## Assessment of Plant Pest Risk for MON 8770 5 Soybean

1 Monsanto Company (Monsanto) has petitioned APHIS for a determination that MON 87705 soybean is unlikely to pose a plant pest risk (Rogan, 2009) and, therefore, should

no longer be a reg ulat ed article under APHIS’ 7 Code of Federal Regulations (CFR) part 34 0. APHIS administers 7 CFR part 340 under the authority of the plant pest provisions of the Plant Protection Act of 2000 1

. This plant pest risk assessment was conducted to determine if MON 87705 is unlikely to pose a plant pest risk.

MON 87 705 was produ ced by transformation of soybean tissue using A. tumefacien s. Because A. tumefaciens is a plant pest and some of the regulatory sequences ( Figwort Mosaic Virus promoter ) used to facilitate expression of these genes in soybean were derived from plant pests

, this soybean has been considered a regulated article under APHIS regulations at 7 CFR part 340.

Potential impacts to be addressed in this risk assessment are those that pertain to the use of MON 87 705 and its progeny in the absence of confinement. AP HIS u s es data and information submitted by the applicant, in addition to current literature, to determine if MON 87 705 is unlikely to pose a plant pest risk

. If APHIS determines that a GE organism is unlikely to pose a plant pest risk , then APHIS has no re gulatory authority over that organis m.
Potential impacts in this Plant Pest Risk Assessment are those that pertain to plant pest risk characteristics. APHIS regulation 7 CFR 340.6(c) specifies the information needed for consideration in a petition for no nregulated status. APHIS will evaluate information submitted by the applicant related to plant pest risk characteristics, disease and pest susceptibilities, expression of the gene product, new enzymes, or changes to plant metabolism, weediness of the regul ated article, any impacts on the weediness of any other plant with which it can interbreed , and transfer of genetic information to organisms with which it cannot interbreed.

An analysis of agricultural or cultivation practices associated with MON 87705 and impacts on the environment will be considered in the Environmental Assessment (EA) for MON 87705 soybean. A thorough assessment of the effects of the determination on nontarget organisms

, beneficial organisms and threatened and endangered species will be considered in the EA.



### H **istory of Development**  **of MON 87705 soybean with an**  **improved fatty acid profile**

Soybean oil is relatively high in saturated fatty acids (15% of total fatty acids) compared
to vegetable oil from other source s . Soybean oil is al so relatively high in polyunsaturated fatty acids (23% of total fatty acids) . These polyunsaturated fatty acids oxidize readily to

1 Section 403 (14) of the Plant Protection Act (7USC Sec 7702(14) ) defines plant pest as: “Plant Pest

- The term “plant pest” means any living stage of any of the following that can directly or indirectly injure, cause damage to, or cause disease in any plant or plant product: (A) A protozoan. (B) A nonhuman animal. (C) A parasitic plant

. (D) A bacterium. (E) A fungus. (F) A virus or viroid. (G) An infectious agent or other pathogen. (H) Any article similar to or allied with any of the articles specified in the preceding subparagraphs.”

## Assessment of Plant Pest Risk for MON 8770 5 Soybean

2 produce off

- flavors and reduce the performance of unprocessed soybean oil. The stability and flavor of soybean oil is impro ved by hydrogenation, which chemically reduces the double bonds. However, hydrogenation has the undesirable consequence of creating trans

- fatty acids. In recent years, trans

- fatty acid s have come under considerable scrutiny by American Heart Association a nd others because of the negative effects by these substances on human health

.

