Massey, R.E. 2009. Glyphosate-resistant waterhemp 

(Amaranthus Rudis) control and economic returns with herbicide programs in soybean. 

Weed Technology 23:54-61. 
Monsanto Company 10-SY-210U 667 of 721 

Loux, M.M., D. Doohan, A.F. Dobbels, W.G. Johnson, G.R.W. Nice, T.N. Jordan and 

T.T. Bauman. 2009. Weed control guide for Ohio and Indiana. Bulletin 789. Purdue 

University and Ohio State University Extension, West Lafayette, Indiana. 

Moechnig, M., D.L. Deneke and L.J. Wrage. 2010. Weed control in soybeans: 2010. 

South Dakota State University, Brookings, South Dakota. 

http://agbiopubs.sdstate.edu/articles/FS525B.pdf Accessed May 12, 2010

Morrison, I.N., and M.D. Devine. 1994. Herbicide resistance in the Canadian prairie 

provinces - 5 years after the fact. Phytoprotection 75:5-16. 

NDSU. 2010a. Weed control guide. http://www.ndsu.edu/weeds Accessed March 26, 

2010. 

NDSU. 2010b. Canada thistle: Cirsium arvense (L.) Scop. North Dakota State University, 

Fargo, North Dakota. http://www.ag.ndsu.nodak.edu/invasiveweeds/canda%20thistle.htm. 

Accessed March 30, 2010

Pedersen, P. 2008. Soybean variety selection. Iowa State University Extension, Ames, 

Iowa. http://extension.agron.iastate.edu/soybean/production_varietyselect.html Accessed 

April 19, 2011. 

Powles, S.B., C. Preston, I.B. Bryan and A.R. Jutsum. 1996. Herbicide resistance: Impact 

and management. Pages 57-93 in Advances in Agronomy. Volume 58. D.L. Sparks (ed.). 

Academic Press, San Diego, California. 

Reicosky, D.C. 2008. Carbon sequestration and environmental benefits from no-till 

systems. Pages 43-58 in No-Till Farming Systems. T. Goddard (ed.). World Association 

of Soil and Water Conservation, Bangkok, Thailand. 

Soltani, N., Vyn, J. D., and Sikkema, P. H. 2009. Control of common waterhemp 

(Amaranthus tuberculatus var. rudis) in corn and soybean with sequential herbicide 

applications. Canadian Journal of Plant Science 89:127-132. 

Thompson, M.A., Steckel, L.E., Ellis, A.T., and Mueller, T.C. 2007. Soybean tolerance 

to early preplant applications of 2,4-D ester, 2,4-D Amine, and Dicamba. Weed 

Technology 21:882-885. 

U.S. EPA. 2005a. EFED Reregistration Chapter For Dicamba / Dicamba Salts. U.S. 

Environmental Protection Agency, Office of Pesticides, Prevention, and Toxic 

Substances, DP Bar Code 317976, August 31, 2005. 

U.S. EPA. 2005b. Dicamba: HED Chapter of the Reregistration Eligibility Decision 

Document, Phase 1. PC Code 029801; DP Barcode: D317720 (Memorandum, C. Olinger 

and Y. Yang, September 13). Health Effects Division, Office of Prevention, Pesticides 

and Toxic Substances, U.S. Environmental Protection Agency, Washington, DC. 
Monsanto Company 10-SY-210U 668 of 721 

EPA. 2008. Dicamba pesticide tolerance. U.S. Environmental Protection Agency, 

Washington, D.C. Vol. 73, No. 64. 

U.S. EPA. 2009a. Amendment to organic arsenicals RED. U.S. EPA U.S. Environmental 

Protection Agency, Office of Pesticides, Prevention, and Toxic Substances, April 22, 

2009. http://www.epa.gov/oppsrrd1/REDs/organic-arsenicals-amended.pdf. 

U.S. EPA. 2009b. Registration eligibility decision for dicamba and associated salts. U.S. 

Environmental Protection Agency, Office of Pesticides, Prevention, and Toxic 

Substances, June 8, 2006, as amended June 17, 2009. 

U.S. EPA. 2009c. Reduced risk/organophosphate alternative decisions for conventional 

pesticides, updated 7/1/09”. Washington, D.C. 

http://www.epa.gov/opprd001/workplan/completionsportrait.pdf. 

U.S. EPA. 2009d. Mesotrione: human-health risk assessment for Section 3 new use on 

soybeans. U.S. Environmental Protection Agency, Office of Pesticides, Prevention, and 

Toxic Substances, Washington, D.C. DP Barcode: D357890, August 12, 2009. 

U.S. EPA. 2009e. Mesotrione; Pesticide Tolerances. U.S. Environmental Protection 

Agency, Washington, D.C. 74 FR 67119 - 67124, December 18, 2009. 

U.S. EPA. 2011. 2011 Edition of the drinking water standards and health advisories. U.S. 

Environmental Protection Agency, Washington, D.C. 

University of Illinois. 2008. Illinois Agricultural Pest Management Handbook.. 

www.ipm.illinois.edu/pubs/iapmh/00contents.pdf Accessed June 14, 2010. 

USDA-NASS. 2007a. Acreage - June 2006. U.S. Department of Agriculture, National 

Agricultural Statistics Service, Washington, D.C. 

USDA-NASS. 2007b. Agricultural chemical usage 2006 field crops summary. U.S. 

Department of Agriculture, National Agricultural Statistics Service, Washington, D.C. 

USDA-NASS. 2009. Crop production 2008 summary. U.S. Department of Agriculture, 

National Agricultural Statistics Service, Washington, D.C. 

Young, B.G., Young, J.M., Matthews, J.L., Owen, M.D.K., Zelaya, I.A., Hartzler, R.G., 

Wax, L.M., Rorem, K.W., and Bollero, G.A.. 2003. Soybean development and yield as 

affected by three postemergence herbicides. Agronomy Journal 95:1152-1156. 

Weed Control Manual for Tennessee. 2010. 

http://www.utextension.utk.edu/publications/pbfiles/pb1580.pdf. 

Zollinger, R. and Rodney, L. 2000. Identification and control of field bindweed. W-802 

(Revised). http://www.ag.ndsu.edu/pubs/plantsci/weeds/w802w.htm Accessed March 26, 

2010. 
Monsanto Company 10-SY-210U 669 of 721 

Zollinger, R., Bradley, K. W., and Massey, R. E. 2006. The glyphosate weeds and crop 

series: biology and management of wild buckwheat. Purdue Extension – Knowledge to 

Go: GWC-10. 

 
Monsanto Company 10-SY-210U 670 of 721 

Alternative Herbicide-Specific References 

The following active ingredient specifi references were used to generate the comparative 

summary information contained in Tables L-13, L-14 and L-15. 

2,4-D

U.S. EPA. 2004. Environmental Fate and Effects Division’s Risk Assessment for the 

Reregistration Eligibility Document for 2,4-Dichlorophenoxyacetic Acid (2,4-D). OPP 

Public Docket (OPP-2004-0167-0003). 

U.S. EPA. 2005. 2,4-D RED Facts. Registration Eligibility Decision for 2,4-D. U.S. 

Environmental Protection Agency, Office of Pesticides, Prevention, and Toxic 

Substances, Document EPA 738-R-05-002, June, 2005. 

2,4-DB

U.S. EPA. 2005. Registration Eligibility Decision for 2,4-DB”. U.S. Environmental 

Protection Agency, Office of Pesticides, Prevention, and Toxic Substances, Document 

EPA738-R-05-001, January, 2005. 

Acifluorfen, Sodium

U.S. EPA. 2003. R.E.D. Facts Sodium Acifluorfen (Blazer®). U.S. Environmental 

Protection Agency, Document EPA-738-F-04-001, September 2003. 

U.S. EPA. 2004. Registration Eligibility Decision for Sodium Acifluorfen, Case No. 

2605. U.S. Environmental Protection Agency, Office of Pesticides, Prevention, and Toxic 

Substances, (undated). See also “Sodium Acifluorfen RED; Notice of Availability”. 69 

FR 4139 - 4138, January 28, 2004. 

U.S. EPA. 2002. Sodium Acifluorfen, HED Chapter for the Reregistration Eligibility 

Decision. U.S. Environmental Protection Agency Office of Pesticides, Prevention, and 

Toxic Substances, DP Barcode: D279497, January 15, 2002. 

U.S. EPA. 2010. OPP Pesticide Ecotoxicity Database. National Information System for 

the Regional IPM Centers. http://www.ipmcenters.org/Ecotox/index.cfm 

http://www.ipmcenters.org/Ecotox/index.cfm Accessed May 28, 2010. 

Chlorimuron

U.S. EPA. 2009. Chlorimuron-ethyl: Revised Human Health Risk Assessment for 

Proposed Uses On Cranberry and Low-growing Berry Subgroup 13-07H, except 

Strawberry, PP# 6E7153. U.S. Environmental Protection Agency, Office of Pesticides, 

Prevention, and Toxic Substances, DP Barcode D361368, February 5, 2009. 

U.S. EPA. 2004. Report of the Food Quality Protection Act (FQPA) Tolerance 

Reassessment Progress and Risk Management Decision (TRED) for Chlorimuron Ethyl. 
Monsanto Company 10-SY-210U 671 of 721 

U.S. Environmental Protection Agency, Office of Pesticides, Prevention, and Toxic 

Substances, September 22, 2004. 

U.S. EPA. 2010. OPP Pesticide Ecotoxicity Database. National Information System for 

the Regional IPM Centers. http://www.ipmcenters.org/Ecotox/index.cfm 

http://www.ipmcenters.org/Ecotox/index.cfm Accessed May 28, 2010. 

Cloransulam

U.S. EPA. 1997. Pesticide Fact Sheet; Name of Chemical: Cloransulam-methyl. U.S. 

Environmental Protection Agency Office of Pesticides, Prevention, and Toxic Substances, 

October 29, 1997. 

U.S. EPA. 1997. Cloransulam-methyl; Pesticide Tolerances. U.S. Environmental 

Protection Agency, 62 FR 49158-49163, September 19, 1997. 

U.S. EPA. 2010. OPP Pesticide Ecotoxicity Database. National Information System for 

the Regional IPM Centers. http://www.ipmcenters.org/Ecotox/index.cfm 

http://www.ipmcenters.org/Ecotox/index.cfm Accessed May 28, 2010. 

Dicamba

Hoberg, James R. “Dicamba technical – Determination of Effects on Seed Germination, 

Seedling Emergence, and Vegetative Vigor of Ten Plant Species”. Springborn 

Laboratories, 1993. 

U.S. EPA. 2009. Registration Eligibility Decision for Dicamba and Associated Salts. 

U.S. Environmental Protection Agency, Office of Pesticides, Prevention, and Toxic 

Substances, June 8, 2006, as amended June 17, 2009. 

U.S. EPA. 2005. Dicamba: HED Chapter of the Reregistration Eligibility decision 

Document (RED) - Phase I. U.S. Environmental Protection Agency, Office of Pesticides, 

Prevention, and Toxic Substances, DP Bar Code D317720, September 13, 2005. 

U.S. EPA. 2005. EFED Reregistration Chapter For Dicamba / Dicamba Salts. U.S. 

Environmental Protection Agency, Office of Pesticides, Prevention, and Toxic 

Substances, DP Bar Code 317976, August 31, 2005. 

Flumiclorac-pentyl

U.S. EPA. 2009. Registration Review - Preliminary Problem Formulation for the 

Ecological Risk Assessment of Flumiclorac-pentyl. U.S. Environmental Protection 

Agency, Office of Pesticides, Prevention, and Toxic Substances, DP Bar Code 362087, 

July 10, 2009. EPA-HG-OPP-2009-0084-0004. 

U.S. EPA. 2009. Flumiclorac Pentyl: Revised Human Health Assessment Scoping 

Document in Support of Registration Review. U.S. Environmental Protection Agency, 
Monsanto Company 10-SY-210U 672 of 721 

Office of Pesticides, Prevention, and Toxic Substances, DP Bar Code D368451, August 

28, 2009. 

U.S. EPA. 2005. Report of Food Quality Protection Act (FQPA) Tolerance 

reassessment Progress and Risk Management decision (TRED) for Flumiclorac pentyl. 

U.S. Environmental Protection Agency, Office of Pesticides, Prevention, and Toxic 

Substances, August 23, 2005. 

U.S. EPA. 2006. Flumiclorac pentyl; Pesticide Tolerance. U.S. Environmental 

Protection Agency, 71 FR 11526-11533, March 8, 2006. 

Flumioxazin

U.S. EPA. 2007. Flumioxazin. Human Health Risk Assessment for the Proposed Food 

Use of the Herbicide Flumioxazin on Alfalfa, Asparagus, Dry Beans, Fruiting Vegetables 

(Group 8, Including Okra), Melons (Subgroup 9A), Bushberries (Subgroup 13B), and 

Tree Nuts (Group 14), and a Request for an Amended Use on Garlic. U.S. 

Environmental Protection Agency, Office of Pesticides, Prevention, and Toxic 

Substances, DP Bar Code 336194, November 28, 2007. 

U.S. EPA. 2008. Flumioxazin; Pesticide Tolerance. U.S. Environmental Protection 

Agency, 73 FR 11826-11831, March 5, 2008. 

U.S. EPA. 2010. OPP Pesticide Ecotoxicity Database. National Information System 

forthe Regional IPM Centers. http://www.ipmcenters.org/Ecotox/index.cfm Accessed 

May 28, 2010. 

Fluthiacet-methyl

U.S. EPA. 1999. Pesticide Fact Sheet; Name of Chemical: Fluthiacet-methyl. U.S. 

Environmental Protection Agency Office of Pesticides, Prevention, and Toxic Substances, 

April, 1999. 

U.S. EPA. 2001. Fluthiacet-methyl; Pesticide Tolerance. 66 FR 65839-65850, 

December 21, 2001. 

U.S. EPA. 2006. Fluthiacet-methyl; Pesticide Tolerance. 71 FR 77620-77625, 

December 27, 2006. 

U.S. EPA. 2010. OPP Pesticide Ecotoxicity Database. National Information System for 

the Regional IPM Centers. http://www.ipmcenters.org/Ecotox/index.cfm Accessed May 

28, 2010. 

Fomesafen

U.S. EPA. 2006. Fomesafen; Pesticide Tolerance. U.S. Environmental Protection 

Agency, 71 FR 25945 - 25951, May 3, 2006. 
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U.S. EPA. 2007. Fomesafen Summary Document; Registration Review - Initial Docket 

March 2007. U.S. Environmental Protection Agency, Office of Pesticides, Prevention, 

and Toxic Substances, Docket No. EPA-HQ-OPP-2006-0239, March 22, 2007. 

U.S. EPA. 2010. OPP Pesticide Ecotoxicity Database. National Information System for 

the Regional IPM Centers. http://www.ipmcenters.org/Ecotox/index.cfm 

http://www.ipmcenters.org/Ecotox/index.cfm Accessed May 28, 2010. 

Glufosinate

U.S. EPA. 2003. Glufosinate Ammonium; Pesticide Tolerance. U.S. Environmental 

Protection Agency, 68 FR 55833 - 55849, September 29, 2003. 

U.S. EPA. 2009. Glufosinate Summary Document; Registration Review - Initial Docket 

March 2008, Case #7224. U.S. Environmental Protection Agency, Office of Pesticides, 

Prevention, and Toxic Substances, Docket No. EPA-HQ-OPP-2008-0190, March 19, 

2009. 

Glyphosate

U.S. EPA 2009. Problem Formulation for the Ecological Risk and Drinking Water 

Exposure Assessments in Support of the Registration Review of Glyphosate and Its Salts. 

EPA-HQ-OPP-2009-0361-0007. 

Imazamox

U.S. EPA. 1997 Pesticide Fact Sheet. Name of Chemical: Imazamox (Raptor 

Herbicide) Reason for Issuance: Conditional Registration. Date Issued: May 22, 1997. 

U.S. EPA. 2010. OPP Pesticide Ecotoxicity Database. National Information System for 

the Regional IPM Centers. http://www.ipmcenters.org/Ecotox/index.cfm Accessed May 

28, 2010. 

Imazaquin

U.S. EPA. 2005. Imazaquin and its Salts: HED Chapter of the Tolerance Reassessment 

Eligibility Decision (TRED) Document DP Barcode: D302939. U.S. Environmental 

Protection Agency, Office of Pesticides, Prevention, and Toxic Substances, October 31, 

2005. 

