### 1 Response to Comments Petition 0 6

- 354

- 01

APHIS reviews a petition for nonregulated status to determine if the genetically engineered (GE) organism should no longer be considered a regulated article under APHIS biotechnology regulations (7 Code of Federal R egulations (CFR) part 340). Prior to reaching a decision, APHIS prepared a plant pest risk assessment to evaluate whether Pioneer 305423 soybean is likely to pose a plant pest risk. After finding that Pioneer high oleic soybean is not likely to pose a pl ant pest risk, and is eligible for nonregulated status, APHIS prepared a draft environmental assessment ( EA ) to evaluate whether there could be significant impacts on the environment arising from a decision to grant a determination of nonregulated status t o Pioneer 305423 soybean

. APHIS prepared the EA as part of its obligation to meet the statutory requirements of the National Environmental Policy Act (NEPA) of 1969. As part of this process, APHIS considered public comments received on the petition for d eregulation and associated draft EA. This document provides APHIS ’ response to these comments

. On September 2, 2009, APHIS published a notice in the Federal Register (74 FR 45413

- 45415, Docket no. 2007

- 0156) announcing the availability of the Pioneer p etition requesting nonregulated status for 305423 soybean and associated draft EA for public review and comment for a 60

- day period. Th is comment period was scheduled to end on November 2, 2009. Because the original docket was not the correct APHIS v ers ion

, a subsequent 60 day comment period was published on October 26, 2009 in the Federal Register (74 FR 54950

- 54951, Docket no. 2007

- 0156) announcing the availability of the corrected EA

. This comment period ended on December 28, 2009. APHIS received a total of 40 comments from various groups and individuals during the two comment periods. Twenty

- two comments supported deregulation, eighteen comments generally opposed the development and use of G E foods (twelve from one individual), one
comment emphasiz ed a need for strict regulation

, and one comment generally disagreed about the overall need for this soybean crop.

Those in favor of non regulated status included five state senators or representatives, five departments of agriculture, three food or ind ustrial companies, three soybean associations, and two soy or food institutes. Those in opposition to granting non regulated status were individual consumers, who as noted, mostly did not offer opposition to this specific product, but were opposed to all genetically modified plants. Supporters of nonregulated status cited a number of benefits if this product were available for commercial production including : (1) the product’s fatty acid ratios are more healthful to the public than conventional soy oil ; (2) oil derived from the product would be useful to the food and non foods industry ; and (3) the growers will be able to market a new product that expands the potential commercial possibilities for soybean because of an improved fatty acid balance. Severa l comments from various industry associations focused on Pioneer/Dupont’s statement of support for adequate risk assessment and risk management programs sponsored by the technology provider, and Pioneer’s acceptance of a total life cycle approach for the p roduct in domestic and export markets. Of those opposed, none provided specific disagreement with the analysis provided by the environmental assessment , but were focused primarily on opposition to control of the seed supply by the federal government, or

c ontrol by large corporations that ostensibly desire to increase the cost of seed . Some were concerned about consequences of gene splicing, such as creating plant pests, introducing dangers from cross

- pollination of wild plants, generating unintended conse quences in the GE plants, producing plants that were poisonous or otherwise hazardous, causing GE plants to become mutagenic, or caus ing organ toxicity, or otherwise generat ing unknown, long

- term risks. Still other comments sta ted that an EIS needs to be done for this proposed product.

### 2 

## 1. Ecological Effects

Two comments suggest that genetically engineered crops will cause chemical harm to the environment or that these GE crops endanger wild and native plants near where they are planted.

APHIS response : Based upon information and analysis presented in the petition, plant pest risk
assessment (USDA 2009) , and EA, APHIS has not identified any potential for chemical harm to the environment deriving from Pioneer 305423 soybean . Aside from the percentages o f poly

- and

mono

- unsaturated fatty acids, the chemical composition of high oleic soybean does not differ significantly from conventionally bred soybeans. O leic acid is a naturally occurring, nontoxic substance present in soy and many other edible oils, ca nola and sunflower oils, especially those enhanced for high oleic acid expression (see EA Section V, C ). Soybean is not a plant native to North America, and as a result, does not have any known wild or weedy relatives in the United States with which it co uld interbreed (see EA Section III, A ). In addition, soybean is primarily self

- pollinating, so gene flow beyond modest isolation distances to surrounding cultivated soybean (beyond 4.6 m) would be rare and not likely (see EA Section III, A ).



## 2. Agricu ltural Impacts

Some comments suggested that inadvertent commingling of the high oleic soybean with commodity soybean will impact the accuracy of data on the Nutrition Facts Panel for food products, because the percentage of saturated fatty acids, mono

- an d poly

- unsaturates would be altered if the product contained as little as 5% of the specialty product.

APHIS response: Pioneer 305423 soybean contains reduced levels of palmitic acid, a saturated fat, and elevated levels of high oleic acid, a monounsatur ed fat (see Section I of the EA). Because of the health benefits of an increase in specific fatty acids in Pioneer 305423 soybean as compared to commodity soybean, manufacturers may preferentially select these oils for manufacturing many food products. F DA requires that product fatty acid ratios be reflected in the Nutrition Facts Panel of the product label . If sufficient amounts of Pioneer 305423 soybeans inadvertently enter the commodity soybean supply stream and not the appropriate identity preserved food chain

, the Nutrition Facts Panel of some product s may not correctly reflect the fatty acid ratios. In public comments submitted to APHIS for this product, Pioneer suggests that commingling of above 5% content of high oleic acid soybean oil from Pione er 305423 in commodity oils could be sufficient to require alteration of the labeled fatty acid content.

