# Syngenta Seeds, Inc. Alpha

- Amylase Maize

Event 3272

OECD Unique Identifier
SYN

- E3272

- 5

Final Environmental

Assessment

February 201 1 United States Department of Agriculture

Marketing and Regulatory Programs

Animal and

Plant Health Inspection
Service

# Syngenta Seeds, Inc. Alpha

- Amylase Maize

Event 3272

Final Environmental Assessment February 201 1

Agency Contact: Cynthia Eck
Document Control Officer Biotechnology Regulatory Services
USDA APHIS 4700 River Road, Unit 147 Riverdale, MD 20737

________________________________ __________________________ The U.S. Department of Agriculture (USDA) prohibits discrimination in all its programs and activities on the basis of race, color, national origin, sex, religion, age, disab ility, political beliefs, sexual orientation, or marital or family status. (Not all prohibited bases apply to all programs.) Persons with disabilities 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, Director, Office of Civil Rights, Room 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 report does not imply recommendation or endorsement by the U.S. Department of Agriculture over others not me ntioned. USDA neither guarantees nor warrants the standard of any product mentioned. Product names are mentioned solely to report factually on available data and to provide specific information.

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

________________________________ __________________________ CAUTION: Pesticides can be injurious to humans, domestic animals, desirable plants, and fish or other wildlife

— if they are not hand led or applied properly. Use all pesticides selectively and carefully. Follow recommended practices for the disposal of surplus pesticides and pesticide containers.

## Table of Contents

I. I. Purpose & Need

................................ ................................ ........ 5

II. II. Affected Environment

................................ ........................... 11

III. III. Alternatives

................................ ................................ .......... 22

A. No Action: Continuation as a Regulated Article

................. 22

B. Preferred Alternative: Determination that Event 3272 Corn Plants are No Longer Regulated Articles, in Whole

............ 22

C. Alternatives Considered but Rejected from Further

Consideration

................................ ................................ ....... 23

D. Comparison of Alternatives

................................ .................. 24

IV. IV. Environmental Consequences

................................ .............. 27

Methodology and Assumptions

................................ ................. 27

Corn Production

................................ ................................ ......... 31

Ethanol Production

................................ ................................ .... 38

Public Health

................................ ................................ .............. 39

Gene Movement

................................ ................................ ......... 45

Water Use in Ethanol Production

................................ .............. 48

Animal and Plant Communities

................................ ................. 48

Soil Biology

................................ ................................ ............... 52

Conservation Reserve Program

................................ ................. 54

Threatened and Endangered Species

................................ ......... 57

Compliance with Statutes, Executive Orders and Regulations

.. 60

V. V. Listing of Agencies and Persons Consulted

......................... 62

VI. VI. References

................................ ................................ ........... 63

VII. Appendix A. Biotech Seed Products Available for the 2011

Planting Season 1,2,3

................................ ................................ ..... 71

VIII. Appendix B. Corn

- producing counties in the 28 states that have

corn ethanol facilities

................................ ................................ . 78

IX. Appendix C. Economic Impact Report submitted by Sygenta

.. 92X. Appendix D. Pollen

- mediated gene flow report submitted by

Syngenta

................................ ................................ ................... 133

XI. Appendix E. Food processing report submitted by Syngenta

.. 138

XII. Appendix F. Food processing report specific to masa submitted

by Syngenta

................................ ................................ .............. 140

XIII. Appendix G. Bryson and Roberts report submitted by Syngenta

................................ ................................ ................................ .. 142

XIV. Appendix H. FDA memo on Event 3272 corn consultation

.... 166

XV. Appendix I. Report on DDGS submitted by Syngenta

............ 180

XVI. Appendix J. Glossary

................................ ............................... 1845 

## I. Purpose & Need

"Protecting American agriculture" is the basic charge of the U.S. Department of Agriculture's (USDA) Animal and Plant Health Inspection Service (APHIS). APHIS provides 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 agricultura l production (conventional, organic, or the use of genetically engineered varieties) can provide benefits to the environment, consumers, and farm

income.