U.S. EPA. 2005. Report of the Food Quality Protection Act (FQPA) Tolerance 

Reassessment Progress and Risk Management Decision (TRED) for Imazaquin. U.S. 

Environmental Protection Agency, Office of Pesticides, Prevention, and Toxic 

Substances, Document No. EPA 738-R-00-009, December, 2005. 

U.S. EPA. 2005. Drinking Water Assessment for Imazaquin and its Salts. 

Environmental Protection Agency, Office of Pesticides, Prevention, and Toxic 

Substances, DP Barcode: D313182, June 21, 2005. 
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U.S. EPA. 2010. OPP Pesticide Ecotoxicity Database. National Information System for 

the Regional IPM Centers. http://www.ipmcenters.org/Ecotox/index.cfm Accessed May 

28, 2010. 

Imazethapyr

U.S. EPA. 2002. Imazethapyr; Pesticide Tolerance. U.S. Environmental Protection 

Agency 67 FR 55323 - 55331, August 29, 2002. 

U.S. EPA. 2010. OPP Pesticide Ecotoxicity Database. National Information System for 

the Regional IPM Centers. http://www.ipmcenters.org/Ecotox/index.cfm Accessed May 

28, 2010. 

Isoxaflutole

U.S. EPA. 1998. Pesticide Fact Sheet; Name of Chemical: Isoxaflutole. U.S. 

Environmental Protection Agency Office of Pesticides, Prevention, and Toxic Substances, 

September 15, 1998. 

U.S. EPA. 1998. Isoxaflutole; Pesticide Tolerance. U.S. Environmental Protection 

Agency, 63 FR 50773 - 50784, September 23, 1998. 

Lactofen

U.S. EPA. 2006. EFED Problem Formulation for Lactofen Review. DP Barcode: 323196. 

December 13, 2006. EPA-HQ-OPP-2005-0287-0004. 

U.S. EPA. 2004. Lactofen; Pesticide Tolerance. U.S. Environmental Protection Agency, 

69 FR 57207 - 57216, September 24, 2004. 

U.S. EPA. 2003. Report of the Food Quality Protection Act (FQPA) Tolerance 

Reassessment Progress and Risk Management Decision (TRED) for Lactofen. U.S. 

Environmental Protection Agency, Office of Pesticides, Prevention, and Toxic 

Substances, Document No. EPA 738-R-04-002, September 24, 2003. 

U.S. EPA. 2000. Lactofen: Preliminary Human Health Risk Assessment for Tolerance 

Reassessment, Incorporating Revised Cancer Unit Risks. U.S. Environmental Protection 

Agency Office of Pesticides, Prevention, and Toxic Substances, DP Barcode: D269621, 

October 12, 2000. 

Mesotrione

U.S. EPA. 2001. Pesticide Fact Sheet; Name of Chemical: Mesotrione. Office of 

Pesticides, Prevention, and Toxic Substances, June 4, 2001. 

U.S. EPA. 2009. Mesotrione; Pesticide Tolerances. U.S. Environmental Protection 

Agency, 74 FR 67119 - 67124, December 18, 2009. 
Monsanto Company 10-SY-210U 675 of 721 

U.S. EPA. 2009. Mesotrione: Human-Health Risk Assessment for Section 3 New Use 

on Soybeans. U.S. Environmental Protection Agency, Office of Pesticides, Prevention, 

and Toxic Substances, DP Barcode: D357890, August 12, 2009. 

U.S. U.S. EPA. 2010. OPP Pesticide Ecotoxicity Database. National Information System 

for the Regional IPM Centers. http://www.ipmcenters.org/Ecotox/index.cfm Accessed 

May 28, 2010. 

Paraquat

U.S. EPA. 1997. Registration Eligibility Decision for Paraquat Dichloride. U.S. 

Environmental Protection Agency Office of Pesticides, Prevention, and Toxic Substances. 

Document No. EPA 738-F-96-018, August, 1997. 

U.S. EPA. 2006. Paraquat Dichloride; Pesticide Tolerance. U.S. Environmental 

Protection Agency, 71 FR 52487 - 52494, September 6, 2006. 

U.S. EPA. 2010. OPP Pesticide Ecotoxicity Database. National Information System for 

the Regional IPM Centers. http://www.ipmcenters.org/Ecotox/index.cfm Accessed May 

28, 2010. 

Sulfentrazone

U.S. EPA. 2009. Revised EFED Problem Formulation for Sulfentrazone Registration 

Review. U.S. Environmental Protection Agency, Office of Pesticides, Prevention, and 

Toxic Substances, DP Bar Code 363522, EPA-HQ-OPP-2009- 0624-0006. September 9, 

2009. 

U.S. EPA. 2009. Sulfentrazone. REVISED Human Health Assessment Scoping 

Document in Support of Registration Review. U.S. Environmental Protection Agency, 

Office of Pesticides, Prevention, and Toxic Substances, DP Bar Code 369064, September 

9, 2009. 

Tembotrione

U.S. EPA. 2007. Pesticide fact Sheet: Tembotrione. U.S. Environmental Protection 

Agency, Office of Pesticides, Prevention, and Toxic Substances, September, 2007. 

Thifensulfuron

U.S. EPA. 2007. Thifensulfuron Methyl; Pesticide Tolerance. U.S. Environmental 

Protection Agency, 72 FR 13179 - 13184, March 21, 2007. 

U.S. EPA. 2008. Ingredient: Thifensulfuron Methyl; Title: Label Amendments and 

petition for Tolerances on Wheat Forage and Hay, Oat Forage and Hay, and Barley Hay. 

U.S. Environmental Protection Agency, Office of Pesticides, Prevention, and Toxic 

Substances, DP Bar Code 350544, April 17, 2008. 
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Appendix M: Potential Impact of Dicamba on Human Health and the Environment 

M.1. Overview 

Dicamba use is regulated at the federal level by EPA, not APHIS. APHIS’s authority under the 

Plant Protection Act does not allow it to specify conditions for the use of herbicides. Instead, 

EPA specifically approves labeling for any herbicide use including uses on agricultural crops. 

Before any dicamba product could be used over MON 87708 it must first be approved by the 

EPA as required by the Federal Insecticide Fungicide and Rodenticide Act (FIFRA); EPA must 

approve the pesticide (herbicide) product labeling for that specific use. Nevertheless, Appendix 

M examines the potential impact of dicamba use on MON 87708 on human health and the 

environment. It first provides a background discussion of U.S. EPA’s authority to regulate 

pesticides. Next, Appendix M addresses the regulatory background for dicamba, including the 

proposed use on MON87708. Appendix M then discusses the potential impact of dicamba on 

human health, including impacts on pesticide applicators. Appendix M concludes with a 

discussion of drift and offsite movement aspects associated with dicamba use. 

M.2. Regulation of Pesticides in U.S. 

M.2.1. Pesticide Registration, Reregistration and Tolerance Setting 

FIFRA requires that before the sale or distribution of a new pesticide or a new use of a registered 

pesticide, a company must obtain a registration, or license, from EPA. The EPA must ensure 

that the pesticide, when used according to its label directions, will not cause unreasonable 

adverse effects on human health and the environment. In order to address this standard, EPA 

evaluates potential risks to humans and the environment, and may require applicants to submit 

more than 100 different scientific studies conducted according to EPA guidelines. According to 

EPA, more than 1000 active ingredients are currently registered as pesticides in the U.S., which 

are, in turn, formulated into many thousands of pesticide products that are available in the 

marketplace (U.S. EPA, 2010). 

Pesticide registration is a scientific, legal, and administrative procedure through which EPA 

examines the ingredients of the pesticide; the particular site or crop on which it is to be used; the 

amount, frequency, method and timing of application, and other conditions of its use; and storage 

and disposal practices. In evaluating a pesticide registration application, EPA assesses a wide 

variety of data indicating the potential human health and environmental effects associated with 

use of the pesticide product. The data required by EPA are used to evaluate a wide array of 

potential impacts, including whether a pesticide has the potential to cause adverse effects on 

humans, wildlife, fish, and plants (including endangered species and other non-target organisms, 

i.e., organisms that the pesticide is not intended to act against). The registration applicant must 

also supply data on the pesticide’s potential impact on surface water and ground water, should 

leaching or runoff occur. The potential human health and safety risks assessed range from shortterm toxicity to long-term effects such as cancer and reproductive system disorders. 

Pesticide label directions are considered as part of EPA’s evaluation to determine whether a 

pesticide can be used safely, as required by FIFRA, and the language that appears on each 

pesticide label has been expressly approved by EPA. A pesticide product can only be used 
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legally according to the directions for use on the labeling accompanying the product at the time 

of sale. 

The registration of a new pesticide is not EPA’s only opportunity to evaluate the product’s safety. 

EPA has recently completed a program to review older pesticides (those initially registered 

before November 1984) under FIFRA to ensure that they meet current scientific and regulatory 

standards. Reregistration, like the initial registration process, considers the human health and 

ecological effects of pesticides and results in actions to reduce risks that are of concern. 

Where pesticides may be used on food or feed crops, EPA also sets pesticide tolerances, i.e., 

maximum pesticide residue levels that can legally remain in or on foods. EPA undertakes this 

analysis under the authority of the Federal Food, Drug, and Cosmetic Act (FFDCA). Under the 

FFDCA, EPA must find that such tolerances will be safe, meaning that there is a reasonable 

certainty that no harm will result from aggregate exposure to the pesticide residue. This finding 

must be made and the appropriate tolerance established before a pesticide can be registered for 

use on the particular food or feed crop in question. EPA must consider several factors before a 

tolerance can be established, including: 

• the aggregate, non-occupational exposure from the pesticide (exposure through diet, from 

using pesticides in and around the home, and from drinking water); 

• the cumulative effects from exposure to different pesticides that produce similar effects in 

the human body; 

• evidence of increased susceptibility to infants and children, or other sensitive 

subpopulations, from exposure to the pesticide; and 

• evidence that the pesticide produces an effect in humans similar to an effect produced by 

a naturally occurring estrogen or produces other endocrine-disruption effects. 

M.2.2. Pesticide Human Risk Assessment 

The process EPA uses for evaluating the health impacts of a pesticide, under either FIFRA or 

FFDCA, is called risk assessment. EPA uses the National Research Council’s four-step process 

to assess potential human health risks. This process involves hazard identification, doseresponse assessment, exposure assessment and risk characterization. Each of these steps is 

discussed below. 

The first step in the risk assessment process is hazard identification to identify potential health 

effects, or hazards, which may occur from different types of pesticide exposure. EPA considers 

the full spectrum of a pesticide’s potential health effects. Hazards are identified through a 

battery of studies that examine the potential toxicity of the pesticide in various tests including, 

where appropriate, tests with laboratory animals. 

To assess human health risk of the pesticide, pesticide companies conduct many toxicity studies 

based on EPA guidelines and the Good Laboratory Practice (GLP) standards. Results are 

evaluated for acceptability by EPA scientists. EPA evaluates pesticides for a wide range of 
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effects, from eye and skin irritation to cancer and birth defects. EPA may also consult the public 

literature or other sources of information on any aspect of the chemical. 

The next step of the risk assessment is dose-response assessment which considers the levels at 

which the pesticide produces adverse effects. Dose levels at which adverse effects were 

observed in test animals are then translated into equivalent doses for humans. 

Step three of the process involves an exposure assessment. People can be exposed to pesticides 

in three ways: 

1. Inhaling pesticides (inhalation exposure), 

2. Absorbing pesticides through the skin (dermal exposure), and 

3. Ingesting pesticides (oral exposure). 

Depending on the situation, pesticides could enter the body by any one or all of these routes. 

Typical sources of pesticide exposure include food (following agricultural uses); home and 

personal use pesticides; pesticides applied to lands that make their way into the drinking water; 

or occupational exposure for agricultural workers or pesticide applicators. 

Risk characterization is the final step in assessing human health risks from pesticides. It is the 

process of combining the hazard, dose-response and exposure assessments to describe the overall 

risk from the use of a pesticide. It explains the assumptions used in assessing exposure as well 

as the uncertainties that are built into the dose-response assessment. The strength of the overall 

database is considered, and broad conclusions are made. EPA’s role is to evaluate both toxicity 

and exposure and to determine the risk associated with use of the pesticide. 

The risk to human health from pesticide exposure depends on both the toxicity of the pesticide 

and the likelihood of people coming into contact with it, i.e., the probability of exposure. At 

least some exposure and some toxicity are required to result in a risk. For example, if the 

pesticide is found to have a high level of toxicity, but people are not exposed to the pesticide, 

there is no risk. Likewise, if there is ample exposure but the pesticide is nontoxic, there is no 

risk. Typically, however, there is some toxicity and exposure, which results in a potential risk. 

EPA recognizes that effects vary between animals of different species and from person to person. 

To account for this variability, a 100-fold uncertainty factor is built into the risk assessment with 

a 10X factor to account for differences between test species and humans, and a 10X factor to 

account for differences between people. This uncertainty factor creates an additional margin of 

safety for protecting people who may be exposed to the pesticides. The Food Quality Protection 

Act (FQPA) requires EPA to use an additional, up to a 10-fold safety factor, if necessary, to 

protect special subpopulations if they show potential increased susceptibility to effects of the 

pesticide, typically infants, children or women of child-bearing age. 

Once EPA completes the risk assessment process for a pesticide, the Agency uses this 

information to determine if there is a reasonable certainty of no harm to human health as a result 

of the use of the pesticide according to label directions as required by FIFRA. Using the 

conclusions of a risk assessment, EPA can then make an informed decision regarding whether to 
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approve a pesticide chemical or use, as proposed by the product label, or whether additional 

protective measures are necessary to limit exposure to a pesticide. For example, EPA may 

prohibit pesticide use on certain crops because consuming the treated commodity may result in 

an unacceptable risk to consumers. Another example of protective measures is requiring workers 

to wear personal protective equipment (PPE) such as a respirator or chemical resistant gloves, or 

not allowing workers to enter treated crop fields until a specific period of time has elapsed 

(Restricted Reentry Interval or REI). If, after considering all appropriate risk reduction measures, 

the pesticide still does not meet EPA’s safety standard, the Agency will not allow the proposed 

chemical or use. Regardless of the specific measures enforced, EPA’s primary goal is to ensure 

that legal uses of the pesticide are protective of human health, especially the health of children, 

and the environment. 

M.3. Dicamba Regulatory Status in U.S. 

M.3.1. Dicamba – Registration History 

Dicamba is a selective broadleaf herbicide belonging to the auxin agonist class, the oldest known 

class of synthetic herbicides, and is a member of the benzoic acids sub-group. Dicamba mimics 

the action of the plant hormone indole acetic acid, and causes rapid uncontrolled cell division 

and growth, leading to plant death. Dicamba has been registered in the U.S. for use on food 

crops since 1967 (U.S. EPA, 2009) and has been widely used in agricultural production for over 

forty years. Dicamba is presently approved for use on asparagus, corn, cotton, grass seed 

production, pastures and rangelands, small cereals (including wheat), sorghum, soybean, and 

sugarcane. Dicamba is also used for industrial vegetation management (e.g., forestry and 

roadsides), professional turf management (e.g., golf courses, sports complexes), and residential 

turf (U.S. EPA, 2009; Durkin and Bosch, 2004). 

Dicamba has a complete and comprehensive regulatory database (toxicity, environmental fate, 

and ecological toxicity) that has been evaluated by the United States Environmental Protection 

Agency (U.S. EPA). A Registration Eligibility Decision (RED) for dicamba was completed by 

EPA in 2006 and subsequently amended in 2008 and 2009 (U.S. EPA, 2009), as required for 

continued registration of all pesticides originally registered prior to 1984. EPA concluded the 

available data submitted for dicamba are complete and adequate to support the continued 

registration of dicamba products. EPA has evaluated the available toxicity data and concluded 

that a high level of confidence exists in the quality of the dicamba data base and in the reliability 

of the dicamba toxicity endpoints for risk assessment. EPA also considered toxicity data and 

available information concerning the variability of sensitive subpopulations, including infants 

and children. The EPA concluded there is reasonable certainty that no harm will result to the 

general population, or to infants and children, as a result of aggregate (all) exposure to dicamba 

residues. Thus, all current dicamba uses were eligible for reregistration (U.S. EPA, 2008a; U.S. 