B ecause of the value

- added traits of this soybean, growers and processers will preserve product identity to the point of sale, thus keeping Pione er 305423 soybean separate from commodity soybean (see Pioneer 305423 p etition

, amend ed S ection X

- XF

- 7, Potential impact on organic and conventional farming)

. Pioneer proposes to use “an identity preserved (IP) system ...from seed development through refin ed oil produced for delivery to end

- use customer s ” (Pioneer Public Comment APHIS Docket 2007

- 0156

- 0045 ; 

### Regulations.gov (2009) . The details would resemble “similar systems ... throughout the vegetable oil industry.” Other IP oil seed management systems inclu de sales of the seed only to growers who sign a contract with the purchaser to produce the oilseed to specified standards, and to sell their soybeans only to the contracted party and to its designated elevator or crushing facility. Oilseed processors woul d likely be required to sign contracts with refiners so that end users would have a trackable product that derived only from identified transgenic seed. I n the same letter submitted as a public
comment , Pioneer notes that analytic al techniques are availab le to qualitatively identify fatty acid

### 3 content in delivered oil seed using NIR (Near infrared) technology . For analysis of fatty acids in derived oils , highly accurate gas chromatograph ic analytic al techniques are available

.

Oils derived from Pioneer 30 5423 soybean will be separately marketed from conventional soybean oils, because these oils contain healthy fatty acids (such as the monounsaturated oil, oleic acid) when consumed in manufactured food products such as salad oils (See EA S ection I. and II. )

. Oleic acid is a natural constituent of a variety of foods, notably for example, olive oil (55

- 83% oleic acid, Vossen, 2010). Olive oil has been shown to be a health

- promoting dietary constituent, and the oleic acid component has been directly shown to be responsible for lowering blood pressure ( Terés et al., 2008).

To monitor product integrity, manufacturers of food products may need to continuously assay fatty acid ratios of constituent soybean oil . If manufacturers are vigilant and employ good quali ty control techniques, it is not likely that high oleic soybean oil would be inadvertently incorporated in sufficient amounts that would change the accuracy of the fatty acid ratio reflected on the Nutrition Facts Panel. In cases of a manufacturer receivi ng shipment of an ingredient oil (ie.

, high oleic soybean oil ) different from that typically used in a specific product, the manufacturer could appropriately blend and dilute the oils before commercial sale and release, and the Nutrition Facts Panels would accurately reflect fatty acid content.

Regulations.gov (2009). Public Comment from Pioneer Hi

- Bred (Natalie Hubbard, Director, Biotechnology, North America). http://www.regulations.gov/search/Regs/home.html#documentDetail?R=0900006480a6ec3a

Terés, S., Barceló

- Coblijn, G., Benet, M., Álvarez, R., Bressani, R., Halver, J.E. and Escribá, P. V. (2008). Oleic acid content is responsible for the reduction in b lood pressure induced by olive oil. Proc Natl Acad Sci U S A 105, 13811

– 13816.

Vossen, P. (2007). International Olive Council (IOC) and California Trade Standards for Olive Oil. University of California, Cooperative Extension. http://ucce.ucdavis.edu/files/filelibrary/2161/34496.pdf

Two comments state that GE soybeans are “dangerous” and that they are “noxious weeds and not food”

APHIS response: As summariz ed in the EA ( Sectio n V

. B. A.) , the products of these GE soybeans are as safe and nutritious as those currently available in the marketplace. Soybean as a crop is not weedy (EA Section III A ) and the modified Pioneer 305423 soybean according to APHIS ’ assessment is also not weedy ( EA Section V. B.B

. )

. The 305423 soybean has been field tested since 2002 in the major soybean growing regions of the continental United States. Under field permits granted by USDA APHIS

, Pioneer conducted comprehensive agronomic perform ance and e cological observations for 305423 soybean in replicated, multi

- site field studies . In 2005 and 2006 growing seasons, Pioneer made observations and measurements of the soybean variety in a total of 13 North American locations

. Characteristics such as emer gence, seedling vigor, plant height, lodging, days to maturity, shattering, seed weight, yield, disease incidence and insect damage were measured. Data for s eed germination and dormancy were also collected in laboratory experiments. From these observatio ns and data submitted in the applicant ’ s petition , APHIS has conclude d in its plant pest risk assessment (USDA 2009) that there are no weed risk issues associated with Pioneer 305423 ( See EA Section V B

. B

. of the EA).

### 4 USDA (2009). APHIS BRS Website. Pet itions for Non

- regulated Status. http://www.aphis.usda.gov/biotechnology/not_reg.html

Two comments stated that genetic engineering methods are less precise and differ from conventional breeding methods, and thus may have unintended harmful effects ; one s tated he had concerns about genetically modified organisms and that they should be treated differently than those plants altered through traditional breeding methods.

APHIS response. The insertion of transgenes is an imprecise process, in the sense that one cannot predict the insertion site within the host genome. Some unintended changes to the plant genome may result and may lead to undesired consequences, but plant breeders place considerable importance in avoiding negative effects on agronomic perform ance and overall health of the transformed plants. Potential unintended changes that might affect plant phenotype include insertions of the transgene into active host genes, insertion of multiple tandem copies of the transgene, and rearrangements of genet ic material at the insertion site. Other genetic changes may occur at locations distant from the insertion site of the transgene of interest, either as a result of the insertion of transgene fragments or deriving from the culture process accompanying the transformation process.

As Pioneer describ ed within the petition (p. 23, Pavely , 2007, Petition for the Determination of Nonregulated Status for High Oleic 305423 Soybean ) , agronomic properties of the plants were assessed over several generations, then backcrossed to elite, well

- performing soybean lines a total of six times

. B ackcrossing with elite lines dilutes the presence of incidental genetic changes not closely linked to the transgene within the transgenic soybean and confers improved agronomic
pro perties to the newly transformed line. As a result of backcrosses , the expect ed transgene is obtained with in a final stable and predictable soybean line that has a reduced likelihood of additional, unexpected genes and traits (see page 20, Figure 2

, P avel y, 2007 ).