APHIS Biotechnology Regulatory Service’s (BRS) mission is to protect America’s agriculture

and enviro nment using a science

- based regulatory framework that allows for the safe development and use of genetically engineered organisms.

R EGULATORY A UTHORITY Since 1986, The United States government has regulated genetically engineered 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 Science and Technology Policy, describes the comprehensi ve federal regulatory policy for ensuring the safety of biotechnology research and products and explains how federal agencies will use existing F ederal statutes in a manner to ensure public health and environmental safety while maintaining regulatory flexi bility to avoid impeding the growth of the biotechnology industry. The Coordinated Framework is based on several important guiding principles: (1) agencies should define those transgenic organisms subject to review to the extent permitted by their respec tive statutory authorities; (2) agencies are required to focus on the characteristics and risks of the biotechnology product, not the process by which it is created; (3) agencies are mandated to exercise oversight of genetically engineered organisms only w hen there is evidence of “unreasonable” risk.

The Coordinated Framework explained the regulatory roles and authorities for the three major agencies involved in regulating genetically engineered organisms: USDA’s Animal and Plant Health Inspection Servi ce (APHIS), the Food and Drug Administration (FDA), and the Environmental Protection Agency (EPA).

The EPA is responsible for regulating the sale, distribution and use of pesticides, including pesticides that are produced by an organism through techniqu es of modern biotechnology.

The FDA is responsible for ensuring the safety and proper labeling of all plant

- derived foods and feeds, including those that are genetically engineered. To help developers of food and feed derived from genetically engineere d crops comply with their obligations under F ederal food safety laws, FDA encourages them to participate in a voluntary consultation process. All food and feed derived from genetically engineered crops currently on the market in the United States

have suc cessfully completed this consultation process.

APHIS is responsible for regulating genetically engineered organisms and plants under the plant 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.6 W HAT IS A R EGULATED O RGANISM ? Under the Plant Protection Act (PPA), the term “plant pest” is defined as “any living stage of any of the following that can directly or indirectly injure, cause damage to, or c ause disease in any plant or plant product: a protozoan; a nonhuman animal; a parasitic plant; a bacterium; a fungus; a virus or viroid; an infectious agent or other pathogen; any article similar to or allied with any of the articles specified in the prece ding paragraphs.” (7 USC § 7702 (14). The PPA gives the Secretary broad discretion to regulate plant pests and prohibits persons from importing, exporting or moving in interstate commerce plant pests, except as authorized under general or specific permit s and in accordance with such regulations as the Secretary may issue to prevent the introduction of plant pests into the United States or the dissemination of plant pests within

the United States. (7 USC § 7711). In enacting the PPA, Congress found that it is the responsibility of the Secretary of Agriculture to facilitate commerce in agricultural products and other commodities that pose a risk of harboring plant pests in a manner that will reduce, to the extent practicable, as determined by the Secretary , the risk of dissemination of plant pests and that decisions affecting imports, exports, and interstate movement of products regulated under

the PPA shall be based on sound science. (7 USC § 7701 (3), (4) )

.

APHIS’ biotechnology regulations at 7 CFR p art 340 (Introduction of Organisms and Products Altered or Produced Through Genetic Engineering Which Are Plant Pests or Which There Is Reason To Believe Are Plant Pests) regulate the importation, interstate movement, or release into the environment (use of a regulated article outside the constraints of a physical confinement that are found in a laboratory, contained greenhouse, a fermenter, or other contained structure) of genetically engineered organisms 1 . A genetically engineered organism is regulated by AP HIS if it is a plant pest or if it or a gene donor or vector used in its construction are plant pests listed in 7 CFR 340.2. In addition, a genetically engineered organism can be considered a regulated article if APHIS has reason to believe it presents a plant pest risk.