Monsanto has developed a transgenic soybean

, event MON 87705 (hereafter referred to as MON 87705)

, that produces soybean seeds with decreased levels of saturated (palmitic and stearic) and polyunsaturated (linoleic) fatty acid s , a nd increase d level s of monounsaturated (oleic) fatty acid

. MON 87705 conta ins DNA segments designed to suppress endogenous d elta

- 12 desaturase ( FAD 2 ) and Acyl

- ACP thioesterase ( FATB ) genes which encode for two enzymes in the soybean fatty acid biosynthetic pathway. MON 87705 also contains the 5

- enolpyruvylshikimate

- 3

- phosphate synthase ( cp4 epsps )
gene encoding the CP4 EPSPS protein . The cp4 epsps gene was used as a selectable marker to identify transge nic plants during the transformation process

. The CP4 EPSPS protein confers tolerance to glyphosate and has been used in many Roundup Ready crops (e.g. canola, corn, cotton, soybean and sugar beet)

. In plants, the microsomal delta

- 12 desaturase

- catalyz ed pathway is the primary route of production of polyunsaturated lipids. The FAD2 gene is strongly expressed in developing seeds

. The seed

- specific expressed FAD2 gene is likely to play a major role in controlling conversion of oleic acid ( a monounsatura ted fatty acid) to linoleic acid ( a polyunsaturated fatty acid) within storage lipids during seed development (Heppard et al.

, 1996). The delta

- 12 desaturase ( FAD2 ) gene in soybean is responsible for the conversion of oleic acid to linoleic ac i d. Sup press i on of the FAD2 gene prevent s linoleic acid from being synthesized and lead s to the accumulation of oleic acid in the soybean seed.

The FATB gene is one of the key enzymes for oil biosynthesis in plants. The suppression of FATB results in a decrease in the transport of saturated fats out of the plastid, thus retaining the availability of those saturate d fats for desaturation to oleic acid ( Petition

, 2009 , page 5, p anel A and B)

. In MON 87705 , t he assembled gene transcripts have an inverted repeat that p roduces double stranded RNA that, via the RNA interference (RNAi) pathway, suppresses endogenous FATB and FAD2 gene expression.

The increase in oleic acid and decrease in saturated fatty acids in MON 87705 is aimed at improving the stability of these soyb ean oils and promoting human heart health

. The commercialization of MON 87705 could be beneficial for consumers. From a nutritional standpoint, the consumption of trans fatty acids results in considerable potential harm with no apparent known b enefit to h uman health (Mo z affarian et al

., 2006). Increased intake of oils high in monounsaturated fatty acids, such as oleic acid

, has been recommended by American Heart Association because of their positive effect on total cholesterol levels when compared to equa l intakes of hydrogenated oils (Lichtenstein et al.

, 2006). Likewise , increased intake of oils high in oleic acid is known to decrease LDL

- cholesterol levels compared to an equal intake of oils high in saturated fats (Mensink et al.

, 1989) an d known to in crease HDL

- cholesterol levels compared to an

## Assessment of Plant Pest Risk for MON 8770 5 Soybean

3 equal intake of polyunsaturated oil s (Mata et al.

, 1992) . Moderate consumption of oil high in oleic acid has also been demonstrated to result in decrease d systolic blood pressure (Bondia

- Pons et al.

, 2007).



### Des **cription of the modification**

MON 88705 was produced by Agrobacterium

- mediated transformation of soybean with a binary vector that contains two T

- DNAs ( T

- DNA I AND T

- DNA II; T

- DNA refer s to the DNA that is transferred to the plant during trans formation) as shown in petition ( Petition

, 2009

, figures IV

- I and IV

- II
, p

. 54

- 55 and Table V

- 2

, p

. 63

- 65 )

.
(1) T

- DNA I contains a cp4 epsps expression cassette and a partial suppression cassette containing sense segments of an FAD2

- 1A intron and FATB1

- A 5 ́ untranslated region (UTR) ( Petition

, Table IV

- 1, p p . 56

- 57 )

.

The cp4 epsps expression cassette contains the following genetic elements:

**•** FMV/TSF1 chimeric promoter: FMV/TSF1 promoter consisting of enhancer sequences from the promoter of the Figwort Mosaic V irus (FMV) 35S RNA combined with the promoter from the Tsf1 gene of Arabidopsis thaliana that encodes elongation factor EF

- 1 alpha ( Axelos et al., 1989 ).

