EPA BIOPESTICIDES AND POLLUTION PREVENTION DIVISION COMPANY NOTICE OF FILING FOR PESTICIDE PETITIONS PUBLISHED IN THE FEDERAL REGISTER

EPA Biopesticides and Pollution Prevention Division contact:

Michael Mendelsohn
Branch Chief
Microbial Pesticide Branch
Biopesticides and Pollution Prevention Division
Office of Pesticide Programs
U.S. Environmental Protection Agency
1200 Pennsylvania Ave. N.W.
Washington, D.C. 20460

SUBMISSION: E-mail the completed template to:

TEMPLATE:

J.R. Simplot Company
5369 W. Irving St.
Boise, ID 83706

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	EPA has received a pesticide petition ###### from J.R. Simplot Company, 5369 W. Irving St., Boise, ID 83706 proposing, 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 to establish an exemption from the requirement of a tolerance for the plant-pesticide, the BLB2 and AMR3 protein products of the Rpi-blb2 and Rpi-amr3 genes, respectively. Pursuant to section 408(d)(2)(A)(i) of FFDCA, as amended, J.R. Simplot Company has submitted the following summary of information, data, and arguments in support of their pesticide petition. This summary was prepared by J.R. Simplot Company and EPA has not fully evaluated the merits of the pesticide petition. The summary may have been edited by EPA if the 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. J.R. Simplot Company Petition Summary
   
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A. Product Name and Proposed Use Practices

	Gen 3 potatoes containing the BLB2 and AMR3 proteins, expressed from the Rpi-blb2 gene from Solanum bulbocastanum and the Rpi-amr3 gene from Solanum americanum, respectively, that confer protection against Phytophthora infestans, the cause of late blight disease, in potato.

B. Product Identity/Chemistry

	1. Identity of the pesticide and corresponding residues.

	The active ingredients that provide late blight protection are the BLB2 and AMR3 proteins produced from the Rpi-blb2 and Rpi-amr3 genes from wild Solanum species.

 Name: BLB2 (from the Rpi-blb2 gene) and AMR3 (from the Rpi-blb3 gene).
 Chemical Identity: Nucleic acid (including promoter, coding sequence, and terminator) and translated amino acid polypeptides.
 Composition: Potato events were generated by Agrobacterium-mediated plant transformation. Confirmation of gene insertion was carried out using PCR, Southern blot, and immunoblot methods based on the gene and protein sequences.

	2. Magnitude of residues at the time of harvest and method used to determine the residue.

	Less than 250 ppb for the BLB2 protein in tubers and less than 500 ppb for the AMR3 protein in tubers as determined by immunoblot detection. It is estimated that the active ingredients, BLB2 and AMR3, are less than 0.000025% and less than 0.00005% of potato, respectively.

	3. A statement of why an analytical method of detecting and measuring the levels of the pesticide residue are not needed.

	The BLB2 protein concentration is less than 250 ppb in tubers. As the expression levels of the BLB2 protein are below detection limits it is impractical to demonstrate methods for detecting and measuring the levels of the pesticide residues.

	The AMR3 protein concentration is less than 500 ppb in tubers. As the expression levels of the AMR3 protein are below detection limits it is impractical to demonstrate methods for detecting and measuring the levels of the pesticide residues.

C. Mammalian Toxicological Profile

	The potential hazard associated with each R-protein was assessed by evaluating i) the source of the gene, ii) the history of safe use of R-genes, iii) any toxic or allergenic potential using bioinformatics, and iv) the mechanism of action.

The weight-of-evidence supporting the minimal hazard associated with BLB2 includes:
::	The Rpi-blb2 gene is from wild potato, Solanum bulbocastanum, which has been used to cross late blight protection into cultivated potato (S. tuberosum) varieties through conventional breeding;
::	The BLB2 protein, encoded by Rpi-blb2, is present in two potato varieties (Toluca and Bionica) cultivated for food use in Europe;
::	BLB2 belongs to a large family of NBS-LRR R-proteins found throughout the plant kingdom;
::	There are hundreds to thousands of R-proteins in potatoes, which have a long history of safe consumption;
::	Bioinformatic analyses confirmed that BLB2 lacks sequence homology to known or putative allergens and toxins; and
::	Like other R-proteins, BLB2 functions as part of the plant immune response to prevent spread of P. infestans but does not directly affect the pathogen.