W HAT IS THIS P ROJECT A BOUT ? APHIS’ regulations provide for developers of genetically engineered plants to file a petition for nonregulated status (7 CFR 340.6). The developer is required to submit scientific data and other

information to demonstrate that the plant does not come within the statutory definition of a plant pest, and, therefore, is no longer subject to APHIS jurisdiction and regulatory oversight.

On October 7, 2005, Syngenta Seeds, Inc. filed a petition for a determination of nonregulated status for a corn variety (Event 3272) genetically engineered to produce a microbial enzyme that facilitates ethanol production. Syngenta requests that APHIS make a determination that these corn plants do not pose a plant pest risk, and, therefore, shall no longer be considered regulated articles under 7 CFR p art 340.

1 A regulated article is a ny org anism which has been altered or produced through genetic engi neering, if the donor organism, recipient organism, or vector or vector agent belongs to any genera or taxa designated in 340.2 and meets the definition of plant pest, or is an unclassif ied organism and/or an organism whose classification is unknown , or an y product which contains such an organism, or any other organism or product altered or produced through genetic engi neering which the Administrator determines is a plant pest or h as reason to believe is a plant pest. Excluded are recipient mic roorganisms w hich are not plant pests and which

have resulted from t he addition of genetic material from a donor organism where the material is well characterized and contains only non

- coding regulatory regions. § 340.17 O BJECTIVES FOR E VENT 3272 C ORN The Federal Energy Policy Act of 2005, signed on August 8, 2005, includes a Renewable Fuels Standard that directs the doubling of the use of ethanol and biodiesel in the U.S. fuel supply by 2012 to 7.5 billion gallons (42 USC 15801). The Energy Independen ce and Security Act of

2007

, passed in December 2007, includes a provision to expand consumption of alternative fuels, including but not limited to ethanol, to 36 billion gallons in 2022. On October 13, 2010, the U.S. Environmental Protection Agency (EPA) waived a limitation on selling gasoline that contains more than 10 percent ethanol for model year 2007 and newer cars and light trucks. The waiver applies to fuel that contains up to 15 percent ethanol

– known as E15. The waiver was extended

to model ye ar 2001 through 2006 passenger vehicles, including cars, SUVs, and light pickup trucks on January 21, 2011. ( http://www.epa.gov/otaq/regs/fuels/additive/e15/ , accessed February 8, 2011). With o ver 194 corn ethanol plants in operation in 28 different states, and at least 7 more under construction ( http://www.ethanolrfa.org/bio

- refinery

- locations/ , accessed February 7, 2011), corn

- bas ed ethanol production may be a feasible way to meet and exceed the ethanol consumption benchmark for 2012 set in the Energy Policy Act of 2005, and 2022 goals set by the Energy Independence and Security Act of 2007.

Ethanol has also become the oxygenate of choice for reformulated gasoline (RFG), due to state

bans of methyl tert

- butyl ether (MTBE) and the increased liability to oil companies for MTBE spills (Hoffman et al. 2007) . As provided in the Federal Clean Air Act amendments of 1990, cities with the worst smog pollution are required to use RFG, while other cities voluntarily adopted the RFG program. The decreased use of MTBE in RFG, and the subsequent benchmark set by the Energ y Policy Act of 2005, and the Energy Independence and Security Act of 2007, along with other Federal programs and policies [see (Hoffman et al. 2007) and (Schnepf 2006) for a thorough discussion], have combined to stimulate the demand for ethanol.

Event 3272 corn is expected to help the U.S. meet its goals for ethanol production . According to the company, the overall efficiency of the dry grind process with using Event 3272 grain is greater than conventional dry ethanol production, due to greater ethanol production per unit of Event 3272 grain

, and greater throughput efficiency during the dry grind process itself.

Event 3272 corn has been field tested in the United States since 2002 as authorized by USDA notifications and permits listed in Table 1

- 1, on page 17 of the Petition. The list compiles a

number of test sites in diver se regions of the U.S. including the major corn growing areas of the

Midwest and winter nurseries in Hawaii and Puerto Rico. Field tests conducted under APHIS oversight allow for evaluation of Event 3272 corn in a n agricultural setting while imposing

meas ures to minimize the risk of persistence in the environment after the completion of the test. Data are gathered on multiple parameters and are used by the applicants to evaluate agronomic characteristics and product performance, and are used by APHIS to d etermine if the new variety poses a plant pest risk (USDA

- APHIS 200 9 )

.