**•** cp4 epsps gene : the cp4 epsps gene is a gene fusion composed of the N

- terminal chloroplast transit peptide (CTP2) sequence from Arabidopsis thaliana, epsps gene and the C

- terminal synthetic 5

- en olpyruvylshikimate

- 3

- phosphate synthase gene (CP4) from an Agrobacterium species, strain CP4 and encoding CP4 EPSPS protein. The CP4 EPSPS protein confers tolerance to glyphosate (Padgette et al., 1996; Barry et al., 1997)

.

**•** E9 3  ́ UTR as transcriptional t ermination: 3  ́ untranslated region of the Pisum sativum ribulose 1,5

- bisphosphate carboxylase small subunit E9 ( RbcS2 ) gene which functions to direct polyadenylation of the cp4 epsps mRNA (Coruzzi et al., 1984)

. The partial suppression cassette contains t he following genetic elements:

**•** 7Sα  ́ promoter : 7Sα  ́ seed specific promoter is from the Sphas1 gene of Glycine

max encoding beta

- conglycinin storage protein (Doyle et al., 1986) that directs transcription in seed.

**•** The sense segment of the FAD2

- 1A intron : Par tial sequence from the intron number 1 of the Glycine max FAD2

- 1A gene that encodes the delta

- 12 desaturase (Fillatti et al., 2003) which forms part of the suppression cas s ette.

**•** FATB1

- A 5' UTR : Partial sequence from the 5' UTR and the plastid targeting

se quence from Glycine max FATB1

- A gene that encodes the palmitoyl acyl carrier protein thioesterase (Fillatti et al., 2003) which forms part of the suppression
cassette

.
(2) T

- DNA II contains a partial suppression cassette that consists of antisense segment s of FAD2

- 1A intron and FATB1

- A 5’UTR

.

## Assessment of Plant Pest Risk for MON 8770 5 Soybean

4 **•** The antisense segment of FAD2

- 1A intron : is an intron from FAD2

- 1A gene that encodes the delta

- 12 desaturase.

**•** The antisense segment of FA TB1

- A 5 ’ UTR: Partial sequence from 5' UTR and the plastid targeting sequence is from Glycine max FATB1

- A gene that encodes the palmitoyl acyl carrier protein thioesterase.

**•** H6 3’ UTR Terminator: H6 3’UTR from Gossypium barbadense is for transcriptional termination and polyadenylation of a fiber protein involved in secondary cell w all assembly.

In addition to the above

- mentioned nucleotide sequences, T

- DNA I and II also contains

non

- coding intervening sequences and elongation factors that facilitate DNA cloning and translational elongation, respectively. The assembled gene trans cript has an inverted repeat that produces double stranded RNA (dsRNA) that, via the RNA interference (RNAi) pathway, suppresses endogenous FATB and FAD2 RNA.

Transformation system

. MON 87705 was developed using a disarmed Agrobacterium

– mediated transf ormation system of soybean, which allows the generation of transformed plants without utilization of callus ( Martinell et al., 2002), . This technique using disarmed Agrobacterium followed by glyphosate selection has a long history of safe use.



### Plant Pes **t Risk Assessment**

Based on information on the biology of soybean ( OECD

, 2000 ) , data presented by Monsanto Company ( Petition , 2009)

, and scientific data relevant to the discussion of plant pest ris k , APHIS makes the following conclusions for MON 87705 soyb ean:
Potential impacts of genetic m odifications on disease and pest susceptibilities

USDA

- APHIS assessed whether MON 87705 is likely to have significantly increased disease and pest susceptibility compared to its unmodified control

. The assessment
encomp asses a consideration of introduced traits and interactions with pest s and disease s