EPA, 2008b; U.S. EPA, 2008c). In 2008, dicamba also successfully completed reevaluation by 

the Pest Management Regulatory Agency of Health Canada (PMRA, 2008) and the European 

Commission Health and Consumer Protection Directorate-General (European Commission, 

2008). Dicamba has been approved by the EPA for a number of food and feed uses, including 

the major agricultural crops of corn, soybean and small grains (e.g., wheat or barley). Dicamba 

presently has 68 food and feed pesticide tolerances (40 CFR §180.227) established in support of 

these uses. 
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Dicamba is presently registered for preemergence (early pre-plant) applications up to 0.5 pound 

acid equivalence per acre (lb a.e./A) and late postemergence (pre-harvest) applications to 

soybean at rates up to 1.0 lb a.e./A, and a pesticide tolerance is established for residues of 

dicamba on soybean seed (10 ppm) in support of these uses. 

M.3.2. Dicamba – Proposed New Use on MON 87708 

An application has been submitted to the EPA to amend Registration Number 524-582 (a 

diglycolamine (DGA) salt formulation) to register a new use pattern for dicamba on MON 87708. 

In support of the new use of dicamba on MON 87708, mammalian toxicity data on the dicamba 

metabolites DCSA (3,6-dichlorosalicylic acid) and DCGA (3,6-dichlorogentisic acid), and crop 

metabolism and residue data on dicamba-treated MON 87708 have been generated. These data 

confirm that existing dicamba food tolerances are sufficient to address any incremental exposure 

to dicamba resulting from its use on MON 87708. Furthermore, the proposed use pattern for 

dicamba on MON 87708 maintains the presently established maximum single and annual 

application rates. Therefore the existing ecological risk assessment conducted by EPA as part of 

the dicamba RED is sufficient to address any incremental ecological effects resulting from this 

new use. 

M.4. Potential Impact of Dicamba on Human Health 

M.4.1. Dicamba Safety Evaluations for Consumers 

On the basis of the risk assessments summarized below, EPA has concluded that there is a 

reasonable certainty that the existing uses of dicamba will not pose a risk to consumers, 

including infants and children. 

Dicamba presently has 68 established food and feed pesticide tolerances in the U.S. (40 CFR § 

180.227). Each time EPA reviews an application to add a new food or feed use to the dicamba 

label, the EPA is required by FFDCA to conduct an aggregate risk assessment. This assessment 

considers potential exposure from the proposed new use with all other existing exposures, 

including non-occupational sources of exposure to the pesticide, and must conclude that 

aggregate exposure to the pesticide will be safe as defined by the statute and regulations. Risks 

associated with potential occupational exposure for each new use are considered under the 

FIFRA standard of no unreasonable adverse effects to the environment, which includes humans 

and workers (hereafter referred to as FIFRA unreasonable risk standard). Over the course of 

numerous reviews, the toxicology of dicamba has been extensively studied. Dicamba does not 

pose any unusual toxicological concerns (U.S. EPA, 2009; Durkin and Bosch, 2004; European 

Commission, 2007a) and is classified by EPA as “Not likely to be Carcinogenic to Humans” 

(U.S. EPA, 2009). PMRA and the European Commission have also classified dicamba as noncarcinogenic (PMRA, 2007, 2008; European Commission, 2007a). 

Dietary exposure, as previously stated, is included in the aggregate exposure assessment, and 

considers pesticide residues that may remain on food from crops on which the pesticide is 

applied (pre- or postemergence), as well as any residue in drinking water as a result of pesticide 

use. Non-dietary exposure is also included in this assessment, and includes exposure to the 

pesticide through residential use, such as on lawns, as well as exposure in a recreational context, 
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such as from a golf course or sports field. Based on these data, EPA must be able to make a 

determination of reasonable certainty of no harm to human health as required by the FFDCA. At 

the time that dicamba was undergoing reregistration review, occupational exposure was not 

considered as part of the aggregate exposure and is evaluated separately under the FIFRA 

unreasonable risk standard. 

In making a determination of whether a reasonable certainty of no harm to human health exists, 

dietary risk assessments are performed that consider both the potential exposure and toxicity of a 

given pesticide. The dietary risk is then described as a percentage of a level of concern. The 

level of concern, which is also referred to as the population adjusted dose (PAD), is the dose 

(level of exposure) predicted to result in no adverse health effects to any human population 

subgroup, including sensitive subgroups, such as infants and children. The PAD may be 

expressed based on acute (aPAD, one day or less) or chronic exposures (cPAD, lifetime 

exposure). The PAD is the reference dose (RfD) for the compound but with any additional 

safety/uncertainty factors to protect sensitive subpopulations, or to address the completeness, 

quality or reliability of the toxicity data. The PAD is an estimate of the amount of daily pesticide 

exposure to the human population that can occur acutely (less than one day) or over a lifetime 

with a reasonable certainty of no harm to human health.69 An estimated exposure less than 100% 

of the PAD is below the level of concern for the EPA. 

The EPA evaluated the potential risks to human health associated with all then-registered 

dicamba uses as part of the reregistration of dicamba, and published the results and conclusion in 

the dicamba RED (U.S. EPA, 2009). Since reregistration, the EPA has approved an additional 

use on sweet corn, and as part of the approval included this new use in an updated dietary risk 

assessment. Since the sweet corn use pattern was within that reviewed during reregistration, 

EPA determined that the ecological and environmental fate assessments did not need to be 

updated and utilized the drinking water concentration from the RED in the updated sweet corn 

dietary risk assessment. The sweet corn use did not result in any noticeable increase in dietary 

exposure compared to the assessment from the RED; therefore, the risk assessment conduced for 

the dicamba RED is representative of all current registered uses of dicamba. EPA has conducted 

acute and chronic dietary (food and water) risk assessments for dicamba based on a theoretical 

worst case exposure estimate. For food, this estimate assumes that dicamba is used on 100 

percent of all the crops on which the pesticide is currently approved for use. It further assumes 

that the resulting pesticide residues found on all harvested food crops and derived animal food 

commodities (e.g., meat and milk) are at the level of the legally established tolerance (i.e., the 

maximum allowable pesticide residue level). Residues of dicamba are defined as dicamba and 

its metabolites 5-hydroxy dicamba and 3,6-dichlorosalicylic acid (DCSA) in soybean 

commodities, and as dicamba and DCSA in animal food commodities, as currently regulated in 

40 CFR § 180.227 (U.S. EPA, 2005a; U.S. EPA, 2005b). For water, EPA assumed that dicamba 

could potentially move offsite to adjacent surface water bodies as a result of drift or runoff. EPA 

also assumed dicamba could move through soil to groundwater; however, estimated 

concentrations in groundwater were significantly lower and therefore surface water estimates 

were used in the worse case dietary assessments. Surface water estimates were generated with 

the conservative screening level models SCIGROW and PRZM/EXAM using an exaggerated 

 

69 RfD is the current terminology used by EPA; however, earlier EPA risk assessment terminology used the term 

Acceptable Daily Intake (ADI). RfD and ADI are synonymous. 
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application rate that is 2.8 times higher than the current 1.0 lb a.e./A maximum single application 

rate established in the dicamba RED (U.S. EPA, 2009, U.S. EPA, 2005b), and the maximum 

single application rate proposed for MON 87708. EPA mandated reductions in dicamba use 

rates as part of the dicamba RED (1 lb a.e./A and 2 lb a.e./A for a single application and for 

annual application, respectively). 

The acute PAD for dicamba is 1 mg per kg body weight per day (mg/kg/day), based on an acute 

neurotoxicity study in rats (U.S. EPA, 2009). Based on the worst-case assumptions outlined 

above, the result of the dietary assessment gave a conservative, high-end (95th percentile) 

estimate of risk, which was well below the Agency’s level of concern for both the U.S. 

population in general and for all population subgroups. As shown in Table M-1, when both food 

and water are combined, infants were the most highly exposed subgroup with 11% of the aPAD, 

or acute level of concern, consumed. Because even this most highly exposed subgroup 

consumes a small percentage of the acute level of concern for dicamba, EPA concluded there is a 

reasonable certainty that acute dietary exposure to dicamba will not pose a risk to human health, 

including that of infants and children (U.S. EPA, 2009). 

The chronic PAD for dicamba is 0.45 mg/kg/day, based on a two-generation reproduction study 

in rats (U.S. EPA, 2009). Based on worst-case assumptions outlined above, EPA developed 

exposure estimates for the general U.S. population and the 8 other subpopulations of consumers 

evaluated by EPA, the major subpopulations are summarized in Table M-1. EPA determined 

that the most highly exposed subgroup for chronic dietary exposure (including both food and 

water) was children aged 1-2 years old, which consumed 6.6% of the cPAD, or chronic level of 

concern. Since even the most highly exposed subgroup consumes a small percentage of the 

chronic level of concern, EPA concluded there is a reasonable certainty that chronic dietary 

exposure to dicamba will not pose a risk to human health, including that of infants and children 

(U.S. EPA, 2009). 

Table M-1. Summary of Dietary Exposure and Risk for Dicamba: Food and Water 

Population 

subgroup 

Acute dietary (95th percentile) Chronic dietary 

Dietary exposure 

(mg/kg/day) % aPAD 

Dietary exposure 

(mg/kg/day) % cPAD 

General U.S. 

population 

0.0435 4.4 0.0118 2.6 

All infants 

(<1 year old) 

0.108 11 0.0199 4.4 

Children 

1-2 years old 0.0756 7.6 0.0297 6.6 

Source: EPA 2005b. 

The EPA also conducted an aggregate risk assessment which included dietary exposure (food 

and water) as well as other non-occupational exposures (e.g., from residential uses such as lawns 

and recreational uses such as golf courses or sports fields). For acute and chronic aggregate risk 

assessment, the aggregate exposure is the same as the dietary exposure. EPA does not typically 

aggregate acute dietary exposures with acute non-occupational exposures because it is unlikely 

that high end dietary exposure will occur on the same day as maximum non-occupational 
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exposures (U.S. EPA, 2005b). Current residential uses of dicamba do not result in long term 

residential exposure scenarios, so chronic aggregate exposure/risk is equal to chronic dietary 

exposure/risk. Therefore, the acute and chronic dietary exposure and risk summarized in Table 

M-1 also represents the acute and chronic aggregate exposure and risk. 

For short-term aggregate risk assessment of dicamba, EPA considered exposures from food, 

water, and residential handling and post-application. As shown in Table M-2, the highest 

exposed individuals in this assessment were adult males mixing, loading and applying dicamba 

using a hose-end sprayer, resulting in an aggregate exposure of 5.1% of the cPAD, and toddlers 

playing on treated turf which results in an aggregate exposure of 9.7% of cPAD. This short-term 

risk aggregate risk assessment was also considered to be protective of intermediate and long term 

exposures to dicamba. A large margin of safety exists for exposure to dicamba, even considering 

all the approved food, feed and non-crop uses of dicamba. 

Table M-2. Aggregate (Short-term) Exposure Assessment for Dicamba 

Population 

Food + Water 

Exposure 

mg/kg/day 

Incidental Oral 

Exposure, 

mg/day 

Dermal 

Dose, 

mg/kg/day 

Combined 

Exposure, 

mg/kg/day %PAD

 Adult Male -

Handler 0.0128 0 0.0102 0.023 5.1 

 Adult Male – 

Post 

Application 

0.0128 0 0.0037 0.0165 3.7 

 Child – 1-2 

years 

0.0297 0.0078 0.0062 0.0437 9.7 

Source: EPA, 2005b. 

In summary, based on the EPA risk assessments discussed above, there is a reasonable certainty 

that the existing uses of dicamba will not pose a risk to consumers, including infants and children. 

M.4.1.1. Dicamba Safety Evaluation for Use on MON 87708

Monsanto has submitted to EPA an application to register the use of dicamba on MON 87708 

(U.S. EPA OPP Decision Number D-432752). The proposed use of dicamba on MON 87708 

will not result in measurable increases in the exposure to dicamba or significant changes in the 

human health risk assessment. Plant metabolism studies have shown that the majority of the 

dicamba residue found in MON 87708 is DCSA, with lesser amounts of 3,6-dichlorogentisic 

acid (DCGA), 5-hydroxydicamba (5-OH dicamba) and parent dicamba (Table M-3). Toxicology 

studies submitted to EPA have demonstrated that the toxicology profiles for both DCSA and 

DCGA are comparable to that of parent dicamba, and that the existing hazard endpoints (RfDs 

and PADs) for dicamba are adequate to assess the potential human health risks from the 

metabolites as well. Studies have shown that the total dicamba residues in soybean seed 

following application of dicamba to MON 87708 will be well below the current 10 ppm soybean 

seed tolerance, an exposure level which has been determined as acceptable by EPA. In addition, 

Monsanto is proposing to establish new feed tolerances for soybean forage (45 ppm) and hay 

(70 ppm), which will allow the feeding of forage and hay to livestock. This practice is not 
Monsanto Company 10-SY-210U 684 of 721 

presently allowed because the preharvest application occurs after the stage where the crop would 

be useful as forage and hay. Note that the maximum residue for forage at 51.2 ppm is above the 

proposed MRL of 45 ppm. This is due to the way the data are distributed and the use of the 

NAFTA MRL calculator, which is the standard method used by EPA to calculate pesticide 

tolerance levels (U.S. EPA, 2011a). The EPA will perform its own calculations based on the 

data summarized in the table below to establish an appropriate feed tolerance for dicamba on 

soybean forage and hay. 

Table M-3. Residues of Dicamba, DCSA, 5-Hydroxydicamba and DCGA in DicambaTolerant Soybean Forage, Hay and Seed 

Commodity 

PPM 

(Expressed as Each Analyte per se) 

PPM 

(Dicamba Acid Equivalents) 

Dicamba DCSA 

5-OH 

dicamba DCGA 

Total as current definition of 

residue (dicamba + DCSA + 

5-OH dicamba)

Forage 

Mean 0.342 15.8 <0.006 2.04 17.3 

 Median 0.068 14.0 <0.005 1.95 15.2 

 Minimum <0.021 8.34 <0.005 0.359 10.0 

 Maximum 2.62 47.9 0.010 5.95 51.2 

Hay 

Mean 0.130 30.1 <0.014 2.68 32.3 

 Median 0.051 29.8 <0.014 2.02 31.9 

 Minimum <0.014 11.4 <0.014 0.169 12.2 

 Maximum 1.16 57.1 <0.014 7.33 61.1 

Seed 

Mean <0.013 0.055 <0.021 0.032 <0.091 

 Median <0.013 0.031 <0.021 0.017 <0.065 

 Minimum <0.013 0.009 <0.021 <0.011 <0.041 

 Maximum <0.013 0.411 <0.021 0.136 0.471 

Human dietary exposure will not increase beyond what has already been evaluated and 

determined acceptable by the EPA because established tolerances for animal food commodities 

(e.g., meat or milk) are sufficient to address livestock consumption of soybean forage or hay. 

This is because the proposed soybean forage and hay tolerances (residues) are lower than 

existing livestock dietary constituents that could potentially be replaced with soybean forage or 

hay in the livestock diet (e.g., grass forage at 125 ppm could be replaced with soybean forage at 

45 ppm). Soybean forage and hay are also not common livestock dietary constituents, as 

concluded by the EPA in its policy to permit label restrictions for livestock feeding of soybean 

forage and hay (U.S. EPA, 1996). 

Furthermore, since existing dicamba food tolerances (soybean seed and animal by-products) are 

inclusive of dicamba exposures arising from its use on MON 87708, the most recent EPA dietary 

and aggregate risk assessments for dicamba also address exposure from the proposed use in 

MON 87708. Therefore, these risk assessments demonstrate that there is a reasonable certainty 

that the use of dicamba on MON 87708, together with all other approved uses of dicamba, will 
Monsanto Company 10-SY-210U 685 of 721 

not pose a risk to human health, including that of infants and children. While the use of dicamba 

is expected to increase as a result of the availability of MON 87708 integrated into the Roundup 

Ready® soybean system, the risk associated with the new use has been adequately assessed by 

EPA through the risk assessment conducted as part of the dicamba RED. Lastly, EPA will 

review and confirm the acceptability of dietary exposure of dicamba residues on MON 87708 as 

part of the review of our pending application. 