Inadvertent, genetic changes closely linked to the site of insertion for the gene of interest would not be so easily removed by backcrossing. For instance, an inserted transgene could conceivably interrupt a gene directing a plant function (“ insertional mutagenesis”) . In many cases however, plants have multiple genes that provide the same function, such as the multiple FAD

- 2 genes of soybean that form linoleic acid (Graef et al.

, 2009). Likewise, plants also may have multiple structural prot eins that contribute to the same plant parts (Barker, et al., 2008). If insertion sites do interrupt essential genes, the resulting plants would most likely be unhealthy and consequently be eliminated by developers.

T he process of transformation c ould potentially lead to new genetic changes by somatic mutation, which may occur spontaneously during a procedure for regenerating plant tissues, which is typically part of the transgenesis process. However, the developer’s normal process of selection, and th en hybridization with conventional, non transgenic lines , along with final product development may also e liminate most of these unintended mutations. For those genetic changes that result in noticeable traits, breeders can assure removal of undesirable tr aits from future product releases.

APHIS requires that any notable changes in the expected plant phenotype, or its agronomic properties, and modifications of pest susceptibility be identified, both after it is released under APHIS permit and later, as a condition of its deregulation. F inally, developers conduct repeated tests to ensure the reproducibility of the trait, as well as its negligible impact on overall plant health . APHIS regulatory actions provide oversight of new genetically modified plants , and the oversight is extensive compared to those for new traits in conventionally bred plants.

### 5 Conventional breeding techniques to gain new traits are not necessarily more precise than biotechnological methods that transpose novel genes into the plant g enome. Conventional seed breeders may gain new material for the host plant by crossing with other species or even genera and by subjecting plants to chemical or radiation mutagenesis . Through much trial and error, the host may acquire genes from the intr ogressed plant genus or species, or by mutation, but these may incorporate regulatory, structural or functional genes not identified or completely understood. The genetic material deriving from “wide crosses” may include large segments of chromosomes, or be associated with translocations or inversions (Osborn et al. 2007)). The unexpected random genetic changes from these crosses and also from mutations may be more extensive than those changes introduced by transgenic technology (Batista et al. 2008).

Reference s

.
Barker, M.S.

, Kane, N.C.

, Matvienko, M.

, Kozik, A.

, Michelmore, R.W.

, Knapp, S.J. , and Rieseberg , L.H. (2008). Multiple paleopolyploidizations during the evolution of the compositae reveal parallel patterns of duplicate gene retention after millions of years. Molecular Biology and Evolution , 25, 2445

- 2455

Batista

, R

. , Saibo

, N

. , Lourenco

, T

. , and Oliveira

, M.M. (2008). Microarray analyses reveal that plant mutagenesis may induce more transcriptomic changes th an transgene insertion. Proceedings of the National Academy of Sciences. 105: 3640

- 3645



### Graef, G., LaVallee, B.J., Tenopir, P., Tat, M., Schweiger, B., Kinney, A.J., Van Gerpen, J.H., and Clemente, T.E. (2009)

. A high

- oleic

- acid and low

- palmitic

- acid soyb ean: agronomic performance and evaluation as a feedstock for biodiesel

. Plant Biotechnology Journal 7 , 411

– 421.

Osborn

, T.C., Kramer

, C

. , Graham

, E

. , and Braun

, C.J. (2007). Insights and Innovations from Wide Crosses: Examples from Canola and Tomato. Cro p Science. 47: S

- 228

- S

- 237.



### P avel y, C. (2007). Petition for the Determination of Nonregulated Status for High Oleic 305423 Soybean , revised version

. Pioneer Hi

- Bred International, Inc.

, Johnston, IA . Available USDA BRS website.

One commenter from a food company asserted that these soybeans will reduce use of pesticides.

APHIS response: Pioneer 305423 is not genetically engineered to be glyphosate tolerant (see EA Section V . B.B. ). W eed management practices for Pioneer soybean will not differ from t hose used in conventional soybean (see EA Section III .B. )

. Although s ulfonylurea tolerant traits are expressed in this Pioneer soybean, these will not be promoted for this Pioneer line; the tolerance

wa s only used for trait selection during product develo pment (see EA Section V).


## 3. Human Health Impacts

Foods derived from biotechnology should not require a different or higher standard of safety than that of conventional foods, but should be examined if appropriate on a case

- by

case basis for safety befo re commercialization.

### 6 APHIS response: Under the Coordinated Framework for biotechnology products, the FDA will continue to assess the safety and quality of G E foods , and the EPA will assess the safety of plant incorporated protectants and herbicide toler ant crop varieties (EA S ection I)

. Th e coordinated deliberative process should continue to reduce concerns by the public about the healthfulness and safety of these genetically engineered products.

One comment stated that b ecause long

- term environmental and health risks of G E crops have not been studied, and because soybean is a commodity crop and ubiquitous in many food products, G E varieties should be strictly regulated, especially soybean.

APHIS response: G E varieties

, including this Pioneer 305432 s oybean are subject to considerable scrutiny by the FDA for food safety and by the USDA for agricultural safety. The Coordinated Framework for Products of Biotechnology intends that both health risks and environmental impacts will be thoroughly analyzed by the appropriate federal agencies. Products with undesirable properties causing extensive impacts greater than those from non

- transgenic products of the same type could be identified by this assessment process

. If these products are not acceptable under the regulations of an agency within the USDA , they will not be granted non regulated status . The FDA is confident that Pioneer 305432 soybean is not materially different from soybean already grown, except for the altered fatty acid content (see FDA Consu ltation Document and Sect ion V.B.A. ). Based upon information and analysis presented in the petition, plant pest risk assessment (USDA 2009) , and EA, USDA

- APHIS has concluded that Pioneer 305432 soybean will have no impacts on the agricultural or natural e nvironment beyond those already arising from other commercial soybean varieties. Pioneer 305423 soybean has received extensive assessment from APHIS and FDA and based upon their assessments

, will likely have only beneficial impacts on stakeholders and the public

.