. The only introduced protein produced in MON 87705 is CP4 EPSP S . The 5

- enolpyruvylshikimate

- 3

- phosphate synthase (EPSPS) family of enzymes is ubiquitous in plants and mic roorganisms. The introduced cp4 epsps gene is from Agrobacterium sp. strain CP4. Th e CP4 EPSPS protein expressed in MON 87705 is similar an d functionally identical to endogenous plant EPSPS enzymes and is identical to the CP4 EPSPSs in other Roundup Ready ® crops including Roundup Ready ® soybean ( 40

- 3

- 2 and MON 89788)

, Roundup Ready ® canola, Roundup Ready ® sugar beet and Roundup Ready f l a x and Roundup Ready ® c otton. The first generation of Roundup Ready ® soybean (40

- 3

- 2) ( USDA

- APHIS, 1993) was deregulated i n 1995. The cp4 epsps gene has been assessed extensively in the last 15 years to confirm that it does not alter the desired agronomic characteristics of transgenic crops , and that it does not present a plant pest risk

. Therefore, i ncorporat ing cp4 epsps g enes into MON 87705 is not likely to alter disease

## Assessment of Plant Pest Risk for MON 8770 5 Soybean

5 and pest susceptibility

. The expression of the epsps gene is controlled by a chimeric promoter wherein a part of the sequence comes from Figwort Mosaic Virus. These plant virus

- derived regulatory sequence s are non

- coding with known function and they are not known , in and of themselves, to cause plant disease

. The introduced endogenous FAD2
gene that supp resses lipid biosynthesis increase s the amount of oleic acid in the seed, a constituent which is presen t in the parent cultivar. Such gene s ha ve been used in field trials previously and are not known to cause plant disease

. There is no indication that inserting the FAD2 gene will result in increased likelihood of introduction or dissemination of a plant pest. The higher concentration of oleic acid is not known to pose a plant pest risk in unconfined releases. APHIS has not identified any plant pest risk

a ssociated with the introduction of the high oleic acid trait in the deregulated DuPont product ( USDA

- APHIS

, 1997 ) and Pioneer product (USDA

- APHIS, 2006)

. The Plant Biosafety Office (PBO) of the Canadian Food Inspection Agency (CFIA

, 2000 ) , which also reviewed a dossier on this product

, also concluded that this novel trait (high oleic acid) will not introduce a ny plant pest characteristics in to MON 87705 soybean. T he introduce d endogenous FATB gene suppresses the ac yl

- acyl carrier protein (ACP) thioesterase and results in a decrease in the transport of saturated fats out of the plastid, thus retaining their avai lability for desaturation to 18:1 oleic acid . As a result, MON 87705 soybean oil is lower in saturated fats. T he information collected from the field trials and literature, there is no indication that by suppressing FATB gene will result in altering plant pest risk.

Field evaluation of MON 87705 an d conventional soybean varieties were carried out at 17 sites covering 10 States and assessed for differences in plant response to abiotic stress, disease damage, and arthropod damage. The p etitioner rated soybea ns for responses to abiotic stress, diseases, and arthropod pests that were present or typically present at each site. For the assessments of the plant

- insect interactions, plant

- disease interactions, and plant responses to abiotic stress, no differences w ere detected between MON 87705 and the control s for 574 of 579 observations at the 17 sites. There were statistical differences noted in the disease and arthropod damage in 5 of 579 observations . In three of the observations

, MON 87705 has less damage fro m bacterial blight. In the other two observations

, MON 87705 has less damage from aphids and leafhoppers ( Petition , 2009

, p. 344

- 346). The 5 noted differences , however, were still within the reference response range of the control plants

.
Based on the analysis of genetic modifications and their functions and field testing data submitted by p etitioner , APHIS conclude s that there are no significant differences between MON 87705 soybean and the non

- transgenic counterparts relative to pest and disease susce ptibility. Soybean ( Glycine max ) is not a plant pest in the United States (USDA

- APHIS, 2000), and the introduced DNA in MON 87705 is unlikely to pose a plant pest risk.