M.4.2. Dicamba Safety Evaluation for Applicator 

Other potential impacts considered by EPA in its human health assessment are occupational 

exposure of the pesticide handler/applicator, and post-application exposure resulting from reentry to treated fields or areas. The occupational exposure scenarios evaluated by the EPA as a 

part of the dicamba RED are also applicable to the proposed use of dicamba on MON 87708 

(U.S. EPA, 2005b; U.S. EPA, 2005d). 

Using exposure data from the Pesticide Handler Exposure Database (PHED), Outdoor 

Residential Exposure Task Force (ORETF), and California Department of Pesticide Regulations, 

the EPA assessed short-term and intermediate-term occupational handler and post-application 

exposures. Handler exposure scenarios included mixer-loader, applicator and flagger activities. 

When exposure assumptions included the wearing of chemical resistant gloves during 

mixer/loader operations involving liquids required by dicamba product labeling, all occupational 

handler and post-application re-entry scenarios exhibited margins of exposure greater than 100 

and did not exceed the EPA level of concern (U.S. EPA, 2009). The use of dicamba on 

MON 87708 does not pose any new exposure considerations for workers beyond those which 

have been previously evaluated by EPA as part of the dicamba RED. Therefore the use of 

dicamba on MON 87708 will not pose a risk to agricultural workers. 

M.5. Potential Impact of Dicamba on the Environment 

As discussed above, environmental effects are carefully considered as a part of the FIFRA 

pesticide registration process. Prior to the approval of a new pesticide or a new use (including a 

change in pesticide application rates and/or timing) and before reregistering an existing pesticide, 

EPA must consider the potential for environmental effects and conclude no unreasonable adverse 

effects to the environment will result from the new pesticide, new use or continued use. 

To make this determination, EPA requires a comprehensive set of environmental fate and 

ecotoxicology data on the pesticide active ingredient (40 CFR Part 158). EPA uses these data to 

assess the pesticide’s potential environmental risk (risk = hazard × exposure). The required data 

include both short and long-term hazard data on representative organisms that are used to predict 

hazards to terrestrial animals (birds, non-target insects, and mammals), aquatic animals 

(freshwater fish and invertebrates, estuarine and marine organisms), and non-target plants 

(terrestrial and aquatic). 

EPA reevaluated the environmental safety of dicamba in 2006 as part of the FIFRA-required 

reregistration of all pesticides. At the end of this evaluation, EPA concluded that all then-

 

® Roundup Ready is a trademark of Monsanto Technology LLC. All other trademarks are the property of their 

respective owners. 
Monsanto Company 10-SY-210U 686 of 721 

registered uses of dicamba were eligible for reregistration, including single terrestrial (i.e., landbased) applications up to 1.0 lb a.e./A dicamba, and annual terrestrial applications up to 2.0 lb 

a.e./A. (U.S. EPA, 2009). This EPA assessment fully supports the use of dicamba on 

MON 87708 as the proposed use pattern is within that assessed in the RED assessment. 

M.5.1. Persistence of Dicamba in the Soil 

Dicamba’s fate in soil has been evaluated to determine the extent of its persistence in soil. While 

dicamba is stable to hydrolysis and photodegrades slowly on soil, dicamba is readily degraded in 

soil by aerobic soil microorganisms (U.S. EPA, 2009). Both laboratory and field studies 

demonstrate that dicamba dissipates quite rapidly in soil and has a low potential for accumulation. 

In a laboratory aerobic soil metabolism study, dicamba was rapidly metabolized to the nonpersistent degradate DCSA with a half-life of six days. DCSA was rapidly degraded at a rate 

comparable to that of dicamba producing carbon dioxide and being incorporated into biological 

natural products (U.S. EPA, 2005c). Other laboratory aerobic soil studies (Krueger et al., 1991) 

have been conducted for dicamba. In these studies, conducted in a number of different soils 

under various moisture and temperature conditions, dicamba degraded with half-lives ranging 

from 17 to 45 days. The European Union did not consider these half-lives indicative of the 

degradation rate of dicamba after evaluating the Krueger and other aerobic soil degradation 

studies (European Commission, 2007b). 

Field dissipation studies conducted with dicamba formulations also demonstrate rapid dissipation 

of dicamba in soil. Application of dicamba formulated as various salts (diglycolamine, 

dimethylamine and sodium salts corresponding to commercial products) to bare-ground plots at 

labeled rates (1 or 2 lb a.e./A) resulted in half-lives for dicamba of 3-20 days. Results also 

confirmed low vertical mobility under the conditions of the studies as demonstrated by only low 

concentrations (<20 ppb) of dicamba and its major degradate DCSA present in soil segments 

deeper than 10 cm (U.S. EPA, 2005c). The proposed maximum use rates of dicamba on 

MON 87708 (single application rate of 1 lb a.e./A dicamba, total annual application rate of 2 lbs 

a.e./A) are within the rates investigated in these dissipation studies. In addition, dicamba 

applications, which can be applied prior to planting up through the R1/R2 reproductive stage to 

MON 87708, should occur when soil temperatures are favorable for aerobic microbial 

degradation of dicamba. 

As with many chemicals, dicamba is metabolized somewhat slowly under anaerobic conditions, 

degrading with a half-life of 141 days (U.S. EPA, 2005c). However, under conditions amenable 

to aerobic degradation of dicamba and where retention of dicamba in aerobic soil layers is 

adequate for microbial degradation to occur, movement of dicamba to anaerobic soil layers 

would be minimal. Under normal soybean growing conditions of adequate soil temperatures and 

soil moisture, dissipation of dicamba through aerobic degradation would be rapid and it is 

unlikely that significant amounts of dicamba would reach anaerobic soil layers. 

Based on these data, EPA concluded that dicamba and DCSA (the primary environmental 

degradate) are “somewhat persistent” under aerobic and anaerobic conditions (U.S. EPA, 2009). 

This statement reflects the difference in degradation rates under aerobic conditions (nonpersistent) and anaerobic conditions (persistent). As discussed above, applications of dicamba 
Monsanto Company 10-SY-210U 687 of 721 

on MON 87708 are likely to occur when environmental conditions are conducive to rapid 

aerobic microbial degradation; therefore, dicamba is not expected to persist in soil as a result of 

the use on MON 87708. Furthermore, EPA concluded, based on the registered use patterns of 

dicamba and product label mitigation statements, no unreasonable adverse effects to the 

environment would result from the (all) registered uses of dicamba. 

M.5.2. Surface Water and Groundwater 

The EPA evaluated potential risks to surface and groundwater from the approved uses of 

dicamba as part of the 2006 Reregistration Eligibility Decision (RED). The EPA estimated 

levels of dicamba in surface and ground water using screening level (high-end exposure) models 

PRZM/EXAMS and SCIGROW, respectively, which estimate concentrations based on the 

physical, chemical and environmental fate properties of dicamba, and high-end use patterns. 

These conservative estimates do not take into account normal variation in environmental 

concentrations that can occur from dilution and dissipation. Based on simulated sugarcane crop 

scenarios for both ground and aerial applications of 2.8 lbs dicamba a.e./A (2.5 to 5 times the 

proposed maximum single application rate for MON 87708),estimated groundwater 

concentrations were 0.016 μg/L dicamba and 0.008 μg/L for DCSA, the major environmental 

degradate (U.S. EPA, 2005b). In addition, EPA measured dicamba as part of its 1971-1991 

pesticides ground water survey. Dicamba was detected at a low level of incidence, less than 3% 

of the 3172 samples, at levels ranging from trace to 44 μg/L(U.S. EPA, 1992). Dicamba also had 

a low incidence of detections in groundwater (less than 3% of the 6571 samples) in the U.S. 

Geological Survey National Water Quality Assessment (NAWQA) monitoring program during 

1993-2003, where dicamba was detected at levels ranging from 0.008 to 2.50 μg/L (U.S. EPA, 

2005b). Only parent dicamba and not the DCSA metabolite was analyzed in these water 

monitoring surveys. 

For drinking water derived from surface water, estimated concentrations (1 in 10 year annual 

mean) using the simulated sugarcane crop scenario for both ground and aerial applications of 2.8 

lbs dicamba a.e./A, ranged from 9.7 to 13 μg/L dicamba a.e. and 0.66 to 0.81 μg/L for DCSA 

(U.S. EPA, 2005b) Estimated surface water concentrations considered for aquatic organism risk 

assessment using simulated soybean crop scenario for ground and aerial applications of 2.0lb 

dicamba a.e./A ranged from 33.3-36.1 μg/L dicamba a.e. (U.S. EPA, 2005c). The NAWQA 

program also analyzed for dicamba in surface water samples in the 1993-2003 survey, where 

dicamba was detected at a low incidence (3% of the 6614 samples) at levels ranging from 0.009 

to 1.76 μg/L (U.S. EPA, 2005b). 

Measured dicamba concentrations in ground and surface water were also evaluated from a 

collection of publicly-available water monitoring data assembled by Monsanto Company from 

across the U.S. during 1990-2010. Data sources include the U.S. EPA (Pesticide Data Program, 

Safe Drinking Water Information System, Storage and Retrieval Data Warehouse and 

Unregulated Contaminant Monitoring Regulation data), the U.S. Geological Survey (NAWQA, 

National Water Information System), and various state water quality databases. From this 

collection of water monitoring data, relevant dicamba samples for ground and surface water were 

identified. First, data were restricted to the major soybean production states in the Midwest (IA, 

IL, IN, KS, KY, MI, MN, MO, ND, NE, NM, OH, SD and WI) and Arkansas. Second, the 

following samples types were excluded from the evaluation: finished drinking water, quality 
Monsanto Company 10-SY-210U 688 of 721 

control samples, urban stream monitoring samples, and alternative purpose sampling (e.g., 

contaminated site investigation). Records for which a sampling date or location was not reported 

were also excluded from the evaluation. Finally, for surface water, the data were limited to those 

samples collected during the months of April – July, which is considered the typical period when 

dicamba would be applied for agricultural uses, and for which maximum concentrations of 

dicamba in surface water would be expected. 

Dicamba concentrations in ground water are presented in Table M-4 for several year ranges and 

for the entire dataset evaluated. Overall, traces of dicamba were detected in less than 6% of the 

samples. For samples with detected dicamba levels, the minimum, median, 90th percentile and 

maximum concentrations are provided. The maximum level of dicamba detected in groundwater 

between 1990 and 2010 was 2.2 μg/L. However the 90th percentile of dicamba detections during 

1994-1998, corresponding to the peak dicamba usage period (see Table VIII-11), and across the 

entire monitoring period was 0.25 μg/L. 

Table M-4. Dicamba Concentrations (μg/L) in Groundwater in Major Soybean Growing 

States 

 

1990-

1993 

1994-

1998 

1999-

2004 

2005-

2010 

1990-

2010 

Number of 

Detects & Nondetects 

n 1796 2781 2558 2568 9703 

Number of 

Detects n 121 413 1 3 538 

Dicamba 

Concentration 

(μg/L) in Ground 

Water Based on 

Detects Only 

Min 0.01 0.01 0.73 0.1 0.01 

Median 0.25 0.25 n/a 0.1 0.25 

90th 

Percentile 0.25 0.25 n/a 0.18 0.25 

Max 1 2.2 0.73 0.2 2.2 

n/a: not applicable due to single data point 

The geographical coverage of the groundwater sampling sites is provided in Figure M-1, which 

shows the locations of the sampling sites associated with all samples (detects and non-detects) 

and for detects only. The location of sampling sites during the 1994-1998 timeframe, which 

corresponds to dicamba’s peak historical use period, provide a geographical representation of the 

areas of dicamba historical agricultural use intensity (see Figure VIII-1) and the major areas of 

soybean cultivation (see Figure VIII-2). 
Monsanto Company 10-SY-210U 689 of 721 

Figure M-1. Ground Water Sampling Sites for Dicamba (Top: Detects only; Bottom: 

detects and non-detects) 

 
Monsanto Company 10-SY-210U 690 of 721 

Dicamba concentrations in surface water are presented in Table M-5 for several year ranges and 

for the entire dataset evaluated. Overall, traces of dicamba were detected in less than 11% of the 

samples. For samples with detected dicamba levels, the minimum, median, 90th percentile and 

maximum concentrations are provided. The maximum level of dicamba detected in surface 

water between 1990 and 2010 was 17 μg/L. However the 90th percentile of dicamba detections 

during 1994-1998, corresponding to the peak dicamba usage period (see Table VIII-11), and 

across the entire monitoring period was 0.74 and 0.883 μg/L, respectively. 

Table M-5. Dicamba Concentrations (μg/L) in Surface Water in Major Soybean Growing 

States 

1990-

1993 

1994-

1998 

1999-

2004 

2005-

2010 

1990-

2010 

Number of 

Detects & Nondetects 

n 588 1,362 2,443 1,033 5,426 

Number of 

Detects n 146 206 222 14 588 

Dicamba 

Concentration 

(μg/L) in Surface 

Water Based on 

Detects Only 

Min 0.01 0.008 0.01 0.03 0.008 

Median 0.2 0.25 0.29 0.2935 0.25 

90th 

Percentil

e 

1.39 0.74 0.944 0.497 0.883 

Max 17 9.4 3.3 0.94 17 

The geographical coverage of surface water sampling sites is provided in Figure M-2, which 

shows the locations of the sampling sites associated with all samples (detects and non-detects) 

and for detects only. The location of sampling sites during the 1994-1998 timeframe, which 

corresponds to dicamba’s peak historical use period, represent a segment of the areas of dicamba 

historical agricultural use intensity (see Figure VIII-1) and the major areas of soybean cultivation 

(see Figure VIII-2). 
Monsanto Company 10-SY-210U 691 of 721 

Figure M-2. Surface Water Sampling Sites for Dicamba (Top: Detects only; Bottom: 

detects and non-detects) 
Monsanto Company 10-SY-210U 692 of 721 

Altogether, the modeling estimates, based on high-end exposure assumptions, and monitoring 

results show that concentrations of dicamba in potential drinking water resources as a result of its 

use are very low, and are well below the lifetime Health Advisory Level (HAL) for dicamba of 4 

mg/L (U.S. EPA, 2011b). Similarly, the estimated and monitoring levels of dicamba in surface 

water are low and do not exceed the EPA level of concern for aquatic plants70 and animals (U.S. 

EPA, 2005c). Therefore, the potential risk of dicamba from leaching or runoff as a result of 

incremental exposure from the use of dicamba associated with the planting of MON 87708 is 

very low and is unlikely to adversely affect human health or aquatic plants or animals. 

M.5.3. Wildlife 

M.5.3.1. Animals 

As a part of the reregistration evaluation under FIFRA, EPA conducted an ecological screening 

assessment for dicamba. This assessment compared the results from toxicity tests with dicamba 

conducted with various plant and animal species to conservative (high end) estimates of the 

concentration of dicamba to which an organism might be exposed in the environment. These 

estimates, called the Estimated Environmental Exposure Concentrations (EECs), are point 

estimates for specific types of exposure (e.g., aquatic or dietary) that assume constant high 

concentrations throughout the lifespan of the organism. These estimates do not take into account 

normal variation in environmental concentrations, the dilution or dissipation of those 

concentrations, or the frequency of exposure of wildlife to the pesticide. Such assumptions 

provide a screening level of assessment where conclusions of no harm can be drawn with high 

confidence; however, it is also possible to reach a conclusion of no harm with more refined 

exposure assumptions. During the reregistration process for dicamba and associated salts, the 

EPA derived exposure estimates for a number of use patterns (U.S. EPA, 2005c), including two 

scenarios that are reasonably close to the proposed use pattern for dicamba on MON 87708. 

These use patterns are: 1) wheat, with two 1.0 lb a.e./A applications; and 2) sorghum, with two 

0.5 lb a.e./A applications. For both cropping scenarios, the two applications were considered to 

occur on May 1st and June 1st. 

Based on the EPA screening-level analysis for relevant use patterns (wheat and sorghum) 

described above, no acute or chronic risk to aquatic animals was identified. Also, no chronic risk 

to birds or acute risk to mammals was identified. Considering maximum and mean residue 

levels assumed from the Kenaga nomogram (Fletcher et al., 1994)71, and application at rates of 

1.0 lb a.e./A and 0.5 lb a.e./A, the screening assessment did determine that the risk quotient (RQ) 

exceeded the Level of Concern (LOC= 0.5) for acute risk to some non-endangered bird species 

based on size and diet (U.S. EPA, 2005c). Using the toxicity endpoints from the avian oral 

gavage72 study, i.e., a study which forces a single high dose of dicamba directly into the birds’s 

crop and is not representative of realistic exposure scenarios for birds, EPA concluded acute 

exposure risks exceeded their level of concern for small (20 g) and medium (100 g) size birds. 

