1 

# Pioneer Hi-Bred International, Inc. Seed Production Technology (SPT)

Process DP-32138-1 Corn

OECD Unique Identifier:

DP-32138-1



## Final Environmental Assessment

May 2011

Agency Contact Cindy Eck Biotechnology Regulatory Services USDA, APHIS Riverdale, MD 20737 Phone: (301) 734-0667 Fax: (301) 734-8669 Cynthia.A.Eck@aphis.usda.gov

The U.S. Department of Agriculture (U SDA) prohibits discrimination in all its programs and activities on the basis of race, color, national origin, sex, religion, age, disability, political beliefs, sexual or ientation, or marital or family statu s. (Not all prohibited bases apply to all programs.) Persons with disabilitie s who require alternative means for communication of program information (Braille, large print, audiotape, etc.) should contact USDA’S TARGET Center at (202) 720–2600 (voice and TDD).

To file a complaint of discrimination, write USDA, Direct or, Office of Civil Rights, Ro om 326–W, Whitten Building, 1400 Independence Avenue, SW, Washington, DC 20250–9410 or call (202) 720–5964 (voice and TDD). USDA is an equal opportunity provider and employer.

Mention of companies or commercial products in this re port does not imply recommendation or endorsement by the U.S. Department of Agriculture over others not mentioned. USDA neither guarantees nor warrants the standard of any product mentioned. Product names are mentioned solely to re port factually on available data and to provide specific

information.

This publication reports research involving pesticides. All uses of pesticides must be regi stered by appropriate State and/or Federal agencies before they can be recommended.

### Table of Contents

2 List of Tables ................................................................................................................ ...................4

List of Figures ............................................................................................................... ...................4

Acronyms and Abbreviations .................................................................................................... ......5

I. Purpose and Need .............................................................................................................. .....7

Regulatory Au thority .......................................................................................................... .............. 7

Regulated O rganism s ........................................................................................................... ............. 8

Petition for Determination of Nonregulated Status: Pioneer 32138-1 Seed Production Technology (SPT) Maintainer ................................................................................................... ....... 8

Purpose of Produc t ............................................................................................................ ................ 9

APHIS’ Response to Petition for Nonregulated Status ................................................................ 10

Coordinated Fram ework Review .................................................................................................. . 10

Public Invol vement ............................................................................................................ ............. 10

II. Affected Environment .......................................................................................................... .12

Agricultural Produc tion of Corn ............................................................................................... ..... 12

Acreage and Areas of Corn Produc tion ................................................................................. 12 Cropping Prac tices ............................................................................................................ ..... 12

Seed Pro duction ............................................................................................................... ....... 13 Organic Fa rming ............................................................................................................... ...... 15 Specialty Corn Systems ........................................................................................................ ... 16

Physical En vironmen t .......................................................................................................... ........... 17

Water Res ources ............................................................................................................... ....... 17

Soil .......................................................................................................................... ................. 17 Air Quality ................................................................................................................... ............ 18 Climate Change ................................................................................................................ ....... 18

Animal and Plan t Communities .................................................................................................. ... 19

Animals ....................................................................................................................... ............. 19

Plants ........................................................................................................................ ............... 20 Biological Di versity .......................................................................................................... ....... 20

Gene Mo vement ................................................................................................................. ...... 20

Public H ealth ................................................................................................................. ................... 21

Socioec onomic ................................................................................................................. ............... 22

III. Alternatives .................................................................................................................. .........24

No Action: Continuation as a Regulated Article .......................................................................... 24

Preferred Alternative: Determination that DP -32138-1 is No Longer a Regulated Article ...... 24

Alternatives Considered but Reject ed from Further C onsiderat ion ............................................ 25

Approve the petiti on in part .................................................................................................. .. 25

Isolation Distance between DP-321 38-1 and Non-GE Co rn Production ............................ 26 Requirement of Testing For Event 3 2138 Corn ..................................................................... 26

### Table of Contents

3 Comparison of A lternatives .................................................................................................... ........ 26

IV. Environmental Consequences ...............................................................................................28

Scope of the Environm ental Anal ysis ........................................................................................... . 28

Other Assu mptions ............................................................................................................. ............. 28

Agricultural Production of Corn ................................................................................................... 29

Organic Production of Corn ................................................................................................... 29 Acreage and Areas of Corn Produc tion ................................................................................. 30 Cropping Practices: Crop Rotation, Tillage, and Pes ticide Use .......................................... 30

Seed Pro duction ............................................................................................................... ....... 32 Organic Fa rming ............................................................................................................... ...... 39 Specialty Corn Systems ........................................................................................................ ... 41

Physical En vironmen t .......................................................................................................... ........... 41

Water Resources ............................................................................................................... ...... 41

Soil .......................................................................................................................... ................. 42 Air Quality ................................................................................................................... ............ 44 Climate Change ................................................................................................................ ....... 45

Animal and Plan t Communities .................................................................................................. ... 46

Animals ....................................................................................................................... ............. 46

Plants ........................................................................................................................ ............... 47 Biological Di versity .......................................................................................................... ....... 48

Gene Mo vement ................................................................................................................. ...... 50

Public Heath .................................................................................................................. ................... 52

Human He alth .................................................................................................................. ....... 52 Worker Safety ................................................................................................................. .......... 54

Animal Feed ................................................................................................................... .......... 54

Socioeconomic Issu es .......................................................................................................... ........... 56

Domestic Economic Envi ronment at Risk .............................................................................. 56

Trade Economic Enviro nment at Risk .................................................................................... 58 Social Environment at Risk from Seed Production Activities ............................................... 59

Other Cumulativ e Eff ects ...................................................................................................... ......... 60

Threatened and Enda ngered Sp ecies ............................................................................................. 60

Consideration of Executive Orders, Standa rds and Treaties Relating to Environmental Impacts ....................................................................................................................... ...................... 62

Impacts on Unique Characteris tics of Geographi c Areas. ................................................... 63

International Implica tions..................................................................................................... .. 64 Compliance with Clean Water Act and Clean Air Act ........................................................... 64

National Historic Preservation Ac t (NHPA) of 1966 as Amended ....................................... 64

V. References .................................................................................................................... .........65

VI. Appendix A .................................................................................................................... .......77

VII. Appendix B .................................................................................................................... .......78

VIII. Appendix C .................................................................................................................... .......79

### Table of Contents

4 

### List of Tables

Table 1: Issues of potential impacts and consequences of alternatives ....................................... 28



### List of Figures

Figure 1. Outline of the SPT Process for Producing Hybrid Seed. .............................................. 35

Figure 2. Production of the 32138 SPT Maintainer Line (Figure 1, Step I) ................................ 36

Figure 3. Visualization of the Red Marker, DsRed2 Expressed by the SPT Cassette ................. 37

Figure 4. Production of the Non-Transgenic Female Sterile Inbred (Step II) . ............................ 375 

### Acronyms and Abbreviations

32138 SPT maintainer Maize line containing the DP-32138-1 event

32138 SPT insertion The DNA from plasmid PHP24597 that has been integrated into the genome of the 32138 SPT maintainer

AMS USDA Agricultural Marketing Service

AOSCA American Organization of Seed Certifying Agencies

APHIS Animal and Plant Health Inspection Service

BRS Biotechnology Regulatory Serv ices (within USDA–APHIS)

Bt Bacillus thuringiensis protein

CAA Clean Air Act

CFR Code of Federal Regulations (United States)

CH 4 methane

CO 2 carbon dioxide

CWA Clean Water Act

DP-32138-1 Event DP-32138-1, also known as 32138 SPT maintainer

DsRed2(Alt1) A specific fluorescent marker gene; a variant of Clontech’s DsRed gene

DsRed2 A specific fluorescent protein expressed by DsRed2(Alt1) gene

EA environmental assessment

EO Executive Order

EPA U.S. Environmental Protection Agency

FDA U.S. Food and Drug Administration

FFDCA Federal Food, Drug, and Cosmetic Act

FIFRA Federal Insecticide, Fungicide, and Rodenticide Act

FR Federal Register

GE genetically engineered

GHG greenhouse gas

IP identity preservation

IPCC Intergovernmental Panel on Climate Change

ISPM International Standard for Phytosanitary Measure

IPPC International Plant Protection Convention

LMO living modified organisms

Ms45 A specific (dominant) male-fertility gene in maize

MS45 A specific protein expressed by the Ms45 gene in maize

ms45 A mutant allele (recessive) of the Ms45 gene in maize6 

### Acronyms and Abbreviations (continued)

NMFS National Marine Fisheries Service

NO 2 nitrogen dioxide

N 2 O nitrous oxide

NEPA National Environmental Policy Act of 1969 and subsequent

amendments

NOP National Organic Program

NPC new protein consultation

OSTP Office of Science and Technology Policy

PRA pest risk analysis

R&D research and development

SPT Seed Production Technology

sq. ft. square feet

TES threatened and endangered species

TSCA Toxic Substances Control Act

U.S. United States

USDA U.S. Department of Agriculture

USDA-ERS U.S. Department of Agricultur e-Economic Research Service

USDA-FAS U.S. Department of Agricultur e-Foreign Agricu ltural Service

USDA-NASS U.S. Department of Agriculture- National Agricultural Statistics

Service

USDA-NOP U.S. Department of Agricult ure-National Organic Program

U.S.C. United States Code

USFWS U.S. Fish and Wildlife Service

ZM-AA1 A specific α -amylase protein expressed in 32138 SPT maintainer and comprised of maize endogenous protei n sequences from the Brittle-1 transit peptide and the ZM-AA1 protein.7 

### I. Purpose and Need

Regulatory Authority

“Protecting American agriculture” is the basic ch arge of the U.S. Department of Agriculture's (USDA) Animal and Plant Health Inspection Serv ice (APHIS). APHIS pr ovides leadership in ensuring the health and care of plants and animals. The agency improves agricultural productivity and competitiveness, and contributes to the national economy and the public health. USDA asserts that all methods of agricultural pr oduction (conventional, or ganic, or the use of genetically engineered (GE) vari eties) can provide benefits to the environment, consumers, and

farm income.

Since 1986, the United States government has regulat ed genetically engineered (GE) organisms pursuant to a regulatory framework known as the Coordinated Framework for the Regulation of Biotechnology (Coordinated Framework) (51 FR 23302, 57 FR 22984). The Coordinated Framework, published by the Office of Scie nce and Technology Policy, describes the comprehensive federal regulatory policy for ensu ring the safety of biotechnology research and products and explains how federal agencies will use existing Federal statutes in a manner to ensure public health and environmental safety wh ile maintaining regulatory flexibility to avoid impeding the growth of the biotechnology indus try. The Coordinated Framework is based on several important guiding principles: (1) agenci es should define those transgenic organisms subject to review to the extent permitted by their respective statutory authorities; (2) agencies are required to focus on the characteristics and risk s of the biotechnology pro duct, not the process by which it is created; (3) agencies are mandated to exercise oversight of GE organisms only when

there is evidence of “unreasonable” risk.

The Coordinated Framework explains the regulato ry roles and authorities for the three major agencies involved in regulating GE organism s: USDA’s Animal and Plant Health Inspection Service (APHIS), the Food and Drug Administra tion (FDA), and the Environmental Protection Agency (EPA).

APHIS is responsible for regulating GE organisms and plants under the plan t pest authorities in the Plant Protection Act of 2000, as amended (7 USC § 7701 et seq .) to ensure that they do not pose a plant pest risk to the environment.

The FDA regulates GE organisms under the author ity of the Federal Food, Drug, and Cosmetic Act. The FDA is responsible for ensuring the sa fety and proper labeling of all plant-derived foods and feeds, including those that are genetical ly engineered. To help developers of food and feed derived from GE crops comply with their obligations under Federa l food safety laws, FDA encourages them to participate in a voluntary consultation process. All food and feed derived from GE crops currently on the market in the Un ited States have successfully completed this consultation process. The FDA policy statemen t concerning regulation of products derived from new plant varieties, including those genetically engineered, was published in the Federal Register on May 29, 1992 (57 FR 22984-23005). Under this policy, FDA uses what is termed a consultation process to ensure th at human food and animal feed sa fety issues or other regulatory issues (e.g., labeling) are resolv ed prior to commercial dist ribution of bioengineered food.

The EPA regulates plant-incorpor ated protectants under the Federa l Insecticide, Fungicide, and Rodenticide Act (FIFRA) and ce rtain biological control organi sms under the Toxic Substances8 Control Act (TSCA). The EPA is responsible fo r regulating the sale, di stribution and use of pesticides, including pesticides that are produced by an organism through techniques of modern biotechnology.

Regulated Organisms

The APHIS Biotechnology Regulat ory Service’s (BRS) mission is to protect America’s agriculture and environment us ing a dynamic and science-base d regulatory framework that allows for the safe development and use of GE organisms. APHIS regulations at 7 Code of Federal Regulations (CFR) part 340, which were promulgated pursuant to authority granted by the Plant Protection Act, as amended (7 Unite d States Code (U.S.C .) 7701–7772), regulate the introduction (importation, interstate movement, or release into the environment) of certain GE organisms and products. A GE organism is no longe r subject to the plant pe st provisions of the Plant Protection Act or to the regulatory requirements of 7 CFR part 340 when APHIS determines that it is unlikely to pose a plant pest risk. A GE organism is considered a regulated article if the donor organism, reci pient organism, vector, or vector agent used in engineering the organism belongs to one of the taxa listed in the regulation (7 CFR 340.2) and is also considered a plant pest. A GE organism is also regulated under Part 340 when APHIS has reason to believe that the GE organism may be a plant pest or APHIS does not have information to determine if the GE organism is unlikely to pose a plant pest risk.

A person may petition the agency that a particular regulated article is unlikely to pose a plant pest risk, and, therefore, is no longer regulated under the plant pest provisions of the Plant Protection Act or the regulati ons at 7 CFR 340. The petitioner is required to provide information under § 340.6(c)(4) relate d to plant pest risk that th e agency may use to determine whether the regulated article is unl ikely to present a greater plant pest risk than the unmodified organism. A GE organism is no longer subject to the regulatory require ments of 7 CFR part 340 or the plant pest provisions of the Plant Protection Act when APHIS determines that it is unlikely to pose a plant pest risk.

Petition for Determin ation of Nonregulated Status: Pioneer 32138-1 Seed Production Technology (SPT) Maintainer

Pioneer Hi-Bred International, Inc. (Pioneer) of Johnston, IA has submitted a petition to APHIS seeking a determination that their “Seed Produc tion Technology” (SPT) co rn maintainer event (hereafter referred to as DP-32138- 1 is unlikely to pose a plant pest risk and, therefore, should no longer be a regulated article unde r regulations at 7 CFR Part 340.

According to Pioneer, DP-32138-1 is engineered to produce male sterile female inbred plants for the generation of hybrid corn seed that is non -transgenic (Pioneer 2009). As detailed in the petition, the carefully cont rolled expression of a seed color ma rker gene and pollen fertility and sterility genes allows for the gene ration of red transgenic seed fo r seed increase of male sterile female inbred lines. The multistep process yields a non-transgenic male-sterile female parent. This non-transgenic material can then be usedfor hybrid seed production (Pioneer 2009). DP32138-1 is currently regulated under 7 CFR part 340. Interstate movements and field trials of DP-32138-1 have been conducted under permits issued or notifications acknowledged by

APHIS.9 DP-32138-1 corn is currently regulated under 7 CFR Part 340. Interstate movements, importations, and field testing of DP-32138-1 co rn have been conducted under notifications acknowledged by APHIS.

DP-32138-1 has been field tested in the U.S. since 2005 as authorized by APHIS. Associated notifications acknowledged and permits issued by APHIS are listed in the petition (Pioneer 2009). The list compiles a total of 11 test sites in diverse regions of the U.S. including the major corn growing area of the Midwest and winter nur series in Hawaii and Pu erto Rico. Field tests conducted under APHIS oversig ht allow for evaluation in agricultural settings under confinement measures designed to minimize the likeli hood of persistence in the environment after completion of the field trial. Under confine d field trial conditions, data are ga thered on multiple parameters and used by applicants to evaluate agronomic char acteristics and product performance. These data are also valuable to APHIS for assessing the potential for a new variety to pose a plant pest risk. The evalua ted data may be found in the APHIS Plant Pest Risk Assessment (USDA-APHIS 2010).

Purpose of Product

Pioneer seeks determination of nonregulated stat us of DP-32138-1 corn that was developed to

facilitate a novel method of seed production to produce male sterile /female inbred plants for the generation of hybrid corn seed that is non-transg enic (Pioneer 2009). As de tailed in the petition, the carefully controlled expression of a seed color marker gene a nd pollen fertility and sterility genes allows for the generation of red transgenic seed for seed increase of male sterile female inbred lines that are used for non-transgenic hybrid commercial seed production. Typically, detasseling is needed in corn seed production, and confers substantial expense, lower seed yield and lower genetic purity. Use of DP-32138-1 would eliminate detasseling a nd lead to increased seed yield and higher genetic puri ty during seed increase operations . As detailed in the petition, the process predictably and reliably results in a commercial product wh ich does not contain the DP-32138-1 transgenes (Pioneer 2009).

Since the 1930’s, corn productivity in the U.S. has been greatly enhanced by the use of hybrid corn seed (Sleper and Poehlman 2006). Corn hybr ids are characterized by increased resistance to diseases and enhanced agronomic characteristics compared with the parental lines (Brewbaker 1964). The production of hybrid corn seed invo lves a cross between two inbred lines 1 , where the pollen from the tassel/male parent is used to fertilize the ear/female parent. Because corn is mostly self pollinated (OECD 2003), hybrid corn seed is typically produced by the removal of male flowers (tassels) from the female parent plant either mechanically or by hand. These methods of detasseling reduce yields and are expe nsive. Mechanical detasseling may result in up to 40 % reduction in seed yield compared to hand detasseling treatments (Wych 1988). Additionally, female plants may escape detasse ling or develop secondary tassels after manual detasseling resulting in female plants that are se lf-pollinated. Thus, the seed of female inbred plants is harvested along with the hybrid seed resulting in redu ced genetic purity and lower seed yield for the final hybrid variety.

To overcome the expense, yield reduction and reduced genetic pur ity associated with detasseling, numerous genetic strategies have been attempted to achieve male sterility 2 as alternative

1 Inbred lines are populations of identical or nearly identi cal plants used as stocks fo r the creation of hybrid lines.

2 Male sterility is the inability to produce functional pollen.1 0 approaches to detasseling (Skibbe and Schnable 2005). Over 40 genetic elements are associated with male sterility; some of which are locate d on the nuclear chromosomes while others are located on the mitochondria chromosomes (Ski bbe and Schnable 2005). A major drawback to male sterile genetic approaches is the difficulty in generating male steril e inbred lines, because no functional pollen is produced in a male sterile plant line.

APHIS’ Response to Petition for Nonregulated Status

Under the authority of the plant pest provision s of the Plant Protection Act and 7 CFR Part 340, APHIS has issued regulations for the safe development and use of GE organisms. As required by 7 CFR 340.6, APHIS must respond to petitione rs who request a determination of the regulated status of GE organisms, including GE plants such as DP-32138-1. When a petition for nonregulated status is submitted, APHIS must make a determinat ion if the GE organism is unlikely to pose a plant pest risk. If APHIS de termines based on its Plant Pest Risk Assessment (PPRA) that the genetically engineered organi sm is unlikely to pose a plant pest risk, the genetically engineered organism is no longer subj ect to the plant pest provisions of the Plant Protection Act and 7 CFR part 340.

APHIS has prepared this environmental assessment (EA) to consider the potential environmental effects of an agency determination of nonre gulated status consistent with Council of Environmental Quality’s (CEQ ) regulations implementing NEPA (40 CFR Parts 1500-1508, 7 CFR 1b, and 7 CFR Part 372), and the USDA and APHIS NEPA implementing regulations and procedures. This EA has been prepared in order to specifically evaluate the effects on the quality

of the human environment 3 that may result from the deregulation of DP-32138-1 corn.

Coordinated Framework Review

DP-32138-1 is not designed for human and animal consumption. Discarded parent seed, whether

it is outdated or discarded from the color sort er, and seed byproducts, such as husks and cobs, will be disposed by placing in landfills, compos ting, or incineration (Pioneer 2009). According to Pioneer policy, seed producers will not feed discard corn to animals (Hubbard, 2011 - Public Comment on EA for DP-32138-1). However, an imals may accidentally gain access to corn fields, to discarded corn seed or by-products and therefore may al so be subject to regulation by FDA. A new protein consultation (NPC) for th e DsRed2 protein color marker was submitted to FDA on October 11, 2006 with the follow up le tter of January 29, 2010 received from FDA (Appendix A). The DsRed2 protein is the only non-corn protein in DP-32138-1. A NPC for the ZM-AA1 protein, normally found in germinating corn seeds, was submitted to FDA on June 18,

2009. The FDA considers Pioneer's consultation on ZM-AA1 alpha-amylase protein to be complete (Appendix B). Because DP-32138-1 does not contain any GE pesticides or the genetic machinery necessary to produce them, or tolera nce to herbicides, EPA consultation is not required.

Public Involvement

APHIS-BRS routinely seeks public comment on dr aft environmental asse ssments prepared in response to petitions for nonregul ated status of GE organisms. APHIS-BRS does this through a

3 Under NEPA regulations, the “human environment” incl udes “the natural and phys ical environment and the relationship of people with that environment” (40 CFR §508.14).11 notice published in the Federal Register. The i ssues discussed in this EA were developed by considering public concerns as well as issues raised in public comments submitted for other environmental assessments of gene tically engineered organisms, c oncerns raised in lawsuits, as

well as those issues that have b een raised by various stakeholders . These issues, including those regarding the agricultu ral production of corn using various production methods, and the environmental and food/feed safety of genetically engineered plants were addressed to analyze the potential environmental impacts of DP-32138-1.

This EA, the petition submitted by Pioneer, and AP HIS’s Plant Pest Risk Assessment, will be available for public comment for a period of 60 days (7 CFR § 340.6(d)(2)). Comments received by the end of the 60-day period will be reviewed and used to inform APHIS’s determination decision of the regulated status of DP-32138-1cor n and to assist APHIS in determining whether an Environmental Impact Statement is required prior to the determination decision of the regulated status of this corn line.

Issues Considered

As stated above, the issues considered in this EA were developed based on APHIS’ determination to deregulate certain genetically en gineered organisms, and for this particular EA, the specific deregulation of DP-32138-1 for seed production.

Management Considerations:

 Acreage and Areas of Corn Production

 Cropping Practices

 Seed Production

 Organic Farming

 Specialty Corn Production

Environmental Considerations:

 Water Use

 Soil

 Air Quality

 Climate Change

 Animals

 Plants

 Biological Diversity

 Gene Movement

Public Health Considerations:

 Human Health

 Worker Safety

Socioeconomic Considerations:

 Domestic Economic Environment

 Trade Economic Environment

 Social Environment1 2 

### II. Affected Environment

Agricultural Production of Corn

Acreage and Areas of Corn Production

Corn ( Zea mays L.) is the world's most widely grown ce real, reflecting its ability to adapt to a wide range of production envir onments (Morris 1998). Corn is an annual plant typically grown

in zones of abundant rainfall and fertile soils (Morris 19 98). In the U.S., the moisture levels and number of frost-free days required to reach mat urity are ideal for corn to be grown within temperate regions. Corn varietie s having a relative maturity of 100 to 115 days are typically grown in the U.S. Corn Belt, which include s Iowa, Illinois, Nebraska, and Minnesota. Approximately 50% of all corn grown in the U.S. is from these four states. The Corn Belt also includes parts of Indiana, South Dakota , Kansas, Ohio, Wisconsin, and Missouri. Approximately 80% of all corn acres grown in th e U.S. are within these top ten corn growing states; however, corn is grown in all states of the U.S. except Alaska (USDA-NASS 2010a).