Potential impacts from new gene products, changes to plant metabolism or composition

## Assessment of Plant Pest Risk for MON 8770 5 Soybean

6 USDA

- APHIS assessed whether changes in plant metabolism or composition in MON 87705 is likely to alter plant pest risk. The assessment encompasses a consideration of the inserted sequences, expressed protein and introduced traits.

There is no new protei n expressed because of the insertion of the FAD2 and FATB gene fragment s

. The only novel protein expressed in MON 87705 is EPSPS (5

- enolpyruvylshikimate

- 3phosphate synthase)

, which imparts glyphosate toleran ce

. The EPSPS protein is found naturally in pl ants, fungi and bacteria , but is absent in mammals, fish, birds, reptiles, and insects (Alibhai et a l., 2001) . The EPSPS in MON 87705 is derived from an Agrobacterium sp . strain CP4

. More than 200 epsps sequences are known ( http://www.ncbi.nlm.nih.gov/nuc core ) . Even though there is significant diversity for amino acid sequence s among EPSPS protein s , they all share a common structure and a conserved active site. The EPSPS protein in Mon 87705 is minimally modified compar ed to the endogenous form in soybean

. The primary mode of action of glyphosate is the competitive inhibition of the plant enzyme 5

- enolpyruvyl

- shikimate

- 3

- phosphate synthase (EPSPS) which catalyses the penultimate step in the shikimate pathway ( Franz et al., 1997 ).

The scientific bas is for glyphosate tolerance in all Monsanto Roundup Ready ® GE crops ( Petition , 2009, page 4) is identical. Th e se products have been grown widely across the U

. S

. since the 19 9 0 ’s and have a long history of safe use. Although EPSPS proteins are ubiquitous in nature, there is no evidence of unexpected plant pest risk from these proteins. Southern blots were used to determine the copy number of each of the genetic elements and to examine the integrity of each fragment inserted into the soybean genome. The p etitioner has concluded that the CP4 EPSPS

- expressing transgene is present in one copy, is integrated at a single locus, segregates as a single dominant Mendelian trait, and is stable within the soybean genome

. The p etitioner also notes that the trait is phenotypically stable over several generations.

The CP4 EPSPS transcript levels were measured in leaf, seed, root and forage tissues

. T he mean protein data for the measured pl ants are summari z ed in Table VI

- 2 ( Petition

, 2009, p

. 89). The Margin of Exposure (MOE) was calculated based on the EPSPS protein expressed in different tissues. The Margin of Exposure (MOE) is the ratio between a defined point on the dose

- response curve (reference point) for the adverse effect of the compound in an animal carcinogenicity study and the estimated human intake of the compound. The MOE is calculated for CP4 EPSPS protein produced by MON 87705 and results indicate that there is no meaningful r isk to human health from dietary exposure

. The conclusions of “no further questions” on nutritional and safety issues were reached by FDA for the CP4 EPSPS protein (FDA , 2000; FDA, 2004; FDA, 2005; FDA

, 2006 )

. Petitioner submitted a summary of its safety and nutritional assessment to FDA for MON 87705 soybean in 2009. A final FDA decision is pending.

The compositional assessment was conducted in accordance with the Organization of Economic Co

- operati o n and Development (OECD) consensus document on
compo sitional considerations for new varieties of soybean (OECD, 2001)

. A total of 38

## Assessment of Plant Pest Risk for MON 8770 5 Soybean

7 fatty acids were analyzed in MON 87705 and control lines. Twenty

- one fatty acid concentrations were near or below the detection limits of the assay. Of the other acids that co uld be statistically analyzed

, concentrations of 13 fatty acids showed statistically significant difference s between MON 87705 and the controls. Four of the 13 differences were intended change s from control soybean lines: palmitic acid (16:0), stearic ac id (18:0), oleic acid (18:1) and linoleic acid (18:2). Nine of the 13 fatty acids were significantly different from the control soybean lines: myristic acid, palmitoleic acid, margaric acid, heptadecenoic, octadecadienoic acid, arachidic acid, eicosenoic acid