 

70 Currently there are no federally listed threatened or endangered nonvascular aquatic plants, so only risk to nonlisted aquatic nonvascular plants is considered. 

71 The Kenaga nomogram was developed by the U.S. Environmental Protection Agency (U.S. EPA) to predict the 

maximum potential pesticide residue levels in the food chain of wildlife for use in risk assessment. 

72 Forcing a single high concentration of dicamba directly into the bird’s crop, and is not representative of realistic 

exposure scenarios for birds. 
Monsanto Company 10-SY-210U 693 of 721 

Despite these predictions of potential effects, no incident reports have been filed with EPA for 

adverse effects on birds from dicamba use (U.S. EPA, 2005c). Furthermore, when exposure via 

the avian diet (the most relevant and realistic route of exposure of birds to dicamba) is 

considered, the EPA classified dicamba acid and its salts as practically non-toxic to birds (U.S. 

EPA, 2005c). Avian toxicity to dicamba (as the DGA salt) via dietary exposure is significantly 

less (> 5-fold) than toxicity from exposure via oral gavage. Therefore, it is unlikely that dicamba 

will pose an acute risk to birds when a more realistic dietary exposure is considered where 

dicamba residues are mixed with other food components and eaten over the entire day as would 

occur in the natural environment. 

The screening level assessment predicts that there will be no chronic risk (RQ < 1) to mammals 

regardless of size or dietary habits when mean residues resulting from application at a maximum 

rate of 1.0 lb a.e./A are utilized in the assessment (U.S. EPA, 2009). Mean initial residues, rather 

than maximum initial residues, provide a more realistic, but still conservative, dietary exposure 

estimate for a chronic exposure which would occur over days to weeks of duration. 

The EPA has classified dicamba as practically nontoxic to honey bees based on an acute contact 

LD50 value of >90.65 μg/bee (the honeybee is used to assess effects on non-target insects in 

general), and practically nontoxic to birds from dietary exposure. Dicamba presents a low 

hazard to aquatic organisms. Consequently, aquatic animals were predicted not to be at risk 

from rates of dicamba up to 2.8 lbs a.e./A, the highest rate assessed (U.S. EPA, 2005c). 

Dicamba has a low octanol-water coefficient (Log P < 3), indicating that it has a tendency to 

remain in the water phase rather than move from the water phase into fatty substances; therefore, 

dicamba is not expected to bioaccumulate in fish or other animal tissues (U.S. EPA, 2005c). 

In conclusion, no acute or chronic risk to aquatic or terrestrial animals, as defined above, is 

anticipated to result from potential incremental exposure of dicamba associated with the use of 

MON 87708. 

M.5.3.2. Plants 

Dicamba is a selective herbicide with more activity on dicotyledonous (dicot) plants than 

monocotyledonous plants (Ashton and Crafts, 1981). The EFED Reregistration Chapter for 

Dicamba / Dicamba Salts concluded that listed and non-listed terrestrial plants are potentially at 

risk from runoff and drift associated with all uses, but no risk to listed and non-listed aquatic 

plants (vascular or non-vascular) is expected as result of dicamba use at single application rates 

up to 1.0 lb a.e./A (U.S. EPA, 2005c). 

In the Reregistration Eligibility Decision (U.S. EPA, 2009), EPA concluded that a reduction in 

the maximum rate for a single application to 1.0 lb a.e./A and reduction in the total annual 

application rate to 2.0 lb a.e./A reduced the risk to terrestrial plants, and further determined that 

all currently registered uses of dicamba were eligible for reregistration provided that these rate 

mitigation measures were adopted on product labels. 

In conclusion, since the application rates for use of dicamba on MON 87708 fall within the 

reduced application rates mandated by the RED, the use of dicamba on MON 87708 will not 

pose a risk to non-target aquatic plants. Further, risks to non-target terrestrial plants are 
Monsanto Company 10-SY-210U 694 of 721 

adequately managed through appropriate application of dicamba as directed on the pesticide 

label and any effects will be mostly limited to non-lethal, temporary visible effects. 

M.5.4. Endangered and Threatened Species 

In the dicamba RED EPA discussed the changes in the registered use pattern of dicamba that 

were required to assure that dicamba meets the regulatory standards for pesticides in the United 

States. The primary use restriction that EPA determined was necessary to provide adequate 

protection of the environment, including non-target species, was the reduction of the maximum 

single dicamba application rate to 1.0 lb a.e./acre and the maximum annual rate of 2.0 lb a.e./A. 

EPA went further in the RED to state that no specific additional mitigations of off-target 

movement due to spray drift were needed (U.S. EPA, 2009). Because these limits were 

mandated by EPA to provide protection to non-target organisms, and because the proposed use 

of dicamba on MON 87708 falls within these limits, the risks to these non-target organisms from 

the use of dicamba on MON 87708 has also been addressed. 

As stated earlier, because the maximum single and annual application rates for the proposed use 

pattern for dicamba on MON 87708 are within the range of maximum application rates 

implemented as the primary use restriction mandated by the dicamba RED, the same conclusion 

can be reached for this use. Monsanto has requested approval of the use of dicamba on 

MON 87708; in its review of this registration request, EPA will address this use of dicamba 

under applicable statutory and regulatory requirements for pesticides. 

In addition, Monsanto has gone beyond the existing restrictions in place for dicamba use under 

FIFRA in order to assure adequate protection of endangered and threatened species under the 

proposed use of dicamba on MON 87708. Specifically, Monsanto’s registration submission for 

dicamba use on MON 87708 requested use only for a low volatile salt form of dicamba 

(diglycolamine or DGA) and requested only ground applications be allowed (i.e., no aerial 

application). 

M.5.5. Potential Effects on Endangered Animal Species Identified in Litigation 

In response to litigation brought by the Washington Toxics Coalition, the EPA conducted an 

assessment for dicamba and its impact on the endangered Pacific anadromas salmonids and their 

critical habitat. In this analysis, the EPA concluded that dicamba does not impact the 

endangered salmonids or their critical habitat (U.S. EPA, 2003). EPA also considered risks to 

endangered species as part of the reregistration of dicamba in 2006. Using an extremely 

conservative deterministic screening level ecological risk assessment (U.S. EPA, 2004), the EPA 

concluded that use of dicamba would have no effect on threatened or endangered freshwater fish, 

estuarine fish, and aquatic invertebrates (U.S. EPA, 2009). 

The EPA has also evaluated the potential effect of dicamba on salmon in eleven areas of 

California and Southern Oregon 73 in response to the consent agreement reached in the 

Washington Toxics lawsuit.74 The conclusion of EPA’s risk assessment is as follows: 

 

73 These areas are called Evolutionarily Significant Units based on the salmonid populations present in these areas. 74 Washington Toxics Coalition v. Environmental Protection Agency, 413 F.3d 1024 (9th Cir. 2005). 
Monsanto Company 10-SY-210U 695 of 721 

“Regardless of the specific dicamba compounds, I conclude that dicamba compounds with 

currently registered uses will have “no effect” on listed Pacific salmon and steelhead and their 

critical habitat...” (U.S. EPA, 2003). 

Since the use rates of dicamba proposed for MON 87708 are within the currently registered and 

evaluated use rates, no further consideration of the risk to Pacific salmon or steelhead is needed. 

This conclusion is consistent with EPA’s determination of no risk to fish or other aquatic animals 

in the dicamba screening assessment in the dicamba RED (U.S. EPA, 2005c). Furthermore, 

soybean production is not commonly practiced in the Pacific Northwest. However, based on the 

analysis above, risk to endangered Pacific salmon or steelhead is not expected from the use of 

dicamba on MON 87708. 

M.5.6. Other Potential Environmental Impacts Associated with Dicamba Use on 

MON 87708 

M.5.6.1. Potential Offsite Movement of Dicamba 

Herbicide drift and offsite movement are actively managed by farmers and applicators specially 

trained to use such products consistent with product labels and other state or local restrictions. 

Depending upon the herbicide being used, factors for managing the potential for spray drift 

include the selectivity and sensitivity of the herbicide, local weather conditions at the time of 

application (wind, temperature, humidity, inversion potential), droplet size distribution, 

application volume, boom height (height of the application equipment above the crop canopy), 

sprayer speed, and distance from the edge of the application area (SDTF, 1997; Felsot et al., 

2010). A variety of measures can be employed to control the potential for spray drift and offsite 

movement, including nozzle selection and application techniques and restrictions. 

The potential for offsite movement is regulated at the federal level by EPA. As indicated, EPA 

specifically approves product labeling for all crop uses of the herbicides. Before any dicamba 

formulation could be used on MON 87708, EPA is required to approve the pesticide label for 

that respective use, including specific directions for the use and application of the herbicide on 

the proposed crop. Label use directions would address not only application rates and timing, but 

as appropriate, could also include other measures to address the potential for offsite movement. 

EPA considers possible effects from offsite movement as part of the pesticide registration 

process required under FIFRA. Specifically, in order to approve the use of a pesticide 

(herbicide), EPA must conclude that no unreasonable adverse effects on non-target vegetation 

will result from potential offsite movement when the pesticide is used according to the product 

label. 

EPA reassessed the potential risks to non-target plants in its analysis in the dicamba RED, 

concluding that no specific additional drift mitigations were needed to support the continued 

registration of all dicamba uses (U.S. EPA, 2009). Since the proposed use pattern for dicamba 

on MON 87708 is consistent with use patterns evaluated and deemed eligible for reregistration in 

the dicamba RED, it is reasonable to conclude that dicamba use on MON 87708 meets the 

FIFRA standards related to drift and offsite movement. Thus, when herbicides are applied in 

accordance with the application instructions on the FIFRA label, offsite impacts can be avoided. 
Monsanto Company 10-SY-210U 696 of 721 

The use of dicamba on MON 87708 does not pose any greater risk to non-target vegetation over 

existing dicamba agricultural uses approved by EPA. Nevertheless, Monsanto has already taken 

additional steps to manage dicamba offsite movement even though EPA stated in the RED that 

no specific drift mitigation measures were needed. In the pending application to EPA, Monsanto 

requested the use of dicamba on MON 87708 on the low volatility DGA salt formulation (U.S. 

EPA Reg. No. 524-582) and limited dicamba applications to ground application equipment. 

Monsanto also plans to further address the specific use of dicamba on MON 87708 with US 

EPA, to evaluate whether additional measures may be appropriate to address potential drift and 

offsite movement. 

In addition, Monsanto will implement a robust stewardship program that will include a strong 

emphasis on grower and applicator training by working with American Association of Pesticide 

Safety Educators (AAPSE) and other stakeholders in applicator training to further facilitate ontarget applications of dicamba. There is an extensive pesticide education system within the 

United States as evidenced by the existence of the AAPSE. Members of this organization train 

and certify 500,000 applicators in agriculture and other pesticide use areas, working in land-grant 

university cooperative extension services and tribal, state, trust, territory, and federal agencies 

across the nation. Furthermore, EPA actively supports pesticide applicator training by funding 

the pesticide safety education program (PSEP). Since 1975, EPA has had an interagency 

agreement with USDA to distribute funds to the state cooperative extension service for the 

purpose of training pesticide applicators. 

M.5.6.2. No-Till Practices 

Conservation (no-till and minimum-till) production practices are used on approximately 40% of 

the U.S. soybean acres (CTIC, 2007). The increase in the number of resistant weed biotypes to 

soybean herbicides, including glyphosate, could adversely affect the sustainability of 

conservation production acres. The availability of MON 87708 and the flexibility to use 

dicamba for both preemergent and postemergent control of resistant and hard-to-control 

broadleaf weeds will provide a tool to help maintain this important agricultural practice. 

Furthermore, the use of dicamba on MON 87708 integrated into the Roundup Ready soybean 

system will help to delay the development of new resistant weed biotypes as a result of a second 

mode-of-action in the weed management system. 

The benefits of conservation tillage are well known and demonstrated, and include soil and water 

conservation, improved water quality and a reduced carbon footprint (CTIC, 2011). No-till 

production is the practice of establishing an agricultural seed bed and controlling weeds without 

mechanically tilling the soil. Instead, the only tillage of the soil is done at the time of planting, 

with the crop being seeded directly into the previous year’s crop residue. Among other 

environmental benefits, no-till production reduces soil erosion and the use of petroleum-based 

fuels for tractors. The practice has been shown to minimize surface water runoff and to improve 

soil quality by increasing the soil organic matter that helps bind soil nutrients and prevent their 

loss to runoff, erosion and leaching (Arriaga and Balkcom, 2005; Leep et al., 2003). Less soil 

erosion into surface waters would positively impact stream dynamics (McVay et al., 2005; 

Reicosky, 2008). 
Monsanto Company 10-SY-210U 697 of 721 

No-till agriculture can provide benefits to water bodies as well. No-till practices reduce soil 

erosion to surface water bodies, decreasing the amount of sediment in rivers and streams. 

Sedimentation increases the turbidity (cloudiness) of surface water bodies, reducing light 

penetration, impairing photosynthesis and altering oxygen levels, which cause a reduction of 

food sources for some aquatic organisms. Sediment can also cover spawning beds and impact 

fish populations. Phosphorus (a major component of fertilizer) bound to soil particles can be 

transferred to rivers and lakes via soil erosion, giving rise to high levels of phosphorus in surface 

waters, which may lead to algae blooms that can impact desirable fish populations (Hill and 

Mannering, 1995). 

No-till practices have also been shown to reduce the carbon footprint from agricultural 

production by decreasing carbon emissions due to fewer cultivation activities. In addition, 

deposits of crop residue on the soil surface are converted to organic matter and humus through 

carbon cycling, preserving soil carbon reserves and increasing fertility and nutrient cycling 

(Reicosky, 2008). 

M.5.6.3. Soil Microorganisms 

Results of standardized tests with dicamba and dicamba formulations performed for submission 

to regulatory agencies indicate no long-term effects on functional processes of soil 

microorganisms (carbon respiration and nitrogen transformation) when exposed to dicamba at 

rates up to 3.8 pound a.e./A. These levels exceed the current maximum single and annual 

dicamba use rates of one lb a.e./A and two lbs a.e./A, respectively (European Commission, 

2007b). 

M.5.6.4. Impact on Beneficial Arthropods 

Dicamba is a broadleaf herbicide that controls a range of dicot plants in the presence of 

monocots and other crops. Its effects primarily arise from direct applications in the field to 

reduce the plant mass and reproductive potential of susceptible weeds. Indirect effects outside of 

the treated areas as a result of spray drift would be significantly less since the amount of dicamba 

that can drift offsite at the field margins is typically less than 1% of the amount directly applied 

to the field (U.S. EPA,2005c). Dicamba has little or no effect on monocot plants and is 

considered by EPA to be practically non-toxic to insects, including honeybees and other 

arthropods (U.S. EPA, 2005c). Most available studies on the effects of herbicide treatment on 

arthropod communities are done with direct applications, and potential indirect effects from 

dicamba outside of the treated area are extrapolations from the direct effects at high exposure 

levels. Within the application area, the expected effects would be reduction in food sources 

(foliage, nectar, seeds) and cover for insects due to the intended weed control in the crop 

provided by dicamba application. 

Indirect effects of dicamba and related herbicides have been studied as potential effects on 

community structure, impacts to butterfly communities and other insect communities in areas 

adjacent to agricultural fields, and in the treated fields themselves. Fuhlendorf et al. (2002) 

studied the impact of direct application of selective broadleaf herbicides on forb78 community 

 

78 Forbs are herbaceous flowering plants that are not graminoids (grasses, sedges and rushes). Examples of forbs are 

clover, sunflower and milkweed. 
Monsanto Company 10-SY-210U 698 of 721 

richness and arthropod composition in a tall grass prairie. The authors reached the following 

conclusions: “This study demonstrates that the application of a selective herbicide can reduce 

the abundance and species richness of forbs that may serve as critical hosts to some arthropod 

species. However, analysis of these tallgrass communities did not yield significant differences in 

arthropod abundance or richness between grasslands treated with an herbicide and grasslands not 

treated with an herbicide... These results suggest that a single application of herbicide in 

tallgrass prairies of North America, although reducing the abundance and richness of forbs, 

neither eradicates the forb growth form nor substantially changes forb composition in the 

grassland community. Hence, neither does the arthropod community change. Relationships 

between forb cover and the arthropod community suggest that single applications of broadleaf 

herbicides may only have an effect on arthropods when forb abundance is dramatically reduced 

(below 5% cover). This study is incapable of concluding that herbicides targeted at forbs do not 

always influence the forb and arthropod communities, because herbicides are applied typically to 

native grasslands on southern Great Plains at 3–5 year intervals (Vallentine, 1989; New, 1997; 

Hanselka et al., 1990).” (Fuhlendorf et al., 2002). 