Once APHIS determines that a GE organism is unlikely to pose a plant pest risk, the GE va riety may be traditionally bred with other conventional varieties or other GE varieties as determined by the applicant or developer , and is not subject to the regulatory requirements of 7 CFR part

340

. Syngenta currently has four GE corn varieties that may be stacked 2

2 “Stacked” refers to t wo or more traits (e.g. he rbicide tolerance and insect resistance) in one plant. with Event 3272 corn:8 t hree varieties that have an insect

- resistance trait (Bt11

, Mir604 , and Mir162 ) and one variety

that has an herbicide

- tolerance trait (GA21). These corn lines (Bt11, MIR604, Mir 162 and GA21) h ave all been granted non

- regulated status, and the environmental assessments conducted by APHIS for each of these products can be found at http://www.aphis.usda.gov/brs/not_reg.html

.

P URPOSE AND N EED FOR APHIS A CTION Under the authority of 7 CFR part 340, APHIS has the responsibility for the safe development and use of genetically engineered organisms under the provisions of the Plant Protection Act.

APHIS must respond to petitioners that request a determination of the non regulated status of genetically engineered organisms, including genetically engineered crop plants such as Event

3272 corn . If a petition for nonregulated status is submitted, APHIS must make a determination

if the genetically engineered organism is unlik ely to pose a plant pest risk.

As a Federal agency subject to compliance with the National Environmental Policy Act (NEPA) (42 U.S.C. 4321 et seq. ), APHIS has prepared this environmental assessment (EA) to consider the potentia l environmental effects of this proposed action (granting nonregulated status) and the reasonable alternatives to that action consistent with NEPA regulations (40 CFR parts 1500

- 1508, 7 CFR 1b, and 7 CFR part 372). This EA has been prepared in order to spe cifically evaluate the effects on the quality of the human environment 3 that may result from the deregulation of Event 3272 corn.

O THER R EGULATORY A PPROVALS Event 3272 corn has successfully completed the consultation process with the FDA concerning

food and feed safety (Appendix H ). Event 3272 corn does not contain any genetically engineered pesticides or tolerance to herbicides; therefore

, EPA consultation is not required for this product . There are no other Federal, state

, or local permits that are n eeded prior to the implementation of this action. A list of the current status of U.S. and international approvals is

found in Table 1

.

Table 1. Status of reviews. Syngenta has submitted documentation on Event 3272 corn to the appropriate officials in the following countries:

Country Type of S ubmission Date Approval

USA FDA Consultation 2005 2007 4

USDA Petition 2005

3 Under NEPA regulations, the “human environment” includes “the natural and physical environment and the relationship of people with the environment” (40 CFR § 1508.14).

4 http://cera

- g mc.org/index. php?action=gm_crop_database&mode=Submit&evidcode=Event%203272 , Date Accessed: February 7 , 201 1

5 http://www2.syngenta.com/en/media/mediareleases/en_100802.html and http://www.bsba.ag/BSBA/Apen.html