Conventional farming, as defined in this do cument, includes any farming system where synthetic pesticides or fertilizer s may be used. This definition of conventional farming includes the use of genetically engineered varieties that have been deregulated by APHIS. Conventional farming covers a broad scope of farming practic es, ranging from farmers who only occasionally use synthetic pesticides and fe rtilizers to those farmers whos e harvest depends on regular pesticide and fertilizer inputs.

U.S. corn production for 2009, including production of non-genetically engineered and genetically engineered corn varieties, was 13. 2 billion bushels from 79. 6 million harv ested acres (USDA-NASS 2009a). In 2007 and 2008, growers harvested 13.0 b illion bushels of grain from

86.56 million acres and 12.1 billion bushels from 78.6 million acres, respectively. Of the total corn acres planted in 2009, 85% were GE co rn varieties (USDA-NASS 2009a) up from 80% in 2008 (USDA-NASS 2009a), 73% in 2007 (US DA-NASS 2009a), and 61% in 2006 (USDANASS 2009a).

Cropping Practices

Today, growers can choose from h undreds of corn hybrids markete d by companies that produce seed including GE varieties (NCGA 2010). Pion eer, the petition applicant, itself offers over 250 varieties (Pioneer 2010), which in turn implies the total number of corn hybrids available industry wide to be approximately 800-1000 since Pioneer has approximately a 30% market share (Pioneer 2008). Another estimate of total nu mber of hybrids available for the industry is 4000 (Monsanto 2010).

Hybrids differ generally in agronomic characteris tics, including disease and pest resistance and length of growing period. The optimum planting date for corn, usually between April and May, is influenced by factors such as the locality, environmental conditions, seed growing period, and seed variety. Harvesting generally occurs fr om mid-to-late September through November. The use of a combine (mechanical harvesting) is th e standard practice for grain production (Olson and Sander 1988).13 Crop rotations (successive planting of different cr ops on the same land) are used to optimize soil nutrition and fertility, and reduce pathogen loads (H oeft, Nafziger et al. 2000). Crops used in rotation with corn vary regionally, but there has been an increase in the number of fields that have a corn-to-corn rotation, as opposed to ro tation to another crop (Erickson and LowenbergDeBoer 2005). In some areas, the corn-to-corn ro tation requires increased levels of fertilizer inputs (Sawyer 2007). Insect pest s may also increase in corn-to- corn rotations as this system may provide a continual host environment for so me insects and diseases. However, in a cornsoybean rotation, continuously growing corn for multiple growing seasons can decrease populations of soybean pests, such as soybean cy st nematode (Hoeft, Nafziger et al. 2000).

Corn production typically involves the extens ive use of agronomic inputs and technology (Rooney and Serna-Saldivar 1987; Shaw 19 88; Pollak and White 1995; White and Pollak 1995), and the main emphasis is placed on obtain ing the best yield (Thomas 2007). Depending on the region and practices used, corn productio n includes inputs such as fertilizer (e.g., synthetic fertilizers, manure, a nd compost containing nitrogen and phosphorus) and pesticides (synthetic or NOP-approved insec ticides, herbicides and fungicides ), as well as irrigation. Each of these inputs can affect segm ents of the environment includi ng, but not limited to, waterways by increases in nutrient pollution, biodiversity b ecause pesticide inputs cause species changes, the water table because of excessive irrigation pr actices or productive fields because irrigation increases salinity (Hoeft, Nafziger et al. 2000).

Weed control methods differ depending on a nu mber of factors including locality, grower resources, and crop trait; the techni ques may be direct (e.g. mechanical 4 and chemical 5 ) or indirect (e.g. cultural 6 ) (Olson and Sander 1988). Pest control (weeds and insects) in corn production is essential in order to obtain good crop yield (Olson and Sander 1988; Hoeft, Nafziger et al. 2000). Generally, growers will manage a range of pests simultaneously. Therefore, growers will likely chose from a number of techniques to effectively and efficiently manage pests in their fields. In 2005, the most prevalent pest management practice was pesticide use (USDA-ERS 2005). Ultimately, the managemen t practices utilized by a grower will depend on the types of pests in their field, level of in festation, cropping system, type of soil, cost, weather, time, and labor. Practices to cope w ith pests, nutrient n eeds, and moisture and temperature requirements vary regionally. All agricultural production systems, including corn production, may affect the surrounding environment.

Seed Production

Once a new variety has been developed by the plant breeder, seed providers need to increase seed so that commercial fields can be planted by farmers that wish to take advantage of new traits. This increase is necessary for all varieties of any plant species, including corn. The increase starts with a single seed, a single plant or a handful of seed. In the case of corn, the seed of all varieties combined must be able to plant over 90 million acres (USDA-NASS 2009a) each year. Since seed companies typically overproduce by 30-40% to assure enough seed for the

4 Mechanical techniques include tillage and mowing.

5 Chemical techniques include herbicide application.

6 Cultural techniques include crop rotation/spot spraying of herbicide/hand removal of weeds

.1 4 coming year, about 50-60 million bushels of seed are needed to meet the projected seed needs of the next year (Wych 1988). The precise number of bushels of seed needed for a given year is not known until all fields have been planted. A num ber of factors may influence the number of final planted acres including: weather, anticipated future commodity prices of corn and competing crops, changing goals for the amount of ethanol to be needed in future years, final crop yields in

the U.S. and other countries that are harvested in the year the seed is produced, changing goals of government programs such as the Conservatio n Reserve Program, etc. (Farnham 2001; Nielsen and Thomison 2002; Brock 2007; Schnitkey 2008; Casselman 2010; Ortiz 2010; UK 2010).

Corn fields in the U.S. are generally planted with hybrid seed because hybrid vigor allows for maximizing grain yield. Beginning in the 1920’s, when the first corn hybrids were introduced commercially, and continuing for 40-60 years, h ybrids were generally double crosses (Wych 1988). If seed of double cross hybrids were used to plant 90 million acres, the 50-60 million

bushels of seed could be harvested from about 500,000 acres since the seed would be harvested from female lines that are single cross hybrids. Th ese 500,000 acres would have to be detasseled or some other method to control fertile pollen production on these female plants (Wych1988).

Starting in the late 1950’s, the hybrids transitioned from double crosses to single crosses. Currently, single crosses are used almost ex clusively (Wych 1988). This transition to single crosses occurred because the single crosses out-yielded double crosses and the farmers demanded the higher yielding single crosse s (Wych1988). To produce 50-60 million bushels of single crosses, the seed is harvested from female parents that are inbred lines. Since inbred lines produce only about 40-60% of the single cross hybri d, then about one million acres would be needed to produce the seed for the 90 million acres of corn grain production. These one million acres of seed production (Pioneer 2009) would have to be detasseled or some other method used to control fertile pollen production on these female plants. To plant these one million acres, about 400,000-500,000 bushels of inbred seed woul d be needed and would be produced on about 10,000-20,000 acres (Pioneer 2009).

A new hybrid variety could potentially be released in about 2-4 years after the inbred lines have been developed. Since inbred lines are maintain ed and reproduced using self pollination, these 10,000-20,000 acres do not have to be detassele d. These 10,000-20,000 acres of seed increase of inbred lines are subdivided by the estimated 800-4,000 conventional corn hybrids with about 5- 15 acres per variety. To plant these 5-15 acre fiel ds requires only 2-5 bushels of seed, which only requires less than 0.1 acres (Pioneer 2009) to pro duce this seed. To produce enough seed to plant these 0.1 acres requires only 2-4 ears from 2-4 plants of the promising inbred line. With good planning and use of favorable growing locations, su ch as winter nurseries in Hawaii and Puerto Rico, 2-3 generations of corn can be grown in one calendar year.

The production of hybrid seed requires stringen t control over the pollen sources. The utilization of hybrid vigor was practical in corn since th e male portion (tassel) of the corn plant was completely separate at the top of the plant from the female portion (ear) in the central region of the plant. In order to assure that no self po llination occurs and obtain hybrid seed in corn, the tassel must be removed (detasseled) from th e seed producing plant (female) while allowing nearby “male” plants with tassels to pollinate th e female plants. Tassels are removed by hand or by machine, but both methods are expensive and result in major plant injury with accompanying lower seed yields. In 1988, detasseling costs ranged from $100 to $13 0 per acre (Wych 1988). Much of the cost of detasseling is personnel costs. Assuming that detasselers earn approximately minimum wages and based on minimu m hourly wage rates of $3.35 in 1988 and1 5 $7.25 in 2009 (US-DOL 2010), it can be estimated that 2009 detasseling costs range from $216 to $281 per acre. Yield reduction due to detasseli ng has been estimated to range from 1.5% to 13.5% for hand detasseling and 2%-45% for mech anical detasseling (Wyc h 1988). To estimate cost of yield losses from detasseling, severa l assumptions can be made. For the production of any seed that requires detasseling, contracts are signed with farmers willing to grow the low yielding inbred plants along with the challenges of growing corn to exact planting, growing and harvesting standards and schedules. In return for these apparent disadvantages of growing seed corn, paying premiums and minimums are expect ed and standard parts of the contract. The minimum standards are likely to be equivalent to the returns that would be expected from growing conventional corn, which would be base d on the average yields in the area and the commodity corn price (Wych 1988). For example, the average yield might be 170 bushels with a commodity price of $4.00 per acre [yields and prices based on a pproximate national averages for 2009 (USDA-NASS 2009a) ] making the minimum standard $680 per acre. A 10% yield reduction due to detasseling is approximately $68 pe r acre. From the calculations here, total costs of detasseling (cost of detasseling plus yield reduction) may be in the range of $280 to $350 per

acre.

To minimize or to overcome impacts and high co sts of conventional detasseling, various means of inducing male sterility have been explored including cytoplasmic male sterility, genic male sterility, and chemically induced male sterility. The cytoplasmic male sterility system, which is the only male sterility system that is widely used throughout the indu stry, tends to have problems with reduced restoration to male fer tility in the commercial field (Wych 1988). The genic male sterility system is complicated, time consuming a nd expensive (Wych 1988). The chemically induced male sterility systems generally had problems with insufficient male sterility or with intolerable levels of female sterility (Wych 1988). Resear ch has been conducted over the last 20 years to develop useful male sterility systems (Albertsen, Fox et al. 1993; Greenland, Bell et al. 1997; Unger, Betz et al. 2001; Unger, Cigan et al. 2002; Havey 2004). Six transgenic male sterility systems – Petition Nos. 95-228-01p, 97-148-01p, 97-342-01p, 98-278-01p, 98- 349-01p and 01-206-01p – have been deregula ted by APHIS (APHIS-BRS 2010) and none of these systems, other than the cytoplasmic male st erility system, have been widely adopted in commercial hybrid corn seed production (Pioneer 2009).

Organic Farming

Organic farming, as defined in this document, includes any production system that falls under the USDA National Organic Program (NOP) defin ition of organic farmi ng and is a certified organic production system. The National Organic Program is administered by USDA’s Agricultural Marketing Service (AMS). Orga nic farming operations, as described by the National Organic Program, require organic produ ction operations to have distinct, defined boundaries and buffer zones to prevent uninte nded contact with excl uded methods from adjoining land that is not under organic management. Organic production operations must also develop and maintain an organic production syst em plan approved by thei r accredited certifying agent. This plan enables the production operati on to achieve and document compliance with the National Organic Standards, including the prohib ition on the use of excluded methods. Excluded methods include a variety of methods used to ge netically modify organisms or influence their growth and development by means that are not possible under natural conditions or processes. In organic systems, the use of synthetic pesticides, fertilizers, and genetical ly engineered crops, such as DP-32138-1 corn, is strictly limited.1 6 An accredited organic certifying agent conducts an annual review of the certified operation’s organic system plan and makes on- site inspections of the certified operation and its records. Organic growers must maintain records to sh ow that production and handling procedures comply with USDA organic standards. Practices growers may use to exclude genetically engineered products include planting only organic seed, planting earlier or later than neighboring farmers who may be using GE crops so that the crops will flower at different times, and employing adequate isolation distances betw een the organic fields and the fields of neighbors to minimize the chance that pol len will be carried between the fields. Although the National Organic Standards prohibit the use of ex cluded methods, they do not require testing of inputs or products for the presence of excluded me thods. The presence of a detectable residue of a product of excluded methods alone does not nece ssarily constitute a violation of the National Organic Standards (USDA-AMS 2010). The uninten tional presence of the products of excluded methods will not affect the status of an organi c product or operation when the operation has not

used excluded methods and has taken reasonable steps to avoid contact with the products of excluded methods as detailed in their approved or ganic system plan. Organic certification of a production or handling operat ion is a process claim, not a product claim.

Certified organic corn acreage is a small perc entage of overall corn production. The most recently available data show 143,000 acres of organ ic corn production in 2007. This is 0.17% of the total 86.2 million acres of corn planted in 2007 (USDA-ERS 2008). In 2005, 131,000 acres of organic corn were planted resulting in a 9% increase in organic corn acreage over the 2 year period from 2005 to 2007 (USDA-ERS 2010a).

Specialty Corn Systems

The vast majority of corn grown in the U.S. is grown as grain for animal feed, ethanol, and industrial uses (USGC 2008). Ho wever, approximately 8% of co rn grown is specialty corn produced for a specific market or use (USGC 20 06). Examples of specialty corn include white corn, waxy corn, hard endosperm, high oil, no n-GE, and organic corn (USGC 2006). The uses of these corn varieties include human consumpti on, food processing (w axy corn) or specialty product (white corn, blue corn). These corn vari eties are specified by buyers and end-users of corn for production, and premiums are paid for delivering a product that meets purity and quality standards for the corn variety. Product differen tiation and market segmentation in the specialty corn industry includes mechanisms to keep tr ack of the grain (tra ceability) for identity preservation (IP) and quality assurance processes (e.g., ISO9001- 2000 certification), as well as contracts between growers and buyers that speci fy delivery agreements (Sundstrom, Williams et al. 2002). Systems used by specialty corn growers and end-users to maintain identity of the production includ e:

 Contracts – written agreem ents detailing responsibilities and duties of both parties, including premiums for reaching goals and penalties for failing to attain specifications.

 Tracking and Traceability System s – correct labeling of all pr oducts (planting seeds and harvested material) and testin g procedures for identifying an d detecting acceptability of

materials.

 Quality Assurance Processes – oversight on handling procedures , testing of planting seeds, and testing of harves ted materials to determine ac ceptability of us e and product requirements, and assuring testing procedures are appropriate.1 7  Closed-Loop Systems – end-user supplies the planting seeds and guarantees to purchase final products. May also include that the end- user conduct intermediate procedures such as planting, provide oversight during the growi ng season, harvesting a nd transportation to processing plant.

 Identity Preservation Systems – us ing systems of identity preser vation that have been shown to be successful in the past, such as the se ed certification systems conducted by members of the Association of Official Seed Certifying Agencies (AOSCA). To maintain the purity of the corn product, this production system is based on controlling, tracking and documenting each step from seed production to end use (processing plants).

Physical Environment

Water Resources

Corn plants use a substantial amount of water du ring growth. About 4,000 gallons of water are needed to produce 1 bushel of corn (NCGA 2007) , and in 2008, 12.1 bill ion bushels of corn were harvested in the U.S (USDA-NASS 2010a). In 2008, about 12.0 million corn acres were irrigated, which is approximately 15% of all corn acres planted for grain and the rest of corn acreage only receive the ambient rainfall in a given area (USDA-NASS 2008b). Since inbred lines are not as vigorous as the hybrid corn plan ts and the seed of the inbreds is more valuable that commercial corn grain, irrigation is used more frequently to lower the risk of drought induced seed reduction for growing inbred plan ts compared to the nearby commercial grain fields (Pioneer 2009).

Soil

Disturbance and exposure of the top soil surf ace layer by certain agronomic practices causes impacts and may leave crop soils prone to de gradation (Hoeft, Nafziger et al. 2000). Two environmental impacts of soil degradation are th e decline in water quality and the contribution to the greenhouse effect (Lal and Bruce 1999). A decline in soil quality and soil resilience 7
enhances the greenhouse effect through emissions of radiatively-active gases 8 (carbon dioxide (CO 2 ), nitrous oxide (N 2 O)) and depletion of the soil carbon pool (Lal 2003; US-EPA 2010). In turn, a decrease in carbon aggregation and sequestration in the soil leads to increased runoff and

soil erosion.

Because inbreds are less vigorous than hybrids, greater care is taken in preparing the seed bed to provide optimum germinating conditions for the inbr ed seed. This usually means that the soil is tilled more frequently, possibly resulting in more carbon loss and exposing the soil to the possibility of increased erosion. In the production of seed corn, when detasseling is required, severe damage to the soil structure can occur f rom the use of detasseler carriers and mechanical detasselers under wet soil conditions, resulting in poor water drainage, increased root damage and ineffective nutrient uptake resulting in lo wer seed yields (Kok et al.1996; Daum 1996)

7 Soil resilience is the ability of a soil to restore itself.

8 Radiatively-active gases are gases that absorb incoming solar radiation or outgoing infrared in turn, affecting the temperature of the atmosphere.1 8 Indirect effects of detasseling, such as soil co mpaction, could also impact the subsurface soil environment. The corn root system is closely as sociated with several mi crobial groups such as bacteria, fungi, protozoa, and mites (Bais, Weir et al. 2006) and probably modifies the ecology because of these associations. Bacteria typically represent the most abundant microbes in the soil followed by fungi (Hoeft, Nafziger et al. 20 00). These microbial groups play an important and particular role in the ecolo gy of the soil, including nutrient cycling and the availability of these nutrients for plant growth. In addition, certain microbial or ganisms may contribute to the protection of the root system ag ainst soil pathogens (OECD 2003).

Air Quality

Many agricultural activ ities affect air quality including sm oke from agricultural burning, tillage, traffic and harvest emissions, pesticide drift from spraying, and nitrous oxide emissions from the use of nitrogen fertilizer (Hoe ft, Nafziger et al. 2000; Aneja, Schlesinger et al. 2009). These agricultural activities individually have potentially adverse environmental impacts on air quality. Tillage contributes to the re lease of greenhouse gases (GHGs ) because of the loss of CO 2 to the atmosphere, and the exposure and oxidation of so il organic matter (Baker , Southard et al. 2005). Emissions released from agricultural equipm ent (e.g., irrigation pumps and tractors) include carbon monoxide, nitrogen oxides, reactive organic gases, particulate matter, and sulfur oxides (US-EPA 2010). Nitrous oxide may al so be released following the use of nitrogen fertilizer (USEPA 2010). Aerial application of pesticides may cause impacts from drift and diffusion. Pesticides may volatilize after application to soil or plant surfaces and move following wind erosion (Vogel, Majewski et al . 2008). Agriculture, including la nd-use changes for farming, is responsible for an estimated 6 % of all hum an-induced GHG emissions in the U.S., and N 2 O emissions from agricultural soil management ar e a large part of this, 68 % of all U.S. N 2 O emissions (US-EPA 2010).

Climate Change

Climate change is possibly interrelated with agri culture in several releva nt ways. Production of agricultural commodities is one of the many huma n activities that could contribute greenhouse gases to the air (Iserman 1993; Hoeft, Nafziger et al. 2000; Aneja, Schlesin ger et al. 2009). First, this may occur through the combustion of fossil fuels to run farm equipment, the use of fertilizers, or the decomposition of agricultu ral waste products includi ng crop residues and animal wastes. Second, the classes of crops pl anted are relevant to climate change, whether trees, grasses or field crops (Cole, Duxbury et al. 1997; Freibauer, Rouns evell et al. 2004). The location and the soil types in which they are planted also affect production of greenhouse gases (Flessa, Wild et al. 1998; Kamp, Steindl et al . 2001). Third, climate change itself may force changes to agricultural practices by extending the ranges of weeds and pests of agriculture (IPCC 2007). The influences that GE agricultural orga nisms may have on global climate change are unclear. Many of the indirect effects of these organisms will be determined by the traits engineered into organisms and the management strategies used in the production of these organisms. APHIS will continue to monitor devel opments that may lead to possible changes in the conventional production system likely to re sult from GE products brought to APHIS for approval. Some of the crops submitted by developers may clearly promote changes that may have impacts on greenhouse gases or the climate.

Climate changing greenhouse gas production will not be significant unless large amounts of crop plantings produce changes in measureable concen trations. The contributio n of agriculture to1 9 climate change is largely dependent on the production practices employed to grow various commodities, the region in which the commoditi es are grown, and the individual choices made by growers. A recent Intergovernmental Panel on Climate Change (IPCC) forecast (IPCC 2007) for aggregate North American impacts on agricultu re from climate change actually projects yield increases of 5-20% for this century. The IPCC repor t notes, however, that certain regions of the U.S. will be more heavily impacted because wate r resources may be substantially reduced. While agricultural impacts on existi ng crops may be significant, North American production is expected to adapt with improved cultivars and responsive farm management (IPCC 2007).

Animal and Plant Communities

Animals

Corn fields have been known to be visited by birds, deer and small mammals (e.g. deer mice), and other types of wildlife species. Although ma ny birds visit row-crop fields such as corn, numbers are low and few nest there (Patters on and Best 1996). The red-winged blackbird ( Agelaius phoeniceus ) is the most abundant bird in Nort h America; they are often initially

attracted to corn fields to feed on insect pests but then feed on the corn. Annually, this bird destroys over 360,000 tons of field corn and su bstantial amounts of sweet corn (Dolbeer 1990); other abundant species of birds that forage an d/or nest on and around corn include the horned lark ( Eremophila alpestris ), the brown-headed cowbird ( Molothrus ater ), and the vesper sparrow ( Pooecetes gramineus ) (Patterson and Best 1996).

Deer, such as the white-tailed ( Odocoileus virginianus ), find field corn attractive because it

functions both as food and cove r throughout the latter half of the growing season (Vercauteren and Hygnstrom 1993). Deer can significantly damag e or completely destroy small corn fields that are surrounded by woody or br ushy areas; however, deer damag e to large corn fields is often limited to a few rows closest to the wooded areas (Neilsen 2005). The deer mouse ( Peromyscus maniculatus ) is the most common small mammal in almost any agricultural field (Stallman and Best 1996; Sterner, Petersen et al. 2003). The deer mouse feeds on a wide variety of plant and animal matter depending on availabi lity, but primarily feeds on seeds and insects.

The deer mouse has been considered benefici al in agroecosystems b ecause it consumes both weed and pest insect sp ecies. The meadow vole ( Microtus pennsylvanicus ) feeds primarily on fresh grass, sedges, and herbs, but also on seeds and grains (Ste rner, Petersen et al. 2003). The meadow vole may also be considered beneficial fo r its role in the consumption of weeds, but can be a significant agricultural pest where ab undant as they rely on cover absent from tilled agriculture. The lined ground squirrel ( Spermophilus tridecemlineatus ) feeds primarily on seeds of weeds and available crops, such as corn a nd wheat (Sterner, Petersen et al. 2003). This species has the potential to damage agricultural cr ops, although this ground squirrel can also be considered beneficial when eating pest in sects, such as grasshoppers and cutworms.