Longley and Sotherton (1997) also reviewed the literature regarding effects of pesticides on 

butterfly populations in agricultural areas. Excerpts from this review follow: “The main impact 

on butterfly populations is accidental drift or intentional direct spraying of broad-spectrum 

herbicides onto field boundaries to reduce certain pernicious perennial weeds (Marshal and 

Smith, 1987). However, the production of annual grass species is encouraged by these herbicide 

applications leading to a species-poor community (Smith and Macdonald, 1992). Consequently, 

herbicides may affect butterfly populations through the direct mortality of their larval food plants 

and flowering-nectar sources such as bramble (Rubus fruticosus), mayweed (Matricaria 

perforata), marjoram (Origanum vulgare) and thistle-like Compositae. The removal of host 

plants or premature senescence of plant parts can lead to larval starvation or the exclusion of 

less-fit adults (Courtney, 1981)”. Later in the review, Longley and Sotherton indicate that 

species vulnerability depends on larval food plants utilized by the species. “With the various 

factors influencing the extent of spray deposition on plants, the degree of pesticide (herbicide) 

exposure encountered by different butterfly families, and hence their vulnerability, will depend 

upon their choice of larval food plants. Members of the Hesperiidae and Satyridae families feed 

mainly on a range of coarse, densely tufted perennial grass species which are commonly found 

growing within many field boundaries”. Application of broadleaf selective herbicides such as 

dicamba that allow for selective growth of grasses would increase the food source for these 

species. Indirect exposures from spray drift at greatly reduced rates would result in substantially 

less impact to plant and arthropod communities surrounding the agricultural fields since 

exposures at drift rates are usually insufficient to reduce the survivability and reproductive 

potential of plants (Al-Khatib and Tamhane, 1999; Auch and Arnold, 1978; Behrens and 

Lueschen, 1979; Andersen et al., 2004). Since control of vegetation other than soybeans within 

the field is a component of soybean production, and since dicamba is anticipated to have little 

effect on forbs outside the field, the use of dicamba on MON 87708 is anticipated to have little 

or no effects on arthropod communities that are dependent on plants for their survival. 
Monsanto Company 10-SY-210U 699 of 721 

References for Appendix M 

Al-Khatib, K. and Tamhane, A. 1999. Dry pea (pisum sativum L.) response to low rates 

of foliar- and soil- applied sulfonylurea and growth regulator herbicides. Weed 

Technology, 13:753-758. 

Andersen, S.M., Clay, S.A., Matthees, D., and Wrage, L.J. 2004. Soybean foliage 

residues of dicamba and 2,4-D and correlation to application rates and yield. Agronomy 

Journal, 96:750-760. 

Arriaga, F.J. and K.S. Balkcom. 2005. Benefits of conservation tillage on rainfall and 

water management. 2005 Georgia Water Resources Conference, University of Georgia, 

Athens, Georgia. 

Auch D.E. and Arnold W.E. 1978. Dicamba use and injury on soybeans (Glycine max) 

in South Dakota. Weed Science 26:471-475. 

Ashton F.M. and Crafts A.S. 1981. Mode of action of herbicides. Second Edition. John 

Wiley & Sons, New York, New York. 

Behrens, M.R., and W.E. Lueschen. 1979. Dicamba Volatility. Weed Science 27:5 486-

493. 

Brown, L. R., Robinson, D. E., Nurse, R. E., Swanton, C. J., and Sikkema, P. H. 2009. 

Soybean response to simulated dicamba/diflufenzopyr drift followed by postemergence 

herbicides. Crop Protection 28:539-542. 

CTIC. 2007. Conservaton Tillage Information Center. http://www.ctic.purdue.edu/ 

Accessed June 1, 2010. 

CTIC. 2011. Top 10 conservation tillage benefits: Conservation for agriculture's future. 

Conservation Technology Information Center, West Lafayette, Indiana. 

http://www.ctic.purdue.edu/resourcedisplay/293/ Accessed September 21, 2011. 

Durkin, P. and S. Bosch. 2004. Dicamba - Human health and ecological risk assessment - 

Final report. SERA TR 04-43-17-06d. U.S. Department of Agriculture, Forest Service, 

Washington, D.C. http://www.fs.fed.us/foresthealth/pesticide/pdfs/112404_dicamba.pdf 

Accessed May 26, 2010. 

European Commission. 2007a. Draft Assessment Report, Volume 3, Annex B6: 

Toxicology and metabolism. Council Directive 91/414/EEC, EU Review Programme. 

European Commission. 2007b. Draft Assessment Report, Volume 3, Annex B9: 

Ecotoxicology. Council Directive 91/414/EEC, EU Review Programme. 

European Commission. 2008. Review report for the active substance dicamba, Finalised 

in the Standing Committee on the Food Chain and Animal Health at its meeting on 14 

March 2008 in view of the inclusion of dicamba in Annex I of Directive 91/414/EEC. 
Monsanto Company 10-SY-210U 700 of 721 

European Commission, Health and Consumer Protection Directorate-General, Paris, 

France. 

Felsot A.S., Unsworth J.B., Lindrs J.B.H.J., Roberts G., Rautman D., Harris C., Carazo 

E. 2010. J Environ Sci Hea. B. 45: 889-911. 

Fletcher, J.S., J.E. Nellessen, and T.G. Pfleeger. 1994. Literature review and evaluation 

of the EPA food-chain (Kenaga) nomogram, an instrument for estimating pesticide 

residues on plants. Environmental Toxicology and Chemistry 13(9): 1383-1391. 

Fuhlendorf, S.D., D.M. Engle, D.C. Arnold and T.G. Bidwell. 2002. Influence of 

herbicide application on forb and arthropod communities of North American tallgrass 

prairies. Agriculture Ecosystems & Environment 92:251-259. 

Hill, P.R. and J.V. Mannering. 1995. Conservation tillage and water quality. Purdue 

University Cooperative Extension Service, West Lafayette, Indiana. 

http://www.ces.purdue.edu/extmedia/WQ/WQ-20.html Accessed July 2, 2009. 

Krueger, J.P., Butz, R.G. and Cork, D.J. 1991. Aerobic and anaerobic soil metabolism 

of dicamba. Journal of Agricultural and Food Chemistry 39:995-999. 

Leep, R., D. Undersander, P. Peterson, D.-H. Min, T. Harrigan and J. Grigar. 2003. Steps 

to successful no-till establishment of forages. Extension Bulletin E-2880. Michigan State 

University, University of Wisconsin, University of Minnesota, and Natural Resources 

Conservation Service, East Lansing, Michigan. 

Longley, M. and N.W. Sotherton. 1997. Factors determining the effects of pesticides 

upon butterflies inhabiting arable farmland. Agriculture Ecosystems & Environment

61:1-12. 

McVay, K.A., D.L. Levlin and J. Neel. 2005. Effects of conservation practices on water 

quality: Sediment. MF2682. Kansas State University Agricultural Experiment Station and 

Cooperative Extension Service, Manhattan, Kansas. 

PMRA. 2007. Re-evaluation – dicamba. The Use of Dicamba in Agricultural and 

Industrial Sites. Pest Management Regulatory Authority, Health Canada. Ottawa, 

Canada. 

PMRA. 2008. Re-evaluation – dicamba. Pest Management Regulatory Authority, 

Health Canada. Ottawa, Canada. 

Reicosky, D.C. 2008. Carbon sequestration and environmental benefits from no-till 

systems. Pages 43-58 in No-Till Farming Systems. T. Goddard (ed.). World Association 

of Soil and Water Conservation, Bangkok, Thailand. 

SDTF. 1997. A summary of ground application studies. Spray Drift Task Force, St. Louis, 

Missouri. http://www.agdrift.com/PDF_FILES/Ground.pdf Accessed August 10, 2010 

and November 26, 2011. 
Monsanto Company 10-SY-210U 701 of 721 

 U.S. EPA. 1992. Pesticides in Groundwater Database: A Compilation of Monitoring 

Studies: 1971-1991. National Summary (EPA 734-12-92-001). Office of Prevention 

Pesticides and Toxic Substances. U.S. Environmental Protection Agency, Washington, 

D.C. 

U.S. EPA. 1996. Residue Chemistry Test Guidelines, OPPTS 860.1000 Background. 

Office of Prevention, Pesticides and Toxic Substances, U.S. Environmental Protection 

Agency, Washington, DC. 

U.S. EPA. 2003. No Effect Determination for Pacific Anadromas Salmonoids 

(Memorandum, L. Turner, Environmental Field Branch, Field and External Affairs 

Division). Field and External Affairs Branch. Division, Office of Prevention, Pesticides 

and Toxic Substances, U.S. Environmental Protection Agency, Washington, DC. 

http://www.epa.gov/espp/litstatus/effects/ne-dicamba.pdf. 

U.S. EPA. 2004. Overview of the ecological risk assessment process in the Office of 

Pesticide Programs. U.S. Environmental Protection Agency, Endangered and Threatened 

Species Effects Determinations, Washington, D.C. 

U.S. EPA. 2005a. Dicamba. Residue Chemistry Considerations for the Reregistration 

Eligibility Decision (RED) Document. Summary of Analytical Chemistry and Residue 

Data. DP Number D317703 (Memorandum, C. Olinger and W. Phang, September 13). 

Health Effects Division, Office of Prevention, Pesticides and Toxic Substances, U.S. 

Environmental Protection Agency, Washington, DC. 

U.S. EPA. 2005b. Dicamba: HED Chapter of the Reregistration Eligibility Decision 

Document, Phase 1. PC Code 029801; DP Barcode: D317720 (Memorandum, C. Olinger 

and Y. Yang, September 13). Health Effects Division, Office of Prevention, Pesticides 

and Toxic Substances, U.S. Environmental Protection Agency, Washington, DC. 

U.S. EPA. 2005c. Environmental Fate and Ecological Risk Assessment for the 

Reregistration of Dicamba and Dicamba Sodium, Potassium, Diglycoamine, 

Dimethylamine and Isopropylamine Salts (Memorandum, W. Erickson, I. Abdel-Saheb 

and S. Borges, August 31). Environmental Fate Effects Division, Office of Prevention, 

Pesticides and Toxic Substances, U.S. Environmental Protection Agency, Washington, 

DC. 

U.S. EPA. 2005d. Dicamba: Occupational and Residential Exposure and Risk 

Assessment for the Reregistration Eligibility Decision (RED) Document. PC Code 

029801, DP Barcode D317701 (Memorandum, T.C. Dole, September 6). Health Effects 

Division, Office of Prevention, Pesticides and Toxic Substances, U.S. Environmental 

Protection Agency, Washington, DC. 

U.S. EPA. 2008a. Dicamba Pesticide Tolerance. U.S. Environmental Protection 

Agency, Washington, D.C. Vol. 73, No. 64. 
Monsanto Company 10-SY-210U 702 of 721 

U.S. EPA. 2008b. Aldicarb, Ametryn, 2,4-DB, Dicamba, Dimethipin, Disulfoton, 

Diuron, et al.; Proposed Tolerance Actions. U.S. Environmental Protection Agency, 

Washington, D.C. Federal Register Vol. 73, No. 108. 

U.S. EPA. 2008c. Aldicarb, Ametryn, 2,4-DB, Dicamba, Dimethipin, Disulfoton, 

Diuron, et al.; Tolerance Actions. U.S. Environmental Protection Agency, Washington, 

D.C. Vol. 73, No. 186. 

U.S. EPA. 2009. Reregistration Eligibility Decision (RED) for Dicamba and Associated 

Salts. Case No. 0065. U.S. Environmental Protection Agency Office of Prevention, 

Pesticides and Toxic Substances, Special Review and Reregistration Division, 

Washington, D.C. 

U.S. EPA. 2010. Registration Review: Conventional Cases Schedule: 2010-2013. U.S. 

Environmental Protection Agency, Office of Prevention, Pesticides and Toxic Substances. 

June 7, 2010. http://www.epa.gov/oppsrrd1/registration_review/2010-13-

conventional.pdf Accessed June 12, 2010. 

U.S. EPA. 2011a. NAFTA maximum residue levels (MRL) calculator. U.S. 

Environmental Protection Agency, Washington, D.C. 

http://www.epa.gov/oppfead1/international/naftatwg/mrl-calculator.html Accessed 

February 22, 2012.

U.S. EPA. 2011b. 2011 Edition of the Drinking Water Standards and Health Advisories. 

U.S. Environmental Protection Agency, Office of Water. EPA 820-R-11-002. 

http://water.epa.gov/action/advisories/drinking/upload/dwstandards2011.pdf
Monsanto Company 10-SY-210U 703 of 721 

Appendix N: Overview of Evaluation of Potential Exposure and Biological 

Effects on Endangered Species for Dicamba Use in MON 87708 

N.1. Introduction 

This overview report provides a summary of a multi step approach that has been utilized 

to evaluate the potential effects on federally listed threatened and endangered (“listed”) 

species from the use of the herbicide dicamba on dicamba-tolerant (DT) soybean. This 

analysis follows the procedures described in the Overview of the Ecological Risk 

Assessment Process in the Office of Pesticide Programs, U.S. Environmental Protection 

Agency (U.S. EPA), Endangered and Threatened Species Effects Determinations, 

published in 2004 (U.S. EPA, 2004), as well as methods utilized in more recent 

threatened and endangered species effects determinations conducted by the U.S. 

Environmental Protection Agency (USEPA) for atrazine (U.S. EPA, 2007), and for a 

number of active ingredients in assessing their potential effects on the California red 

legged frog (U.S. EPA, 2008a). A four step process is described in more detail below. 

N.2. Endangered Species Exposure and Effects Analysis 

N.2.1. Screening Level Analysis 

An initial analysis of the potential for exposure and effects to all taxa from dicamba use 

in dicamba-tolerant soybean based on the use pattern pending at EPA was conducted 

(Wright et al., 2011) using the EPA deterministic risk quotient approach (U.S. EPA, 

2004). 79 The use pattern for dicamba utilized in this analysis was a pre-emergence 

application at a rate of 1.0 lb dicamba acid equivalents per acre (a.e/A) followed by two 

post-emergence applications each at a rate of 0.5 lb a.e./A with a 6-day interval between 

the pre-emergence application and the first post-emergence application, and a 6-day 

interval between the two post-emergence applications, with all applications being made 

using ground application equipment.80 The 6-day application intervals utilized in this 

analysis are expected to be shorter than the intervals actually used in practice, since a 

grower is expected to wait at least 7 days before deciding to make a subsequent 

application in order to allow evidence of dicamba efficacy to develop. 

Initial exposure estimates using the above described use pattern were based on standard 

EPA exposure models and default assumptions; toxicity effects endpoints were taken 

from the EPA Environmental Fate and Effects Division (EFED) Science Chapter for the 

Reregistration Eligibility Decision for dicamba (U.S. EPA, 2005) or from more recent 

EPA guideline studies conducted by BASF under an EPA data call-in (Porch et al., 

 

79 This approach calculates a risk quotient (RQ) by dividing the Estimated Environmental Concentration 

(EEC) by the appropriate toxicity endpoint, and then compares that value with the appropriate Level of 

Concern (LOC). The LOC is established by EPA policy as the criteria used by EPA in comparison to the 

calculated risk quotient (RQ) to assess the potential for a pesticide use to cause adverse effects to nontarget organisms. 

80 The dicamba label for DT soybeans will not permit aerial application. 
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200981), when these endpoints were more consistent with current EPA guidelines. The 

conclusion from this initial analysis, based on risk quotients (RQs) being less than the 

EPA Levels of Concern (LOCs), is that dicamba use on dicamba-tolerant soybean would 

not affect threatened and endangered species in the following taxa:

82

 Fish, aquatic phase amphibians, and aquatic invertebrates (acute or chronic 

exposure) 

 Birds, terrestrial-phase amphibians, and reptiles (chronic exposure) 

 Mammals (acute exposure) 

 Insects, aquatic vascular plants and monocotyledonous terrestrial plant species83,84

Although the RQ for aquatic nonvascular plants exceeded the LOC additional analysis 

was not conducted because there are no federally listed nonvascular aquatic plants. 

