Pioneer Hi - Bred International, Inc. Page 30 of 34 Study Num ber: PHI - 2012 - 183 Appendix I i EPA B IOPESTICIDES AND POLLUTION PREVENTION DIVISION COMPANY NOTICE OF FILING FOR PESTICIDE PETITIONS PUBLISHED IN THE FEDERAL REGISTER EPA Biopesticides and Pollution Prevention Division contact: [insert name and telephone number with area code] SUBMISSION: Email the completed template to: hollis.linda@epa.gov . TEMPLATE: Pioneer Hi - Bred International, Inc. [Insert petition number] EPA has received a pesticide petition ( [insert petition number] ) from Pioneer Hi - Bred International, Inc.(DuPont Pioneer) , 7100 NW 62 nd Avenue, P.O. Box 1000, Johnston, Iowa, 50131 requesting, pursuant to section 408(d) of the Federal Food, Drug, and Cosmetic Act (FFDCA), 21 U.S.C. 346a(d), to amend 40 CFR part 180 (Option s (pick one) 1. by establishing a tolerance for residues of - NA - Remove 2. to establish an exemption from the requirement of a tolerance for - GM - HRA 3. to establish an amendment/expansion of an existing tolerance exemption for the NA - Remove (Options (pick one ) 1. m icrobial pesticide NA - Remove 2. biochemical pesticide NA - Remove 3. plant - pesticide inert GM - HRA in or on soybean. Pursuant to section 408(d)(2)(A)(i) of FFDCA, as amended, DuPont Pioneer has submitted the following summary of information, data, and argumen ts in support of their pesticide petition. This summary was prepared by DuPont Pioneer and EPA has not fully evaluated the merits of the pesticide petition. The summary may have been edited by EPA if thePioneer Hi - Bred International, Inc. Page 31 of 34 Study Num ber: PHI - 2012 - 183 terminology used was unclear, the summary contained extraneous material, or the summary unintentionally made the reader conclude that the findings reflected EPA’s position and not the position of the petitioner. I. DuPont Pioneer Petition Summary [Insert petition number] A. Product Name and Proposed Use Pra ctices

GM - HRA is a protein obtained from a genetically modified endogenous soybean gene, als I , that provides tolerance against specific classes of herbicides allowing it to be used as a selectable herbicide - tolerant marker in soybean transformation event s. As part of a genetic construct introduced into a plant’s genome, GM - HRA itself does not have insectical activity and is therefore functionally inert as part of a plant - incorporated protectant. B. Product Identity/Chemistry 1. Identity of the pesticide a nd corresponding residues. The gene encoding the GM HRA protein is designated gm - hra because it is derived from Glycine max and is an herbicide - resistant allele. The herbicide - tolerant gm - hra gene was made by isolating the herbicide - sensitive soybean gm - a ls I gene and introducing changes in the DNA sequence that result in two specific amino acid changes in the protein. These amino acid substitutions are analogous to those in herbicide - tolerant tobacco (Creason and Chaleff, 1988; Lee et al., 1988), and add itionally represent commonly found natural tolerance mutations. Ultimately, this two - amino acid modification of the endogenous GM - ALS I protein to GM - HRA protein yields an enzyme that is insensitive to ALS - inhibiting herbicides, allowing its use as a sele ctable marker during transformation and potentially a herbicide - tolerant trait in soybeans. 2. Magnitude of residues at the time of harvest and method used to determine the residue. Because an exemption from the requirement of a tolerance is requested for GM HRA protein as expressed in soybean, this section of the petition is not applicable. However, ranges of GM - HRA protein expression in events containing the gm - hra gene are quite low. 3. A statement of why an analytical method of detecting and measurin g the levels of the pesticide residue are not needed. Because an exemption from the requirement of a tolerance without numerical limitation is requested for GM - HRA protein as expressed in soybean, this section of the petition is not applicable . C. Mammalian Toxicological Profile 1) The donor organism is soybean ( Glycine max .), which has a long history of human use. The GM - HRA protein is only two amino acids different from the endogenous soybean ALS I protein from which it was derived (Volume 3, PHI - 2012 - 020). ALSPioneer Hi - Bred International, Inc. Page 32 of 34 Study Num ber: PHI - 2012 - 183 enzymes have also been part of the human diet by virtue of their presence in soybeans. 