, behenic acid and 18:2 other trans isomer fatty acids ( Petition , 2009, p p. 311

- 312, Table E15 ). However, the levels of these fatty acids were biologically not significant , as most of them are relatively minor fatty acids and are common at similar leve ls in vegetable oils ( USDA

- ARS

, 2006)

. Therefore, APHIS conclude s that MON 87705 is compositionally equivalent to conventional soybean with the exception of intended changes in fatty acid levels.

Compositional and nutritional data w ere collected on MON 87705 and comparisons were made to a conventional control li n e and a set of reference soybean varieties. Based on the results of comparative analysi s APHIS conclude s that MON 87705 is compositionally and nutritionally equivalent to conventional soybean va rieties currently in commerce

, except for the intended changes

.

Based on all the consideration s above

, APHIS concludes that MON 87705 poses no more of a plant pest risk from new gene products, changes to plant metabolism or composition than conventional s oybean.

Potential impacts from outcrossing of MON 87705 to wild relatives

Soybean is a self

- pollinated species propagated by seed (OECD

, 2000)

. In its papilionaceous flower, the anthers mature in the bud and directly polli n ate the stigma of the same flo wer. As a result, soybean s exhibit a high percentage of self fertilization. Natural or artificial cross

- pollination can only take place during the short time when the pollen is viable. The c ross pollination rate ( with and without pollinators) is less th an 1.5% beyond one meter from the pollen source ( Garber, et al., 1926; Carviness, 1966; Ahrent et al., 1994 ; Ray, et al., 2003; Yoshimura, et al., 2006 )

. At greater distances from the pollen source , cross pollination rates decrease rapidly. Based upon th ese factors , it is unlikely that MON 87705 will naturally outcross or hybridize to a significant extent with other soybean varieties in agricultural settings.

In assessing the risk of gene introgression from MON 87705 into its sexually compatible relative s, APHIS considers two primary issues: 1) the potential for gene flow and introgression and

, 2) the potential impact of introgression.

Cross pollination with wild species The genus Glycine is divided into two subgenera, G lycine and S oja

.

## Assessment of Plant Pest Risk for MON 8770 5 Soybean

8 The subgenus S oja consists of three annual species: G. soja Sieb. and Zucc., the form of soybean, G. gracilis Skvortz., the weedy form of soybean and G. max, the cultivated soybean. They grow wild or semi

- wild in Asia. Fertile hybrids between G. max and G. soja (Broic h

, 1978), and between G. max and G. gracilis (Karasawa, 1952) occur. G. soja and G. gracilis grow naturally only in Asia and Australia, not in the United States (Skvortzow, 1972)

. The subgenus G lycine consists of twelve wild perennial species. These s pecies grow wild in Australia, South Pa cific Islands and Asia (Newell et al.

, 1978) , and do not exist naturally in the U.S. Hybrids between perennial Glycine species are fertile.

G. max is the only Glycine species located in the United States , thus the re are no other plant species with which G. max can interbreed

. G. max has never been found in the wild (Hymowitz et al., 1987) without human intervention

. T herefore

, it is highly un likely that soybean plants in the United States will be found outside of an agricultural se t ting. It is also highly unlikely that gene flow and introgression will occur between MON 87705 and soybean plants in a natural environment

. USDA has therefore determined that any adverse consequences of gene flow from MON 87705 to wil d or weedy species in the United States are highly unlikely

. Potential impacts based on the relative weediness of MON 87705.

APHIS assessed whether MON 87705 is any more likely to become a weed than the

n on

- transgenic recipient soybean line, or other soy bean currently cultivated. The assessment encompasses a thorough consideration of the basic biology of soybean and an evaluation of unique characteristics of MON 87705

. In the U.S., soybean is not listed as a weed in the major weed references (Crockett

, 1 977; Holm et al.