Although many of the invertebrate organisms found in corn-producing areas are considered pests, such as the European corn borer ( Ostrinia nubilalis ) and the corn rootworm ( Diabrotica spp.), many others are considered beneficial (H oeft, Nafziger et al. 2000). Numerous insects and related arthropods perform valuable functions: th ey pollinate plants, contribute to the decay of organic matter, cycle soil nutrients, and attack ot her insects and mites that are considered to be

pests.2 0 Plants

The landscape surrounding a corn field varies de pending on the region. In certain areas, corn fields may be bordered by other corn (or any ot her crop); fields may also be surrounded by woodland and/or pasture/grassland areas. Therefore, the types of weeds in and around a corn field depend on the immediate area in which the corn is planted. Those weed species present will also vary depending on the ge ographic region where the corn is planted. Weeds compete with crops for water, nutrients, light, and other gr owth factors. Each year in the U.S., corn yields are threatened by more than 200 weed species (H eap 2008). Weed species such as giant foxtail and barnyardgrass have been s hown to reduce corn yields by up to 13 and 35%, respectively (Bosnic and Swanson 1997; Fausay, Kells et al. 1997). Common weeds that cause problems in corn fields include velvetleaf, common cock lebur, common lambsquarters (annuals) and quackgrass and Johnsongrass (perennials) (Hoeft, Nafziger et al. 2000).

Biological Diversity

Species diversity and abundance in corn agro-e cosystems may differ between the three corn production methods: conventional w ith GE varieties, conventional with non-GE varieties, and organic. Many studies over the last 10 years ha ve investigated the differences in biological diversity and abundance between GE and non-GE fields, particularly those GE crops that are resistant to insects (e.g., Bt crops) or herbicides (e.g., glyphosate-tolerant or glufosinate-tolerant crops.) Different studies have indicated potential decreases in biological diversity or abundance due to GE crops, or the presence of a pesticidal protein in some GE crops (Bt) (e.g., (Hansen Jesse and Obrycki 2000; Ponsard, Gutierrez et al . 2002; Pilcher, Rice et al. 2005). Some studies investigating decreases in weed populations fo llowing the use of herbicides and herbicidetolerant crops have observed decreases in animal populations using the weeds as a food or refuge source. Herbicide inputs in some cases may reduce overall biological diversity in farmed fields (Marshall, Brown et al. 2003). Other st udies of GE crops, such as Bt corn, when compared to non-GE crops sprayed with insectic ides demonstrate that GE crops do not cause any changes in arthropod abundance or diversity (e.g., (Bitzer, Rice et al. 2005; Torres and Ruberson 2005; Romeis, Meissle et al. 2006; Marvie r, McCreedy et al. 2007; Chen, Zhao et al. 2008; Wolfenbarger, Naranjo et al. 2008)). So me reports show that GE crops may even increase biological diversity (e.g., (Romeis, Me issle et al. 2006; Marvier, McCreedy et al. 2007)) in agro-ecosystems. Herbicide-tolera nt corn, when compared to non-GE corn production, may not result in changes in arthr opod abundance and may ev en increase species diversity during different times of the year (e.g., (Brooks, Bohan et al. 2003; Haughton, Champion et al. 2003; Hawes, Haughton et al. 2 003; Roy, Bohan et al. 2003; Wolfenbarger, Naranjo et al. 2008)). Since biological diversity can be defined and measured in many ways, APHIS considers determining the level of biologi cal diversity in any crop to be complex and hard to achieve concurrence. Another difficulty with biodiversity studies is separating expected impacts from indirect impacts. For example, reductions of biological control organisms are seen in some Bt-expressing GE crops, but are caused by reduction of the pest host population following transgenic pes ticide expression in the tr ansformed crop plant.

Gene Movement

Corn plants are pollinated through wind movement of pollen to other receptive corn plants. In the U.S., there are no other species that can be pollinated by corn pollen without human intervention (e.g., manually forcing reproduct ion in the laboratory) (USDA-APHIS 2010).21 Thus, the public concern surrounding gene m ovement for GE corn is between GE and non-GE corn plants. Of the total corn acres planted in 2009, 85% were GE co rn varieties (USDA-NASS 2009a). Specialty corn, those with traits of partic ular interest to various markets such as blue corn, waxy corn or organic corn, are typically grown with various management practices that intend to limit corn pollen from reaching the sp ecialty corn crop during the period of time that the specialty corn crop is receptive to polle n. For example, the NOP has requirements for organic plans to address pollen flow from GE crops (Kuepper 2002; Kr ueger 2007; Kuepper, Born et al. 2007). The Association of Official Seed Certifying Agencies (AOSCA) also has information for specialty corn crops, and a protocol for grow ing non-GE corn (AOSCA 2008). There is a price premium associated with growin g these types of specialty crops in conjunction with the extra regimens in place to maximize the pur ity of these specialty crops. For example, in 2007, conventional corn averaged $4.19/bushel (USDA-NASS 2010b), whereas organic corn averaged $7.08/bushel (USDA-NASS 2010c).

A recent paper reviewed studies investigating gene flow and cross-fertili zation studies in corn grain production fields, and using the data from these studies recommended 50m (approx. 164ft)

as the distance needed to isolate GE corn from non-GE corn (Sanvido, Widmer et al. 2008). The authors limited their analysis to studies that confirmed fertilization in the non-GE corn plants, and excluded studies on pollen dispersal (e.g., (Raynor, Ogden et al. 1972; Di Giovanni, Kevan et al. 1995; Aylor, Schultes et al. 2003)) that only measured pollen flow, because pollen flow does not necessarily result in fertilization. Suc cessful cross-fertilizatio n requires many different biological and physical factors, such as synchrony of flowering between corn fields, viability of pollen, and presence of physical ba rriers, and thus pollen dispersal is not equivalent to cross fertilization. Sandivo et al. (2008) analyzed exis ting studies and fo und that the cross-fertilization rate in non-GE corn typically remained below 0. 5% at this distance. This result was validated when large scale studies were analyzed for cross- fertilization events (e.g. (Henry, Morgan et al. 2003; Weber, Bringezu et al. 2007).

However, there are studies that refute the success of distance as an isolation strategy. Jones and Brooks (1950) found cross-fertilizat ion to be as high as 2.5 % at 660ft, which is the isolation distance used by AOSCA to isol ate corn fields for seed production (AOSCA 2004). One potential reason for the discrepancy between this study and many other gene flow studies in corn may be due to the type of corn used in the Jones and Brooks study. Jones and Brooks (1950) investigated the appropriate isolation distance for seed production in open-pollinated varieties, and not in hybrid varieties. Due the biology of open-pollinated varieties, these types of plants may be more receptive to pollen over a longer period of time than hybrid corn plants (Sanvido, Widmer et al. 2008), allowing for a gr eater chance of pollination events. Thus the results from Jones and Brooks (1950) may be an overestimation of cros s-fertilization potential for hybrid corn plants.

Public Health

Public health concerns surrounding GE field corn, like DP-32138-1 corn, focus primarily on human and animal consumption. Non-GE corn va rieties, both those developed for conventional use and for use in organic pr oduction systems, do not requi re routine evaluation by any regulatory agency in the U.S. for food or feed safety prior to release in the market. Under the Federal Food, Drug, and Cosmetic Act (FFDCA), it is the responsibility of food and feed manufacturers to ensure that the products they market are safe and pr operly labeled. Food and feed derived from DP-32138-1 must be in compliance with all applicable legal and regulatory2 2 requirements

. GE organisms for food and feed may und ergo a voluntary consultation process with the FDA prior to release onto the market . Although a voluntary proc ess, applicants who wish to commercialize a GE variety that will be included in the food supply invariably complete

a consultation with the FDA. In a consultation , a developer who intends to commercialize a bioengineered food meets with the agency to identify and discuss relevant safety, nutritional, or other regulatory issues regarding the bioengin eered food and then submits to FDA a summary of its scientific and regulatory assessment of the food; FDA evaluates the submission and responds to the developer by letter (US-FDA 2010).

Socioeconomic

The last three decades have been marked by significant transition in the seed industry (Fernandez-Cornejo 2004). Before the 1970’s, seed development occurred in the public domain. Most private seed companies focused on cleaning, handling, storing packaging, and selling seed (Fernandez-Cornejo 2004). However, following th e passage of the 1970 Plant Variety Protection Act, more than 50 seed companies were acquired by pharmaceutical, petrochemical, and food firms (Fernandez-Cornejo 2004)

. Mergers and acquisitions created a new seed industry structure and, by the early 1980s, several international firm s were among the top seed sellers worldwide (Fernandez-Cornejo 2004). The seed company can be viewed in terms of different functions including plant breeding research and devel opment (R&D), seed production, seed conditioning, and seed marketing and distribut ion. The high costs associated w ith large-scale R&D limit this activity to Federal Government agencies, land-grant colleges and universities, and a small number of large companies. Given the size of thei r R&D investments, these plant breeders play a central role in managing the entire production, di stribution, and marketing processes in the seed industry (Fernandez-Cornejo 2004) . In 1999-2002, approximately 300 firms were involved with seed corn production with the top five compan ies having 64% market share, the next 10 companies having an additional 16% market share, and the rest of the companies having the remaining 20% market share (K alaitzandonakes and Magnier 2004).

The top five companies involved in seed producti on are national or multina tional companies with their own R&D departments developing their ow n genetics and their own production, processing

and distribution faciliti es. The next 10 largest companie s are regional with a dependence on foundation seed firms and university developed lines in addition to their own proprietary genetics along with their own production and dist ribution facilities. The remaining smaller firms depend almost solely on purchased genetics and others for production and concentrate on distribution (Kalaitzandonakes and Magnier 2004). These smaller seed companies with a marketable seed product typically contract out the production and multiplication processes to farmers, farmers’ associations, and private firm s such as Illinois Crop Improvement Association (www.ilcrop.com ) , Indiana Crop Improvement Associati on (http://www.indianacrop.org/), and Iowa Crop Improvement Association (h ttp://www.agron.iastate.edu/ICIA).

In all cases, seed companies closely manage th e production of seed by contract growers. Seed companies want to ensure that the desirabl e plant characteristics are carried through to subsequent generations and to prevent problems that could aff ect the quality of the product (Fernandez-Cornejo 2004). After harvesting certified seed, the seed is conditioned, which includes the drying, cleaning, and sor ting of seed. Seeds are also treated with insecticides and fungicides and packaged for distribution and sa le to farmers (Fernandez-Cornejo 2004). Large seed companies play a more direct role in assuring their seed meets at least the minimum

standards for certified seed as we ll as in marketing and distributi ng their end produc t to regional,23 national, and international markets. Many comp anies also license or outsource marketing and distribution to private firms and individuals to improve access to local markets (FernandezCornejo 2004).

Technical developments in the genetic engin eering of plants has b een one of the prime

motivations for the consolidation of the seed corn companies (Kalaitzandonakes and Hayenga 1999; Fernandez-Cornejo 2004; Kalaitzandonake s and Magnier 2004) with high biotechnology regulatory/compliance costs being the major r eason for limiting the development and marketing of new biotech corn varieties to the larg e national/multinational companies (Fulton and Giannakas 2001; Kalaitzandonake s and Magnier 2004). The greater the regulatory requirements in an industry, the more concentr ated the industry is expected to be (Fulton and Giannakas 2001) with accumulating evidence suggesting that biot echnology compliance costs have increased over time (Kalaitzandonakes, Alston et al. 2007).

In the production of hybrid seed co rn, the task that is one of the most difficult but essential to the overall success is the detasseling of the cor n. Manual detasseling of corn is a physically demanding job consisting of pulling tassels out of th e top of thousands of tall corn plants by each detasseler each day. Detasseling op erations are conducted under cha llenging conditions, such as those environmental conditions that persist in July in the U.S. Midwest. Detasseling is a temporary job lasting only one to four weeks in mid-summer. The job is time sensitive because tassels need to be pulled within a day of emergence before any po llen is released; a high level of flexibility for the task is required because the emergence of tassels depends on the weather. Hot humid weather speeds up the emergence of tassels by one to more weeks and cold wet weather slows down the emergence of tassels by one or mo re weeks. Thousands of people are needed to complete the operation in timely basis. Because of the short duration and physical demands of the job, the detasseling operation mainly attracts high school st udents during the summer break. Thousands of high school students are hired fo r detasseling each year (Byron 2002; Pioneer 2007; Gustafson 2008; Holloway 2010; Schernikau 2010).

There are many challenges for the farmers whose land is contracted to grow the low yielding inbred plants including growing corn to meet exacting planting, growing and harvesting standards and schedules. Additionally, pay premiu ms and minimums are standard parts of the

contract. The minimum standards are likely to be equivalent to the returns that would be expected from growing conventional corn, and ba sed on the average yields in the area and the commodity corn price. For example, the averag e yield might be 170 bushels with a commodity price of $4.00 per acre making the minimum sta ndard $680 per acre. These contract conditions also require that the farmers n eed to accept the large ruts that are left by detasseling machines used in muddy conditions. These ruts destroy soil structure, increas e tillage requirements, increase erosion, result in major yield reduction in the corn crop and possibly in the crop the following year as well, and result in da mage to harvesting and tillage machinery (Daum 1996; Kok, Taylor et al. 1996; Gelderman 2009; Tilg hman 2009; Roegge 2010)

.

For the company that wants to produce and sell the hybr id seed corn at a profit, detasseling is an expensive operation whether it is manual or mechan ical. Mechanical detasse lers reduce the need for manual (human) detasselers, but they tend to reduce yields even more than manual detasseling operations and manual detasseling is stil l required to completely finish the job (Wych 1988).2 4 The United States is the largest seed mark et worldwide, followed by China and Japan (Fernandez-Cornejo 2004). Seed expenditures by U.S. farmers rose from about $500 million per year in 1960 to nearly $7 bill ion in 1997 (Fernandez-Cornejo 2004). The United States is a net exporter of seed. In 1996, the U.S. seed trad e surplus was $384 million: $698 million in seed exports were mainly sent to Mexico, Canada, Italy, Japan, and Argentina. About $314 million in seed imports, were received mainly from Cana da, Chile, the Netherlands, and China (FernandezCornejo 2004).



### III. Alternatives

This document analyzes the pot ential environmental consequences of a determination of nonregulated status of DP-32138-1 corn. To re spond favorably to a petition for nonregulated status, APHIS must determine that DP-32138-1 corn is unlikely to pose a pl ant pest risk. Based

on its risk assessment APHIS has concluded that DP-32138-1 corn is unlikely to pose a plant pest risk (USDA-APHIS 2010) . Therefore APHIS must determine that DP-32138-1 corn is no longer subject to 7 CFR part 340 or the plan t pest provisions of the Plant Protection Act.

Two alternatives will be evaluated in this EA: (1) no action and (2 ) determination of nonregulated status of DP-32138-1 corn. APHIS has assessed the potential for environmental impacts for each alternative in the “Environmental Consequences” section

No Action: Continuation as a Regulated Article

Under the No Action Alternative, APHIS w ould deny the petition. DP-32138-1 corn and progeny derived from DP-32138-1 corn would con tinue to be regulated articles under the regulations at 7 CFR Part 340. Permits issued or notifications acknowledged by APHIS would still be required for introducti ons of DP-32138-1 corn and meas ures to ensure physical and reproductive confinement would continue to be implemented. APHI S might choose this

alternative if there were insuffici ent evidence to demonstrate the lack of plant pest risk from the

unconfined cultivation of DP-32138-1 corn.

This alternative is not the Preferred Alterna tive because APHIS has concluded through a Plant

Pest Risk Assessment that DP-32138-1 corn is unlikely to pose a plant pest risk (USDA-APHIS 2010) . Choosing this alternative would not satisfy the purpose and need of making a determination of plant pest risk status and resp onding to the petition for nonregulated status.

Preferred Alternative: Determination that DP-32138-1 is No Longer a Regulated

Article

Under this alternative, DP-32138-1 corn and pr ogeny derived from them would no longer be regulated articles under the regul ations at 7 CFR Part 340. DP-32138-1 corn is unlikely to pose a plant pest risk (USDA-APHIS 2010) . Permits issued or notifications acknowledged by APHIS would no longer be required for introductions of DP-32138-1 corn and progeny derived from this event. This alternative best meets the purpose an d need to respond appropriately to a petition for nonregulated status based on the requirements in 7 CFR part 340 and the agency’s authority under the plant pest provisions of the Plant Prot ection Act. Because the agency has concluded that DP-32138-1 corn are unlikely to pose a pl ant pest risk, a determination of nonregulated status of DP-32138-1 corn is a response that is c onsistent with the plant pest provisions of the PPA, the regulations codified in 7 CFR part 340, and the biotechnology regul atory policies in the

Coordinated Framework. Under this alternative, growers may have future access to DP-32138-12 5 corn and progeny derived from this event if the developer decides to commercialize DP-32138-1

corn.

Alternatives Considered but Rejected from Further Consideration

APHIS assembled a list of altern atives that might be considered for DP-32138-1 corn. The agency evaluated these alternativ es in light of the agency's authority under the plant pest provisions of the Plant Protection Act, and the regulations at 7 CFR pa rt 340, with respect to environmental safety, efficacy, and practicality to identify which alternatives would be further

considered for DP-32138-1 corn. Based on this ev aluation, APHIS rejected several alternatives. These alternatives are discussed briefly below al ong with the specific reasons for rejecting each.

Prohibit any DP-32138-1 from Being Released

In response to public comments that stated a preference that no GE organisms enter the marketplace, APHIS considered prohibiting the release of DP-32138-1 , including denying any permits associated with the field testing. AP HIS determined that this alternative is not appropriate given that AP HIS has concluded that DP-32138-1 is unlikely to pose a plant pest

risk (USDA-APHIS 2010)

.

In enacting the Plant Protection Act, Congress found that

[D]ecisions affecting impor ts, exports, and interstate movement of products regulated under (the Plant Protection Act) shall be based on sound science... §402(4) (see 7 U.S. C. §7701(4)).

On March 11, 2011, in a Memorandum for the H eads of Executive Departments and Agencies, the White House Emerging Technologies Interage ncy Policy Coordination Committee developed broad principles, consistent with Execu tive Order 13563, to guide the development and implementation of policies for oversight of emergi ng technologies (such as genetic engineering) at the agency level. In acco rdance with this memorandum, agen cies should adhere to Executive Order 13563 and, consistent with that Executive Order, the followi ng principle, among others, to the extent permitted by law, when regulating emerging technologies:

“[ D]ecisions should be based on the best reasonably obtainable scientific, technical, economic, and other information, within the boundaries of the authorities and mandates of each agency”

Based on our Plant Pest Risk Assessment (USDA-APHIS 2010) and the scientific data evaluated therein, APHIS concluded that DP-32138-1 are unlikely to pose a plant pest risk

. Accordingly, there is no basis in sc ience for prohibiti ng the release of DP-32138-1

.

Approve the petition in part

The regulations at 7 CFR 340.6(d) (3)(i) state that APHIS may "a pprove the petitio n in whole or in part." For example, a determination of nonregulated st atus in part may be appropriate if there is a plant pest risk associat ed with some, but not all lin es described in a petition. Because

APHIS has concluded that DP-32138-1 corn is unlikely to pose a plant pest risk, there is no2 6 regulatory basis under the plant pest provisions of the Plant Protection Act for considering approval of the petition only in part.

Isolation Distance between DP-32138-1 and Non-GE Corn Production

In response to public concerns of gene m ovement between GE and non-GE plants, APHIS considered requiring an isolation distance of DP-32138-1 corn and non-GE corn production. However, because APHIS has concluded that DP-32138-1 corn is unlikely to pose a plant pest risk (USDA-APHIS 2010), an alternative based on requiring isolation distances would be inconsistent the statutory authority under the plant pest provisions of the Plant Protection Act and regulations in 7 CFR part 340. APHIS also considered geographically restri cting the production of DP-32138-1 corn based on the location of produc tion of non-GE corn in organic production systems or production systems for GE-sensitive mark ets in response to public concerns regarding possible gene movement between GE and non-GE plants. However, as presented in APHIS’ plant pest risk assessment for DP-32138-1 corn, there are no geogr aphic differences associated with any identifiable plant pe st risks for DP-32138-1 corn (USDA-APHIS 2010) . This alternative was rejected and not analyzed in detail because APHIS has concluded that DP-32138- 1 corn does not pose a plant pest risk, and will not exhibit a greater plant pest risk in any geographically restricted area. Therefore, such an alternative would not be consistent with APHIS’ statutory authority unde r the plant pest provisions of the Plant Protection Act and regulations in Part 340 and th e biotechnology regulatory policies embodied in the Coordinated

Framework.

Based on the foregoing, the imposition of isolation di stances or geographic restrictions would not meet APHIS’ purpose and need to respond appropri ately to a petition for nonregulated status based on the requirements in 7 CFR part 340 and the agency’s authority under the plant pest provisions of the Plant Protection Act. Neverthele ss, APHIS is not expecting significant effects. However, individuals might choose on their ow n to geographically isolate their non-GE corn production systems from DP-32138-1 corn or to use isolatio n distances and other management practices to minimize gene movement between corn fields. Information to assist growers in making informed management decisions for DP-32138-1 corn is available from Association of Official Seed Certifying Agencies (AOSCA 2010).

Requirement of Testing For Event 32138 Corn

During the comment periods for other petitions for nonregulated stat us, some commenters requested USDA to require and provide testing for GE products in non-GE production systems.

APHIS notes there are no nationa lly-established regulations involvi ng testing, criteria, or limits of GE material in non-GE systems. Such a requirement would be extremely difficult to implement and maintain. Additionally, because DP-32138-1 corn does not pose a plant pest risk (USDA-APHIS 2010) , the imposition of any type of testing requirements is inconsistent with the plant pest provisions of the Plant Protection Ac t, the regulations at 7 CFR part 340 and the biotechnology regulatory policie s embodied in the Coordinated Framework. Therefore, imposing such a requirement for DP-32138-1 corn would not meet APHIS’ purpose and need to respond appropriately to the petition in accord ance with its regulatory authorities.

Comparison of Alternatives

Table 1, below, briefly summarizes the results for each of the issues raised in the Environmental2 7 Consequences (Section IV) by each of the alternatives described in the Alternatives section (Section III).

Table 1. Issues of Potential Impact s and Consequences of Alternatives.

Attribute/Measure Alternative A: No

Action Alternative B: Determination of Nonregulated Status
Meets Purpose and

Need and No Yes

Unlikely to Pose a Plant

Pest Risk Satisfied through use of regulated field

trials Satisfied—plant pest risk assessment (USDA-APHIS 2010)

Management Practices Acreage and Areas of

Corn Production Unchanged Unchanged

Cropping Practices Unchanged Unchanged Pesticide Use Unchanged Unchanged Seed Corn Production Unchanged Minimal - may decrease need for seed corn acreage due to increased yields

efficienc y Organic Farming Unchanged Unchanged

Im p act to S p ecialt y

Corn Unchan g ed Unchan g ed

Environment Water use Unchanged Unchanged Soil Unchanged Unchanged Air Quality Unchanged Unchanged Climate Change Unchanged Unchanged Animals Unchanged Unchanged Plants Unchanged Unchanged Biological Diversity Minimal Minimal

Gene Movement Minimal Minimal

Human and Animal Health

Public Health: Risk to

Human Health Unchanged Unchanged

Public Health: Risk to Worker Safety Minimal – detasseling

carries some risk to

workers Minimal – fewer safety risks with less detasseling

Public Health: Risk to

Animal Feed Unchanged Unchanged

Socioeconomic

Domestic Economic

Environment Unchanged Minimal – seed producers save 4% of retail sales price; growers – possible small

seed cost decrease Unchanged – immeasurably small impact

on cost of commodit y corn
Trade Economic

Environment Unchanged Unchanged2 8 Attribute/Measure Alternative A: No

Action Alternative B: Determination of Nonregulated Status

Social Environment Unchanged Minimal – fewer part time detasselers

hired for summer work

Other Regulatory Approvals U. S. Completed new protein

consultations with FDA Completed new protein consultations

with FDA
Compliance with Other Laws

CWA, CAA. EOs Fully compliant Fully compliant



### IV. Environmental Consequences

This analysis of potential environmental conseque nces addresses the potential impact to the human environment from the alternatives analyzed in this EA, namely taking no action and a determination by the Agency th at DP-32138-1 corn does not pose a plant pest risk.