2) Bioinformatic analyses revealed GM - HRA to be similar to other ALS proteins found in bacteria, fungi, algae and plants (Friden et al., 1985; Falco et al., 1985; R eith and Munholland, 1995; Mazur et al., 1987). 3) There was no evidence of acute toxicity in mice at a target dose of 2000 mg purified protein preparation per kg of body weight (equivalent to approximately 436 mg of full - length GM - HRA protein per kg of body weight) (Volume 4, PHI - 2006 - 008). 4) Bioinformatic analyses revealed no biologically relevant amino acid sequence identities between known or putative protein allergens and the GM - HRA protein sequence. These data indicate the lack of both amino acid identi ty and immunologically relevant similarities between the GM - HRA protein and known protein allergens (Volume 5, PHI - 2006 - 070/070). 5) No biologically relevant amino acid sequence identities were observed between known protein toxins and the GM - HRA protein seq uence (Volume 6, PHI - 2006 - 071/070). 6) The GM - HRA protein is rapidly digested in both simulated gastric and intestinal fluids (less than 30 seconds in simulated gastric fluid containing pepsin at pH 1.2 as demonstrated by SDS - PAGE analysis and Western blot, and less than 30 seconds in simulated intestinal fluid containing pancreatin at pH 7.5 as demonstrated by SDS PAGE/Coomassie analysis or less than 1 minute as demonstrated by Western blot analysis) (Volume 7, PHI - 2006 - 199; Volume 8 , PHI - 2006 - 074). 7) The G M - HRA protein is heat labile (i.e., heating at 50 °C for 15 min completely inhibits the enzyme activity of the GM - HRA protein) (Volume 9 , PHI - 2006 - 135). 8) The plant - expressed GM - HRA protein is not glycosylated as demonstrated by glycoprotein staining (Volum e 2, PHI - 2011 - 225). 9) As part of two other transgenic soybean events (DP - 3Ø5423 - 1 and DP - 356Ø43 - 5), the GM - HRA protein has been the subject of safety reviews by both USDA and FDA (Rood et al., 2006; Pavely et al., 2007; US - FDA, 2007a; US - FDA, 2009a). Neith er agency identified safety concerns with the protein (USDA - APHIS, 2008; USDA APHIS, 2010; US - FDA, 2007b; US - FDA, 2009b). Additionally, a safety conclusion regarding GM - HRA exists in the scientific literature published by Mathesius et al (2009). 10) Based on the identical nature of the gm - hra cassette used in 82117 soybean and the two other transgenic soybean events expressing the GM - HRA protein (DP - 3Ø5423 - 1Pioneer Hi - Bred International, Inc. Page 33 of 34 Study Num ber: PHI - 2012 - 183 and DP - 356Ø43 - 5), expression ranges and dietary exposure of GM - HRA from 82117 soybean are expected to be similarly low. D. Aggregate Exposure 1. Dietary exposure. Expression ranges and dietary exposure calculations for GM HRA as expressed in DP - 3Ø5423 - 1 and DP - 356Ø43 - 5 are very low. Based on the identical genetic elements used in 82117 soybean, GM - HRA expr ession and subsequent exposure are expected to be similarly quite low. i. Food. Expression ranges and dietary exposure calculations for GM - HRA as expressed in DP - 3Ø5423 - 1 and DP - 356Ø43 - 5 are very low. Based on the identical genetic elements used in 82117 soybean, GM - HRA expression and subsequent exposure are expected to be similarly quite low. ii. Drinking water. While not likely, oral exposure at very low levels may occur through drinking water; however, a lack of mammalian toxicity has been demonstrate d. 2. Non - dietary exposure . Exposure to GM - HRA via inhalation or dermal contact is not likely as GM - HRA is contained within plant cells. E. Cumulative Effects There is no indication of mammalian toxicity potential for GM - HRA. It is therefore reasonable to conclude there will be no cumulative toxic effects for this protein. F. Safety Determination 1. U.S. population. Based on the available data for the GM - HRA protein, there are no indications of mammalian toxicological effects. GM - HRA was derived from a