, Date Accessed: February 7, 20119 China Food, Feed, Processing including

Environment Currently in review

Australia/

New Zealand Food 2006 200 8 4

Taiwan Food 2006 2010 5

Korea Environment 2006

Food 2006

Canada Food 2006 2008 4

Feed 2006 2008 4

Environment 2006 2008 4

Japan Food 2006 2010 5

Feed 2006 2010 5

Environment 2006 2010 5

Switzerland Food 2006

Feed 2006

Russia Food 2006 2010 5

Feed 2006 2010 5

Philippines Food, feed, processing 2007 2008 4

Mexico Food and feed 2007 2008 4

EU Food and feed Currently in review

Indonesia Food, feed, processing Currently in review Republic of South

Africa Food and feed 2006 Not approved 6

P UBLIC I NVOLVEMENT
APHIS

- BRS routinely seeks public comment on draft environmental assessments. The issues discussed in this EA were developed by considering the public concern for ethanol production, as well as issues raised in public comments submitted for other environmental assessments of genetically engineered organisms, concerns raised in lawsu its, as well as those issues of concern that have been raised by various stakeholders. These issues, including those regarding the agricultural production of corn u sing various production methods and the environmental and food/feed safety of genetically e ngineered plants were addressed to analyze the potential environmental impacts of Event 3272 corn.

This EA and the petition submitted by Syngenta were available for public comment for a period of 60 days (7 CFR § 340.6(d)(2))

. Subsequently, APHIS reopened the public comment period

for an additional 30

- day comment period to allow interested persons additional time to submit comments on the petition, environmental assessment, and the revised PPRA to clarify APHIS’ definition of a plant p est in respon se comments received during the initial comment period.

Comments received during these two comment periods were used to analyze and inform APHIS ’

6 APHIS Docket 2007

- 0016; APHIS

- 2007

- 0016

- 0221.1 Attachment to public comment from Bill Freese, May 12,

200910 determination of whether to grant nonregulated status , and also whether there is a requirement to

prepare an Environmental Impact Statement in response to Syngenta’s petition for a determination of nonregulated status of Event 3272 corn.

I SSUES As stated above, the issues considered in this EA were developed based on APHIS’ review of a petition deregulate certa in genetically engineered organisms, and for this particular EA, the specific deregulation of Event 3272 corn for ethanol production.

Management Considerations :

• Corn Production

• Cropping Practices

• Tillage Practices

• Pesticide Use

• Specialty Corn and Processing

• Ethanol Production

Public Health Considerations

• Human Health

• Worker Safety

• Animal Feed and DDGS

Environmental Considerations

• Gene Movement (Pollen flow)

• Water Use in Ethanol Production

• Animals

• Plants

• Biodiversity

• Soil

• Conservation Reserve Program

D ECISIONS TO B E M ADE APHIS will use the information from this EA, and the comments it received, to make a

determination of whether to grant nonregulated status to Event 3272 corn and also whether to

prepare an Environmental I mpact Statement in connection with its determination of whether to grant nonregulated status to 3272 corn

.11 

## II. Affected Environment

C ORN P RODUCTION Corn ( Zea mays L.), otherwise known as maize, is the world's most widely grown cereal, reflecting its ability to adapt to a wide range of production environments (Morris 1998) . Corn is an annual plant typically grown in zones of abundant rainfall and fertile soils (Morris 1998) . In the U.S., corn is grown in temperate regions due to the moisture level and number of frost

- free days required to reach ma turity. Corn varieties having a relative maturity of 100 to 115 days are typically grown in the U.S. corn belt, which includes Iowa, Indiana, Illinois, and Ohio

— approximately 50% of all corn grown in the U.S. is from these four states. The Corn Belt also includes parts of South Dakota , Nebraska, Kansas, Minnesota, Wisconsin, Michigan, Missouri, and Kentucky.

Agricultural production systems may affect the surrounding environment, a nd corn production is no different. Depending on the region and practices used, corn production includes inputs such as fertilizer (e.g., synthetic fertilizers, manure, and compost containing nitrogen and phosphorus) and pesticides (synthetic or NOP

- appro ved insecticides, herbicides and fungicides), as well as irrigation. Each of these inputs can affect the environment including, but not limited to, increasing nutrient pollution in waterways, alterations in biodiversity due to pesticide inputs, and deplet ion of the water table or increased salinity in the fields due to irrigation.