The weight-of-evidence supporting the minimal hazard associated with AMR3 includes:
::	The Rpi-amr3 gene is from the wild Solanum species, Solanum americanum, which is consumed by humans and used for both food and medicinally;
::	AMR3 belongs to a large family of NBS-LRR R-proteins found throughout the plant kingdom;
::	There are hundreds to thousands of R-proteins in potatoes, which have a long history of safe use and consumption;
::	Bioinformatic analyses confirmed that AMR3 lacks sequence homology to known or putative allergens and toxins; and
::	Like other R-proteins, AMR3 functions as part of the plant immune response to prevent spread of P. infestans but does not directly affect the pathogen.

D. Aggregate Exposure

	1. Dietary exposure.

	Acute consumption data were used to calculate conservative (i.e., high end) R-protein exposure estimates. The exposure estimate for BLB2 was 0.00076 mg/kg bw/d, and 0.0015 mg/kg bw/d for AMR3 and VNT1, for the total U.S. population. For children age 1-2, the exposure estimates were 0.002 mg/kg bw/d for BLB2 and 0.0041 mg/kg bw/d for AMR3 and VNT1.

This dietary exposure assessment was overestimated based on a number of conservative assumptions:
1)	The amount of each R-protein in tubers was assumed to be at the limit of quantification (250 ppb for BLB2 and 500 ppb for AMR3 and VNT1), which is greater than the actual amount;
2)	United States potato consumption data was used, which is higher than other global regions, except Europe;
3)	Potato consumption values of consumers at the 95th percentile were used;
4)	100% of the potatoes consumed in the diet were assumed to be Gen 3 potatoes; and
5)	There was no degradation of the R-protein during cooking and processing.
Even with conservative assumptions, the human dietary exposure to each of the three R-proteins in Gen 3 potatoes remains negligible.
	i. Food. 

	The BLB2 and AMR3 proteins are expressed in potato. Human exposure is expected from eating potato. The BLB2 and AMR3 proteins are not similar to any known allergens or toxins. The BLB2 and AMR3 proteins are expressed at low levels. The low hazard and low exposure minimizes risk associated with the BLB2 and AMR3 proteins.

	ii. Drinking water.

	Human exposure is not expected through drinking water.

	2. Non-dietary exposure.

	The BLB2 and AMR3 proteins are plant-incorporated protectants. Other uses where human exposure might occur are not intended or expected.

E. Cumulative Effects

	Human exposure to the BLB2 and AMR3 proteins from diet, food, drinking water, or other occupational or residential use is negligible.

F. Safety Determination

	1. U.S. population.

	The BLB2 and AMR3 proteins are not likely to pose risks to human health. Study reports assessing the safety of the BLB2 and AMR3 proteins demonstrate a non-toxic mechanism of action for all R-proteins, including BLB2 and AMR3, a history of safe use for homologous proteins in potato and other crops, and no significant sequence similarities between BLB2 and AMR3 with known allergens or toxins. The protein concentrations were shown to be less than 250 ppb and 500 ppb for BLB2 and AMR3 in tubers, respectively. A minimal hazard and negligible exposure associated with R-proteins indicates a low level of risk to human safety from consumption of potatoes expressing the BLB2 and AMR3 proteins. Therefore, Simplot is petition for an exemption from the requirement of a tolerance for the BLB2 and AMR3 proteins.

	2. Infants and children.

	The BLB2 and AMR3 proteins are unlikely to pose risk to infants or children.

G. Effects on the Immune and Endocrine Systems

	The BLB2 and AMR3 proteins are unlikely to cause effects on the immune and endocrine systems.

H. Existing Tolerances

	A permanent exemption from the requirement of tolerance has been established for the similar protein, VNT1, expressed from the Rpi-vnt1 gene (40 CFR §174.534). The exemption was established for residues of the VNT1 protein when used as a plant-incorporated protectant in potato on February 24, 2017 [82 FR 11515].

	There are no existing tolerances for the BLB2 or AMR3 proteins.

I. International Tolerances

	None.