Potential environmental impacts from the No Ac tion Alternative and the Preferred Alternative for DP-32138-1 are described in detail throughout this section. A cumulative effects analysis is also included for each environmental issue. Certain aspects of this product and its cultivation

would be no different between the alternatives: those are described below.

Scope of the Enviro nmental Analysis

Although the preferred alternative would allow for new plantings of DP-32138-1 to occur anywhere in the U.S., APHIS will limit the environmental analysis to those areas that currently support corn production. To determine areas of corn produ ction, APHIS used data from the National Agricultural Statistics Service (NASS) 2007 Census of Agriculture to determine where corn is produced in the United States (USD A-NASS 2010d). Corn grain was produced in all states except for Alaska.

The scope of possible impacts is limited in some ways by the relatively small area of potential use. DP-32138-1 lines containing transgenes only have utility in seed production and are not intended to be a commercial product to be used to plant conventional co rn acres (Pioneer 2009). The total acreage of DP-32138-1 planted in the U.S. each year under close supervision is expected to be less than 5,000 acres (Pioneer 2009). If the DP-32138- 1 is licensed to third parties and adopted across the entire U.S. seed industry, the total acreage is no t expected to exceed 20,000 acres each year (Pioneer 2009).

Other Assumptions

The environmental consequences of the different alternatives described above will be analyzed under the assumption that seed producers who produce corn seed using DP-32138-1 corn and farmers who produce conventional corn or corn using organic methods are using reasonable, commonly accepted best management practices fo r their chosen system and varieties during agricultural corn production. Howe ver, APHIS recognizes that not all farmers follow these best management practices for corn. Thus, the analyses of the environmental effects will also include the assumption that some farmers do not follow these best management practices.2 9 Agricultural Production of Corn

One of APHIS’ missions is to improve Ameri can agricultural productivity. Best management practices, such as planting dates, seeding rates, and harvest times are commonly accepted, practical ways to grow corn, regardless of whether the corn farmer is using conventionalpractices with non-GE or GE varieties, or organic practices. These well-established, widelypracticed means to produce corn can be obtai ned through local Cooperative Extension Service offices and their respective websites (NSF C-IPM 2010).

GE and non-GE corn varieties are continually unde r development. From 2004 to 2009, the corn acreage ranged from a low of 78.3 million acres planted in 2006 to a high of 93.5 million acres planted in 2007. Most of the corn acreage in the U.S. is planted to GE varieties. Of the 86.3 million acres of corn planted in the U.S. in 2009, 85% of the acreag e was planted with GE varieties (USDA-NASS 2009a). Base d upon these trends, conventiona l production practices that

use GE varieties will li kely continue to dominate in terms of acreage, or perhaps increase in acreage, with or without a determination of nonregulated status of DP-32138-1.

Organic Production of Corn

APHIS recognizes that producers of non-GE corn, particularly producers who sell their products to markets sensitive to genetically engineered tr aits (e.g. organic or some export markets) can be reasonably assumed to be using practices on thei r farm to protect their crop from unwanted substances and maintain their price premium. For example, the National Organic Program (NOP) has recognized the feas ibility of protecting organi cally-produced crops, and the investment farmers put into their production prac tices, by requiring that organic production plans include practical methods to pr otect organically-produced crops.

“Organic crops must be protected from cont amination by prohibited substances used on adjoining lands (for example, dr ifting pesticides, fertilizer-laden runoff water, and pollen drift from genetically engineered...)” (NCAT 2003).

Typically, growers use more than one method under organic practices to prevent unwanted material from entering their fiel ds including: isolation of the fa rm, physical barriers or buffer zones between organic production and non- organic production, as well as formal communications between neighborin g farms (NCAT 2003). The organic plan used as the basis for organic certification should in clude a description of practices used to prevent or reduce the likelihood of unwanted substances, like GE pollen or seed, at each step in the farming operation, such as planting, harvesting, storing and transp orting the crop (Riddl e 2004; Krueger 2007; Kuepper, Born et al. 2007). Or ganic plans should also include how the risk of GE pollen or comingling of seed will be monitored (Kuepper, Bo rn et al. 2007). Farmers using organic methods are requested to let neighboring farmers know that they are using organi c production practices and request that the neighbors also help the or ganic farmer reduce contamination events (NCAT 2003; Krueger 2007). Recommended organic production practices for field corn are also readily available (Kuepper 2002). Thus, commonly used production practices for corn, and the practical methods typically used by organic corn farmers to protect their crop and maximize their profits and price premiums from corn under organic pr oduction, currently provide many measures that greatly reduce the likelihood of accidental gene flow between DP-32138-1 corn and non-GE corn fields. APHIS will use the assumption that fa rmers are already using, or have the ability to use, these common, reasonable pr actices as its baseline for th e analyses of the following

alternatives below.3 0 Acreage and Areas of Corn Production

No Action: Acreage and Areas of Corn Production

The amount of GE corn planted in conventional systems in the U.S. is increasing. Since 2000 when 25% of the total corn acreage was devoted to GE varieties, the share of total corn acres devoted to GE corn varieties has risen each year to the present 85% level in 2009 (USDA-NASS 2009a).

Conventional corn production with GE varieties will likely continue to increase without a determination of nonregulat ed status of DP-32138-1 under the No Action Alternative, based on current acreage trends. Seed for currently available conventional and GE varieties w ill

remain the same under the No Action Alterna tive, except progeny of DP-32138-1 will be unavailable. Corn is currently produced commercia lly in 49 states, excluding only Alaska according to the 2007 Census of Agricultur e (USDA-NASS 2009b) and under the No Action Alternative, this range of production will be unchanged.

Preferred Alternative: Acreage and Areas of Corn Production

In 2009, GE corn was planted on 85% of all corn acre s currently in produc tion in the U.S., and the use of GE corn has been steadily increasing over the last 10 ye ars (USDA-NASS 2009a). Most corn is planted in fiel ds that have been in crop production for many years. A determination of nonregulat ed status of DP-32138-1 under the Preferred Alternative is not expected to alter the range of corn cultivation as the new GE tr ait (DP-32138-1) does not change the growth habits compared to conventional varieties (USDA-APHIS 2010). Additionally,

because DP- 32138-1 is a technique f or more efficiently producing seed that will be used to plant conventional corn production acres without introducing ne w transgenes to th ese production acres, even widespread use of DP-32 138-1 will have no significant effect on increasing or decreasing the use of GE corn or on total corn production acreage.

Thus, under the preferred alternative, a determ ination of nonregulated status of DP-32138-1 would not increase corn production, either by its availability alone or accompanied by other factors, or cause an increase in overall GE corn acreage. Impacts would be similar to the No

Action Alternative.

Cumulative Effects: Acreage and Areas of Corn Production

Cumulative effects of a determination of nonre gulated status of DP-32138-1 are unlikely.

Neither the No Action Alternative nor a determinat ion of nonregulated st atus of DP-32138-1 will directly cause an increase in agricultural acreage devoted to corn pr oduction, or those corn acres devoted to GE corn cultivation. DP-32138-1 will also not change cultivation areas for corn production in the U.S. There are no anticipatedchanges to the availability of GE and nonGE corn varieties on the market under either alternative.

Cropping Practices: Crop Rotation , Tillage, and Pesticide Use

The current economics of corn production are driv ing the change or perceived change in crop rotation practices (such as corn-to-corn cro pping, Erickson and Lo wenberg-DeBoer, 2005). Growers make choices to plant certain corn va rieties and use certain crop rotation practices based on factors such as yield, weed and diseas e pressures, cost of seed and other inputs,31 technology fees, human safety, potential for crop injury, and ease and flexibility of the production system (Olson and Sander 1988; Giannessi 2005) . Therefore, when taking into account these factors, growers w ill ultimately base their choice of inputs and agronomic practices

on individual wants and needs. Th ere may be more tillage and use of pesticides involved with the production of seed corn than with commercial corn grain. The low vigor of seed and plants of inbred lines compared to hybrid corn generally means a better seed bed is needed (generally requiring more tillage) along with the protection from diseases a nd insects provided by pesticide

used on the seed and the plants (Pioneer 2009).

No Action: Cropping Practices: Crop Rotation, Tillage, Production, and Pesticide Use

Producing seed corn requires ta king into account th e crop rotation on the farm on which the seed production is taking place as well as the crop rotations on su rrounding farms. Adequate isolation must be assured to maintain high genetic purity. Corn after corn rotation is discouraged to help avoid the po ssibility of lowering genetic purity from vo lunteer corn plants arising from a previous corn crop (Wych 1988).

Conventional tillage and the removal of plant residue from soil ar e considered agriculture practices that accentuate loss of soil organic carbon (Lal and Bruce 1999). This loss has negative impacts on the atmosphere and increases soil eros ion, among other effects. Because of numerous benefits to growers and the agricultural environm ent (Fawcett and Caruan a 2001), it is not likely that DP-32138-1 will require a change from th e increasingly prevalent practice of low-till production of corn. Under the No Action Alternative, the use of tillage methods in U.S. agricultural production of s eed corn and commercial corn will remain unchanged.

Under the No Action Alternative, corn production a nd pesticide use will remain as it is practiced today by the farming community. Growers make choi ces to use certain pesticides based on weed, insect and disease pressures, cost of seed and other inputs, technology fees, human safety, potential for crop injury, and ease and flexib ility of the system (Olson and Sander 1988; Giannessi 2005). Growers will ultimately base th eir choice of these inputs on individual wants and needs. As an example of th e pesticides used duri ng the production of field corn, the Pesticide Action Network has an online database, including a detailed description of a ll the pesticides used in corn agriculture in California (Kegley, Hill et al. 2010). It lists the top 50 pesticides (e.g., herbicides, insecticides, fungicides) used in Ca lifornia corn production. Any effects due to crop rotation, tillage and pesticide use in the agricultural production of seed corn and commercial

corn will remain the same under the No Action Alternative.

Preferred Alternative: Crop Rotation, Ti llage, Production, and Pesticide Use

As stated above, the current economics of corn production are driving the change or perceived change in crop rotation practices (such as corn-to-corn cropping, Erickson and LowenbergDeBoer, 2005). A determination of nonregulated st atus of DP-32138-1 is unlikely to change the entire pricing scheme of corn commodities in the U.S. Prices will continue to be set by market demand, without regard to the number or type of corn varieties available on the market, nor to decreased costs of seed production. Based on in formation provided by the applicant, DP-32138-1 lines containing transgenes are expected to reach an industry wide maximum of only 20,000 acres in seed corn production, which is only 0.02% of the approxim ately 90 million acres planted to corn each ye ar. DP-32138-1 is not likel y to affect a farmer’s decision to either stop using a corn-to-corn rotation or other type of rota tion, or to increase th e overall use of corn-tocorn rotation or other type of rotation as a cro pping strategy with the U.S. farming community.3 2 Adoption of DP-32138-1 will not change current producti on or cultivation practices. Crop rotation schemes that assure adequate isolation to maintain high genetic purity and tillage for seed corn production will remain the same. In those cases where seed producers employ conventional tillage for the inbred crops used in seed production, planting of DP-32138-1 will not change those practices, eith er. A determination of nonregulat ed status of DP-32138-1 will not change the loss of soil organi c carbon due to tillage in seed corn production systems or in commercial production of corn in which DP-32138-1 was used in the production of hybrid seed.

DP -32138-1 seed corn production uses the same ag ricultural inputs (e.g., pe sticides, fertilizers) as seed corn currently grown. DP -32138-1 does not require different types of pesticides than are currently used in seed corn produc tion. A determination of nonregulated status of DP-32138-1 will not have any effect on the pesticides used in the production of seed corn or commercial corn production in the U.S., compared to the No Action Alternative.

Cumulative Effects: Crop Rotation, Tillage, Production, and Pesticide Use

A determination of nonregulated status of DP-32138-1 will not have any cumulative effect on the crop rotation, tillage and crop production practices or on the pe sticides used in the production

of seed corn or commercial corn production in the U.S., compared to the No Action Alternative. The limited number of seed corn production acres that may use DP-32138-1 lines containing transgenes is expected to have no cumulative e ffect on crop rotations, tillage and crop production practices, or pesticide use. The requirement s for crop rotation, tillage and crop production practices, and pesticide use for the non transg enic hybrid progeny re sulting from DP-32138-1 seed production that will be used to plant convent ional corn fields, will be exactly the same as those used for conventionally produced hybrids.

Seed Production

Seed corn production differs from commercial grain production because seed companies impose strict requirements to maintain seed identity and high levels of genetic purity of the final product. This purity is accomplished using contracts, track ing and traceability systems, quality assurance processes, record maintenance, auditing, proper labeling, appr opriate sampling and testing and identity preservation systems (Sundstrom, Willia ms et al. 2002). An example of a successful identity preservation system are the Seed Certif ications programs as used by the AOSCA which have maintained seed purity standards for national and international trade since the 1920s (Sundstrom, Williams et al. 2002).

Seed increase for a new corn variety encompasses larger and larger acreage for each successive generation. The initial generation of the new inbred line consists of one or only a few plants. These plants will produce a few ears. The seeds from these few ears are used to plant a small plot, approximately 0.1 acres (3000-6000 sq. ft.). Ear ly generation plants are self pollinated and pollen control is generally achieved by using bags over the tassels and ears. Using this system of pollen control allows many lines to be grown in a small area with an extremely low possibility of cross pollination with other lines grown in the immediate area. The 2-10 bushels of seed harvested are used to plant 2-20 acres for the next increase. Plants at this stage are also self pollinated to increase the seed; however, the ar ea is likely too large for bagging as a means of pollen control. Pollen control is achieved by mean s of temporal or physical isolation from other lines. This increase produces 100 to 1000 bushels of inbred line seed.33 In the next stage, which produces single cross hy brid seed to be used for planting commercial corn grain fields, two different inbred lines (both are from the previous stage increase) are planted in the same field. One inbred line is considered the male line providing the pollen and the second line is the female line, from which the hybrid seed will be harvested. Procedures must be taken to prevent these plants of the female line from producing viable fertile pollen. Presently, detasseling or cytoplasmic male sterilit y systems are used to prevent self pollination of this stage female plants.

The Pioneer Hi-Bred SPT proce ss is designed to produce non-tra nsgenic male-sterile female inbred parent plants for hybr id corn seed production without the need for tassel removal or bagging. As a process, SPT uses two types of in bred corn lines: 1) nontransgenic male-sterile inbred lines (referred to as ms45 ) that cannot produce pollen but can produce fertile egg cells, and 2) a transgenic male-fertile line containing a DP-32138-1 SPT maintainer cassette within the

ms45 genetic background (referred to as DP-32138- 1 corn) that permits pollen development in which half the pollen is non-transgen ic and viable and the other half is transgenic but not viable. The maintainer can also produce fertile eggs wh ich allows for production of new transgenic seeds of the maintainer line (see below). Th e DP-32138-1 SPT maintainer cassette contains three linked genes that are differentially expre ssed over time, including Ms45 (expressed in the anther),  -amylase (expressed in the pollen grain), and dsRed (expressed in the kernel).

The Pioneer Hi-Bred SPT process consists of th ree major steps (Figure 1): 1) DP-32128-1 seed increase, 2) non-transgenic ms45 male sterile corn increase, a nd 3) hybrid seed production using non-transgenic seed from use of the SPT process. The steps of this process are described in additional detail, below.

Step 1: 32138 SPT Maintainer Seed Increase. The first major step of the Pioneer Hi-Bred SPT process propagates DP-32138-1 corn (see Step 1, Figure 1, and Figure 2). Propagation involves both the self-pollination of DP-3213 8-1 corn in the field and mechan ical sorting to select for the

desired seed. In contrast to a male sterile ms45 corn line, DP-32138-1 co rn is able to produce pollen by means of the fertility trait MS45 which was engineered into the DP-32138 SPT maintainer cassette. Half of the pollen pr oduced by DP-32138-1 corn contains the DP-32138-1 SPT maintainer cassette and is not viable (p ollen expresses a starch -consuming enzyme, ZMAA1); however, the remaining half of th e pollen does not contain the DP-32138-1 SPT maintainer cassette and is fully viable.

Both transgenic (where the DP-32138-1 SPT mainta iner cassette is present) and non-transgenic (SPT absent) kernels are produced in 1:1 ratio s, following the self-fer tilization of DP-32138-1 corn (Figure 2). Seed that is normal (yello w) in color is non-transgenic for SPT (does not

contain the DP-32138-1 SPT inse rtion). Pinkish-red seed that fluoresces a bright red color under appropriate illumination is transgenic fo r SPT (contains the DP-32138-1 SPT insertion). In order to separate these two ke rnel types, mechanical color sor ting is used to detect the the 32138 SPT maintainer cassette (containing the dsRed fluorescent protein) (s ee Figure 3). The remaining kernels, non-transgeni c due to the absence of the 3 2138 SPT maintainer cassette, are discarded after mechanical sorting. Pure seed of DP-32138-1 SPT maintain er is then available for the propagation of non-transgenic male-sterile female inbred parent seed (Figure 1, Step II). Yellow, non-transgenic seed collected from the first color sort will be discarded.3 4 Figure 1. Outline of the SPT Process for Producing Hybrid Seed.3 5 Source: (Pioneer 2009)

Figure 2. Production of the 32138 SPT Maintainer Line (Figure 1, Step I).

Step II: Male-Sterile Female Inbred Parent Seed Increase. The second major step of the Pioneer Hi-Bred SPT process produces male-sterile female inbred plants for use in hybrid seed production (Figure 1, Step II). This process i nvolves the interplanting of both DP-32138-1 and

ms45 corn (non-transgenic). Within this cro ss, DP-32138-1 represents the only pollen donor. Consequently, DP-32138-1 corn may cross with itself or with ms45 corn. Both transgenic (where the 32138 SPT mainta iner cassette is present) and non-tr ansgenic kernels (SPT cassette is absent) are produced following thes e crosses (Figure 4). Only th e nontransgenic pollen is viable and can pollinate the male-sterile female inbred line.

Similar to the previous step of DP-32138-1 propa gation, mechanical color sorting is used to distinguish transgenic kernels from non-transgenic kernels; in contrast, however, two rounds of mechanical color sorting are used to select fo r non-transgenic kernel s (absence of the dsRed fluorsence protein, and thus, absence of the 32138 SPT maintainer cassett e) for use in hybrid seed production (see Sort B, Figure 3). Kernel s expressing DsRed, and thus containing the 32138 SPT maintainer cassette, are discarde d after mechanical color sorting.

Kernels produced from the second major component of the Pioneer Hi-Bred SPT process are non-transgenic. Consequently, any hybrid corn lin es produced from these kernels are also nontransgenic. Additionally, propagation of both the DP-32138-1 (Stage I) and ms45 corn lines (Stage II) precedes commercial corn grain produc tion by two generations. The final stage of of hybrid seed production is likely accomplished at a distant location rela tive to Stage I or II production, at seed production site s not directed by Pioneer (which handles the first stages of production). Consequently, because of the geogra phic separation of any seed that may contain3 6 Figure 3. Visualization of the Red Marker, DsRed2 Expressed by the SPT Cassette.

Photograph A shows a corn ear with both the tra nsgenic SPT cassette kernels/seeds (red coloring) and nontransgenic seeds (yellow coloring). Photograph B shows the SPT and the nontransgenicseed under standard illumination. Photograph C shows the SPT seed (+ DsRed2) and the nontransgenic seed (- DsRed2) under fluorescent illumination (Pioneer 2009).

Figure 4. Production of the Non-Transgeni c Female Sterile Inbred (Step II).3 7 Source: (Pioneer 2009)

the 32138 SPT maintainer cassette, admixture by mechanical means of commercial corn grain with DP-32138-1 corn is unlikely. This unlikel y potential for admixture with DP-32138-1 corn is further reduced by the relatively small cultiv ation acreage that will use the SPT technology; fields containing DP-32138-1 are not expected to exceed 20,000 acres or approximately 0.023 percent of total U.S. corn gr ain producing acreage.

Step III: Commercial Corn Hybrid Seed Produc tion. Commercial corn hybrid seed production (Figure 1, Step III) using the male-sterile seed (nontransgenic) prepar ed using SPT technology will be completed as it is for conventional pr oduction methods. The purity of the male-sterile female inbred plants produced from the SPT proce ss is insured in Step II because of the built-in biological feature in which transgenic pollen is non-viable and because the seed was passed twice through mechanical colo r sorters (refer to Pioneer 200 9, Appendix 3 for details on the color sorter). Therefore, the hybrid seed harves ted and sold to growers for commercial grain production is non-transgenic for SPT and the resu lting hybrid plants are fully fertile. The hybrid seed grown for grain is non-transgenic for SPT and therefore, the grain harvested is also nontransgenic for SPT.

The field sizes that will be used for any one corn variety at each step of increase can vary considerably depending on the present or expected market share of each variety, the level of testing to be conducted during the increase phase, and the expected yield of the inbred lines. In the case of corn, based on current production data, the amount of seed of all varieties combined must be sufficient to plant over 90 million acres each year (USDA-ERS 2010a) . Since Pioneer, the applicant in this petition, has approximately a 30% market share and has approximately 250 different corn hybrids available, this implies the number of corn varieties available industry-wide is about 800-1000 (Pioneer 2008; Pioneer 2010) with other estimates of over 4000 hybrids available in the U.S. in 2009 (Monsanto 2010). The development of new inbreds along with the Steps I and II seed increases are the stages when the transgenic plants of DP-32138-1 are expected to be in the field. The 250 Pioneer va rieties multiplied by the 2-20 acre Step II increase per variety is the “up to 5000 acres” maximum ac reage of DP-32138-1 to be used by Pioneer (Pioneer 2009). The industry-wid e assumed total of 1000 to 4000 varieties multiplied by the 2 to 20 acre Step II increase per vari ety equates to the “up to 20,000 acres” maximum acreage of

DP-32138-1 to be utilized if a dopted industry-wide (Pioneer 2009). In the Step III seed increase, DP-32138-1 confers its largest be nefits through the planting of th e non-transgenic, male sterile female inbred produced with DP-32138-1. Pion eer may avoid detasseling up to 300,000 acres, while other seed industry organizations may pot entially have up to 1 million acres that would not require detasseling.

No Action: Seed Production

The availability of methods used to produce seed corn would be the same as currently used in seed corn production systems under the No Action Alternative.

Preferred Alternative: Seed Production

A determination of nonregulated status of DP-32138-1 corn under the Preferred Alternative would not change the use of presently available systems for seed corn production. DP-32138-1 will be produced in a manner similar to other seed corn inbreds and resulting hybrids. As discussed above, these inbred s and resulting hybrid s are typically produced under identity3 8 preservation systems that include contracts w ith growers, traceability, product tracking, and process verification since Pioneer and other seed co rn companies take precautions to insure that inbred parent lines are not misappropriated by th ird parties. These proc edures greatly minimize any chances of commingling of the DP-32138-1 seed with other seed and, ultimately, commercial grain.