These conclusions for aquatic plants and animals, birds, mammals and insects are 

consistent with the risk conclusions presented in the EFED science chapter for dicamba 

reregistration (U.S. EPA, 2005; 2006). 

The conclusion for monocots is based on more recent nontarget plant studies with a 

typical dicamba end use product that were required by EPA during the reregistration 

process (Porch et al., 2009).85 Using the no-observed-effect endpoints from these 2009 

studies and the EPA model TerrPlant to calculate exposure and risk quotients, the RQs 

for monocots were below the LOC, and thus result in a conclusion of no effect. The 

levels of concern considered by EPA for threatened and endangered (listed) species risk 

assessments are given in Table N-1. 

 

81 New studies were required by EPA because previous studies did not meet current regulations requiring 

the use of formulated product for these studies. This new testing resulted in a more conservative endpoint 

for the vegetative vigor study and confirmed field observations of relative sensitivity for the seedling 

emergence study. 

82 “If assumptions associated with the screening level action area result in RQs that are below the listed 

species LOCs, a “no effect” determination conclusion is made with respect to listed species in that taxa, 

and no further refinement of the action area is necessary.” EFED Reregistration Chapter for 

Dicamba/Dicamba Salts,. 

83 According to EPA methodology (U.S. EPA, 2004), risk to non-target plants is assessed only outside the 

application area. 

84 Exposure for insects and plants is not divided into acute and chronic durations. 

85 The new study was conducted under the OPPTS 850.4225 draft guideline which is very similar to the 

current OECD nontarget plant guideline, and the results are considered to be more representative of 

effects that would be expected to occur in the field because of an improved study design. 
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Table N-1. EPA Levels of Concern for Threatened and Endangered Species 

Risk Presumption Calculation for Risk Quotient Level of Concern 

Birds:

 Acute Risk EEC/LC50 (application of a liquid) 

or LD50/ft2 or LD50/day 

(application as a granule, bait, or 

treated seed) 

0.1 

 Chronic Risk EEC/NOAEC 1.0 

Wild Mammals

 Acute Risk EEC/LC50 (application of a liquid) 

or LD50/ft2 or LD50/day 

(application of a granule, bait, or 

treated seed) 

0.1 

 Chronic Risk EEC/NOAEC 1.0 

Aquatic Animals

 Acute Risk EEC/LC50 or EC50 0.05 

 Chronic Risk EEC/MATC or NOAEC 1.0 

Terrestrial and Semi-Aquatic Plants

 Acute Risk EEC/EC05 or NOAEC 1.0 

Aquatic Plants

 Acute Risk EEC/EC05 or NOAEC 1.0 

Source: U.S. EPA, 2004 

N.2.2. Refinement to the Screening Level Analysis - Use of Dicamba-specific Foliar 

Residue Values 

Consistent with EPA guidance found in the Overview Document (U.S. EPA, 2004), if 

screening-level assessments do not result in a “no effect” determination, EPA does not 

then conclude that an herbicide “may affect” threatened and endangered species; rather, 

more refined assessments must be conducted to ascertain if any effects are expected to 

occur. Accordingly, for threatened and endangered animal species and the exposure 

durations for which the risk quotient exceeded the LOC in the screening level analysis 

(acute exposure for birds, amphibians and reptiles, and chronic exposure for mammals), 

refined exposure estimates were developed utilizing measured dicamba residues on 

pasture grasses and soybean forage86 (as representative residues for short grass and broadleaf 

foliage87 components of animal diets) (Wright et al., 2011). These data were from residue 

 

86 Residue values from these studies were converted to values expressed as parts per million per pound 

dicamba acid from residues values for application rates of 0.5, 1.0, and 2.0 lb a.e./A for grasses and 0.5 

and 1.0 lb a.e./A for soybeans. 

87 EPA assumes that broadleaf foliage and small insects have the same residue values, so these dicambaspecific values were used both for broadleaf foliage and small insects. 
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studies conducted under Good Laboratory Practices by dicamba registrants and by 

Monsanto, respectively, and are used to provide dicamba-specific residues for 

components of animal diets, instead of using the EPA default residue values based on the 

Kenega nomogram (Hoerger and Kenega, 1972) as revised by Fletcher (1994). Both the 

soybean residue study and one of the grass residue studies, as well as a wheat residue 

study, included sites with several sampling points for forage which occurred soon after 

application so that the rate of decline of dicamba residues on soybean, grass, and wheat 

foliage could be determined, and the time required for dissipation of fifty percent of the 

residues (DT50) calculated.88 The Overview Document (U.S. EPA, , 2004)89) indicates that 

chemical-specific foliar dissipation values can be used for multiple application exposure 

modeling for wildlife; accordingly, as a conservative assumption, the longest of the 

representative dicamba-specific foliar DT50 values for these three crops (5.63 days, for 

pasture grass) was used in the calculation of the dicamba-specific residues for chronic 

exposure. For grass and broadleaf dietary items, both mean and upper bound residue 

values90 were considered in the evaluation; these levels are suitable to estimate realistic 

(mean) and worst case (upper bound) levels of exposure. A comparison of the default 

upper bound residue values and dicamba-specific upper bound residue values is given in 

Table N-2. 

 

88 For the pasture grass and wheat residue studies, there were over 50 treatments per study for which a DT50

value could be calculated. With such a large number of data points, the 90th percentile upper confidence 

limit of the mean DT50 value was selected as an appropriate DT50 value to use for calculation of residue 

decline for these crops. For the soybean residue study with only six treatments available to calculate DT50

values, the maximum value was considered representative of residue decline in soybean forage. 

89 Overview Document (U.S. EPA, 2004), 

90 A 90th centile value was used for pasture grass and the maximum residue value was used for soybean 

forage due to the difference in numbers of residue values available to calculate the upper bound residue. 
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Table N-2. Comparison of Default Kenaga Residues and Dicamba-specific Residues 

in Food-Items 

Maximum dicamba residues in food items (ppm a.e.) based 

on a 1 lb a.e./A pre-emergence application followed by two 

sequential post-emergence 0.5 lb a.e./A applications 

Short 

Grass 

Tall 

Grass 

Broadleaf 

Plants/ 

Small Insects 

Fruits/Pods/Seeds/

Large Insects 

Using upper bound Kenaga residues & default foliar half-life (35 days) 

Day 0 (after 1st

application) 240.0 110.0 135.0 15.0 

Maximum residuea (after 

3rd application) 415.8 190.6 233.9 26.0 

Using Dicamba-specific residues & foliar half-life (5.63 days) 

Day 0 (after 1st

application) 131.0 60.0 103.9 15.0 

Maximum residuea (after 

1st application) 131.0 60.0 103.9 15.0 

 a Using the default foliar half-life, the highest residue occurs immediately after the third 

application. Using the dicamba-specific half-life, the highest residue occurs immediately after the 

first application

These refined residue values for dietary items were considered when using the EPA 

model T-REX (v1.4.1) for the calculation of risk quotients and subsequent comparison to 

LOCs for acute exposures to birds, terrestrial amphibians, and reptiles, and for chronic 

exposures to mammals.91 Figure N-1 describes the process for identifying the taxa that 

had risk quotients exceeding the EPA’s LOC after the screening level analysis. 

 

91 The T-REX model is not designed to allow modification of the dietary food item residues. Thus, the 

dicamba-specific residues reported in Table N-2 were considered in the risk quotient calculations by 

applying a correction factor to the estimated environmental concentrations (EEC) values computed using 

the standard T-REX model for a single 1-lb/A application. 
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Figure N-1. Identification of Taxa Exceeding Endangered Species Levels of 

Concern 

a

 The basis for this conclusion is described in U.S. EPA, 2004. Overview of the Ecological Risk 

Assessment Process in the Office of Pesticide Programs, U.S. Environmental Protection Agency, 

Endangered and Threatened Species Effects Determinations 

Risk quotients for chronic exposure to mammals calculated using upper bound residues 

slightly exceed the LOC for small and medium-sized mammals consuming short grass; 

however, utilizing mean residue values as a further refinement, risk quotients are all 

below the LOC (Table N-3) leading to a conclusion of no effect on the species. This is 

consistent with the finding in the EFED Science Chapter for dicamba (U.S. EPA, 2005) 

and the revised RED (U.S. EPA, 2006) for a single application rate of 1.0 lb a.e./A. 

Because there will be at most only three applications per year and dicamba dissipates 

rapidly, an individual small or medium-sized mammal will not chronically consume 

foliage containing maximum residue levels from dicamba-treated fields. Therefore, 

Exposure Estimate: 

Worst Case 

Estimated Environmental Concentration 

(EEC) Using Standard EPA Models Or 

Analysis Methods

Toxicity Endpoint (TE): 

Lowest Acute Or Chronic Value 

Meeting EPA Quality Standards 

RQ > Level Of Concern 

(LOC)? 

Refine EEC based on 

experimental data 

Determine Risk Quotient 

(RQ = EEC/TE) 

Yes

For Each 

Taxon 

No Direct Effect 

Expected on the Taxaa

No 

RQ > Level Of Concern 

(LOC)? 

No Direct Effect 

Expected on the Taxaa 

No

Further Analysis 

Yes 
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reliance on dicamba-specific Day 0 mean residue values is also a conservative dietary 

exposure estimate for chronic exposure that is expressly countenanced by EPA.92 

Table N-3. Risk Quotients for Chronic Exposure a for Mammalian Species Based 

on Dicamba-Specific Residue Values and DT50. 

Diet Mammal 

Size (g) 

Risk Quotient using 

Upper Bound 

Residuesa,b

Risk Quotient using 

Mean Residues

Short Grass 

15 1.26 0.75 

35 1.08 0.64 

1000 0.58 0.34 

Tall Grass 

15 0.58 0.32 

35 0.49 0.27 

1000 0.27 0.15 

Broadleaf Foliage / 

Small Insects 

15 1.00 0.75

35 0.86 0.64 

1000 0.46 0.34 

Fruits/Pods/Seeds/Large 

Insects 

15 0.14 0.07 

35 0.12 0.06 

1000 0.07 0.03 

Grain 

15 0.032 0.015 

35 0.027 0.013 

1000 0.015 0.007 

a Dicamba-specific refined residue values based on a 1 lb a.e./A application rate were 131, 60.0, 103.9, and 

15.0 mg a.e./kg diet, respectively, for upper bound values for short grass; tall grass; broadleaf foliage / 

small insects; and fruits/ pods/ seeds/ large insects, and 77.9, 33.0, 77.7, and 7.0 mg a.e./kg, respectively, 

for mean residues. The dicamba-specific residue decline half-life (DT50) of 5.63 days instead of the EPA 

default value of 35 days was also used in determining the peak residue value resulting from the three 

sequential applications. 

b

 Numbers with Bold font indicate the RQ exceeds the LOC (1.0). 

With respect to birds, analyses were performed using measured dicamba residue values 

(Table N-2) for RQ calculations for acute exposure. The following could be excluded 

from concern (and, therefore, no effects would be expected to occur): birds of all sizes 

consuming only grain or fruits/pods/seeds/large insects, and large birds consuming 

broadleaf foliage/small insects or tall grass. Small and medium-sized birds consuming 

broadleaf foliage/small insects or grasses and large birds consuming short grass required 

further analysis. RQ values are presented in Table N-4. Because birds are the surrogate 

 

92 Overview Document (USEPA, 2004) 
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species for terrestrial phase amphibians and reptiles, further analysis was also conducted 

to ensure that amphibians and reptiles could be excluded from concern for adverse effect. 

Table N-4. Risk Quotients for Acute Exposure to Birds Using Measured Dicamba 

Residue and Residue Decline Values 

Diet 

Acute RQs: (Dose-based EEC/adjusted LD50)a 

Upper Bound Residues 

20 g 

Small 

100 g 

Medium 

1000 g 

Large 

Short grass 0.79 0.35 0.11 

Tall grass 0.36 0.16 0.05 

Broadleaf plants/small 

insects 0.63 0.28 0.09 

Fruits/pods/seeds/large 

insects 0.09 0.04 0.01 

Granivore 0.02 0.01 0.003 

a

 Bold numbers indicate the RQ exceeds the LOC (0.1). 

N.2.3. Refined Analysis Considering Species County-Level Locations 

Certain sizes of threatened and endangered birds, amphibians, reptiles, and mammals had 

risk quotients exceeding the LOC as a result of the use of dicamba on dicamba-tolerant 

soybean based on a default exposure analysis or an analysis using refined residues. 

Consistent with guidance set forth in the EPA Overview Document, a more detailed 

evaluation of the locations of these threatened and endangered species relative to 

potential areas of soybean production was undertaken. 

First, the co occurrence of threatened and endangered species of these remaining 

taxa93and the production of soybeans was determined at the county level. Listing status94,, 

species habitat and proximity data to soybean production at the county level were 

evaluated for these identified species to determine which species in which counties can 

be excluded from further evaluation and which require further evaluation. This process is 

referred to as the “county-level analysis” (Frank and Kemman, 2012) and is discussed in 

more detail in Section N.3 below. 

Next, the Overview Document provides that – for those threatened and endangered 

species (avian, reptilian, amphibian (terrestrial-phase), and mammalian) that require 

further evaluation – each species be considered individually to determine whether it can 

 

93 For completeness, all threatened and endangered avian, amphibian, reptilian, and mammalian species 

were included in the county-level analysis. 

94 Listing status refers to whether the species is classified as threatened, endangered, no longer considered 

threatened or endangered (delisted), etc. 
Monsanto Company 10-SY-210U 711 of 721 

be excluded from concern for potential effects based on body weight or dietary 

considerations. Accordingly, for these animal species, refined risk quotient calculations 

were performed considering individual species body weight, food type and food intake 

rate, as described in Section N.4 and in Schuler and Leopold (2012), and consistent with 

EPA guidance found in the Overview Document (U.S. EPA, 2004). Based on these 

refined county-level analyses, as discussed below, animal species were excluded from 

concern, and, therefore, would not be affected by dicamba use on DT-soybean. 

N.3. County-level Analysis: Co-occurrence of Listed Species in Crop Production 

Areas 

The procedures used in the county-level analysis to identify counties containing 

threatened or endangered species that require further evaluation are depicted as a flow 

chart in Figure N-2 and are described in detail in Frank and Kemman (2012).95. The U.S. 

counties where soybean production was reported were identified using available data 

from the U.S. Census of Agriculture. Census data from 1997, 2002 and 2007 were 

utilized to identify these counties. This information was supplemented with soybean 

production data available from the California Department of Pesticide Regulation. 

Counties without soybean production, but adjacent to counties with soybean production 

were also identified. In total, there were 2,728 counties considered in this analysis (2,198 

soybean counties, 530 adjacent counties). The counties considered at the beginning of 

county-level analysis are shown graphically in Figure N-3. 

 

95 Overview Document (US EPA, 2004) 
Monsanto Company

Figure N-2. County-Level Analysis Process

a Information from the California Pesticide Use Reporting databas

herbicide applications to soybean fields in California.

10-SY-210U

Level Analysis Process

Information from the California Pesticide Use Reporting database was also used to identify counties with 

herbicide applications to soybean fields in California.

712 of 721 

e was also used to identify counties with 
Monsanto Company 10-SY-210U 713 of 721 

Figure N-3. Counties with Soybean Farms and Adjacent Counties 

In the identified counties, available county-level presence information for threatened or 

endangered avian, reptilian, amphibian, and mammalian species was evaluated, using 

county-level location information compiled by the FIFRA Endangered Species Task 

Force (FESTF) in the FESTF Information Management System (IMS).96 Of the 2,728 

counties initially considered, there were 2505 counties with listed species in the taxa of 

concern and where soybeans are produced (including counties adjacent to counties where 

soybeans are produced). These counties are depicted graphically in Figure N-4 and 

contain 140 distinct species in 49 states (all states except Alaska) and the District of 

Columbia. There were 7,048 species/county co-occurrence records considered across all 

animal taxa evaluated. In these counties, each species was evaluated with respect to the 

current listing status, county-level locations, species biology, and species habitat 

requirements, in order to determine whether exposure to dicamba from use on DTsoybean could potentially result in adverse effects to the species. Some listed species 

could be removed from concern for adverse effects based on exclusions that currently 

 

96 The FIFRA (Federal Insecticide, Fungicide, and Rodenticide Act) Endangered Species Task Force 

(FESTF) Information Management System 2.7 (IMS) (referred to as the “FESTF IMS”) was developed 

in order to meet the legal obligations of its member companies to submit data required by EPA/OPP 

under FIFRA (as described in Pesticide Registration Notice 2000-2) in support of the members’ 

registration and re-registration actions. The purpose of the IMS is to meet the data requirements in a 

manner that significantly improves the consistency, quality, availability and use of existing information 

on threatened and endangered species and pesticide use. http://www.festf.org/visitors/default.asp 
Monsanto Company 10-SY-210U 714 of 721 

exist and are documented. Exclusions that have been employed include change in species 

listing status, not present due to extirpation97, habitat not in proximity to agriculture, 

product properties98, and species not in proximity to agriculture for other reasons. 