Some comments stated that GE soybeans are harmful because they may have as yet unrecognized adverse long term effects on human health

. APHIS response: Beginning in December 2006, Pioneer provided data to the FDA establishing the food safety an d nutrition al properties of this product . The FDA concurred with Pioneer’s determination that the 305423 soybean and downstream products from it are not materially different from commercial soybeans in composition, safety or any other relevant parameter ( FDA Consultation Document)

. I n animal feeding tests using mice and broiler chickens ( S ee studies cit ed in EA Section V . B.C. ) no short term impacts could be observed from consumption of Pioneer 305423 soybean. No evidence of nutritional deficiencies were found after analysis of proteins, fats (except for the intentionally changed fatty acids) or fiber ( EA Section V . B.C. ) . No differences were found in antinutrient content, when compared with average antinutrient content of similar commercial soybean vari eties.

Under FDA supervis ion of test s of food ingredients, the chemical structure of a proposed additive and its expected concentration within foods must be considered in order to assign a concern level; the level will determine which types of toxicity tests will be done (NRC, 2006; FDA, 2000) . For additives of low concern, short term feeding studies are required , and if of highest level of concern then lifetime studies and even some multi

- generation studies, or studies contingent on certain results bas ed on tests results from lower level s of concern are required (NRC, 2006)

. If new food components are rated as high risk to consumers, then successively more lengthy and demanding tests are required. FDA’s level of concern is related to hazard and risk and FDA has required of Pioneer the types and term s of analys es appropriate to risk for 305423 soybean.

References.

### 7 FDA (2000). Redbook 2000. Revised July 2007

. Guidance for Industry and Other Stakeholders Toxicological Principles for the Safety Assessme nt of Food Ingredients. http://www.fda.gov/Food/GuidanceComplianceRegulatoryInformation/GuidanceDocuments/Foo dIngredientsandPackaging/Redbook/default.htm

NRC (2006). Toxicity Testing for Assessment of Environmental Agents. Interim Report. Committee on Toxicity Testing and Assessment of Environmental Agents National Research Council of the National Academies. The National Academies Press. Washington, D.C.

One commenter asserts that G E crops have decreased nutritional value

. Another comment states that G E food should be separated from “natural”, non - genetically modified food.

APHIS response: Under the Federal Food, Drug, and Cosmetic Act, the FDA issued a policy statement concerning genetically engineered food and animal feed . Following the announced procedure, FDA employs a consultation process for products derived from biotechnology to ensure that regulatory issues such as food and feed safety and labeling concerns are addressed and resolved (see EA Section I

. ). Based upon this FDA consultation proc ess, c omparisons with commercial soybean show that Pioneer 305423 is equally safe and nutritious as other varieties currently marketed (FDA P remarketing C onsultation : see attachment to EA ) From FDA’s consultation on foods derived from this soybean

, and A PHIS’ assessment of the laboratory evidence provided by Pioneer (Pavely, 2007) and accompanying scientific literature , APHIS concludes that there is no scientific basis to require separate storage, marketing or handling of this variety and commodity soybea ns.

B ecause of the value

- added traits of this soybean, growers and processers will preserv e product identity to the point of sale (Pioneer, see Summary in Pavely, 2007) as is commonly done for many specialty oils (Lee and Herbek, 2004 ) , thus keeping Pio neer 305423 soybean separate from commodity soybean (see Petition amendment X

- XF

- 7, Potential impact on organic and conventional farming)

. The operational details for Identity Preserved programs with oilseed
crops are discussed in th is Response to Public Comments under “2. Agricultural Impacts.” O ils derived from Pioneer 305423 soybean will also be separately marketed from conventional soybean oils (Pioneer HiBred, See Summary in Pavely, 2007) , because these oils contain healthy fatty acids (such as the mo nounsaturated oil, oleic acid) for manufactured food products such as salad oils (See EA S ection I. and II. )

. Oleic acid is a natural constituent of a variety of foods, notably for example

, olive oil (55

- 83% oleic acid, Vossen, 20 1 0). Olive oil has been shown to be a health

- promoting dietary constituent, and the oleic acid component has been directly shown to be responsible for lower ing blood pressure ( Terés 2008)

.
Other non transgenic varieties of soybean also express increased oleic acid from extract ed oils (Fehr, 2007 ; Clemente and Calhoon, 2009) . Some of these have been in commercial production (J.J.G. Minot, Iowa State University Research Foundation, personal communication)

. Soybean expressing a nontransgenic trait to decrease linolenic acid has also been combined with a trait for increased oleic acids (>50% oleic; i.e., IA2028 available from Iowa State University Research Foundation )

. These non

- transgenic l ines of soybean with altered fatty acids have been derived from mutagenized soybean and

, h ighly selected lines

. Pioneer’s high oleic soybean varieties are not significantly different from these varieties derived by conventional means, and the non

transgenic soybeans have already been used commercially without reported environmental impacts (J.J .G. Minot, Iowa State University Research Foundation, personal communication)

### 8 Reference s

.
Clemente, T.E. and Calhoon, E.B. (2009). Soybean oil: Genetic approaches for modification of functionality and total content. Plant Physiology 11 : 1030

- 1040.



### Fe hr, W.R. ( 2007 ). Breeding for modified fatty acid composition in soybean

. Crop Science

, 47, ( SUPPL. DEC. ) S72

- S87

Lee, C and Herbek, J. (2004). Specialty Soybean Production and Management in Kentucky, University of Kentucky, Cooperative Agricultural Extension Service. AGR

- 182. http://www.ca.uky.edu/agc/pubs/agr/agr182/agr182.pdf

Terés, S

., Barceló

- Coblijn, G

., Benet, M

., Alvarez, R

., Bressani, R

., Halver, J .E., and Escribá, P .V. (2008). Oleic acid content is responsible for the reduction in blood pressure induced by olive oil

. Proceedings of the National Academy of Sciences 105 (37): 13811.