With the price premiums involved with production of any seed corn variety, including the DP32138-1 corn, there is no benefit for modifyi ng any of the well known procedures to maintain high genetic purity. Using DP-32138-1 in the increase of corn varieties could reduce the yield losses associated with detasseling and the cons equence would be fewer acr es of increase needed per variety. Additionally, there could be a d ecrease in the number of people required for detasseling resulting in reduced expenses and fewer safety hazards during the seed corn

increase. The assumed maximum use of 20,000 acres industry wide for DP-32138-1 is only

about 0.02% of the 80-90 million acres of annual corn produc tion. The agronomic practices used for these seed production acres would be al most identical to the agronomic practices and

locations used under the No Action alternativ e, so no overall effects are anticipated.

Discarded parent seed, whether it is outdated or di scarded from the color sorter, and seed byproducts, such as husks and cobs, will be di sposed by placing in landfills, composting, or incineration (Pioneer 2009). According to Pione er policy, seed producers will not feed discard corn to animals (Public Co mment on EA for DP-32138-1).

Since the DP-32138-1 insertion has three separate ge nes, each of which is required for the production system to work correctly, APHIS consider ed the effects if one or two of these three genes were lost through mutation or recombin ation, which was addressed and discussed in

detail in the petition (Pioneer 2009). DP-32138-1 will maintain the propagation of the male

sterile female inbred as well as e liminate detasseling. All of the pl ants in Steps I, II, and III are inspected for off-type characteris tics and any off-types will be rogu ed out. Since Step I and II fields are very small, these fields can be expect ed to be inspected more heavily than the larger Step III fields. In conventional seed corn produc tion, fields under Steps I, II, and III of seed production are similarly inspected for plants w ith off-type characteristics, so the inspection process is not an additional step required for DP-32138-1 corn. Th ere is the possib ility that the mutation or recombination could take place at a very low frequency during Step II allowing a very low frequency of female inbred plants that would also have male fertility in Step III. This frequency would be expected to be much lower than the incomplete detasseling that happens during conventional seed production. None of these plants would produce seeds with the DsRed2 color marker protein, the only non-native corn protein in DP-32138-1 corn. Please refer to the petition for the complete analysis (Pioneer 2009).

There is always the remote possibility for misdir ection of DP-32138-1 in th e handling, planting, harvesting, transportation or processing stream. Because DP-32138-1 has successfully completed the new protein consultation process for the DsRed2 color marker protein with FDA (Appendix A), there are no human or animal health concerns if DP-32138-1 enters the food or feed supply (see section on Public Hea lth below for more discussion).

Cumulative Effects: Seed Production

Because the acreage, agronomic practices, and locations for se ed production using DP-32138- 1 are expected to be the same or similar to the No Action alternative, no cumulative effects

have been identifie d for this issue.3 9 Organic Farming

Certified organic corn acreage is a small perc entage of overall corn production. The most recently available data show 143,000 acres of organ ic corn production in 2007. This is 0.17% of

the total 86.2 million acres of corn planted in 2007 (USDA-ERS 2008). In 2005, 131,000 acres of organic corn were planted resulting in a 9% increase in organic corn acreage over the 2 year period from 2005 to 2007 (USDA-ERS 2010a). GE co rn has been in production since the mid1990’s, and in 2009 accounted for 85% of the 86.7 million acres of corn planted (USDA-NASS 2009a). GE corn production and organic corn pro duction in the U.S. are both increasing. Based upon trend data, it is anticipate d that both production practices w ill likely continue to increase with or without a determination of nonregulat ed status of DP-32138-1. APHIS also recognizes that producers of non-GE corn, particularly pr oducers who sell their products to markets sensitive to genetically engineer ed traits (e.g. organic or some export markets) can be reasonably assumed to be using practices on their farm to protect their crop from un wanted substances and maintain their price premium. For example, the NOP has recognized the practicality of protecting organically-produced crops, and the investment farmers put into their production practices, by requiring that or ganic production plans include me thods to protect organicallyproduced crops. “Organic crops must be protecte d from contamination by prohibited substances used on adjoining lands (for example, drifting pest icides, fertilizer-laden runoff water, and pollen drift from genetically engineered...)” (NCAT 2003)

.

Typically, there is more than one method for farms under organic practices to prevent unwanted material from entering thei r fields including: isolation of th e farm, physical barriers or buffer zones between organic production and non-org anic production, as well as formal communications between neighboring farms (NCAT 2003). The organic plan used as the basis for organic certification should include a descri ption of practices used to prevent or reduce the likelihood of unwanted substances, like GE polle n or seed, at each step in the farming operation, such as planting, harvesting, stori ng and transporting the crop (Riddle 2004; Krueger 2007; Kuepper, Born et al. 2007). Organic plans should also include mechanisms to monitor the risk of GE pollen or seed co-mingling with the organic crop (Kuepper, Born et al. 2007). Farmers using organic methods are requested to let neighboring farmers know that they are using organic production practices and request that the neighbors also help the organic farmer

reduce contamination events (NCAT 2003; Krueger 2007). Thus , commonly used production practices for corn, and the practical methods typically used by corn farmers using organic methods currently provide many measures that gr eatly reduce the like lihood of accidental gene

flow between DP-32138-1 and non-GE corn fields. These practices protect organic crops and thus maximize profits and price premiums accorded to corn u nder organic production. APHIS

will assume that farmers are alrea dy using, or have the ability to use, these common practices as APHIS’ baseline for the analyses of the following alternatives below. Recommended organic production practices for field corn ar e also readily av ailable (Kuepper 2002). Without any requirements in place for GE corn varieties pr eviously deregulated pursuant to Part 340 and the Plant Protection Act, the production of corn using organic methods continues to increase.

In agricultural systems, growers may choose to grow GE or non-GE co rn, and obtain price premiums for growing varieties of corn for particular markets (e.g., using organic methods for corn production or producing a specia lty corn variety for particul ar processing needs). For example, in 2007, conventional corn averaged $4.19/bushel (USDA- NASS 2010b), whereas organic corn averaged $7.08/bushel (USDA- NASS 2010c). USDA asserts that agricultural4 0 practices that use conventional means, organic production systems, or genetically engineered varieties can all provide benefits to the environment, consumers, and farm income.

No Action: Organic Farming

Current availability of seed for conventional (both GE and non-GE) corn varieties, and those corn

varieties that are developed for organic production, are expe cted to remain the same under the No

Action Alternative. Commercial production of conventional and or ganic corn will not change

and will remain the same under the No Action Alte rnative. Planting and production of GE corn varieties and organic corn have both increased due to market de mands over the last ten years, and these markets are likely to continue to increase under the No Ac tion Alternative (USDANASS 2010a; USDA-NASS 2010c).

Preferred Alternative: Organic Farming

It is not likely that organic farmers who choose not to plant transgenic variet ies or sell transgenic grain will be impacted from the use by the developer of DP-32138-1 corn or by other developers

licensed to use the corn . C ommonly used production practices for corn, and the practical methods typically used by corn farmers using organic methods to protect their crop under organic production (NCAT 2003) pr ovide many measures that gr eatly reduce the likelihood of accidental gene flow between DP-32138-1 and n on-GE corn fields. Pollen from DP-32138-1 corn containing SPT transgenes is not functional and as a result will not fertilize corn plants. Therefore, DP-32138-1 corn is not likely to affect neighboring fa rms. Similar to the No Action Alternative, the use of GE corn varieties and the use of orga nic corn production systems are likely to continue a trend toward increasi ng production under the Pr eferred Alternative.

According to the petition, no transgenes will be present in the seed fo r planting the Step III increase acres (Pioneer 2009). Likewise, after the seed harvest of Step III, the seed for planting Step IV will also contain no transgenes in either commercial corn grain fi elds or in the harvest from the grain fields (Pioneer 2009).

A determination of nonregulated status of DP-32 138-1 will not impact the availability of seed corn for organic production sy stems. In accordance with NOP production standards (7 CFR 205.204a) “the producer must us e organically grown seed, annual seedlings, and planting stock: except, that (1) Nonorganically produced, untreated seed s and planting stock may be used to produce an organic crop when an equivalent organically produced variety is not commercially available.” Organic seed corn is commercially available and therefore in accordance with NOP organic seed corn must be used in certified or ganic corn production systems. The use of DP32138-1 will have no impact on a grower’s ability to obtain seed corn for organic production systems. The Organic Trade Asso ciation provides availabl e resources to locate suppliers that sell organic seeds including corn (OTA 2010).

Cumulative Effects: Organic Farming

Under the No Action Alternative and under the Preferred Alternative, a determination of nonregulated status of DP-32138-1 will not cha nge the market demands for GE corn or corn produced using organic methods. A determination of nonregulat ed status of DP-32138-1 will

not add another GE corn variety to the c onventional corn market, since the SPT genes are confined to the seed production process, a nd are not found in the commodity corn. The SPT technology will not change the amount of GE co rn available for production, because it will only41 replace detasseling operations. Additionally, based upon recent trend information, the market changes of conventionally farmed corn varietie s are not related to the ability of organic production systems to increase their market share. Between 2002 and 2008, acreage of conventionally farmed corn varieties increase d by 12% (USDA-ERS 20 10d) while the acreage associated with the organic production of corn rose 50% during this same time period (USDAERS 2008).

Specialty Corn Systems

Specialty corn, such as waxy corn, white corn, blue corn, and organic corn, comprises 8% of the U.S. market (USGC 2006). With 85% of conventional corn being GE variet ies in 2009, a

market exists for non-GE corn wi th low or no le vel of GE corn. The size of this non-GE corn market, generally for Japan and Europe, was 300, 000 to 550,000 acres in 2005 with a flat projected growth (USGC 2006). These specialty corn products intensively use systems to maintain the purity of the corn product, based on the dema nds of the end-us er (e.g., food processing plants).

No Action: Specialty Corn Systems

The availability of methods used to separate specialty corn products from corn used as grain

would remain the same.

Preferred Alternative: Specialty Corn Systems

A determination of nonregulated status of DP -32138-1 under the Prefer red Alternative would not change the availability of using contract s, tracking and traceability systems, quality assurance processes, closed-loop systems, a nd identity preservation systems to maintain product purity for specialty co rn production. Impacts woul d be similar to the No Action

Alternative.

Cumulative Effects: Specialty Corn Systems

The availability of methods used to separate specialty co rn products from corn used as grain would be the same as currently used in corn producti on systems, and no changes are foreseeable. No

cumulative effects have been identified for this issue.

Physical Environment

Water Resources

Since inbreds are much less vigo rous than hybr ids, irrigation can be preferred in the seed production of inbreds to help assure desired seed yields (Wych 1988), es pecially in those areas subject to freque nt droughts.

No Action: Water Use

Under the No Action Alternative, DP-32138-1 intera ctions with water would be limited to the areas that were approved for regulated releases by APHIS. Irrigation practices associated with conventional seed corn production would not be affected. Under the No Action Alternative, there is no change in the conventional seed produc tion procedures of irrigating corn inbred lines in drought prone areas and no change in irrigatio n practices of commercial corn production.4 2 Preferred Alternative: Water Use

Impacts would be similar to the No Action Alte rnative. DP-32138-1 does not change cultivationpractices for seed production of corn inbreds. A determination of nonregulated status of DP32138-1 will not change the use of irriga tion practices in s eed corn production or commercial corn production . The very small number of acres involved is an additional reason that the use of DP-32138-1 in seed corn production is exp ected to have no effect on water use. Since hybrid corn

will not contain the DP-32138-1 transgenes or non-native corn pr oteins at the commercial stage of planting and harvesting, the consequences of the Preferred Action Alternative on commercial corn production are the same as the No Action Alternative.

Cumulative Effect: Water Use

No cumulative effects ha ve been identified for adetermination of nonregulated status of DP32138-1 corn. A determination of nonregulated stat us of DP-32138-1 will not change the current irrigation practices used in seed corn production or commercial corn production

.

Soil

Certain agronomic practices such as cultivatio n cause disturbance and exposure of the top surface layer of soils and allow some soils to be prone to degradation. Two environmental impacts of soil degradation are the decline in water quality and the possible contribution of

additional CO 2 towards climate change (Lal and Bruce 1999). One production step for corn seed that may have detrimental effects on soil are detasseling operations. Detasseling must take place, rain or shine, when the tassels start to appe ar on the plant. If the so il is wet, using heavy implements, such as the detasseler carriers and the mechanical detasselers, produce large ruts in

the soil. These ruts result in dama ge to the corn roots and negatively impact soil structure which in turn change water movement and nutrient uptake by pl ants (Daum 1996; Kok, Taylor et al. 1996; Gelderman 2009; Tilghman 2009; Roegge 2010).

Detasseling operations resulting in soil compac tion could also potentially affect subsurface soil and constituent organisms. The soil environm ent in and around corn fields is complex, and rich in microorganisms a nd arthropods. The corn root system modifies soil through its

close association with se veral microbial groups such as ba cteria, fungi, pr otozoa, and mites (Bais, Weir et al. 2006). Bacteria typically represent the most abundant microbes in the soil followed by fungi. These microbial groups play an im portant and particular role in the ecology of the soil, including nutrimental cy cling and the availability of nutrients for plant growth (Hoeft, Nafziger et al. 2000; OECD 2003)

No Action: Soil

DP-32138-1 interactions with the soil would be limited to the ar eas that are approved by APHIS for regulated releases. Cultivation practices asso ciated with conventiona l seed corn production

would not be affected. The soil environment woul d be modified by corn roots and crop soils would still be affected by agronomic practices associated with conventional seed corn cultivation, including the negativ e impacts associated with de tasseling when soils are wet. Cultivation practices associated with commercial production of corn would not be affected.

Preferred Alternative: Soil43 In the event of a determinati on of nonregulated status of DP-32138-1 under the Preferred Alternative, soil interactions with DP-32138-1 would occur at a large r scale than under the regulated releases. DP-32138-1 seed corn producti on could have three possible effects on soils as compared to conventional seed corn production: no effects from detasseling, possible presence

of additional ZM-AA1 alpha-amylase protein, and possible presence of DsRed2 color marker protein

.

Cultivation practices associated with DP-32138-1 seed corn production would be the same as conventional seed corn production during seed bed preparation, planting and early plant development. The soil environment would be m odified by corn roots an d crop soils would still be affected by agronomic practices associated with conventional seed corn cultivation. However, for DP-32138-1 seed corn production, detasseling would no longer be requ ired and the negative consequences of detasseling would be avoided, su ch as compaction and large ruts in the soil. Generally these seed corn produc tion fields, including DP-32138- 1 seed corn fields, are in rotation with other crops. These rotations with other non-corn crops generally result in averaging out any positive or negative impacts that may ha ve resulted from the seed corn production. These positive or negative effects cannot be quantified because the DP-32138-1 seed corn fields are a very small proportion of commercial corn production . In comparison to commercial corn grain production in which no detasseling takes place, DP-3 2138-1 seed corn would be no different.

DP-32138-1 expresses small amounts of ZM-AA1 alpha-amylase protein during plant development and only at low levels in the seed at physiological maturity (Pioneer 2009). In both conventional and DP-32138-1 seed corn production, the rows of male plants are generally destroyed to aid in increasing seed yield of the female rows by decreasing plant competition and to decrease the chances of commingling of male and female seeds at harvest (Pioneer 2009). In DP-32138-1 corn, the plants that contain the tr ansgenes (male fertile) would be destroyed following pollination, generally by chopping up the plant. These immature plants contain low levels of ZM-AA1 alpha-amylase protein thro ughout plant tissues. Consequently, there is a potential for ZM-AA1 alpha-amylase protein to be incorporated into agricultural soils. However, alpha-amylases are ubiquitous enzymes found in micr oorganisms, plants, and animals (Jane č ek, Lévêque et al. 1999).

If the DP-32138-1 plants are allowed to reach physiol ogical maturity, these plants are generally harvested with most, if not all, of the seed rem oved from the field. For seed that may remain in the field after harvest, physiologically mature seed of DP-32138-1 will contain low levels of ZMAA1 alpha-amylase protein. However, if soil moisture and temperatures are adequate to stimulate germination, seeds of both conventional and DP-32138-1 would produce similar amounts of ZM-AA1 alpha-amylase protein. Since the levels of ZM-AA1 alpha-amylase protein are essentially the same in conventional corn and DP-32138-1 corn, impacts of DP-32138-1 seed corn production on the soil would be similar to those in conventional corn seed production.

DP-32138-1 expresses small amounts of the DsRed2 color marker protein during plant development with the highest leve ls in the seed detected at ph ysiological maturity (Pioneer 2009). During the production of non transgenic male sterile inbred for seed increase, plants that contain the transgenes may be chopped and left on the soil surface (Pioneer 2009). These immature plants express low levels of DsRed2 color marker protein throughout the plant. The DsRed2 color marker protein will be susceptible to degradat ion and adsorption to humus or minerals as are other proteins in agricultural soils (Bastida, Moreno et al. 2009). Soils contain numerous organisms and enzymatic activity from decomposed organisms that degrade a wide4 4 array of molecules including proteins (Bastida, Moreno et al. 2009). These processes would metabolize or sequester the DsRed2 color marker protein with no likely effect on the soil.

If the DP-32138-1 plants are allowed to reach physiol ogical maturity, these plants are generally harvested with most, if not all, of the seed re moved from the field. In the few seeds that may remain in the field after harvest, physiologica lly mature seed of DP -32138-1 will contain DsRed2 color marker protein. Similar to DsRed2 th at originated in whol e plants, normal soil degradation and adsorption would be e xpected to remove this protein.

For the preferred alternative of deregulating DP-32138-1 corn, the three possible effects of no detasseling, possible presence of additional ZM-AA1 alpha-amylase protein and possible presence of DsRed2 color marker protein combined with the very low presence in the environment (a maximum 20,000 acres), impacts on soil are expected to be similar to those from conventional corn seed production. Sin ce hybrid corn will not contain the DP-32138-1 transgenes or non-native corn pr oteins at the commercial stag e of planting and harvesting, the consequences of the Preferred Action Alternativ e on commercial corn prod uction are the same as

the No Action Alternative.

Cumulative Effects: Soil

APHIS has not identified any cumulative effects for this issue. The fields devoted to seed corn production, conventional as well as DP-32138-1 s eed corn, are normally rotated with other crops with the maximum potential for 20,000 acres expected to be devoted to DP-32138-1 seed corn.

DP-32138-1 seed corn acres would be only 0.02% of all corn production acres. APHIS

concludes that adverse effects of the ZM-AA1 alpha-amylase protein or of the DsRed2 color marker protein are not likely, effects of avoiding detasseling are slightly positive, if any, and exposure of 0.02% of all annual corn production acres is small, and therefore with adverse impacts on soil unlikely, no cumulative impacts are li kely. Since hybrid corn will not contain the

DP-32138-1 transgenes or non-native corn proteins at the commercial stage of planting and harvesting, the effects of the the Preferred Actio n Alternative on commercia l corn production are not significant and are therefore no cumulative effects are expected.

Air Quality

Tillage, traffic and harvest emissions, pesticide drift from spraying, and nitrous oxide emissions from the use of nitrogen fertilizer may affect air quality (Aneja, Schlesinge r et al. 2009). Tillage

contributes to the release of GHGs because of the loss of CO 2 to the atmosphere, and the exposure and oxidation of soil organic matter (Baker , Southard et al. 2005). Emissions released from agricultural equipment (e.g., irrigation pumps and tractors) include carbon monoxide, nitrogen oxides, reactive organic gases, part iculate matter, and sulfur oxides (US-EPA 2010). Nitrous oxide may also be released following th e use of nitrogen fert ilizer (US-EPA 2010). Aerial application of pesticid es may cause impacts from drift and diffusion. Pesticides may volatilize after application to soil or plant su rfaces and move following wind erosion (Vogel, Majewski et al. 2008). Agriculture, including land-u se changes for farming, is responsible for an

estimated 6% of all human-induced GHG emissions in the U.S., and N 2 O emissions from agricultural soil management are a larg e part of this, 68 % of all U.S. N 2 0 emissions (US-EPA 2010).

No Action: Air Quality4 5 Under the No Action Alternative, DP-32138-1 interactions with the air would be limited to the areas that were approved for regulated releases by APHIS. Cultivation practices associated with conventional seed corn pr oduction or commercial corn production would not be affected. Air quality would still be affected by agronomic pra ctices associated with conventional seed corn cultivation and commercial corn pr oduction such as tillage, pestic ide application, and use of agricultural equipment.

Preferred Alternative: Air Quality

DP-32138-1 seed corn production does not change cultivation practices for seed production of corn inbreds. A determination of nonregulated status of DP-32138-1 will not change the use of tillage, use of agriculture equipment, irrigation, pe sticide applications and fertilizer applications in seed production or commercial corn produc tion. With DP-32138-1, no detasseling would be required, subsequently reducing the combusti on gas emissions from detasseler carriers,

mechanical detasselers and moveme nt of detasselers between homes and seed fields resulting in a beneficial effect on air qualit y. Considering the very small nu mber of acres involved, the use of DP-32138-1 in seed corn pr oduction is expected to have mi nimal effects on air quality.

Since hybrid corn will not contain the DP-32138-1 transgenes or non-native corn proteins at the commercial stage of planting and harvesting, the consequences of the Preferred Action Alternative on commercial corn production are the same as the No Action Alternative.

Cumulative Effects: Air Quality

APHIS has not identified any cumulative effects for this issue. The use of DP-32138-1 in seed corn production is expected to have no cumulative effect on air qua lity because of the very small

number of acres that would be involved in DP -32138-1 seed corn production and there are no changes in cultivation practices. Since hybrid corn will not contain the DP-32138-1 transgenes or non-native corn proteins at the commercial stage of planting and harvesting, the effects of the

Preferred Action Alternative on commercial corn production were not significant, and consequently there are unlikely to be any cumulative effects.

Climate Change

Production of agricultural commodities is one of the many human activities that could possibly GHGs that affect climate (see discussion in Affected Area, Physical Environment). CO 2 , nitrogen dioxide (NO 2 ), and methane (CH 4 ) may be produced through the combustion of fossil fuels to run farm equipment, the use of fertilizers, or the decomposition of agricultural waste products including crop residues and animal wastes. Classes of cr ops planted are relevant to climate change, as are the locations and the soil types in which they are planted. Climate change itself may force changes to agricu ltural practices by extending the ranges of weeds and pests of agriculture (IPCC 2007). Indirect effects of new crops will be determined by the traits engineered into organisms and the management strategies used in the production of these organisms.

No Action: Climate Change

Under the No Action Alternative, environmental re leases of DP-32138-1 would be under APHIS regulation. There would be no measurabl e effect from these confined environmental releases. Cultivation practices associated with conventional seed corn production or4 6 commercial corn production would not be affected. Agronomic practices associated with

conventional seed corn cultivatio n or commercial corn production such as tillage, pesticide application, and use of agriculture equipment would remain the same.

Preferred Alternative: Climate Change

DP-32138-1 seed corn production does not change cultivation practices for seed production of corn inbreds. A determination of nonregulated st atus of DP-32138-1 will not change the use of tillage, use of agriculture equipment, irrigation, pe sticide applications and fertilizer applications in seed production on commercial corn production . With DP-32138-1, no detasseling would be required, consequently reducing the energy re quirements for detasseler carriers, mechanical detasselers and for movement of detasselers between homes and seed fields resulting in a reduced level of greenhouse gases. However, wi th the relatively small number of acres that would be involved with seed corn production using DP-32138-1 a slight decrease of GHG production is likely too small to calculate any meaningful impact for climate change. Since hybrid corn will not contain the DP-32138-1 transgenes or non-native corn proteins at the commercial stage of planting and harvesting, the consequences of the Preferred Action Alternative on commercial corn production are the same as the No Action Alternative.

