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CBI-Deleted Petition for the Determination of Nonregulated CBI-deleted Petition for the Determination of Nonregulated Status for MON 88017 Corn The undersigned submits this petition under 7 CFR Part 340.6 to request that the Administrator make a determination that the article should not be regulated under 7 CFR Part 340 Submitted by: Ravinder S. Sidhu, Ph.D. Monsanto Company 800 North Lindbergh Blvd. St. Louis, MO 63167 Submitted April 30, 2004 Revised, December 15, 2004 Monsanto Petition # 04-CR-108U OECD Unique Identifier: MON-88Ø17-3 Prepared by: Ravinder S. Sidhu, Ph.D. and Shazia Brown Contributors: Brenda M. Bader, Kim A. Beazley, Natalia N. Bogdanova, D.V.M., Heather K. Bonner, Ph.D., Nina S. Bhakta, Austin Burns, Ph.D., Brett Bussler, Carolyn W. Carrera, Timothy R. Coombe, Natalie DiNicola, Ph.D., Jian J. Duan, Ph.D., Melinda C. McCann, Mike Mckee, Ph.D., Thomas Nickson, Ph.D., Jay Pershing, Todd A. Pester, Ph.D., Elena A. Rice, Ph.D., Eric W. Rosenbaum, Niki K. Scanlon, Ty T. Vaughn, Ph.D. Revised Confidential Business Information-deleted Monsanto Company 04-CR-108U Page 1 of 277 CBI-deleted Release of Information Monsanto is submitting the information in this petition for review by the USDA as part of the regulatory process. By submitting this information, Monsanto does not authorize its release to any third party. In the event the USDA receives a Freedom of Information Act request, pursuant to 5 U.S.C., § 552 and 7 CFR Part 1, covering all or some of this information, Monsanto expects that, in advance of the release of the document(s), USDA will provide Monsanto with a copy of the material proposed to be released and the opportunity to object to the release of any information based on appropriate legal grounds, e.g., responsiveness, confidentiality, and/or competitive concerns. Monsanto expects that no information that has been identified as CBI, will be provided to any third party. Monsanto understands that a CBI-deleted copy of this information may be made available to the public in a reading room and by individual request, as part of a public comment period. Except in accordance with the foregoing, Monsanto does not authorize the release, publication or other distribution of this information (including website posting) without Monsanto's prior notice and consent. Monsanto Company 04-CR-108U Page 2 of 277 CBI-deleted Petition for the Determination of Nonregulated Status for MON 88017 Summary The Animal and Plant Health Inspection Service (APHIS) of the U.S. Department of Agriculture (USDA) has responsibility, under the Plant Protection Act (Title IV Pub. L. 106-224, 114 Stat. 438, 7 U.S.C. § 7701-7772) and the Plant Quarantine Act (7 U.S.C. § 151-167), to prevent the introduction and dissemination of plant pests into the United States. The APHIS regulation 7 CFR Part 340.6 provides that an applicant may petition APHIS to evaluate submitted data to determine that a particular regulated article does not present a plant pest risk and should no longer be regulated. If APHIS determines that the regulated article does not present a plant pest risk, the petition is granted, thereby allowing unrestricted introduction of the article. Monsanto Company is submitting this request to APHIS for a determination of nonregulated status for MON 88017 corn, any progeny derived from crosses between MON 88017 and other corn, and any progeny derived from crosses of MON 88017 with other genetically-enhanced corn that also has been granted nonregulated status under 7 CFR Part 340. MON 88017 plants, developed by the use of recombinant DNA techniques, are tolerant to the action of the Roundup® family of agricultural herbicides and are protected from damage caused by corn rootworm (CRW) larval feeding. MON 88017 produces a 5- enolpyruvylshikimate-3-phosphate synthase protein from Agrobacterium sp. strain CP4 (CP4 EPSPS), which confers tolerance to glyphosate, the active ingredient in Roundup agricultural herbicides, and a modified Bacillus thuringiensis (subspecies kumamotoensis) Cry3Bb1 protein that selectively controls CRW species. The U.S. Food and Drug Administration (FDA) and the U.S. Environmental Protection Agency (EPA) have previously reviewed the safety of the CP4 EPSPS and Cry3Bb1 proteins present in other biotechnology-enhanced corn products. A synopsis of these safety assessments is provided to the USDA in this petition. Corn (Zea mays L.) is the largest crop grown in the United States in terms of acreage planted and net value. In 2003, corn production covered 79.1 million acres that yielded 10.1 billion bushels and had a net value of $24.8 billion. Weed control is essential in cornfields, as weeds compete with the crop for sunlight, water and nutrients. Failure to control weeds results in decreased yields and reduced crop quality. Introduction of MON 88017 will offer U.S. farmers the ability to apply Roundup agricultural herbicides over the top of corn for broad-spectrum weed control with minimal risk of crop injury. In addition, the use