, 1979; Muenscher

, 1980) nor is it listed as a noxious weed species by the U.S. Federal Government ( USDA

- APHIS

, 20 10 a ) and States (USDA

- APHIS 2010 b )

. Soy beans are not frost tolerant

, do no t survive freezing winter condition s (OECD

, 2000)

, a nd d o not reproduce vegetative ly. After crop harvest, soybean may germinate as a volunteer weed in the succeeding crop (Padgette et al.

, 1996)

. If any seeds remain viable and germinate the following year , mechanical methods or herbicides can be used to

c ontrol volunteers

. Soybean is one of the largest acreage crops in the worl d

. Soybean has never been reported as a serious weed . In the U.S.

, soybean is grown on over 7 7 million acres (USDA

- NASS

, 200 9 a )

. Based on the familiarity of soybean as the paren t plant , there has been no report of soybean escaping cultivation and becoming established as a weed in United State s (Holm et al.

, 1979).

Weediness for the purposes of this part of the p lant p est r isk assessment is an attribute, which causes a crop to act as a weed due to the addition of genes ( compar ed to the non

GE plant)

. If the fitness of MON 87705 improves in natural or agricultural ecosystems due to the inserted DNA , the potential for weediness could increase. The following analysis of the inser ted DNA is intended to document that MON 87705 has a negligible likelihood of increased weediness.

## Assessment of Plant Pest Risk for MON 8770 5 Soybean

9 MON 87705 differs from conventional soybeans only in the expression of the EPSP S protein and silencing of the endogenous FAD2 and FATB gene s

. T he RNA

- based suppression of FATB and FAD2 soybean genes are mediated by double stra nded RNA (dsRNA) molecules and do not code for any protein. Double stra nded RNAs are commonly used in plants for endogen ous gene suppression and pose no novel risks from a plant pest pe rspective

. The only introduced protein in MON 87705 is CP4 EPSPS. This protein has the same functional and enzymatic acti vity as the C P4 EPSPS in other Roundup Ready ® crops

. The large scale commercial cultivation of glyphosate tolerant soybean crop acrea ge has steadily increa sed from 1996, accounting for 91 % of soybean acreage in 2009 (USDA

- NASS, 2009 b )

. No data of which APHIS is aware indicate that the presence of the cp4 epsps gene improves the ability of this soybean line to survive without human int ervention, nor is there any foreseeable reason to conclude that this gene would facilitate MON 87705’s survival in unmanaged areas

.

In the 2007 growing season

, Monsanto evaluated 14 phenotypic and agronomic characteristics in 17 field locations in the U nited States ( Petition , 2009, p

. 124, Table VIII

- 3) . Data such as emergence, seedling vigor, plant height, lodging, days to maturity, shattering, seed weight, yields, disease incidence and insect damage were used to assess whether there was an increase in weediness potential. Monsanto observed four statistically significant differences between MON 87705 and the control: (1) e arly stand count, (2) f inal stand count, (3) d ays to 50% flowering, and (4) s eed weight. MON 87705 was lower than control for both e arly sta nd count and final stand count ( Petition , 2009, p

. 349

- 350, Table G

- 3, Appendix G). MON 87705 flowered about one day later than the control and the weight of 100 seeds was lower for MON 87705 compared to the control (15.6g vs. 16.1g)

. APHIS is no t aware of any information indicating that decreased stand counts, seed weight or differences in days to 50% flowering would contribute to an increased weed iness potential of soybean

.