There are available data for GE corn acreage (in parentheses for 20 10 ) for the following corn belt

states that also have ethanol facilities: Illinois (8 2 %), Indiana ( 83 %), Iowa ( 90 %), Kansas (90%), Michigan ( 80 %), Minnesota ( 92 %), Missouri (7 9 %), Nebraska ( 91 %), North Dakota ( 93 %), Ohio ( 71 %), South Dakota (95%), Texas ( 85 %) and Wisconsin ( 80 %) ( USDA

- NASS 20 10 b )

. GE corn is currently being grown for use in ethanol plants, altho ugh data is unavailable as to the percent of GE compared to non

- GE corn used in ethanol facilities.

Farming with GE and non

- GE Varieties Conventional farming covers a broad scope of farming practices, ranging from farmers who only occasionally use synthet ic pesticides and fertilizers to those farmers whose harvest depends on regular synthetic pesticide and fertilizer inputs. The use of GE varieties may be used in conventional farming, but there is also a smaller segment of conventional farmers that may gr ow

non

- GE corn for GE

- sensitive markets.

United States corn production for 20 10 , including production of conventional and genetically engineered corn varieties, was 8 1 million harvested acres (USDA

- NASS 20 10b ) . Of the total corn acres planted in 2010 , 8 6 % were GE corn varieties (USDA

- NASS 20 10b ) , slightly up from the 85% of 2009, and up from 73% in 2007 (USDA

- NASS 2007a) , and 61% in 2006 (USDA

NASS 2006)

. About 5 billion bushels of corn grain, which includes both GE and non

- GE varieties, were used for ethanol production in 2010 (USDA ERS 2011 ) , which corresponds to approximately 4 0% of U.S. corn production.

In organic systems, the use of synthetic pesticides and fertilizers is limited 7

7 7 CFR § 205.601 Synthetic substances allowed for use in organic crop production , and genetically engineered crops are prohibited . For t his EA, only organic farming systems produced using12 methods that fall under the USDA National Organic Program definition of organic farming and are certified organic production systems will be considered. Event 3272 corn is not approved for use in organic systems because it is genetically engineered. 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 flo wer at different times and not pollinate each other, and employing adequate isolation distances between the organic field and the fields of neighbors to minimize the chance that pollen will be carried between the fields. Organic growers must also maintain records to show that production and handling procedures comply with USDA organic standards.

Certified organic corn acreage is a small percentage of overall corn production. Extrapolating from the most recent certified organic corn acreage data published in 2010 (USDA

- NASS 2010) ; the estimated harvested acreage for certified organic corn in 2008 was 1 68,303 acres, representing 0.21% of the total corn acreage harvested for grain in the U.S. (USDA

- NASS 2009a). APHIS was unable to locate any ethanol production facilities that solely use organic corn; if organic corn is used exclusively for any ethanol pr oduction it is at a very small scale.

Agronomic Practices Today, conventional and organic growers can choose from hundreds of corn hybrids marketed by companies that p roduce seed (refer to Appendix A for examples of available GE varieties for use in conventional systems ). Hybrids differ generally in agronomic characteristics, including disease and pest resistance and length of growing period (Olson and Sander 1988)

. The optimum planting date for corn is influenced by factors such as the locality, environmental conditions, seed growing period, and seed variety, and it usually occurs in April or May. Harvesting generally occurs from mid

- to

- late September through November; the use of a combine (mechanical harvesting) is the standard practice for grain production in both conventional and organic systems

.

Crop rotations (successive planting of different crops on the same land) are used to op timize soil nutrition and fertility, and reduce pathogen loads. 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 rotation to another crop besides corn (Erickson and Lowenberg

- DeBoer 2005) . The increase in corn

- to

- corn rotations , mainly in conventional systems, has been

attrib uted to the increase in corn prices due to higher demand, mainly for ethanol production (Hart 2006, Stockton et al. 2007) . In some areas, the corn

- to

- corn rotation causes increased levels of fertilizer inputs (Sawyer 2007) . Insect pests may also increase in corn

- to

- corn rotations as this system may provide a continual host environment for some insects and diseases. However, in a corn

- soybean rotation, continuously growing corn for multiple growing seasons can decrease populations of soybean pests, such as soybean cyst nematode. Thus, corn

- to

- corn rotations may be used in s ituations outside of growing corn for ethanol production, and corn

- to

corn rotation has been used prior to the relatively higher increase in corn for ethanol (Erickson and Lowenberg

- DeBoer 2005)

.