Vossen, C. (2010 download). International Oli ve Oil Council (I OOC) Trade Standard for Olive Oil. University of Californian Cooperative Extension, UC Davis, Sonoma County Extension. http://cesonoma.ucdavis.edu/hortic/pdf/iocc_standards_purity_grade.pdf

One commenter says that G E has caused greater amounts of disease s in consumers since the introduction of G E crops.
APHIS response : Although the rate of certain diseases or conditions in consumers may have indeed increased since the introduction of G E crops in the mid

- 1990’s (for example, see Branum and Lukacs, 2009), a trend cannot be attributed directly to use of any food item, including G E crops or their products. A n apparent increase may o ften result from increases in incidence reporting (Branum and Lukacs, 2009). Increased incidence of disease could also be attri buted to any number of environmental or medical causes. The Pioneer 305423 soybean does not contain any other constituent likely to enhance disease incidence. For example, measured content of antinutrients is not greater than conventional soybean (see E A Section V . B.C. ) , nor is the constitutive GM

- HRA protein a known human allergen (see EA Section V . Potential impact on nontarget organisms ) . Pioneer submitted th is data to the FDA who reviewed the conclusions to complete the FDA consultation. No conseq uences for other animals consuming products from this soybean line are expected on the basis of these tests, and none would be expected from the nature of the transformation in Pioneer 305423.

References:

Branum A

. M

., and Lukacs S

. L . (2009). Food allergy among children in the United States. Pediatrics. 124, 1549

- 55.


## 4. Animal Impacts

One comment suggests that gene transformation is highly mutagenic and could lead to multi

- organ toxicity and impacts on blood and immune systems. One commenter states that there are many animal studies that show a link between adverse health risks and

### 9 ingestion of G E foods an d asks for a moratorium on G E food approvals. One commenter says that animals fed with G E crops show acute signs of aging, and decreased fertility.

APHIS response : Based upon information and analysis presented in the petition, plant pest risk
assessment (USDA 2009), an EA and an FDA consultation process , APHIS has concluded that Pioneer 305423 soybean, and the foods and feeds derived from it are not materially different in safety, composition, or any other relevant parameter from soybeans now grown, marke ted, and consumed.

Pioneer 305423 soybean contains a modified acetolactate synthase gene

, which confers herbicide resistance for the purpose of selection during transformation (see EA Section I

. ). As part of the petition, Pioneer submitted studies show ing that mice fed with 2000 mg of purified GM

- HRA protein (derived from the Gm

- hra gene, a modified version of the soybean acetolactate synthase gene) per kilogram of body weight showed no acute toxicity. Expected concentrations of ingested GM

- HRA protein soybean grain are far lower, at 2.5 ng/mg, than the purified dosage used in the mice trial ( See Section VII C., Taveley, 2007 ). It is not likely that Pioneer 305423 soybean will attain the high concentration to which these mice were exposed (see EA Secti on V .B.C. ). A second transformation in Pioneer 305423 soybean does not produce a novel protein, but suppress es the conversion of oleic acid into linoleic acid by an endogenous gene normally present in soybean (see Summary in Tavely, 2007 )

. T h e sequence gm

- fad2

- 1 derived from an existing soybean gene, if expressed, would not have any novel nutritional impacts because it does not express a novel protein ( Tavely, 2007 )

. In addition, APHIS evaluated evidence submitted by Pioneer of a 42

- day study showing th at broiler chickens, when fed 305423 soybean, did not have statistically significant differences in mortality, weight gain, mortality

- adjusted feed efficiency, or carcass yields from those fed control (non

- GE) soybean (see Petition, page 8 to 9). As a res ult, Pioneer 305423 soybean should not adversely affect wildlife feeding in soybean fields planted to 305423 (see EA Section V . Potential Impact on Nontarget Organisms ). As far as the general assertion of transgenesis causing unhealthy mutations, the proc ess of tissue culture used to regenerate a plant from an undifferentiated single cell to a mature organism after transformation may cause some somatic mutations (see discussion above)

. Selection and removal of plants with undesirable agronomic characteris tics after transformation will ex clude such mutations in the final commercializ ed product (Filipecki and Malepszy 2006). The issue of mutagenesis by gene disruption ( insertional mutagenesis ) was discussed in the response to a comment above about the preci sion of transgenic methods used to improve crop plants. M.W. Ho and J. Cummins have asserted that certain types of promoters used in transgenesis may be mutagenic but this opinion has received no confirmation or support from molecular biologists (Hodgson, 2000)

. Evidence for impacts of transgenic foods on tissues, organs or blood, while asserted by a limited number of authors, is unlikely . For example, an initial report was shown to be inadequate, poorly controlled and further evidence was contradictory ( Royal Society, 1999; Chassy, 2002)

. Although Ewen and Pusztai 1999b endeavored to show pathology of rat intestinal tissue in rats fed lectin transformed potatoes, ( a line never seriously considered for commercialization or release) there are several late r studies that contradict these findings of toxicity or pathology for a substantial number of histological benchmarks (Hashimoto et al. 1999; Pusztai et al., 1999a ; Teshima et al., 2002; El

- Sanhoty et al., 2004)

. APHIS asserts from considerable evidence that GE plants in general do not have any general toxic properties (as noted above, and in the detailed studies submitted in support of all petitions to grant nonregulated status for GE crop varieties). L ectin s

are known to have toxicities when animal cel ls are exposed to them (Vasconcellos and Oliveira

### 10 2004) and if express ed in potatoes could possibl y induce pathologies, although even these were
not substantiated in the case of Ewen and Puszai (1999 ; see Royal Society, 1999)

.

Reference s

. Chassy, B.M. (2002). Food Safety Evaluation of Crops Produced through Biotechnology Journal of the American College of Nutrition 21, 166S

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.