common dietary source (soybean), was not acutely toxic in mice, and the amino acid sequences of the GM - HRA protein exhibited no significant homology to the amino acid sequences of proteins known to be food allergens or toxins; and the protein is not glyc osylated. GM - HRA was also highly digestible in vitro , as well as thermally inactivated at 50  ̊C. Further, t he dietary risk for other transgenic soybean events expressing GM - HRA ( DP - 3Ø5423 - 1 and DP - 356Ø43 - 5) using genetic elements identical to those conta ined in 82117 soybean to drive GM - HRA expression, have been calculated and are quite low. 82117 soybean is expected to express GM - HRA at ranges comparable to other GM - HRA expressing events, to have a similarly low dietary risk, and is not anticipated to c ause any harm to the U.S. population, including any sensitive sub - populations or vulnerable age groups. 2. Infants and children . Based on the available data for the GM - HRA protein, there are no indications of mammalian toxicological effects. GM - HRA was d erived from a common dietary source (soybean), was not acutely toxic in mice, and the amino acid sequences of the GM - HRA protein exhibited no significant homology to the amino acid sequences of proteins known to be food allergens or toxins; and the protein is notPioneer Hi - Bred International, Inc. Page 34 of 34 Study Num ber: PHI - 2012 - 183 glycosylated. GM - HRA was also highly digestible in vitro , as well as thermally inactivated at 50  ̊C. Further, t he dietary risk for other transgenic soybean events expressing GM - HRA ( DP - 3Ø5423 - 1 and DP - 356Ø43 - 5) using genetic elements identical to those contained in 82117 soybean to drive GM - HRA expression, have been calculated and are quite low. 821 17 soybean is expected to express GM - HRA at ranges comparable to other GM - HRA expressing events, to have a similarly low dietary risk, and is not anticipated to cause any harm to the U.S. population, including any sensitive sub - populations or vulnerable ag e groups. G. Effects on the Immune and Endocrine Systems As required under FFDCA section 408(p), EPA has developed the Endocrine Disruptor Screening Program (EDSP) to determine whether certain substances (including pesticide active and other ingredients) may have an effect in humans or wildlife similar to an effect produced by a “naturally occurring estrogen, or other such endocrine effects as the Administrator may designate.” Between October 2009 and February 2010, EPA issued test orders/data call - ins fo r the first group of 67 chemicals, which contains 58 pesticide active ingredients and 9 inert ingredients. This list of chemicals was selected based on the potential for human exposure through pathways such as food and water, residential activity, and cert ain post - application agricultural scenarios. The GM - HRA protein is not among the group of 58 pesticide active ingredients on the initial list to be screened under the EDSP. H. Existing Tolerances There are no existing tolerances or exemptions from the r equirement of a tolerance for the GM - HRA protein and the genetic material necessary for its production. However, GM - HRA as encoded by the gm - hra gene has been previously evaluated for its safety by the U.S. Food and Drug Administration (FDA) and the U.S. Department of Agriculture (USDA) in two transgenic soybean events, one designed to offer high oleic soybean oil and one that offered glyphosate and ALS - inhibiting herbicide tolerance. Based on the information provided, FDA concluded that the safety profil es of the new soybean events including the GM - HRA protein were not materially different from that of other marketed soybean varieties, and FDA required no further premarket review or approval activities related to the events or the proteins. Upon its revi ew of information regarding these events and the GM - HRA protein, USDA deregulated both transgenic soybean events. I. International Tolerances International tolerances do not exist for this protein, however safety information related to the GM - HRA protein has been reviewed by international food safety authorities as part of the high oleic and herbicide - tolerant event submissions.