Monsanto also did c omparative assessments of seed dormancy and germina tion characteristics on MON 87705 and A3525 (a conventional variety) , where A3525 served as a control because it has background genetics similar to MON 87705 but does not possess the transgenes

. T he da ta related to s eed germination characteristics were us ed to assess whether there was an increase in weediness potential

. Four statistically significant differences were detected between MON 87705 and the control : (1) MON 87705 had lower percent viable hard seed (0.0 vs. 0.3%) at 20 ° C, (2) MON 87705 had lower percent germinated seed at 30 ° C (92.8 vs. 95.5%), (3) MON 87705 had higher percent dead seed at 30 ° C (7.3 vs. 4.5%), and (4) MON 87705 had higher percent dead seed at 20/30 oC (2.7 vs. 1.3%). A decrease in hard seed, a l ower percent germinated seed and hi gher percent dead seed would not contribute to increased weediness. Furthermore, all values were well within the recommended standards for cert ified soybean seed (AOSCA, 2009 ).

Th ese data provide an evaluation of the phenotypic and agronomic characterist ics of MON 87705 compared to the conventional A3525 control. Results from the phenotypic and agronomic assessments indicate that MON87705 does not possess characteristics that would confer a plant pest risk compared to conventional soybean. T hese data in dicate

## Assessment of Plant Pest Risk for MON 8770 5 Soybean

10 that the engineered plant is not different in any fitness characteristics from its parent that might cause MON 87705 to become weedy or invasive

. Therefore, soybean is unlikely to become a weed through the introduction of the glyphosate tolerance tr ait.

A few earlier investigations (Kodama et al., 1994 and Kodama et al.,1995) indicated that the increases in levels of trienoic fatty acids such as hexadecatrienoic acid and linolenic acid c ould enhance cold tolerance in model plants such as Arabidopsi s and tobacco. In MON 87705, the levels of linolenic acid are significantly decreased. The decreased levels of linolenic acid in MON 87705 would, therefore, not be expected to enhance cold tolerance. If the seed overwintering capacity improved, the pote ntial for a successful weed could increase. There is no indication that MON 87705 possesses a selective advantage that would result in increased weediness potential. Therefore, the chances for MON 87705 to behave as a weed are negligible.

From these obse rvations and da ta submitted in the petition, APHIS conclude s that there

are no weed iness risk issues associated with MON 88705

.

Potential impacts from transferring genetic information from MON 87705 to organisms with which it cannot interbreed.

Horizonta l gene transfer and expression of DNA from a plant species to bacteria is unlikely to occur. First, many genomes (or parts thereof) have been sequenced from bacteria that are closely associated with plants including Agrobacterium and Rhizobium (Kaneko et al.

, 2000 ; Wood et al.

, 2001 ; Kaneko et al.

, 2002)

. There is no evidence that these organisms contain genes derived from plants. Second, in cases where review of sequence data implied that horizontal gene transfe r occurred, these events are inferred to occur on an evolutionary time scale on the order of millions of years (Koonin et al.

, 2001 ; Brown
, 2003)

. Third, transgene DNA promoters and coding sequences are optimized for plant expression, not prokaryotic bacterial expression. Thus even if horizontal gene transfer occurred, proteins corresponding to the transgenes are not likely to be produced. Fourth, the FDA has evaluated horizonta l gene transfer from the use of antibiotic resistance marker genes, and concluded that the likelihood of transfer of antibiotic resistance genes from plant genomes to microorganisms in the gastrointestinal tract of humans or animals, or in the environment, is remote ( FDA, 1998). Therefore APHIS concludes that horizontal gene transfer is unlikely to occur and thus poses no significant environmental or plant pest risk.



### Conclusion

APHIS has reviewed the potential of MON 87705 to pose a plant pest risk and conducted

an assessment based on the information provided by the p etitioner on MON 87705

. APHIS determined no plant pest risk would derive from the inserted genetic material, found no atypical responses to disease or plant pests in the field

, assessed th at weediness of other plants with which MON 87705 can interbreed would be unlikely

, discerned no

## Assessment of Plant Pest Risk for MON 8770 5 Soybean

11 weediness characteristics of MON 87705 , and found no expectation for horizontal gene transfer, so that APHIS concludes that MON 87705 is unlikely to pose a pla nt pest risk.

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