For animal species that have been identified in the county-level analysis as requiring 

further analysis, species-specific refinements as described in Section N.4 have been 

considered. T-HERPS was utilized to calculate risk quotients for amphibians of similar 

body weight as the California red-legged frog. Diet information was utilized for 

individual species of birds, reptiles, and mammals to determine if actual diet 

considerations can justify removing a species from concern. 

 

97 An extirpated species is defined as “a species no longer surviving in regions that were once part of its 

range” (http://www.fws.gov/midwest/endangered/glossary/index.html). 98 A product properties exclusion is based on a quantitative analysis of dicamba properties beyond the 

screening level analysis in the EFED Science Chapter. Specifically, this analysis involves assessing use 

of measured foliar residues and foliar decline rates following dicamba applications, adjusted to reflect the 

use pattern for DT-soybeans. The size and diet of the animal species are also considered. (See Sections 2 

and 4) 
Monsanto Company 10-SY-210U 715 of 721 

Figure N-4. Counties with Soybean Farms (or Adjacent Counties) with Listed 

Species in the Taxa of Interest 

N.4. Species-specific Risk Quotient Calculations 

Consistent with EPA’s Overview Document, species-specific dietary exposure was 

assessed considering animal body weight and diet (Schuler and Leopold, 2012) for birds 

and mammals of certain sizes because risk quotients exceeded the level of concern after 

the initial refinements described above. A similar assessment was conducted for 

amphibians and reptiles because birds serve as the surrogate for terrestrial phase 

amphibians and reptiles. Biological information was gathered from a number of sources, 

including U.S. Fish and Wildlife Service (USFWS) species recovery plans, primary 

literature, and other technical sources (e.g., NatureServe99)

Mammals

For mammals, 38 species were examined for the potential for chronic adverse effects 

based on biological characteristics (i.e., dietary preferences and body size). The analysis 

progressed in a step-wise fashion: 

 

99 NatureServe is a non-profit conservation organization that collects detailed local information on plants, 

animals, and ecosystems. NatureServe has a public website available at http://www.natureserve.org. 
Monsanto Company 10-SY-210U 716 of 721 

1) From the baseline analysis, assuming the maximum-use pattern (as described 

Section N.2.1.) and using the upper bound dicamba-specific foliar residue and 

foliar half-life values, the LOC for wild mammals was exceeded only for chronic 

exposures to small and medium sized mammals consuming short grasses (Section 

N.2, Table N-3). 

2) Using species-specific information, 27 species were given product property 

exclusions based on size (e.g. >1kg, large-sized mammals) and/or diet (e.g. 

carnivorous, frugivorous, etc.). 

3) The remaining 11 mammals, small and medium-sized species that consume 

grasses, were examined using the U.S. EPA T-REX model with species-specific 

weights used for the model input parameters. The predicted EECs were 

determined based on the upper bound dicamba- specific foliar residue and 

half-life values, as described in Section N.2.2. 

4) For 4 of these 11 species, RQs were below the LOC considering upper bound 

dicamba-specific residues and species-specific body weights, For another 3 of the 

11 species, examination of the diet indicated that seeds were the primary 

constituent of the diet with grasses or other items being a very small component, 

and with this consideration RQs were below the LOC. For the remaining 4 

species, considering the rapid rate of dicamba foliar residue dissipation, chronic 

RQ’s calculated with upper bound residues exceed the RQ for such a short 

exposure duration (i.e., < 1 day to < 4 days) that it should not be considered a 

chronic exposure. 

Furthermore, all mammalian species can be excluded from further consideration based on 

chronic RQ’s being below the LOC when RQs are calculated with initial mean residues. 

As indicated earlier, initial mean residues are considered more appropriate for assessing 

chronic risk. Thus, for these 38 species of mammals, chronic exposure to dicamba from 

application to DT-soybean can be excluded from concern and, therefore, would not affect 

these species. 

Birds 

For birds, of the 43 species evaluated in this refined analysis: 

1) Using species-specific information, 24 species were excluded from the need for 

further consideration because their refined T-REX acute RQs were less than the 

LOC based on characteristics of diet and body weight considering dicambaspecific residues. 

2) The 19 remaining species were excluded from the need for further consideration 

because the upper bound EECs calculated for these species were lower than dosebased No Observed Effect Levels (NOELs) calculated from acute dietary 

exposure studies. 
Monsanto Company 10-SY-210U 717 of 721 

The potential for acute adverse effects as determined in the analysis in Table N-4 is based 

on an oral gavage, a route of administration that can overestimate the expected exposure 

compared to more environmentally relevant methods with a slower rate of delivery (e.g., 

dietary exposure). Therefore, the use of the acute oral gavage toxicity test to predict the 

potential for adverse effects to birds from dicamba use on DT-soybean should be 

considered a screening-level approach that is expected to overestimate the actual 

potential for effects. A more appropriate method to determine the acute risk to birds from 

consuming prey or food items containing dicamba residues is to estimate the dose-based 

effect levels from dietary toxicity studies (Durda and Preziosi, 2000). This approach is 

consistent with the EPA Overview Document (U.S .EPA, 2004) which indicates that for 

avian risk assessments, dietary residues are compared with dietary toxicity endpoints 

based on dietary concentrations (e.g. LC50s for acute effects)100. This method was used 

for the 19 remaining species described above, with the conservative assumption that the 

avian species under evaluation are passerine species (highest metabolic/ingestion rates; 

U.S. EPA, 1993), and the toxicity endpoint selected from the dietary studies was the 

NOEL value rather than a LD50 value. Even with the high ingestion rate relative to body 

weight, these birds cannot ingest enough food to achieve doses that would exceed the 

NOEL (highest dose tested). Also, it is likely that the birds will not be in soybean fields 

actively foraging, but rather foraging in the adjacent habitats (e.g., tree canopy) outside of 

the fields. Threatened and endangered species are “strongly associated” with their 

specific habitat type and, therefore, it is improbable that 100% of the bird’s diet will 

consist of dietary items exposed in the field. Thus, for these 43 species of birds, acute 

exposure to dicamba from application to DT-soybean can be excluded from concern and, 

therefore, would not affect these species. 

Amphibians and Reptiles

For amphibians and reptiles, a total of 25 species (10 amphibians and 15 reptiles) were 

evaluated. The allometric equations used in the T-REX model assume that the ingestion 

rates for amphibians and reptiles are equal to those of birds, when in fact they are lower. 

Thus, the dose-based estimates of exposure calculated by T-REX are an over-estimation 

of exposure for amphibians and reptiles. Therefore, the risk evaluation for amphibians 

and reptiles for dicamba from dietary exposure was refined using the U.S. EPA THERPS model Version 1.0. Following U.S. EPA guidance, the T-HERPS model is to be 

used only after the RQs exceed the LOC for endangered species using the standard TREX model (U.S. EPA, 2008b). T-HERPS is currently approved for use for terrestrialphase amphibian and reptile species with diets similar to the federally threatened 

California red-legged frog, since the exposure values are considered more representative 

of potential exposure to herpetofaunal species. 

The T-HERPS model was used to generate upper bound EECs resulting from the 

maximum application scenario using refined residue and half-life data (Section N.2). The 

avian acute and chronic toxicity data was used as a surrogate for the amphibian and 

reptilian species. A default scaling factor of 1.0 was used in the model calculations, as 

the relationship between body weight and toxicity has not been examined for amphibian 

 

100 Overview Document (U.S. EPA, 2004) p. 40. 
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and reptile species (U.S. EPA, 2008b). Given the limited availability of morphometric 

data for many species, similar species were analyzed collectively as a group (e.g., frogs 

and toads) using the weight data from a single representative species as a proxy for all 

species within the group. The exposure analysis only considered dicamba residues within 

prey items typical for each species group and animal size. With these considerations, the 

T-HERPS model was used to calculate RQs for frogs, toads, salamanders, lizards and 

skinks. For these species, the acute, subacute, and chronic RQ values based on refined 

residue and half-life values do not exceed the LOC for endangered species. Thus, for 

frogs, toads, salamanders, lizards, and skinks, exposure resulting from the dicamba 

maximum use pattern can be excluded from concern and, therefore, would not affect 

these species. 

For snakes, turtles and tortoises, a species-specific exposure analysis was conducted 

using allometric equations for each specific dietary component (U.S. EPA, 1993). 

Exposure estimates based on refined dicamba-specific residue data and residue decline 

data were compared to acute and chronic avian toxicity values, and the calculated risk 

quotients were all determined to be below the level of concern, indicating that these 

species can be excluded from concern and, therefore, would not be affected. 

After considering species-specific body sizes and dietary components, all the amphibian 

and reptilian species evaluated were determined to have risk quotients below the level of 

concern resulting in the conclusion that dicamba use on DT-soybean will not affect these 

species. 

N.5. Indirect effects on Threatened and Endangered Species of Other Taxa 

resulting from Direct Effects on Plant Species 

To assess the potential for indirect effects on threatened and endangered species, the U.S. 

EPA evaluates the risk of direct effects on non-endangered species from relevant 

taxonomic groups to make inferences concerning the potential for indirect effects upon 

threatened or endangered species that rely on non-endangered species in that category for 

critical resources (U.S. EPA, 2004). The RQ values used to assess the potential for 

indirect effects from dicamba applications to DT-soybean indicate that indirect effects to 

other taxa resulting from effects on monocot plant species from application of dicamba at 

rates up to 1.0 lb a.e./A would not occur. Further, indirect effects to other taxa as a result 

of effects on dicot plant species from a combination of sheet runoff and spray drift to soil 

would also not occur (Wright et al., 2011). In addition, utilizing a refined exposure 

assessment, indirect effects to other taxa resulting from effects on dicot plant species 

exposed via a combination of channelized runoff and spray drift to soil would not occur 

(Wright et al., 2011). When RQ values are calculated for all six of the dicot species 

tested in the vegetative vigor study, RQs exceed the LOC for only two of the six species 

indicating that habitat and food sources from dicot plants will be present adjacent to areas 

treated with dicamba (Wright et al., 2011). Furthermore, field studies demonstrate that 

even very sensitive plants at the edge of a field sprayed with dicamba are not affected to 

the extent that impacts on habitat would result. Additionally, effects from drift can be 

further minimized through the use of best management practices such as drift reducing 

technologies (e.g., low drift spray nozzles, boom height, deposition aides). Considering 
Monsanto Company 10-SY-210U 719 of 721 

these factors, indirect effects on threatened and endangered animals resulting from spray 

drift onto nontarget plants can be excluded from concern. 

N.6. Conclusions 

In the EFED ecological risk assessment conducted for dicamba reregistration (U.S. EPA, 

2005), using a screening level assessment, EPA scientists concluded that a “no effect” 

determination can be made for all dicamba uses for listed fish, aquatic invertebrates, and 

vascular aquatic plant species. The potential for adverse effects to listed birds and 

mammals, non-vascular aquatic plants and terrestrial and semi-aquatic plants could not be 

excluded based on the EPA screening level assessment, the initial phase of analysis. 

However, since there are currently no listed nonvascular aquatic plants, this taxon does 

not require further investigation at this time. Recognizing that screening level analysis is 

only the first step in evaluating impacts on threatened and endangered species, the EFED 

Science Chapter (U.S. EPA, 2005) indicates that “additional information on the biology 

of listed species, the locations of these species, and the locations of the use sites ... could 

be considered along with available information on the fate and transport properties of the 

pesticide to determine the extent to which screening assumptions regarding an action area 

apply to a particular listed organism.” 

This report summarizes the conclusions of three other reports (Wright et al., 2011; Frank 

and Kemman, 2012, and Schuler and Leopold, 2012) that utilize some of the refinements 

described by EPA in the previous paragraph to evaluate the potential for adverse effects 

to listed birds and mammals as well as amphibians, reptiles, and terrestrial invertebrates. 

When the properties of dicamba and species-specific information are taken into account, 

these reports demonstrate that listed birds, mammals, amphibians, reptiles and terrestrial 

invertebrates would not be affected by dicamba use in DT-soybean. In addition, based on 

non-target plant studies conducted as a requirement of dicamba reregistration, 

monocotyledonous terrestrial and semi-aquatic plants outside the treated area also would 

not be affected (Wright et al., 2011). 

 
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References for Appendix N 

Durda JL, Preziosi DV. 2000. Data Quality Evaluation of Toxicological Studies Used to 

Derive Ecotoxicological Benchmarks. Human and Ecological Risk Assessment 6(5): 

747-765. 

Fletcher JS, Nellessen JE, and Pfleeger TG. 1994. Literature Review and Evaluation of 

the EPA Food-Chain (Kenaga) Nomogram, an Instrument for Estimating Pesticide 

Residues on Plants. Environ. Tox. Chem. 13(9): 1383-1391. DOI: 

10.1002/etc.5620130902. 

Frank AR, Kemman R. 2011. Information to Support an Endangered Species 

Assessment for Dicamba Use in Dicamba-Tolerant Soybeans: A County-Level Analysis 

for Animal Taxa. Monsanto Technical Report MSL-22547. MRID 48644204. 

Hoerger F, Kenaga EE. (1972) Pesticide residues on plants: Correlation of 

representative data as a basis for estimation of their magnitude in the environment. In: 

Environmental Quality and Safety: Chemistry, Toxicology, and Technology, F. Coulston 

and F. Korte, eds. Georg Thieme Publishers, Stuttgart, West Germany, pp. 9-28. 

Schuler LJ, Leopold VA, Mortensen SR. 2011. Information to Support an Endangered 

Species Assessment for Dicamba Use in Dicamba-Tolerant Soybeans: Refined Analysis 

for Terrestrial Animals. Monsanto Technical Report MSL-22546. MRID 48644203. 

U.S. EPA. 1993. Wildlife Exposures Handbook. Office of Health and Environmental 

Assessment. Office of Research and Development. EPA/600/R-93/187a. 

U.S. EPA. 2004. Overview of the Ecological Risk Assessment Process in the Office of 

Pesticide Programs, U.S. Environmental Protection Agency, Endangered and Threatened 

Species Effects Determinations. Washington, D.C. 

U.S. EPA. 2005. Environmental Fate and Ecological Risk Assessment for the 

Reregistration of Dicamba and Dicamba Sodium, Potassium, Diglycoamine, 

Dimethylamine and Isopropylamine Salts. 

U.S. EPA. 2006. Reregistration Eligibility Decision for Dicamba and Associated 

Salts. 

U.S. EPA. 2007. Risks of atrazine use to federally listed endangered Pallid Sturgeon 

Scaphirhyncus albus Retrieved January 1, 2008, from 

http://www.epa.gov/oppfead1/endanger/litstatus/effects/pallid_sturgeon_eff_deter_08-31-

07.pdf 

U.S. EPA. 2008a. Risks of Glyphosate Use to the Federally Threatened California Red 

legged Frog. Retrieved October 8, 2010, from 

http://www.epa.gov/espp/litstatus/effects/redleg-frog/glyphosate/determination.pdf
Monsanto Company 10-SY-210U 721 of 721 

U.S. EPA. 2008b. T-HERPS (Terrestrial Herpetofaunal Exposure Residue Program 

Simulation). Version 1.0. 

Wright JP, Schuler LJ, Carr KH, Honegger JL. 2011. Information to Support an 

Endangered Species Assessment Dicamba Use in Dicamba-Tolerant Soybeans: An 

Evaluation of Potential Exposure and Biological Effects. Monsanto Technical Report 

MSL-22545. MRID 48644202.