Cumulative Effects: Climate Change

APHIS has not identified any cumulative effects for this issue. The use of DP-32138-1 in seed corn production is expected to have no cumulative effect on climate change b ecause of the very small number of acres that would be involved in DP-32138-1 seed corn production and there are no changes in cultivation practices. Since hybrid corn will not contain the DP-32138-1 transgenes or non-native corn proteins at the commercial st age of planting and harves ting, the effects of the

Preferred Action Alternative on commercial corn production were not significant, and consequently there are no cumulative effects expected.

Animal and Plant Communities

Animals

Corn production systems in agricu lture are host to many animal species. Mammals and birds may seasonally use grain, and invertebrates can feed on the plant during the entire growing season. The cumulative effects analysis for this issue is found below at “Cumulative Effects: Plants, Animals, Biodiversity.”

No Action: Animals

Under the No Action Alternative, environmental re leases of DP-32138-1 would be under APHIS regulation. Animal incursions would be limite d to regulated field trials. The new protein consultation with FDA was successfully completed for DsRed2 color marker protein, the only non native corn protein in DP-32138-1 (Appendix A), whic h addressed any concerns of composition. The consultation demonstrated a lack of toxicity and allergenicity of DP-32138-1 for human and animal consumption. Based upon the FDA’s new pr otein consultation, APHIS supports Pioneer’s conclusions th at corn produced using Pioneer’s SPT technology is considered safe for animal consumption.

Preferred Alternative: Animals4 7 APHIS has reviewed and accepts th e data submitted by the appli cant, which are similar to the data submitted during the FDA new protein consultation process for DsRed2 color marker protein, the only non native corn protein in DP-32138-1 (Appendix A). Impacts would be

similar to the No Action Alternative. The agronomic practices used to produce DP-32138-1

will be the same as those used to produce c onventionally grown seed corn. Therefore, the discussion of effects on animals of DP-32138-1 will focus solely on the introduced proteins in DP-32138-1 corn, the ZM-AA1 alpha-amylase protein and the DsRed2 color marker protein.

Pioneer DP-32138-1 has no t been genetically engi neered to produce any pesticides. DP-32138-

1 contains the ZM-AA1 protein in the developing pollen and in other parts of the plant at low levels except at physiological maturity when it has declined below the limits of quantitation (Pioneer 2009). The ZM-AA1 alpha-amylase pr otein is normally found predominantly in germinating kernels (sprouts) of conventional corn. Amylases may also be found in germinating pollen (Bhadula and Sawhney 1989; Castro and Clement 2007), including corn (Brewbaker 1971; Agarwala, Sharma et al. 1981; Wakhle, Phadke et al. 1983). In DP-32138-1, ZM-AA1 concentration is 400 times greater in pollen than in seed and 64 tim es greater than in R4 stage leaves. The alpha-amylases are ubiquitous enzyme s present in many organisms including plants and insects (Campos, Xavier-Filho et al. 1989; Raim baud, Buleon et al. 1989; Silva, Terra et al. 1999; Cristofoletti, Ribeiro et al. 2001). The al pha-amylases from multiple sources including plants, fungi and bacteria have a long history of safe consumption in foods (Pariza and Johnson 2001). Animals that feed primarily on corn kernels are seed-fee ding insects and rodents found in agricultural fields. During field trials, the a pplicant found no differences in insect feeding damage (Pioneer 2009), indicating similar ins ect susceptibility of DP-32138-1 tissues and

conventional corn.

DP-32138-1 contains the DsRed2 color marker protein in the seed (410 ng/mg tissue dry weight) but not in leaves or pollen of physiologically mature plants, and at lower levels in leaves (87 ng/mg during the vegetative growth stage a nd 160-210 ng/mg during th e early reproductive stages of growth) of the growing plant (Pioneer 2009). Rodents, such as mice or squirrels, in some seasons may feed exclusively on corn kernels. Thus, these animals are most likely to have a diet containing large amounts of corn kernels. DsRed2 showed no signs of toxic ity when used as a vital marker in mice (Figueiredo, Fernandes et al . 2008). In a feeding study of mice to determine acute toxicity level of the DsRe d2 protein, a wide margin of sa fety was observed (Pioneer 2009). Other mammals, such as deer, would have even lower exposure to DsRed2 color marker protein

and ZM-AA1 alpha-amylase protein because of feeding habits; for example, deer nibble on tips of corn ears as opposed to kernels (Steffey, Rice et al. 1999). Finally, transg enic plants stably expressing DsRed variants did not show any ab normalities in growth habits, development, fertility, germination or morphogenesis (Jach, Binot et al. 2001; Dietrich and Maiss 2002; Stuitje, Verbree et al. 2003; Wenck, Pugi eux et al. 2003; Mirabella, Fra nken et al. 2004; Pioneer 2009).

APHIS has reviewed this information and has determined that there would be no negative effects to animals that forage on DP-32138-1 corn (USDA-APHIS 2010).

Plants

Corn production acreage is host to many plant species as well. The landscape surrounding a

corn field varies depending on the region. In certain ar eas, corn fields may be bordered by other corn (or any other crop); fields may also be surrounded by wooded or past ure/grassland areas. Therefore, the types of vegetation, including weeds, around a corn field depend on the area where the corn is planted. A variety of weeds dw ell in and around corn fi elds; those species will4 8 also vary depending on the geographic region wher e the corn is planted. Corn itself is not sexually compatible with any other plant speci es found in the U.S. (USDA-APHIS 2010). The cumulative effects analysis for this issue is found below at “Cumulative Effects: Plants, Animals, Biodiversity.”

No Action: Plants

Under the No Action Alternative, environmental releases of DP-32138-1 would be under APHIS regulation. Plant species that t ypically inhabit corn production systems will be managed as in conventional corn production, incl uding the use of mechanical, cu ltural, and chemical control

methods.

Preferred Alternative: Plants

In the event of a determination of nonregulated status of DP- 32138-1, agricultural practices used for conventional corn would be used for plant management during the cultivation of DP-32138-1 corn. Impacts would be similar to the No Action Alternative. DP-32138-1 does not exhibit characteristics associated with weedy growth an d will not compete with pl ants found outside of agricultural production (USDA- APHIS 2010). Weeds within fi elds of DP-32138-1 will be managed using mechanical, cultural, and chemi cal control, as weeds are now managed in conventional corn systems. As there are no to xic effects on animals (see Animals discussion above), there are no toxic effects on animals that could be pollinator s of other plants in or around

fields cultivated with DP-32138-1 corn. DP-32138-1 has not been genetically engineered to be tolerant to any herbicides.

Biological Diversity

Biological diversity, or the variation in species or life forms in an area, is highly managed in agricultural systems. Farmers typically plant crops that are genetically adapted to grow well in a specific area of cultivation and have been bred for a specific market. In the case of corn agriculture, varietie s have been developed for food proces sing needs (e.g., waxy corn), consumer qualities (e.g., blue corn or wh ite corn), or for use as a ve getable (e.g. sweet corn). In conventional agriculture, farmers want to enco urage high yields from their corn crop, and will intensively manage the ‘plant communities,’ or weeds, found in corn crops through chemical, cultural or mechanical means. Animals, particular ly insect and other pest species will also be managed through chemical and cultural controls to protect the crop from damage by certain animal pests. Therefore, the bi ological diversity in agricultural systems (the agro-ecosystem) is highly managed and may be lower than in th e surrounding habitats. The cumulative effects analysis for this issue is found below at “Cumulative Effects: Plants, Animals, Biodiversity.”

No Action: Biological Diversity

Under the No Action Alternative, environmen tal releases of DP-32138-1 would be under APHIS regulation. Animal and plant species th at typically inhabit seed corn and commercial production systems will be continue to be affected by the management plan of typical conventional corn production, whic h includes the use of mechanical, cultural, and chemical control

methods.

Preferred Alternative: Biological Diversity4 9 The importance of corn as a food crop, and its dependence on human management, has produced a long history of effort to protect the integrity of the germplasm lines of corn. Decades prior to the introduction of transgenic corn products, the corn industry developed effective methods and means to maintain produc t segmentation and standa rds of genetic purity. Specialty corn crops, for example, were succe ssfully isolated over long term cultivation and continue to be grown today, even with transgenic commodity corn widely adopted in the U.S. Moreover, with respect to both conventional and transgenic corn, the ability to protect and maintain the genetic purity of breeding lines is critical to seed companies and developers of new technologies such as DP-32138-1 . Consequently, seed companies routinely apply standard breeding techniques, including physi cal and temporal isolation, wh ich have proven effective at maintaining the genetic purity of breeding lines (Wych 1988).

Genetically engineered corn lines have b een available on the market since 1994 and the preponderance of evidence in peer-reviewed literatu re does not establish any consistent adverse effect of its production on biodiversity (Bitz er, Rice et al. 2005; Torres and Ruberson 2005; Romeis, Meissle et al. 2006; Marvier, Mc Creedy et al. 2007; Chen, Zhao et al. 2008; Wolfenbarger, Naranjo et al. 2008 ). APHIS’ review and analys is of Pioneer’s data (USDAAPHIS 2010) indicate that the line DP-32138-1 exhibits no traits that would cause increased weediness, that its unconfined cultivation shoul d not lead to increased weediness of other cultivated corn or other sexually compatible relatives, and that it is unlikely to harm non-target organisms common to the agricultu ral ecosystem or threatened or endangered species recognized by the U.S. Fish and Wildlife Service.

Cultivation of DP-32138-1 seed corn requires the same agrono mic practices as conventional seed corn production. Anim al and plant species that typically inhabit seed corn production systems will be managed as in conven tional corn production, likely with th e use of mechanical, cultural,

and chemical control methods. Ov erall impacts would be similar to the No Action Alternative. Since hybrid corn will not contain the DP-32138-1 transgenes or non-native corn proteins at the commercial stage of planting and harvesting, the consequences of the Preferred Action Alternative on commercial corn production are the same as the No Action Alternative.

Cumulative Effects: Animals, Plants, Biodiversity

APHIS has determined that there are no impacts from past, present, or reasonably foreseeable actions that would aggregate with effects of the proposed action to create cumulative impacts or reduce the long-term productivity or sustainability of any of the resource s associated with the ecosystem in which the DP-32138-1 is planted. DP-32138-1 has not been genetically engineered to produce a toxin or pesticide, and has not been genetically engineered to be tolerant to an herbicide. Although some studies have found both increases and decreases in animal and plant diversity and abundance in the agro-ecosystem due to the use of GE crops (Hansen Jesse and Obrycki 2000; Ponsard, Gutierrez et al. 2002; Brooks, Bohan et al. 2003; Haughton, Champion et al. 2003; Hawes, Haughton et al. 2003; Marshall, Brown et al . 2003; Roy, Bohan et al. 2003; Romeis, Dutton et al. 2004; Sister son, Biggs et al. 2004; Bitzer, Ri ce et al. 2005; Pilcher, Rice et al. 2005; Torres and Ruberson 2005; Romeis, Meissle et al. 2006; Marvier, McCreedy et al. 2007; Chen, Zhao et al. 2008; Wolf enbarger, Naranjo et al. 2008), DP-32138-1 is unlikely to affect the animal or plant communities found in conventional corn production systems because of the lack of toxicity and allergenicity, and because there will be no change to agronomic practices caused by the cultivation of DP-32138-1 corn.5 0 Gene Movement

An environmental impact to consider as a result of planting this corn variety is the potential for gene flow (the transfer of genetic information between different individuals or populations). Pollen flow, or the movement of genes from one plant to another, occurs between plants that are sexually-compatible, or able to receive po llen at the appropriate time during the a ppropriate plant stage. Corn does not have sexuall y-compatible relatives found in ‘natural’ ar eas; in the U.S., corn is only able to reproduce with other cu ltivated corn plants (USDA-APHIS 2010).

Corn pollen moves by the wind (OECD 2003). Succes sful gene movement from one plant to another requires many different biological and physical factors, such as synchrony of flowering between corn fields, viability of pollen, and presence of physical barriers (OECD 2003); thus pollen movement is not eq uivalent to gene movement. A recent paper (Sanvido, Widmer et al. 2008) reviewed studies investigating gene flow and gene movement studies in corn grain production fields, and using the data found that the gene movement

from GE corn to non- GE corn typically remained below 0.5% at 50m (approx. 164ft). In large scale studies, this same frequency of cross-fertil ization events was similarly validated (Henry, Morgan et al. 2003; Weber, Bringezu et al. 2007).

One study found cross-fertilization rates higher at comparable distances than other studies (Jones and Brooks 1950). Jones and Brooks (1950) found successful gene movement to be as high as 2.5 % at 660ft. One potential reason for the discrepancy between this study and most other gene flow studies in corn ma y be a result of the type of co rn used in this study. Jones and Brooks (1950) investigated the appropriate isolation dist ance for seed production in open-pollinated varieties, and not for hybrid variet ies. Due the biology of open-pollinated varieties, these types of plants may be more receptive to pollen over a longer period of time than hybrid corn plants (Sanvido, Widmer et al. 2008), allowing fo r a greater chance of pollination events. Thus, the results from Jones and Brooks (1950) may overestimate the potential for cross-fertilization of hybrid corn plants.

No Action: Gene Movement

Under the No Action Alternativ e, DP-32138-1 would remain a re gulated article and would require an APHIS permit or notification for release into the environment. Under regulated releases, regulated GE corn is typically separated from non-regulated corn by a distance of 660ft, based on distance s set for seed production (AOSCA 2004), if distance is the only method used to prevent movement of pollen or genes. APHIS has concluded this separation distance is sufficient to substantially limit gene movement to other corn crops.

Preferred Alternative: Gene Movement

In 2009, GE corn was planted on 85% of all corn ac res currently in prod uction in the US, and has been steadily increasing (USDA-NASS 2009a). Concurrently, organic corn acreage has also increased by 9% over the 2 year period from 2005 to 2007 (USDA-ERS 2010a). Separation between the two types of corn varieties has been maintained by means of only recommended practices for organic growers and no mandated requirements for GE corn. P roduction of both organic and conventional corn will likely c ontinue to increase with or without a determination of nonregulated status of DP-32138-1.51 In agricultural systems, growers may choose to grow GE or non-GE co rn, and obtain price premiums for growing these varieties of corn for particular markets (e.g., using organic methods for corn production or producing a specialty corn variety for particular processing needs). For example, in 2007, conventional corn averaged $4.19/bushel (USDA- NASS 2010b), whereas organic corn averaged $7.08/bushel (USDA- NASS 2010c). USDA asserts that agricultural practices that use conventional means, with GE or non-GE vari eties, and orga nic production systems can all provide benefits to the environment, co nsumers, and farm income. Gene

movement into and out of thes e specialized corn production systems have been managed using various types of buffer zones or isolation prac tices, such as differen ces in planting (which results in differences in flowering) or making sure fields are suffici ently distant from other compatible crops (such as usi ng appropriate isolation distances).

In addition to the typical identity preservation and closed loop systems in place for specialty crops, those farmers using organi c production also put into pla ce measures to maintain purity of their crops. Typically, more than one method is used by farmers under organic production rules to prevent unwanted material from enteri ng their fields including: isolation of the farm, physical barriers or buffer z ones between organic production and non-organic production, and formal communications between neighboring farms (NCAT 2003). The plan used as the basis for organic certification should include a description of practices used to prevent or

reduce the likelihood of unwante d substances, like GE pollen or seed, at each step in the farming operation, such as planting, harvesting, storing and transporting the crop (Riddle 2004; Krueger 2007; Kuepper, Born et al. 2007). Practical methods typically used by corn farmers will protect their crop and maximize thei r profits and premiums. The financial benefits accrue to organic corn produced using a pproved plans that are also su ccessfully accomplished. For example, farmers using organic me thods are requested to let ne ighboring farmer s know that they are using organic pr oduction practices and request that th e neighbors also help the farmer reduce contamination events (NCAT 2003; Krue ger 2007). Thus, commonly used organic production practices for corn cu rrently provide many measures th at greatly reduce the likelihood of accidental gene flow. In addition, pollen contai ning the SPT transgene is not fertile, and gene

flow between DP-32138-1 and non-GE corn fields is highly unlikely.

DP-32138-1 would be an additional GE corn va riety that may be available to the seed production industry. The production area for DP-3213 8-1 seed is likely limited to less than

5000 acres if SPT is utili zed only by Pioneer or no more than 20,000 acres if SPT is licensed to all seed corn pr oducers in the U.S. Overall impacts of gene movement would be similar to

the No Action Alternative.

Cumulative Effect: Gene Movement

DP-32138-1 would be an additiona l technology that may be available to the seed corn industry. The production area for DP-32138-1 seed is likely limited to less than 5000 acres if SPT is used only by Pioneer or no more than 20,000 acres if SPT is licensed to all seed corn producers in

the U.S. The production of conventional, GE and organi c corn is expected to continue. GE and organic acreage currently coexist, and there is no change in this coexistence with DP-32138-1 corn. The potential for movement of transgenes from GE crops into non GE varieties is not expected to change as a result of approving DP-32138-1.5 2 Public Heath

Human Health

Under the FFDCA, it is the responsibility of food and feed manufacturers to ensure that the products they market are safe an d properly labeled. Food and feed derived from DP-32138-1 must be in compliance with all applicable legal and regulatory requirements. GE organisms for food and feed may undergo a voluntary consultation process with the FDA prior to release

onto the market.

No Action: Human Health

The FDA considers Pioneer's consultation on DsRe d2 color marker protein, the only non-native corn protein in DP-32138-1 corn, and the consultation on the al pha-amylase ZM-AA1 to be complete (Appendix A). The status of the FDA consultation will not change under the No

Action Alternative.

Preferred Alternative: Human Health

DP-32138-1 is genetically engineered to contain a sequence of DNA with three genes, Ms45

, zm-aa1 , and DsRed2 (Alt1). Ms45 is a corn gene that encodes the MS45 protein required for the production of fertile pollen and is controlled by a corn an ther preferred promoter 5126.
zm-aa1 is a corn gene that encodes the ZM-AA1 alpha-amylase protein, which breaks down starch (Jane č ek 1994). zm-aa1 is controlled by a corn pol ygalacturonase promoter ( Pg47 ) that targets expression to pollen (Allen and Lonsda le 1993). When the ZM-AA1 protein is expressed in immature pollen, the depletion of starch reserves renders the pollen infertile. The DsRed2 gene is from a marine coral-like anemone Discosoma sp. (Matz, Fradkov et al. 1999; WassonBlader 2001) that was modi fied by a single base pa ir substitution to produce DsRed2(Alt1).

The lipid transfer protein ( Ltp2 ) promoter from barley targets e xpression to the aleurone layer of the seed (Kalla, Shimamoto et al. 1994). DP- 32138-1 seeds expressing the DsRed2 color marker protein are colored pinkish-red, which will enable detection and sorting of transgenic red corn seeds from non-transgenic yellow seeds using a color sorting machine (Wasson-Blader 2001; Pioneer 2009).

The MS45 protein is a native corn fertility gene. The MS45 protein in DP -32138-1 was found only in the developing anther tissues during early pollen formation, the same location as conventional corn, and was not found in later stages of pollen formation or in other parts of the plant (Pioneer 2009) . Pioneer also submitted to APHIS information on identity, function, characterization of genes, expre ssion levels of gene products, as well as information on the potential allergenicity a nd toxicity of the expr essed MS45 protein. APHIS’ assessment of the safety of this product for human s and animals focuses on plant pest risk (USDA-APHIS 2010)

and effects on wildlife and threatened and en dangered species (sec tion on Animals and Threatened and Endangered Species), and thos e analyses are based on the comparison of the GE-corn to its non-GE counterpart . No new issues appear to be associated with the Ms45 transgene in DP-32138-1 corn.

In conventional corn, the alpha-amylase protein is expressed predominantly in the scutellum of germinating seed and minimal expression is observed in the developing endosperm. The endogenous corn alpha-amylase protein cannot be detected in non transformed corn pollen (Pioneer 2009). Alpha-amylases ar e ubiquitous in the environmen t, being naturally present in53 microorganisms, plants and animals (Jane č ek, Lévêque et al. 1999). Many types of commercial food processing, feed ingredient ap plications, and industrial applications also utilize alphaamylase enzymes, including the production of fu el and potable alcohol (brewing, distillation processes), and corn syrups (Jane č ek, Lévêque et al. 1999; Lévêque, Jane č ek et al. 2000; Pariza and Johnson 2001; Olempska-Beer, Merker et al. 2006).

In DP-32138-1 corn, the ZM-AA1 protein is additionally f ound in the developing pollen and at low levels in other parts of the plant except at physiological matur ity when it is below levels of quantitation (Pioneer 2009). Since the ZM-AA1 protein in DP-32138-1 is found in a new tissue location in corn, a new protein consultation wa s submitted to the FDA. Pioneer provided the FDA with information on identity , function, and characterization of the genes, as well the expression levels of the gene products. They al so provided information on the potential allergenicity and toxicity of the expressed proteins. The FDA considers Pioneer's consultation on ZM-AA1 alpha-amylase protein to be complete (Appendix B). Pioneer also submitted information on identity, func tion, characterization of genes, expression levels of gene products, as

well as information on th e potential allergenicity and toxici ty of the expressed proteins to APHIS. APHIS’ assessment of the safety of this product for humans and animals focuses on plant pest risk (USDA-APHIS 2010) and effects on wildlife and threatened and endangered species (section on Animals and Threatened and Endangered Species). Thes e analyses are based on the comparison of the GE-corn to its non-GE c ounterpart. No new issu es appear to be

associated with the zm-aa1 transgene in DP-32138-1 corn.

The DsRed2 color marker protein is th e only expressed transgenic protein in DP-32138-1 not derived from corn. In physiologically mature plants of DP-32138-1 corn, the DsRed2 protein is found in the seed, but not in leav es or pollen. In the growing plan t, however, it is found in leaves (Pioneer 2009). Pion eer provided the FDA with inform ation on identity, function, and characterization of the genes, as well the expr ession levels of the gene products. They also provided information on the potential allergenicity and toxicity of the expressed proteins. The

FDA considers Pioneer's cons ultation on DsRed2 protein in DP-32138-1 to be complete (Appendix A). Pioneer also submitted information on iden tity, function, characterization of genes, expression levels of gene products, as well as information on the potential allergenicity and toxicity of the expressed protei ns to APHIS. APHIS’ assessment of the safety of this product

for humans and animals focuses on plant pest risk (USDA-APHI S 2010) and effects on wildlife and threatened and endangered species (section on Animals a nd Threatened and Endangered Species), and those analyses are based on the comparison of the GE-corn to its non-GE counterpart. No new issues appear to be associated with the DsRed2 (Alt1) transgene in DP32138-1 corn.

Based on the assessment of laboratory data provided by Pioneer in the submitted petition and an analysis of the scientific literature (USDA-APHI S 2010), along with the completion of the consultation process with FDA regarding DsRed2 color marker protein in DP-32138-1 (Appendix A), APHIS has concluded that under this alternative, the proposed action to approve nonregulated status for DP-32138- 1 would have no significant impacts on human or animal

health.

Cumulative Effects: Human Health5 4 There are no significant impacts on human or animal health related to the No Action Alternative

or a determination of nonregulated status of DP-32138-1 corn, and no cumulative effects have

been identified.

Worker Safety

No Action: Worker Safety

During agricultural production of corn, farmers may be exposed to pesticides during application of these chemicals to crops. Under the No Acti on Alternative, exposure to these agricultural chemicals during corn production would remain the same. Under the No Action Alternative, workers continue to be exposed to safety ha zards associated with farm equipment during detasseling operations.