El

- Sanhoty, R., El

- Rahman, A.A.A., and Bögl

, K.W. (2004). Quality and safety evaluation of genetically modified potatoe s Spunta with Cry V gene: compositional analysis, determination of some toxins, antinutrients compounds and feeding study in rats. Nahrung: Food

, 48

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Ewen, S.W.B. and Pusztai, A. (1999b) Effects of diets containing genetically modified potatoes
exp ressing Galanthus nivalis lectin on rat small intestine. The Lancet 354, 1353

- 1354

Filipecki, M

. and Malepszy

, S . (2006)

. Unintended consequences of plant transformation:a molecular insight

. J Appl Genet 47: 277

– 286

.

Hashimoto

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. , Ohkawa

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. , Ishige

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. , Kito

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. , Utsumi

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, K . (1999). Safety Assessment of Transgenic Potatoes with Soybean Glycinin by Feeding Studies in Rats. Biosci

. Biotechnol

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John Hodgson (2000). Scientists avert new GMO crisis. Nature Biotechnology 18, 13 doi:10.1038/71838

.

Palombo

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. , Bistrian

, B

. R

. , and Huang

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. S . (2000). Comparison of growth and fatty acid metabolism in rats fed diets containing equal levels of gamma

- linolenic acid from high gamma

- linolenic acid canola oil or borage oil. Lipids 9

, 975

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Pusztai

, A

. , Grant

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. , Bardocz

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. , Alonso

, R

. , Chrispeels

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. , Schroeder

, H

. E

. , Tabe

, L

. M

. , and Higgins

, T

. J

. V . (1999a). Expression of the Insecticidal Bean

- Amylase Inhibito r Transgene Has Minimal Detrimental Effect on the Nutritional Value of Peas Fed to Rats at 30% of the Diet. Journal of Nutrition

. 129

, 1597

- 1603.

Royal Society (1999) Review of Data on Possible Toxicity of GM Potatoes (Ref. 11/99). London: The Royal Soc iety

. http:// royalsociety.org/WorkArea/DownloadAsset.aspx?id=5890

Teshima

, R

. , Watanabe

, T

. , Okunuki

, H

. , Isuzugawa

, K

. , Akiyama

, H

. , Onodera

, H

. , Imai

, T, Toyoda

, M

. , and Sawada

, J . (2002). Effect of subchronic feeding of genetically modified corn (CBH3 51) on immune system in BN rats and B10A mice. Shokuhin Eiseigaku Zasshi. 43

, 273

- 9.



## 5. Economic Impacts

One comment asked whether nonregulated status for Pioneer 305423 was necessary if naturally occurring oleic acid can be found in other sources, su ch as grapeseed oil

.

APH IS response : Under the authority of 7 CFR part 340, APHIS has the responsibility for the safe development and use of genetically engineered organisms under the provisions of the Plant Protection Act. APHIS must respond to petiti oners that request a determination of the regulated status of genetically engineered organisms, including genetically engineered crop plants such as

### 11 Pioneer 305423 soybean. If a petition for nonregulated status is submitted, APHIS must make a determinatio n if the genetically engineered organism is unlikely to pose a plant pest risk.

For economic reasons, modified soybeans may provide the best solution for a food manufacturer’s needs as well as for grow ers’ crop choices. H igh oleic oils can be produced in other crops , such as sunflower (“NuSun”), safflower and canola (EA, Section V.B.B

. ) However, soybean oil is highly economical for use in food s , and s oybean production may be more profitable for grower s than either sunflower or canola in permissive growin g regions such as eastern North Dakota (Metzger, 2009).

Reference:

Metzger, S. (2009) ND2010 Projected farm cash flow by crop, NDSU Carrington Research Extension Center, http://www.ndfarmmanagement.com/2010BEP121509.xls

One trade organization said that APHIS has a responsibility to consider scientific data assessing the risks to food functionality, risk mitigation and risk responsibility in its review of all special use traits like 305423.

APHIS response : APHIS ’ pest risk assessment for non

- regulated status does not require that risks to food functionality or risk mitigation for inappropriate product use be considered ( See 7 CFR part 340.6)

. For products developed using nonregulated GE organisms

, APHIS does not have the statutory authority to mandate that product developers be assigned r isk responsibility or provide compensation for adverse consequences to injured parties resulting from the use of a nonregulated GE organism (See Title 4. Plant Protection Act, June 20, 2000)

. A GE organism is no lon ger subject to the regulatory requirements of 7 CFR part 340 when APHIS determines that it is unlikely to pose a plant pest risk.

As appropriate, APHIS does consider potential food and industrial use impacts in their NEPA documents. The EA prepared fo r this petition and response to comments above

, notes that the ratios of the typical soybean fatty acids have been altered by this Pioneer 305423 line, and that inadvertent entry of Pioneer 305423 into the commodity soybean supply stream is possible. Howe ver, b ecause of the value

- added traits of this soybean, growers and processers will preserve product identity to the point of sale (See Petition Amendment: X

- XF

- 7, Potential impact on organic and conventional farming) , thus keeping Pioneer 305423 soybean s eparate from commodity soybean . The methods that are chosen to prevent oilseed mixing into the commodity stream of soybean seed is discussed in Section 2 , Agricultural Impacts, of this Response to Comments document

. APHIS supports the initiatives of ind ustry associations in addressing issues of risk mitigation and responsibility

. APHIS understands that the biotechnology industry consortium, BIO

, is currently discussing stewardship plans with their constituent biotech companies that would address policie s

on these types of issues. When complete, the steward ship plans would likely assist the industry in addressing issues associated with risks to food functionality, risk mitigation and risk responsibility.

One food processing trade organization said th at any precommercial release must include commercial assurances for downstream stakeholders relative to comingling of the product into the supply chain.