of glyphosate in conjunction with MON 88017 will allow the grower to utilize reduced tillage techniques, which provide significant environmental benefits, such as reduced soil erosion, reduced use of fossil fuels, and improved soil quality. ® Roundup is a registered trademark of Monsanto Technology LLC. Monsanto Company 04-CR-108U Page 4 of 277 CBI-deleted Corn yields are also negatively impacted by a number of insect pests. One of the most pernicious in the U.S. corn belt is corn rootworm. Corn rootworm larvae damage corn by feeding on the roots, reducing the ability of the plant to absorb water and nutrients from soil, and causing harvesting difficulties due to plant lodging. Corn rootworm is the most significant insect pest problem for corn production in the U.S. corn belt from the standpoint of chemical insecticide usage, as over 14 million acres of corn were treated with organophosphate, carbamate, and pyrethroid insecticides to control corn rootworm in 2000. Corn rootworm has been described as the billion-dollar pest complex based on costs associated with the application of soil insecticides and crop losses due to pest damage. Introduction of MON 88017 will offer U.S. farmers a safe and effective alternative to the use of synthetic chemical insecticides for control of the corn rootworm pest. Corn hybrids containing both the corn rootworm protection and glyphosate tolerance traits have been developed previously by crossing two inbreds containing the individual traits by traditional breeding techniques to produce the stacked trait product. However, the traditional breeding process can be inefficient, requiring long development times. MON 88017 was therefore developed using a transformation vector that contained both the cp4 epsps and cry3Bb1 genes. This approach enables the simultaneous introduction of both traits into corn hybrids, thereby providing growers access to a variety of elite corn germplasms containing both traits. MON 88017 was produced by Agrobacterium-mediated transformation of corn cells with plasmid vector PV-ZMIR39. This plasmid contains a gene encoding the CP4 EPSPS protein that provides tolerance to the action of Roundup agricultural herbicides and a gene encoding the Cry3Bb1 protein that has activity against corn rootworm. The Agrobacterium tumefaciens transformation vector is a disarmed, binary vector that contains both left and right transfer-DNA (T-DNA) border sequences to facilitate transformation. The DNA region that was integrated into the corn genome during the transformation process contains the cp4 epsps and cry3Bb1 gene expression cassettes. Molecular analyses of MON 88017 confirmed that single copies of the cp4 epsps and cry3Bb1 genes are integrated at a single locus in the corn genome with all expression elements intact and no plasmid bacterial backbone present. Segregation analysis across ten generations confirmed the heritability and stability of the cp4 epsps and cry3Bb1 coding sequences. The results of this analysis are consistent with the finding of a single active site of insertion that segregates according to the Mendelian laws of genetics. The CP4 EPSPS protein in MON 88017 is structurally homologous to EPSPSs naturally present in food crops (e.g., soybean and corn) and in microbial food sources such as bakers yeast. The amino acid sequence of the CP4 EPSPS protein produced in MON 88017 is the same or >99% identical to the CP4 EPSPS protein produced in commercial Roundup Ready® crops, such as soybean, NK603 corn, cotton, and canola, which have been granted deregulated status by the USDA. ® Roundup Ready is a registered trademark of Monsanto Technology LLC. Monsanto Company 04-CR-108U Page 5 of 277 CBI-deleted A panel of analytical tests was used to characterize and confirm the identity of the CP4 EPSPS protein produced in MON 88017. An evaluation of the CP4 EPSPS protein based on its similarity to natural EPSPS proteins that are ubiquitous in the environment, lack of toxicity in laboratory investigations of Roundup Ready crops against nonpest and pest species, and no adverse effects to human or animal health indicates that the CP4 EPSPS protein produced in MON 88017 will have a no significant impact on the environment. Bacillus thuringiensis (B.t.) Cry proteins also have a long history of safe and widespread use. The Cry3Bb1 protein produced in MON 88017 is a member of the Cry3Bb class of proteins and shares >99% amino acid sequence identity with the wild type Cry3Bb1 protein contained in the topically applied commercial microbial product, Raven® Oil Flowable Bioinsecticide. The amino acid sequence of the Cry3Bb1 protein produced in MON 88017 is >99.8% identical (a difference of a single amino acid) to that of the Cry3Bb1 protein produced in YieldGard® Rootworm corn (MON 863), which has been deregulated by the USDA, completed the FDA consultation process, and been granted registration by the EPA. The Cry3Bb1 proteins have been exempted from the requirement of a tolerance in corn by the EPA.
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