Preferred Alternative: Worker Safety

Worker safety issues related to the use of pestic ides during agricultural pr oduction of DP-32138-1

would remain the same as the No Action Alte rnative. As discusse d under the issue of “Cropping Practices: Crop Rotation, Tillage, Produc tion, and Pesticide Use”, DP-32138-1 does not change the agronomic practices, or use of chemical s such as pesticides, associated with seed corn production. With DP-32138-1, detassel ing would no longer be needed and therefore workers would no longer be exposed to safety hazards associated with farm equipment during detasseling operations.

Cumulative Effects: Worker Safety

Worker safety issues related to agronomic practices and the use of pesticides during agricultural production of DP-32138-1 would continue and remain the same under both alternatives. As

discussed under the issue of “Pesticide Us e”, DP-32138-1 does not change the agronomic practices, or use of chemicals such as pesticides, associated with seed corn production. With DP-32138-1, detasseling would no longer be need ed and therefore, work ers would no longer be exposed to safety hazards associated with fa rm equipment during detasseling operations. There

are no cumulative effects identified for this issue.

Animal Feed

More than 60 % of the corn produced in the US and much of the exported corn is used for animal feed (Hoeft, Nafziger et al. 2000). Under the FFDCA, it is th e responsibility of feed manufacturers to ensure that the products they market are safe and pr operly labeled. Feed produced with DP-32138-1 technol ogy must be in compliance with all applicable legal and regulatory requirements. GE organisms may unde rgo a voluntary consultation process with the FDA for feed safety prior to release onto the market.

No Action: Animal Feed

The FDA considers Pioneer's consultation on DsRe d2 color marker protein, the only non-native corn protein in DP-32138-1 corn, to be complete (Appendix A). The FDA also considers Pioneer's consultation on ZM-AA1 alpha-amylase protein to be complete (Appendix B). This native corn protein is normally expr essed in germinating seeds but in DP-32138-1 corn the5 5 modified amylase is expressed in the pollen. The status of the FDA consultations will not change under the No Action Alternative.

Preferred Alternative: Animal Feed

DP-32138-1 is genetically engineered to contain a sequence of DNA with three genes, Ms45

, zm-aa1 , and DsRed2 (Alt1). Ms45 is a corn gene that encodes the MS45 protein required for the production of fertile pollen and is controlled by a corn an ther preferred promoter 5126.
zm-aa1 is a corn gene that encodes the ZM-AA1 alpha-amylase protein, which breaks down starch (Jane č ek 1994). For a detailed description of each of gene and its impacts, please refer to

the Human Health section above.

The FDA considers Pioneer's consultati on on ZM-AA1 alpha-amylase protein in DP-32138-1 to be complete (Appendix B). Pioneer also submitted inform ation on identity, function, characterization of genes, expre ssion levels of gene products, as well as information on the potential allergenicity and toxicity of the expressed proteins to APHI S. APHIS’ assessment of the safety of this product fo r animals focuses on plant pest risk (USDA-APHIS 2010) and effects on wildlife and threatened and endangere d species (section on Animals and Threatened and Endangered Species), and th ose analyses are based on the comparison of the GE-corn to its non-GE counterpart. No new issues appear to be associated with the zm-aa1 transgene in DP32138-1 corn (Pioneer 2009).

The FDA considers Pioneer's c onsultation on DsRed2 protein in DP-32138-1 to be complete (Appendix A). Pioneer also submitted information on iden tity, function, characterization of genes, expression levels of gene products, as well as information on the potential allergenicity and toxicity of the expressed protei ns to APHIS. APHIS’ assessment of the safety of this product for animals focuses on plant pest risk (U SDA-APHIS 2010) and effects on wildlife and threatened and endangered species (section on Animals and Threatened and Endangered Species), and those analyses are based on the comparison of the GE-corn to its non-GE counterpart. No new issues appear to be associated with the DsRed2 (Alt1) transgene in DP32138-1 corn.

Based on the asse ssment of laboratory data provided by Pioneer in the submitted petition and an analysis of the scientific literature (USDA-APHIS 2010), along with the completion of the consultation process with FDA regarding DsRed2 color marker protein in DP-32138-1 (Appendix A), APHIS has concluded that under this alternative, a determination of nonregulated status of DP-32138-1 would have no significant im pacts on animal feed, nor on animal health. Pioneer has communicated to APHIS (Weber, pe rsonal communication) that transgenic seed produced with the DP-32138-1 line is not intended for and is not expected to enter the general corn commodity supply. The procedures esta blished by Pioneer will effectively prevent appearance of the SPT genes in corn commodities unless willful actions are taken to contradict company policy and practice.

Cumulative Effects: Animal Feed

There are no significant impacts on animal health related to the No Action Alternative or a determination of nonregulated status of DP-321 38-1 corn, and no cumula tive effects have been

identified.5 6 Socioeconomic Issues

Domestic Economic En vironment at Risk

The U.S. domestic market for seed corn is estim ated to be $4.6 billion in 2007 and $5.2 billion in

2008 based on the estimated seed costs per acre of $49 per acre in 2007 and $60 per acre in 2008 (USDA-ERS 2010b) and 93.5 million acres planted in 200 7 and 86.0 million acres planted in

2008 (USDA-NASS 2010a).

The production of hybrid seed requires substan tial control over the pollen sources. Detasseling is the most common procedure fo r pollen control. Tassels are r emoved by hand or by machine; both methods are expensive and result in major plant injury with accompanying lower seed yields. In 1988, detasseling costs ranged from $100 to $130 per acre (Wych 1988). Much of the cost of detasseling is personnel costs. Assumi ng that detasselers earn approximately minimum wages and based on minimum hourly wage rate s of $3.35 in 1988 and $7.25 in 2009 (US-DOL 2010), it can be estimated that 2009 detasseli ng costs range from $216 to $281 per acre. Yield reduction due to detasseling has been estimated to range from 1.5% to 13.5% for hand detasseling and 2%-45% for mechanical detasse ling (Wych 1988). To estimate cost of yield losses caused indirectly by detasseling, severa l assumptions can be made. For the production of any seed that requires detasseling, strict contracts are put in place with farmers who are willing to grow the low yielding inbred plants along with the challenges of growing corn to exacting planting, growing and harvesting protocols and sc hedules. In return for the complexity of growing seed corn, paying premiums and minimu ms are standard parts of the contract. The minimum standards are likely to be equivalent to the returns that would be expected from growing conventional corn, which would be base d on the average yields in the area and the commodity corn price. For example, the averag e yield might be 170 bushels with a commodity price of $4.00 per acre [yields and prices based on approximate national averages for 2009 (USDA-NASS 2009a) ] making the minimum standard $680 per acre. A 10% yield reduction from detasseling is approximately $68 per acre. To tal costs of detasseling (cost of detasseling plus yield reduction) may be in the range of $280 to $350 per acre. The total industry wide cost of detasseling of up to 1 million acres is estimated at approximately $280-350 million. Spreading

this total cost over 90 million acres of conventiona l corn production indicates that the detasseling costs are approximately $3 to $4 per planted acre of conventional corn production which is 6%-

8% of the 2007 seed costs and 3%-7% of the 2008 seed costs.

No Action: Domestic Economic Environment

Under the No Action Alternativ e, DP-32138-1 would remain a re gulated article and would require an APHIS permit or notification for re lease into the environment. Seed corn production methods would be the same as currently used under the No Action Alternative. Manual and mechanical detasseling are the major methods for controlling pollen in the hybrid seed corn production fields. No effects are anticipated in the commercial production of corn under the No

Action Alternative.

Preferred Alternative: Dome stic Economic Environment

Under the Preferred Alternative, the availabili ty of seed corn would be determined by the market demand for commercial corn grain as it is under the No Action Alternative. However, under the Preferred Alternative, DP-32138-1 could be used to reduce the need for detasseling along with the associated cost of detasse ling. This could lead to a yield increase due5 7 to a reduction of plant injury caused by mechanical detasseling. The yield increases would reduce the total number of acres devoted to seed corn production and would lower overall seed production costs. If all 1 million acres of seed increase used DP-32138-1 corn, the cost savings from avoiding detasseling operations and associated increased yields would most likely exceed $200 million. The estimated cost of detasseling is about $3-$4 per planted acre of conventional corn production which is equivalent to yield of about one bushel per acre in conventional corn production.

The cost savings from eliminating detasseling operations would provide seed companies with several options. One option is to keep their seed sales price unchanged from previous seasons to enhance their profit margin. Another option woul d be to pass along the entire cost savings by reducing the selling price of seed to the farmer. This option may help to gain company market share for the company. One other option is to keep the cost savings and invest the money in

research to make better varieties. The method of dealing with these cost savings, whether it is a choice of one of these options or a combination is a business decision. The cost savings for DP32138-1-produced seed corn will potentia lly be small ones for conventional corn growers, but the impact on conventional grain production levels or prices using DP-32138-1 would be smaller still

and difficult to measure.

No estimate is available for the level of accepta nce, if any, of usi ng DP-32138-1 in seed corn production for the applicant or for the industry as a whole. Since 1995, three GE male sterility systems for corn have been proposed and deregul ated by APHIS, but have not been used commercially (Pioneer 2009). Therefore the potential implementation of the commercial use of

DP-32138-1 technol ogy by the seed corn industry would be speculative.

The Seed Production Technology is a process that uses the DP-32138-1 SPT insertion to produce hybrid seed to contain no transgenes and without need for detasseling. To make male sterile inbreds (the female parent for hybrid production), the male fertile SPT maintainer line contributes pollen to a male sterile inbred line; virtually all SPT containing pollen grains are inviable, and thus will not be transferred to the seed for the male sterile female parent. 99.999% of seed from this cross is lacking the SPT insert. To eliminate any possible transgenic SPT seed, this seed is passed twice through a seed sorter (with an accuracy of detection of 99.95%). Two steps, non-transgenic pollen shed and seed sortin g, result in a process that produces male sterile female inbreds that are predictably and reliably transgene free. These plants are then used by contract seed growers to produce commercial seed.

Production of the DP-32138-1 SPT maintainer line and production of the female inbred parent lines (Step I and II of Figure 1,) would be acco mplished at Pioneer-controlled field sites or on those sites of other companies having rights to the technology (should Pioneer choose to license it). Thus, the transgenic lines would be under the continuous control of a seed technology provider. After each of these two steps, seed would be sorted by Pioneer into transgenic and non-transgenic seeds, with the discarding of all tran sgenic, marked seeds after Step II (Figure 1). Pioneer demonstrated that the efficiency of the SPT system in producing non transgenic seed was >99.999% (Pioneer 2009). The accuracy of the co lor-based sorting system for transgenic seed was >99.95%. Consequently, the likelihood that a transgenic seed would not be removed from the Step II selection would be the product of both the accuracy of detection and seed removal with the efficiency of the process, which woul d occur with a frequency approaching zero. One additional means of analyzing transgenic content was undertaken. When non transgenic

machine-identified seed was sorted from marked tr ansgenic seed, and the putative non transgenic5 8 seed was used to grow corn, leaf tissue was an alyzed for the presence of transgenes. When DP32138-1 was used as a pollen donor for male sterile female inbred production, seed was collected

from this cross and sorted twice to remove any remaining DP-32138-1 corn seed. Plants grown from the 15,000 collected seed were analyzed using PCR techniques; none of the plants contained genes derived from the SPT insertion (Pioneer 2009).

The non-transgenic seed derived from the Step II increase (Figure 1) would be used to provide the male-sterile female plants for hybrid producti on. Although the DP-32138-1 line is genetically engineered, the progeny of DP-32138-1 that would be grown in Step III (last major increase before planting commercial grain fields) along with Step IV (commercial grain production) would reliably and predictably yield non-transgeni c seed. The final phase of seed production, Step III, would typically be accomplished by c ontract growers, whose non-transgenic seed would be sold to seed suppliers for commercial production.

No information was found to determine whether growers and consumers of non-GE or organic corn would accept corn derived through GE t echnology, but now containing no transgenes. Therefore, for buyers who desire non-GE corn seed or commodity corn in the US, the marketing potential for seed produced from DP-32138-1 is unclear and speculative.

Cumulative Effects: Dome stic Economic Environment

Assuming complete acceptance of the DP-32138-1 technology, cumulative domestic economic effects on seed corn producers would be cost savings of an estimated $200 million per year. Because the cost savings for DP -32138-1 seed corn are small with respect to all conventional corn production, using DP-32138-1 may have little measurable cumulative impact on conventional grain production levels or prices.

Trade Economic Envi ronment at Risk

Seed corn and commercial corn grain are both major exports for the U.S. In 2005 (the most recent date with specific information on seed corn) seed corn exports totaled $136 million (USDA-FAS 2005). Corn is exported in bulk, as co rn by-products, and feed. Since corn is used heavily as an animal feed for poultry, hogs and cattle, export of animal products could also be considered as related to corn production. Expor t of corn grain in bulk was $8.8 billion in 2009, $13.4 billion in 2008, and $9.8 billion in 2007 (U SDA-ERS 2010c). Foreign customers in Europe and Japan may have concerns with a new variety of GE grain a nd new GE traits must undergo extensive regulatory assessment befo re they are approved for import (USGC 2006).

No Action: Trade Economic Environment

The availability of methods used to produce seed corn would be the same as currently used in seed corn production systems w ith detasseling being a major method for controlling pollen in the hybrid seed corn production fields. Since crop production practices are not anticipated to change, no effects are anticipated in the commer cial production of corn under the No Action

Alternative.

Preferred Alternative: Trade Economic Environment

Under the Preferred Alternative, the total supp ly of seed corn would be determined by the

international market demand for commercial corn grain as it is under the No Action5 9 Alternative. However, under th e Preferred Alternative, DP -32138-1, savings would accrue to the seed producers in reducing resource outlays, and total cost of productio n and increasing seed crop yields (see discussion in preceding section, Preferred Alternative: Domestic Ec onomic Environment). The level of acceptance of th e technology is not easily predicted (see discussion in preceding section, Preferred Alternative: Domestic Economic Envi ronment) which makes potential for impacts on trade unclear. Another difficulty is determining how international consumers and regulatory framewor ks would categorize corn produced with GE technologies but now containing no transgenes. If DP-32138-1 is used to produce seed corn for export to those regions and countri es with rigorous regulatory regi mes, the potential impacts for

international marketing of seed produced with DP-32138-1 is unclear and acceptance would be speculative. Because use of the DP-32138-1 cannot be detected or otherwise established in commercial corn seed, it is possible that use by s eed providers would cause no impacts. Because export of seed corn is an important market for US companies, these providers would likely need to negotiate seed availability using the techno logy with international customers and regulatory systems well in advance of future sales abroa d. Because use of the technology is elective, and seed providers could negotiate acceptance of th e technology with these customers and regulatory systems, the potential impact of DP- 32138-1 is not likely to be significant.

Cumulative Effects: Trade Economic Environment

There are no likely cumulative effects identified for this issue. Because the cost savings for

DP-32138-1 seed corn are so small w ith respect to total conventional corn production, using DP32138-1 may have little measurable impact on conventional grain production levels or prices for the international commodity markets.

Social Environment at Risk fr om Seed Production Activities

Conventional seed corn producti on relies heavily on de tasseling as the method for pollen control and often rely heavily on thousands of field wo rkers to accomplish this operation (Pioneer 2007). Detasseling jobs provide a source of income, but also has the pot ential to expose workers to safety risks associated with the use of h eavy machinery used for seed corn detasseling operations.

No Action: Social Environment

The availability of agricultural methods used to produce seed corn would be the same as currently used in seed corn production systems under the No Action Alternative with detasseling being a major method for controlling pollen in the hybrid seed corn production fields. Detasseling operations would continue to primarily rely on thousands of field workers . Potential economic impacts would be unchanged.

Preferred Alternative: Social Environment

Under the preferred alternative, DP-32138-1 woul d be deregulated. If this technology is accepted commercially, all seed corn production acres that use this technology would no longer require detasseling as the method for pollen control . Depending on the level of acceptance by the seed corn production industry, field workers w ould no longer have detasseling as a possible source of income. The possible economic effects of losing these temporary jobs are unclear, but it is reasonable to assume that it would have a potential negative impact until another source of

income could be secured.6 0 Cumulative Effects: Social Environment

Depending on the level of acceptance by the seed corn production industry, thousands field workers would no longer have detasseling as a possible source of income. This impact is expected to be only a short term one as these workers secure other sources of income to replace

those incomes lost.

Other Cumulative Effects

All potential cumula tive effects regarding specific issues have been anal yzed and addressed above. No further potential cumulative effects ha ve been identified. Stacked varieties, those crop varieties that may c ontain more than one trai t, are currently found in the marke tplace and in agricultural production. In the event of a determination of nonregulated status of DP-32138-1

, DP-32138-1 may be used to combine with non-GE and GE corn varieties by traditional breeding techniques, resu lting in hybrids that, for example, may also be resistant to herbicides or insects. The resulting hybrids w ould not contain the SPT transgene.

APHIS’ regulations at 7 CFR Part 340 do not provi de for Agency oversight of GE corn varieties previously deregulated pursuant to Part 340 and the Plant Protection Act, nor over stacked varieties combining deregulated GE varieties unless it can be positively shown that such stacked varieties were to pose a likely plant pest risk. To date, none of the GE corn varieties that have been deregulated pursuant to Part 340 and the Plant Protec tion Act and used for commercial breeding pr ogram have been subseq uently found to pose a plant pest risk.

There is no guarantee that DP-32138-1 will be stacked with any partic ular deregulated GE variety, as company plans and market demands play a significant role in those business decisions. Moreover, DP-32138-1 could even be combined with non-GE corn varieties. Thus, predicting all potential combinations of stacked varieties that could be created using both deregulated GE corn varieties and also non-GE corn varieties is hypothetical and purely speculative.

Threatened and E ndangered Species

APHIS analyzed the potential for effects from cultivation of DP-32138-1 and its progeny on federally listed threatened and endangered species (TES) and species proposed for listing, as well as designated critical habitat and habitat propose d for designation, as required under Section 7 of the Endangered Species Act. Di rect effects are analyzed by considering the response that TES could have if exposed to DP -32138-1 corn. Indirect effects are those that co uld result from

the use of DP-32138-1 in corn production, would occur later in time, but are still reasonably certain to occur. Consideration is given for the potential of DP-3 2138-1 to change the baseline

habitat of TES incl uding critical habitat. If the analysis dete rmines that a determination of nonregulated status of DP -32138-1 may affect listed species or critical habita t, consultation

with the United States Fish and Wildlife Se rvice (USFWS) or the Na tional Marine Fisheries Service (NMFS) is required.

DP-32138-1 was deve loped solely to be used to produce male sterile inbred plants for the generation of hybrid corn seed that is not tran sgenic (Pioneer 2009). Th e carefully controlled expression of a seed color marker gene and polle n fertility and sterility genes allows for the generation of red transgenic seed for seed increas e of male sterile-female inbred lines that are used in non transgenic h ybrid commercial seed. DP-32138-1 contains a DNA insert with three61 genes- a male fertility gene, Ms45 , from corn; an alpha-amylase gene, zm-aa1 , from corn; and

a

fluorescent color marker protein gene, DsRed2(Alt1) , from a marine coral Discosoma sp ., the only gene not native to corn.

DP-32138-1 is not genetically engineered to produce a toxin or pesticide, and is not genetically engineered to be tolerant to an herbicide. A NPC for the DsRed2 protein color marker was submitted to FDA on October 11, 2006 with the follow up letter of January 9, 2010 received from FDA (Appendix A). The FDA considers Pioneer's consultation on ZM-AA1 alpha-amylase protein to be complete (Appendix B).

If the seed corn industry completely converts to using this technology, t he DP-32138-1 would be grown on a maximum of 20,000 acres (the tota l area currently used fo r corn seed production and 0.02% of all corn production acres in the U.S.). These 20,000 acre s would likely be the same 20,000 acres used in conventional seed corn production that have been in agricultural production for many years. The same agronomic practices for conventional seed corn production would be us ed for DP-32138-1 production - in cluding the same fertilizers, herbicides, fungicid es, insecticides, irrigatio n, crop rotations, a nd tillage. Corn seed for planting produced using DP-32138-1 and the plants grown from th ese seed no lo nger have any of the transgenes. Therefore, the maximum expo sed area is up to 20 ,000 acres in locations where agricultural seed corn production has historically o ccurred. There is no hazard associated with producing corn seed using the DP-321 38-1 technology that would be different from production of conventional corn.

Corn is a feed commonly provided to many livestock and wild life (e.g., birds, deer, and rodents). To identify the direct effects on listed animal species that could result from feeding,APHIS compared the composition and nutritional quality of DP-32138-1 corn, with a nongenetically engineered control corn line a nd the natural variation found in two commercial corn inbred lines. If the composition of DP-32138-1 inbred is similar to other commercial corn inbred plants, it is unlikely that DP-32138-1 would affect listed animal species, regardless of

exposure.

The data presented in the petition suggests ther e is no difference in compositional and nutritional quality of DP-32138-1 compared to conventiona l corn, apart from the presence of DsRed2 protein (MS45 protein and ZM-AA1 alpha-amylase protein are both found in conventional corn). Although some of the variables measured by th e applicant showed st atistically significant

differences between DP-32138-1 corn and the nontransgenic inbred controls (Pioneer 2009), none of the values for the grain compos ition characteristics were out side the range of natural variability of conventional corn as found in the International Life Sciences Institute Crop Composition Database (OECD 2003; Ridley, Shillito et al. 2004; ILSI 2 008) or in the OECD consensus document on corn composition (OECD 2003). DP-32138-1 does not express additional proteins, natural toxicants, allelochem icals, pheromones, hormones, etc. that could directly or indirectly affect a listed TES or sp ecies proposed for listing. A toxicological safety

assessment conducted on DsRed2 protein showed that it is un likely to be a toxin or have any

relevant similarities to known or putative allergens (Pioneer 2009). Thus, the composition and nutritional quality of DP-32138-1 is not biologically different than conventional corn, and would not be expected to affect TES differently.

Corn itself is not sexually comp atible with any liste d plant species; therefore there is no potential for a direct effect of DP-32138-1 on TES plants. Indi rect effects of DP-32138-1 on6 2 listed plant species were also evaluated. C orn does not have sexuall y-compatible relatives found in ‘natural’ areas; corn is only able to reproduce with other corn pl ants in the U.S. (OECD 2003). As stated above, DP-32138-1 will have no effect on animals, including animals such as insects, bats or birds that may be pollinators of TE S plants. Because corn cannot

naturalize and would not affectpollinators, there are no expected indirect effects of DP32138-1 on TES plants.

Cultivation of DP-32138-1 is not expected to differ from practices normally used for seed corn production or commercial corn produc tion. DP-32138-1 corn plan ts with transgenes are expected to be grown on ly on a maximum of 20,000 acres (0 .02% of all corn produced in the USA) and is expected to replac e only conventional corn varieties currently grown for seed. The potential environmental impacts on TES from this product are those associated with typical corn seed production and commercial co rn production, in areas where corn seed and commercial corn are typically pr oduced, and therefore would not a ffect the baseline habitat of any listed species.

After reviewing possible effects of a determinati on of nonregulated stat us of DP-32138-1 corn, APHIS has not identified any stressor that could affect the reproduction, numbers, or distribution of a listed TES or species proposed for listing. As a result, a detailed exposure analysis for individual species is not necessary. APHIS has cons idered the effect of DP-32138- 1 production on designated critical habitat or habitat proposed for designation and could identify no difference from effects that would occur from the seed production of other corn varieties. Therefore, APHIS has reached a conclusion that the release of DP -32138-1 corn, following a determinati on of nonregulated status , would have no effect on federally listed threatened or endangered species or species proposed for listing, nor would it affect designated critical habitat or habitat proposed for designation. Consequently, consultation with the USFWS or NMFS is not require d for this action.