### 12 APHIS response : APHIS does not have regulatory authority or responsibility for validating the specif ic properties of a commodity nor mandating commercial assurances. Pioneer in public comments to APHIS (2007

- 0156

- 0045 ) plans to establish industry standard oilseed IP management procedures, and encourage use of analytic techniques to substantiate oil iden tity O il derived from this variety may be beneficial for many downstream users, but may not be optimal for all users (See Pioneer public comment Docket No. APHIS

- 2007

- 0156, December 21, 2009, “Identity Preserved Systems” and “Comingling Implications)

. M arketing a greements put in place by product developers may influence acceptance and use of these oils by soybean industry stakeholders. Although many adverse impacts following widespread use of oil deriving from Pioneer 305423 soybean are not likely, i ncr eased surveillance may be warranted in specific instances ( See Pioneer public comment Docket No. APHIS

- 2007

- 0156, December 21, 2009, “Comingling Implications ” ) to detect novel fatty acid ratios of commodity soybeans and in soy oil

. For example, characteri stics of product stability, taste, and texture depend on fatty acid ratios (Warner et al., 1988). Distinction between input soybean varieties would be a necessary part of manufacturer procedure when multiple types of soybean fatty acid contents are in the marketplace. Soybean varieties can be distinguished by scanning cotyledons for differences in certain lipid classes using scanning spectroscopy ( with F ourier Transformed Infra Red techniques (F TIR ) ) , a fast and straightforward assessmen t (Caires, 2008).

To alleviate concerns surrounding inadvertent mixing of Pioneer 305423 high oleic soybean with soybeans in the commodity stream, the Grocery Manufacturers Association ( GMA; Jeff Barach, personal communication, 2009 ) ha s indicated that its constituent memb ers desire more information about use and functionality of the high oleic oil. The GMA expects that s ome of the technology providers of seeds with altered fatty acid oils will commit to extensively compar ing physical properties of inadvertent oil mixtures deriving from the altered product and the commodity oil

. By informing producers of the oil’s compatibility with existing food processing and manufacturing procedures , this testing could avert po tential economic losses sustained by producers.

References :

Ca ires, A.R.L

. , Teixeira, M.R.O., Suarez, Y.R. , Andrade, L.H.C. and Lima, S.M. (2008). Discrimination of transgenic and conventional soybean seeds by F ourier transform infrared photoacoustic spectroscopy. Applied Spectroscopy, 62

, 1044

- 1047.

Warner, K.

, Orr

, P. and Glynn, M. (1988). Effect of altered fatty acid composition on soybean oil stability. Journal of the American Oil Chemists' Society 65, 624

- 628.

Some comments expressed concerns that w ithout rigorous marketing agreements, or stewardship plans from technology companies, and a “higher lev el of commercial responsibility,” commercial disruptions in the export market are possible, and these concerns need to be addressed if ‘pre

- commercial release” is the chosen production and marketing pattern . P recommercial release is the commercial producti on of a product which does not have international regulatory acceptance as a novel G E crop, such as within EU countries or Japan. Since some countries maintain a zero threshold for unapproved products, a high level of marketing policy measures, and strong enforcement and compliance with the policy is necessary for a pre - commercial release program to be successful

. APHIS response: APHIS does not have regulatory authority or responsibility for validating the specific properties of a commodity nor mandating commercial assurances for purchasers or

### 13 distributors of nonregulated products

. However, b ecause this soybean variety expresses value

added traits

, growers and processers will voluntarily preserve product identity to the point of sale, thus keeping Pionee r 305423 soybean separate from commodity soybean ( S ee Pavely , 2007, Pioneer Petition A mendment : X

- XF

- 7)

. While undesired mixing of this specialty oilseed with standard commodity soybean oil is possible, several market forces will minimize its occurrence. Pioneer has committed itself to using mechanisms similar to industry standards to maintain product identity, which have “established effectiveness” for maintain ing the integrity of the export and domestic market for soybean oils (Pioneer Public Comment APH IS 2007

- 0156

- 0045 ). Furthermore, APHIS supports the initiatives of industry associations in addressing issues of risk mitigation and responsibility as part of corporate stewardship of special crop production

.

Some comments note d that Pioneer has propos ed taking responsibility for the product beyond point of sale for seed, but that “further dialogue” and commitment to sharing of information for all stakeholders will be necessary throughout the lifecycle of the product. Three trade associations expressed concerns about the details of a Pioneer management plan to mitigate the risk of G E soybeans comingling with commodity soybeans and being sold abroad where GE is not accepted

. APHIS response: APHIS understands that the industry association

, Biotechnology Industry Organization ( BIO ; http://www.bio.org/ ) , has encouraged a common stewardship approach by all companies producing GE crop varietie s , especially when properties of a standard food commodity are altered by a new trait

. Pioneer may choose to partici pate in this stewardship approach for Pioneer 305423 soybean

. APHIS has no role in these discussions , and advises stakeholders to work closely with these providers and BIO, who are both committed to the success of these products in domestic and export mar kets

. Another comment addressed the possible unintended impacts on crop uses or processing streams because of higher oleic acid, lower linoleic and linolenic acid content in the modified soybean oil deriving from this soybean variety.

APHIS response: As described in the EA, this product improves the qualities of soybean oil, decreasing the need to hydrogenate the soybean oil, which produces unhealthy trans fats

. Pioneer has indicated that the primary use of Pioneer 305423 soybean will be for food oil as well as for industrial oil product s (See P avely, 2007 , Section I .B

. , Benefits of 305423 Soybean). B ecause of the value

- added traits of this soybean, growers and processers will preserve product identity to the point of sale, thus keeping Pioneer 305423 soybean separate from commodity soybean ( S ee Taveley, 2007, Pioneer Petition A mendment : X

- XF

- 7, and Section I.B., P avely, 2007 )

. Pioneer in public comments to APHIS (2007

- 0156

- 0045 ) plans to establish industry standard oilseed IP management procedures, and encourage use of analytic techniques to substantiate identity of constituent fatty acids

. The high oleic soybean would also be useful for deriving environmentally friendly, renewable, and cost effective industrial oil s , such as lubricants and biodiesel oi ls (see Summary and I.B. in Taveley, 2007 )

. Increased oleic acid at the expense of linoleic and linolenic would improve cetane numbers (time between injection into a cylinder and autoignition ; Refaat, 2009)

, and improve lubricity because of higher number s of unsaturated fatty acids and other factors (see I

- B. Benefits of 305423 Soybean in Pavely , 2007 and Refaat, 2009 )

. To determine which fatty acid ratios are useful for specific products, a use

- by

- use analysis would be required for each industry employin g soybean oils

. Pioneer notes that “Oil functionality considerations for [high oleic soybean] are the same as those for existing modified fatty acid oilseed crops, such as high oleic canola and mid/high oleic sunflower oil ( Pioneer public comment Docket N o. APHIS

2007

- 0156, December 21, 2009 ).”