Consideration of Executive Or ders, Standards and Treaties Relating to Environmental Impacts

Executive Order (EO) 12898 (US-NARA 2010), “Federal Actions to Address Environmental Justice in Minority Populations and Low-Income Populations,” requires Federal agencies to conduct their programs, polic ies, and activities that substantially affect

human health or the environment in a manner so as not to exclude persons and populations from participation in or benefiting from such prog rams. It also enforces existing statutes to prevent minority or low-income communities from being subjecte d to disproportionately high

and adverse human health or environmental effects. EO 13045 (US-NARA 2010), “Protection of Children from Environmental Health Risks and Safety Risks,” acknowledges that children may suffer disproportionately from environmental health and safety risks because of their developmental stage, greater meta bolic activity levels, and behavi or patterns, as compared to adults. The EO (to the extent permitted by la w and consistent with the agency’s mission) required each Federal agency to identify, assess, and address environmental health risks and safety risks that may disproport ionately affect children.

Each alternative was analyzed with respect to EO 12898 and 13045. Based on the information submitted by the applicant and assessed by APHIS, DP-32138-1 is not significantly different than conventional corn pr oduction. FDA has re viewed the introduced proteins ( DsRed2 and ZM-AA1) in the unlikely case that the Event may inadvertently enter63 the commodity grain stream. Pioneer concluded th at the proteins were safe for food and feed use (Pioneer 2009) and FDA agreed that the presen ce of the proteins would not lead to safety concerns when present in low levels in th e food supply. Therefore, DP-32138-1 is not expected to have a disp roportionate adverse e ffect on minorities, low-income populations, or

children. Based on historical experience with corn production a nd the data submitted by the applicant and assessed by APHI S, DP-32138-1 should eliminate the need for detasseling,

which in turn would increase worker safety by significan tly reducing the number of seasonal workers exposed to the hazards of working around heavy farm machinery.

EO 13112 (US-NARA 2010), “Invasive Species” , states that Federal agencies take action to prevent the introduction of invasive species, to provide for their contro l, and to minimize the economic, ecological, and human health impacts that invasive species cause. Both non-GE and GE corn varieties that have been previously deregulated pursuant to Part 340 and the Plant

Protection Act are widely grown in the U.S. Based on historical experi ence with corn and the data submitted by the app licant and assessed by APHIS , DP-32138-1 plants ar e very similar in

fitness characteristics to other corn varieties currently grown and are not expected to become weedy or invasive (USDA-APHIS 2010).

EO 13186 (US-NARA 2010), “Respons ibilities of Federal Agencies to Protect Migratory Birds”

, states that Federal agencies taking actions that have, or are likely to have, a measurable negative effect on migratory bird populations are directed to develop and implement, within 2 years, a Memorandum of Understanding (MOU) with the Fi sh and Wildlife Service that shall promote the conservation of migratory bird populations . Data submitted by the applicant has shown no difference in compositional and nutritional quali ty of DP-32138-1 compared to conventional corn, apart from the presence of DsRed2 protein. Th e migratory birds that occasionally forage in corn fields are unlikely to c ontain high amounts of DP-32138-1 as corn grain availability is limited by seed germination and harvest. Base d on APHIS’ assessment of DP-32138-1 it is unlikely that a determination of nonregulated stat us of this corn variety will have a negative effect on migrator y bird populations.

Impacts on Unique Characteristics of Geographic Areas.

There are no unique characteristics of geographic areas such as park la nds, prime farm lands, wetlands, wild and scenic areas, or ecologically critical areas th at would be adversely impacted by a determination of nonregulated status of DP-32138-1 corn. The common agricultural practices that would be carried out under the proposed action will not cause major ground disturbance, do not cause any physical destruct ion or damage to property, do not cause any alterations of property, wildlife ha bitat, or landscapes, and do not involve the sale, lease, or transfer of ownership of any property. This action is limited to a determination of nonregulated

status of DP-32138-1 corn. The product will be deployed on a lim ited number of acres of agricultural farm land used for co rn seed production which may be focused in a small number of sites where Pioneer and future potential licensees may produce seed corn, but potential for such production may exist where corn is grown in all US states, except Alaska. Progeny of this variety that express the identifie d traits of the DP-32138-1 corn will be retained by Pioneer or licensed users, and those progeny derived from production of hybrid commercia l corn will be predictably and reliably without transgenes. This action w ould not convert land use to nonagricultural use and therefore would have no adverse impact on pr ime farm land. Standard agricultura l practices for land preparation, planting, irrigation, and harves ting of plants would be used on agricultural lands planted to DP-32138-1 corn including the use of EPA register ed herbicides. Applicant’s6 4 adherence to EPA label use restrictions for all pesticides will mitigate potential impacts to the

human environment. In the event of a determin ation of nonregulated stat us of DP-32138-1 corn, the action is not likely to affect historic or cultural resources, park lands, prime farmlands, wetlands, wild and scenic rivers, or ecologically critical areas that may be in close proximity to corn production sites.

International Implications

EO 12114 (US-NARA 2010), “Environmental Effects Abroad of Ma jor Federal Actions”

, requires Federal officials to take into consideration any potent ial environmental effects outside the U.S., its territories, and possessions that result from actions being taken. APHIS has given

this due consideration and does not expect a significant environmental impact outside the U.S. in

the event of a determination of nonregulated status of DP-32138-1 corn. It should be noted that all the considerable, existing na tional and international regulator y authorities and phytosanitary regimes that currently apply to introductions of new corn cultiva rs internationally, apply equally to those covered by an APHIS determination of nonregulated status under 7 CFR part 340. Any

international trade of DP-32138-1 subsequent to a determination of nonregulated status for the product would be fully subject to national phytosa nitary requirements and be in accordance with phytosanitary standards developed under the International Plant Protection Convention (IPPC 2010).

The purpose of the IPPC “is to secure a common and effective ac tion to prevent the spread and introduction of pests of plants and plant products and to promote appropriate measures for their control” (IPPC 2010); the protection it affords ex tends to natural flora and plant products and

includes both direct and indirect damage by pests, including weeds. The IPPC set a standard for the reciprocal acceptance of phyt osanitary certification among the nations that have signed or acceded to the Convention (172 countries as of Ma rch 2010). In April 2004, a standard for pest risk analysis (PRA) of living modified organi sms (LMOs) was adopted at a meeting of the governing body of the IPPC as a supplement to an existing standard, Inte rnational Standard for Phytosanitary Measure No. 11 (ISPM -11, Pest Risk Analysis for Qu arantine Pests). The standard acknowledges that all LMOs will not present a pest risk and that a determination needs to be made early in the PRA for importation as to whether the LMO poses a potential pest risk resulting from the genetic modification. APHIS pe st risk assessment procedures for genetically engineered organisms are consistent with th e guidance developed under the IPPC. In addition, issues that may relate to commercialization and transboundary move ment of particular agricultural commodities produced through bi otechnology are being addressed in other

international forums and through national regulations.

Compliance with Clean Wate r Act and Clean Air Act

This EA evaluated the changes in corn production due to the unrestricted use of DP-32138-1 corn. DP-32138-1 will not lead to the increased produc tion of corn in U.S. a griculture. There is no expected change in water use due to the pro duction of DP-32138-1 compared to current corn seed production regimes, nor is it expected that air quality will change because of production of DP-32138-1.

National Historic Preservation Act (NHPA) of 1966 as Amended6 5 The NHPA of 1966, and its implementing regulations (36 CFR 800), requires Federal agencies to: 1) determine whether activities they propose constitute "undertakings " that has the potential to cause effects on historic propert ies; and 2) if so, to evaluate the effects of such undertakings

on such historic resources and consult with the Advisory Council on Hist oric Preservation (i.e.,

State Historic Preservation Offi ce, Tribal Historic Preservati on Officers), as appropriate.

A determination of nonregulated status of DP-3 2138-1 corn will not adve rsely impact cultural resources on tribal properties. Any farming activities that may be taken by farmers on tribal lands are only conducted at the tr ibe’s request; thus, the tribes have control over any potential

conflict with cultural resources on tribal properties.

A determination of nonregulated status of DP-3 2138-1 corn would have no impact on districts, sites, highways, structures, or objects listed in or eligible for listing in the National Register of Historic Places, nor would they likely cause any loss or destruc tion of significant scientific, cultural, or historical resources . This action is limited to a determination of non-regulated status of DP-32138-1 corn. Standard agricultural pract ices for land prepara tion, planting, irrigation, and harvesting of plants would be used on thes e agricultural lands including the use of EPA registered pesticides. Applicant’ s adherence to EPA label use rest rictions for all pesticides will mitigate impacts to the human environment.

A determination of nonregulated status of DP -32138-1 corn is not an undertaking that may directly or indirectly cause alte ration in the character or use of historic pr operties protected under the NHPA. In general, common agricultural activ ities conducted under this action do not have the potential to introduce visual, atmo spheric, or audible elements to areas in which they are used

that could result in effects on the character or use of historic properties. For example, there is potential for audible effects on the use and en joyment of a historic property when common agricultural practices, such as the operation of tractors and other mechanical equipment, are conducted close to such sites. A built-in mitigating f actor for this issue is th at virtually all of the methods involved would only have temporary effect s on the audible nature of a site and can be ended at any time to restore the a udible qualities of such sites to their original condition with no

further adverse effects. Add itionally, these cultivation practic es are already being conducted throughout the corn production regions. The cult ivation of DP-32138-1 corn does not inherently change any of these agronomic practices so as to give rise to an impact under the NHPA.



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### VI. Appendix A7 8 

### VII. Appendix B7 9 

### VIII. Appendix C

Description of Seed Pro duction Technology (SPT)
( from Pioneer 2009 ; all figure and table references are to the Petition figures and tables)

I. Introduction to Seed Production Technology (SPT)

I-B. Maize Hybrid Seed Production

Hybrid maize ( Zea mays L.) is grown on more than 80 million acres in the United States (USDANational Agricultural St atistics Service (NASS), 2008), making it the largest pr oducer of hybrid maize. The seed used to produce hybrid maize is generated by crossing two unique inbred parental lines referred to as the ‘male inbred pare nt’ (the plant that is the pollinator) and ‘female inbred parent’ (the plant on wh ich the seed is produced).

For inbred parent seed increase, male and female inbred parental lines are grown separately and allowed to self-pollinate in Pione er fields that are re productively isolated from other maize plants in order to maintain genetic purity and increase the quantity of the inbred parents (Figure 1). In order to produce hybrid seed, male and female inbr ed parent lines are inte r-planted in adjacent rows ( e.g., 2 rows male, 4 rows female, 2 rows male) in reproductively isolated contractor fields and allowed to open pollinate.

To produce pure hybrid seed, the male inbred pare nt must cross pollinate the female inbred parent and the female inbred pa rent must be prevented from self-pollinating. The most common practice to accomplish this is to cut off the pollen-bearing tassels ( i.e., detasseling) on the female inbred parent (Figure 1).

In the hybrid seed production fields , the ears of the detasseled fe male inbred parent can only be fertilized with pollen from the male inbred pa rent planted in adjacent rows. The male inbred parent is removed from the fields before the ea rs reach maturity. Thus, the seed produced on the female seed parent will be uniformly hybri d. The F1 hybrid seed is harvested, cleaned, conditioned, and bagged for sale to growers. When planted in growers’ fields, the hybrid seed will produce a uniform F1 crop. The grain harvested from growers’ fields is used for feed and fuel production, processed into food products and industrial materi als, or exported.

I-C. Maize Fertility Control in Hybrid Maize Production

I-C.1. Detasseling

The physical removal of the pollen-producing tass els (detasseling) has been the most widely used method of preventing female inbred parent self-pollination since maize hybrids were first produced in the 1920s (Wych, 1988). Less comm on or experimental methods include the application of chemical sterila nts, the use of biological syst ems based on cytoplasmic malesterility, and systems to contro l male-fertility using biotechno logy (Refer to Appendix 1 for additional information on these systems).

Detasseling can be accomplished manually or mechan ically with tassel-cutting or tassel-pulling machines. Thousands of temporary workers are employed to detassel maize hybrid seed production fields. The detasseling process is not en tirely reliable, and on occasion a female plant8 0 will escape complete detasseling. For example, natu ral variation in plant development can result in the emergence of tassels after the detasseling process is complete or a mechanical detasseler can fail to remove a tassel completely. Incomp lete detasseling results in pollen shed by the female inbred parent plant and self-pollination. When this occurs, seed of the female inbred parent is harvested along with th e hybrid seed which re sults in reduced seed quality, lowered trait purity, and lost yield potential. M echanical detasseling is faster than hand detasseling, but causes more damage to the plants and results in up to 40% reduction in seed yield (Wych, 1988). Today,

a combination of mechanical and hand detasseli ng is often used; no form of detasseling is entirely satisfactory. Therefore, a need exists fo r an alternative method of pollen control that is safe, efficient, and highly effective.

I-C.2. Genetic Male-Sterility

Male-sterility in maize refers to the failure to produce functional pollen. Male-sterile maize plants can neither self-pollinat e, nor sib-pollinate neighboring pl ants and can serve as female inbred parents in the production of hybrid maize seed. Male-sterility traits display either nuclear (also known as genetic) or cytoplasmic inher itance (Refer to Appendix 1 for details).

Genetic male or pollen sterility in maize was first described by Eyster (19 21) that it is due to a single recessive gene, designated as ms . Since then, many different ge netic male-sterility loci

have been identified in maize. The chromosome number and map position of nearly all presently identified male-sterility gene s are well characterized (Neuffer et al ., 1997). Sterility phenotype is determined by a single gene and the alle le for male-sterility is recessive ( ms ), although a small

number of dominant male-sterile a lleles have been described (Neuffer et al ., 1997). In maize, a male-sterility plant trait is expressed onl y by maintenance of a homozygous recessive ( ms/ms ) condition for the fertility gene.

Limitations to the Use of Homozygous Recessive ( ms/ms ) Lines as Male-Sterile Parents

Increase of male-sterile ( ms/ms ) inbred parent seed is not possible via self-pollination as the homozygosity of the recessive gene ( ms ) results in male-sterility.

One way to increase the seed of the male-sterile inbred parent is to cross-pollinate male-sterile plants ( ms/ms ) with male-fertile plants which are he terozygous for the ma le-sterility gene ( Ms/ms ). The progeny from such a cross-breeding ar e approximately fifty percent male-sterile ( ms/ms ) and fifty percent male-fertile ( Ms/ms) . Since the male-sterile seed can not be separated from the male-fertile seed, this is not a practical way to increase the male -sterile parent seed.

Recovery and maintenance of pure male-sterile ( ms/ms ) inbred populations using traditional breeding methods requires: a) additional maize li nes for the maintenance of the male-sterility trait, and b) several intermed iate crosses and selection met hods that are time consuming. Therefore, it is not practical for use in commercial hybrid maize production.

Thus, a need exists for a conve nient and effective process for generating male-sterile inbred female parent lines.

I-C.3. Basis for the Development of Seed Production Technology81 Section I-C.2 described how cross-pollinating the male-sterile ( ms/ms ) plants with heterozygous ( Ms/ms ) male-fertile plants yields a mix of male-s terile and male-fertile genotypes. However, if all pollen containing the fertility restoration gene ( Ms ) could be eliminated from the heterozygous male parent ( Ms/ms ) thereby ensuring pollination of male-sterile ( ms/ms ) parent only with pollen carrying the recessive ms allele, the male-sterile progeny would retain their homozygous ( ms/ms ) condition and could be used as male-ste rile female inbred parent in hybrid seed production. This is the basis for the Pi oneer Seed Production Technology (SPT) process utilizing a transgenic 32138 SPT maintainer (Figure 2).

Pioneer’s novel SPT process generates male-sterile female inbred parent lines that maintain the homozygous recessive state ( ms/ms ) without any intermediate cr osses. Male-sterile female inbred parent seed can be multiplied, while maintaining the homozygous recessive state of the male-sterility gene and providing fo r full restoration of fertility in the F1 hybrid plants grown in

commercial fields.

Figure 2. Schematic Diagram of the Basis for SPT

‡ = Represents 32138 SPT maintainer.

The SPT construct containing three gene cassettes [ Ms45, zm-aa1, and DsRed2(Alt1 )] was

inserted into a male-sterile maize line that is homozygous recessive for ms45 ( ms45/ms45 )

shown as ms/ms in Figure 2. The SPT insertion was integrated in the genome at a locus that segregates independently from the endogenous ms45 locus (data not shown). Therefore, 32138 SPT maintainer is homozygous for the ms45 recessive allele and hemizygous for the

recombinant Ms45 gene from the SPT insertion, indicated as Ms/- in Figure 2. A single copy of

Ms45 gene from the 32138 SPT insertion restor es fertility in the male-sterile ( ms45/ms45 ) line (Figure 7B). 32138 SPT maintainer sheds two diffe rent types of pollen in a 1:1 ratio: a) nontransgenic fertile pollen (with the ms45 allele) that do not contain the SPT insertion, and b) transgenic infertile * pollen (with both the ms45 allele and the transgenic Ms45 allele) that contain the 32138 SPT insertion (Figure 5). The transgenic pollen carrying Ms45 are rendered infertile

due to the α -amylase gene (Refer to Sections 1-D and 1-H.2 for more details) in the SPT insertion (Figure 9).

* = Pollen infertility refers to the inability of pollen to germinate and fertilize the ovule.8 2 Therefore, homozygous recessive ( ms45/ms45 ) male-sterile female inbr ed parent seed can be increased by utilizing the pollen from an isoge nic SPT maintainer (Figure 3, Step II and Figure 5).

I-D. Description of the SPT Process

Seed Production Technology is a pr ocess rather than a trait or a product. The SPT process works across all germplasm, requires no de tasseling, and produces hybrid seed that are fully fertile. The male-sterile female inbred parent increase proces s is entirely controlled by Pioneer and the total acreage in the US planted to th e transgenic 32138 SPT maintainer and SPT derived male-sterile female inbred parent seed increase e ach year will be less than 5,000 acres.

32138 SPT Maintainer

The SPT process uses a transgenic 32138 SPT maintainer in an ms45/ms45 genetic background (Figure 3, Step I). The 32138 insertion in SPT maintainer contains three gene expression

cassettes essential for the f unctioning of the SPT system: Ms45

, zm-aa1 , and DsRed2(Alt1) (Figure 6).

The 32138 SPT maintainer is homozygous for the endogenous ms45 recessive mutant allele and hemizygous for the SPT insertion ( i.e

., ms45/ms45; SPT/- ) and therefore cont ains a single copy

of the Ms45 gene. Expression of a single copy of Ms45 gene in the ms45/ms45 genetic background restores male-fertili ty and enables pollen production in the 32138 SPT maintainer.

The 32138 SPT maintainer sheds two di fferent types of pollen in a 1: 1 ratio: a) fertile, does not contain SPT insertion, and b) infertile, doe s contain 32138 SPT insertion (Figure 5). Pollen containing the 32138 SPT insertion is rendered infertile due to the action of ZM-AA1 α -amylase protein. Expression of ZM-AA1 α -amylase in 32138 SPT maintain er pollen results in the depletion of starch (Figure 9) and depriving the SPT pollen of th e energy reserves required for successful pollen germination and fertilizati on. Hence, any pollen containing the 32138 SPT

insertion is made infertile.

A fluorescent color marker ( DsRed2(Alt1 )) gene is linked to the other two SPT genes ( Ms45 and
zm-aa1 ) and it confers a pinkish red phenotype to any seed expressing the 32138 SPT insertion (Figures 5 and 10). The DsRed2 pr otein exhibits a high fluorescent intensity and emits a strong red fluorescence under appropriate illumination so seeds containing the SPT insertion can be readily detected and separated from the pr ogeny seeds that do not contain the 32138 SPT insertion in a highly efficient manner using mechan ical color sorting (Figure 3, Color Sort A and B; Refer to Appendix 3).

Step I: 32138 SPT Maintainer Seed Increase

32138 SPT maintainer seed is replicated via self-pollination (Figure 3 St ep I and Figure 4). The SPT insertion is transmitted maternally. Upon self-pollination, 32138 SPT maintainer produces two different types of seed (yello w: pinkish red) in a 1:1 ratio (Figures 5 and 11A). Yellow seed is non-transgenic for SPT (does not contain th e 32138 SPT insertion). Pinkish red seed that83 fluoresces a bright red color unde r appropriate illumina tion is transgenic for SPT (contains the 32138 SPT insertion).

Color Sort A

Self-pollinated 32138 SPT maintainer seed contai ning a mixture of yellow and pinkish red (SPT maintainer) seed in a 1:1 ratio is passed thru m echanical color sorters tw ice (Refer to Appendix 3 for details) to separate the 32138 SPT maintainer s eed from the yellow seed (Figure 3, Color Sort A and Figure 4). Color sorters detect the fluoresce nt red 32138 SPT maintain er seed and separate it from the yellow/non-transgenic for SPT seed. Pure seed of 32138 SPT maintainer is then available for the propagation of non-transgenic male -sterile female inbred parent seed (Figure 3, Step I). Therefore, color sort A is for the pur ification of 32138 SPT mainta iner seed. Yellow seed

collected from color sort A will be discarded (Refer to Appendix 3 for more details).

Step II: Male-Sterile Female Inbred Parent Seed Increase

For seed increase of male-sterile ( ms45/ms45 ) female inbred parent, 32138 SPT maintainer will be planted in either a 2:2 or 4:4 or 4:2 planting pattern (male-sterile female: 32138 SPT maintainer rows). Fertile pollen (that does not contain the 32138 SPT insertion and carry the

ms45 recessive allele) from the 32138 SPT maintainer plants will cross-pollinate and fertilize nearly isogenic male-sterile ( ms45/ms45 ) female inbred parent plants (Figure 3, Step II and Figure 5). The inbred seed produced on the male-ste rile female inbred plants will maintain their homozygous recessive ( ms45/ms45 ) state and genetic purity. A ll male-sterile progeny seed harvested will be yellow in color and will not contain the 32138 SPT insertion (Figure 5 and Refer to Appendix 4 for more details).

Color Sort B

The SPT system is highly genetically efficient (R efer to Appendix 4 and 5) and virtually all of the fertile pollen from 32138 SPT maintainer shou ld be non-transgenic for SPT and therefore all of the male-sterile inbred progeny seed made using the pollen from 32138 SPT maintainer should also be non-transgenic for SPT. The purpose of the color sort B is to ensure the purity of the male-sterile female inbred parent by re moving any seeds of 32138 SPT maintainer. Each batch of male-sterile female inbred parent seed is passed twice through th e color sorter (Figure 3, Color Sort B and Figure 5). This step helps to assure that no seed of the 32138 SPT maintainer will be planted in the F1 hybrid seed fields that pr oduce seed sold to growers (Figure 3, Step III).

Steps III. Commercial Maize Hy brid Seed Production

Commercial maize hybrid seed pr oduction (Figure 3, Steps III) using the male-sterile progeny seed derived using SPT technology will take pl ace normally. The purity of the male-sterile female inbred progeny derived from the SPT pr ocess is insured by passing the seed twice through the mechanical color sort ers (Refer to Appendix 3 for details on the color sorter). Therefore, the F1 hybrid seed ha rvested and sold to growers for commercial grain production is non-transgenic for SPT and the resulting F1 hybrid plants are fully fertile.

Steps IV. Commercial Maize Grain Production8 4 Commercial maize grain production using the F1seed generate d using the male-sterile female inbred parent will take place normally. The F1 hybr id plants grown in the growers’ fields are non-transgenic for SPT and therefore, the grai n harvested from growers’ fields is also nontransgenic for SPT (Figure 3, Step IV).