### 14 Reference:

Refaat, A.A. (2009). Correlation between the chemical structure of biodiesel and its physical properties. Int. J. Environ. Sci

. Tech. 6, 677

- 694.

Three comment er s desired Pioneer to establish whether it will cover the costs of regulatory interceptions if these soybeans were comingl ed and then exported to countries in which the events we re not approved

. APHIS responds: No requirement exists that APHIS should enforce mitigation standards for users of the technology, nor provide for consequences of their failure. This type of action is outside APHIS regulatory authority. Although s ome commenters s uggest that Pioneer will cover costs of trade disruption caused by the possible presence of the new fatty acid soybean in export supplies , APHIS makes no interpretation of intention and takes no position on appropriateness of this policy . If Pioneer agree s to comprehensive product liability, they will do so without APHIS guarantee s or support for the guarant ee

. One commenter suggests that large agribusinesses monopolize seed production, and force farmers to use G E seeds.

APHIS response

. As far as APHIS can determine, while large seed companies sell limited lines of non

- transgenic soybean seed, some nontran sgenic lines are available and can also be provided by smaller suppliers and state university foundations

. Despite buyers ’ displeasure at the high technology fee s of G E seed

, they continue to purchase such G E seed , because growers benefit from reduced pro duction expenses and require fewer farm operations for growing soybean

. Based on USDA survey data, adoption of genetically engineered herbicide

- tolerant (HT) soybeans increased from 17 percent of U.S. soybean acreage in 1997 to 68 percent in 2001 and 92
p ercent in 2008 (Fernandez

- Cornejo, 2008). As Traxler and Falck

- Zepeda show ( 1999; for G E cotton), the re are benefits to both producers and technology providers alike

. Substantial competition exists for s oybean seed and for differing oil product s , includi ng conventional soybean varieties that may serve many of the marketplace needs

.

Reference s

.

Fernandez

- Cornejo, J. (2008) Adoption of Genetically Engineered Crops in the U.S. (Data Sets). USDA

- Economic Research Service. http://www.ers.usda.gov/Data/BiotechCrops/ (Accessed 1/4/2009)

.
Traxler, G., and Falck

- Zepeda, J. (1999). The distribution of benefits from the introduction of transgenic cotton varieties. AgBioForum 2, 94

- 98.

Several commente rs said that US growers of soybean have been losing extensive soybean

acr e age to foreign vegetable oil production because US manufacturers have needed to replace hydrogenated soybean oil in food products, following imposition of FDA labeling requirements. The proposed soybean line would provide an economic benefit to US soybean producers.

Pioneer 305423 will help soybean growers produce a desirable oil for food manufacturers ( S ee EA Section I . Introduction ) , and potentially recapture market share that was previously lost (Balvanz, C., Iowa Soybean Association, public comment on APHIS 2007

- 0156

- 0018)

. When

### 15 FDA deemed the desired functionality of hydrogenated soybean oil a health risk , food manufacturers turned to foreign food grade oils (for example, incr eased use of palm oils: ( Unnevehr and Jagmanaite, 2008 ; Soyatech, 2010) )

. High oleic oils such as Pionee r 305423 will have features of increased oil stability and useful blending properties (Warner and Gupta, 2005) and could replace imported oils such as palm oils for the necessary functional and quality properties needed by manufacturers

.

Reference s :

Soyatech (2010 download). Transfat facts. Soyatech LLC, Maine. http://www.soyatech.com/trans_fats.htm

Unnevehr, L.J. and Jagmanaite, E. (2008) Getti ng rid of trans fats in the US diet: Policies, incentives and progress

. Food Policy 33

, 497

– 503

. Warner, K. and Gupta, M. (2005). Potato chip quality and frying oil stability of high oleic acid soybean oil. J. Food Sci. 70. Supplement. Sensory and Nutrit ive Qualities of Food. S395

- S400.



## 6. Process Comments

One comment stated that USDA is not adequately assessing the environmental impact of the genetically modified soybean.

APHIS response. APHIS carefully considered the possible environmental impact s of the proposed product, and is satisfied that the EA developed for Pioneer 305423 soybean is adequate and sufficient . The EA follows all applicable laws, regulations, and guidelines in analyzing potential impacts of this action, including those establi shed by NEPA. In making an informed decision of potential environmental impacts, APHIS used the best available scientific information, data and expert advice.

One comment stated that an environmental impact assessment is needed.

APHIS response: 

### APHIS has determined that the analysis in its EA showed no significant impact on the quality of the human environment if APHIS was to grant the petitioner’s request to deregulate Pioneer 305423 soybean and that APHIS did not have to prepare an environmental imp act statement (EIS). The EA took a hard look at the need for action, the issues, alternatives, and environmental consequences. APHIS also reviewed the assessment of plant pest risk for Pioneer 305423 soybean and carefully considered all comments submitte d by respondents to the public involvement efforts. As a result of this analysis, APHIS prepared a final EA, from which came the NEPA decision document and a find ing of no signification impact ( FONSI) that discussed, under each of the t en Council for Envi ronmental Qua lity (CEQ) points of significance, why ea ch point was not significant, and why an EIS was not required. The agency followed CEQ NEPA regulations and Agency NEPA implementing procedures.