Pesticides Use Agronomic practices for conventional or GE corn used in food/feed production and ethanol production for fuel are similar. Corn production typically involves the extensive use of inputs and technology (Rooney and Serna

- Sald ivar 1987, Shaw 1988, Pollak and White 1995, White13 and Pollak 1995) , and the main emphasis is placed on obtaining the best yield (Thomas 2007)

. Organic farming systems allow the use of some synthetic pesticides, and a list these can be found

at http://ecfr.gpoaccess.gov/cgi/t/text/text

- idx?c=ecfr&tpl=/ecfrbrowse/Title07/7cfr205_main_02.tpl.

Weed control methods differ depending on a number of factors including locality, grower resources, crop trait , and farming system ; the techniques may be direct (e.g. mechanical 8 and

chemical 9 ) or indirect (e.g. cultural 10 ) (Olson and Sander 1988)

. (Synthetic herbicides are not allowed in organic systems; a list of approved herbicides for use in organic systems may be

found at http://ecfr.gpoaccess.gov/cg i/t/text/text

- idx?c=ecfr&tpl=/ecfrbrowse/Title07/7cfr205_main_02 .tpl ) Pest control (weeds and insects) in corn production is essential in order to obtain good crop yield. Generally, growers will manage a range of pests simultaneously. Therefore, growers will likely

cho o se 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)

. Ulti mately, the management practices utilized by a grower will depend on the types of pests in their field, the level of infestation, the cropping system, the type of soil, cost, weather, time, and

labor. Practices to cope with pests, nutrient needs, and mois ture and temperature requirements vary regionally.

Specialty Corn Uses and Processing

C orn produced in the United States has a wide variety of end

- use functions. For the 2009/10 Market Year, of the almost 15 billion bushels of corn in U.S. supply, 11 billion bushels were consumed domestically (approximately 2 billion bushels were exported and the rest remained in supply). Of the 11 billion bushels used domestically

- 5.1 billion were used for animal feed and residual purposes, 4.7 billion were used for ethanol production, 770 million were used for the production of corn syrup and other sweeteners, 250 million were used to produce starch, about 194 million were used to produce cereals and other products, and 22 million were used to produce seed for futur e cropping (USDA

- ERS 2011). Approxim ately 8% (about a billion bushels) of the U.S. grown corn is specialty corn, with unique properties that are produced for a specific market or use (USGC 2006) . Some examples of specialty corn include popcorn, white corn, blue corn, sweet corn, waxy corn, hard endosperm, higher ferm entable corn, high protein and modified protein corn including corn with high amylose levels, high oil corn, non

- GE for

GE

- sensitive markets, and organic corn (USGC 2006)

.

These corn varieties are specified by buyers including end

- users of corn for specialized production, and premiums are paid for delivering a produ ct that meets purity and quality standards for the corn variety Although some specialty corn grain is easy to tell apart (e.g., popcorn, white corn, blue corn), most specialty corn varieties look similar to traditional corn but have compositional differenc es that allow for specified uses. For example, waxy corn has been bred to be 99% amylopectin (one component of starch), compared to 75% in typical food grade corn. High amylose corn is 55%

- 70% amylose, compared to 25% in common corn. These two types of specialty corn, waxy and high amylose, are used to produce specialty starches for use in various applications such as production of plastics, textiles, adhesives, food thickeners, and candy (Thomison, no date).

8 Includes tillage and mowing.

9 Herbicide application.

10 Crop rotation/spot spraying of herbicide/hand removal of weeds.14 Both high amylose corn and waxy corn are g rown using closed loop or identity preservation systems (similar to other specialty corn) as each of the corn varieties needs to meet the requirements and specifications of buyers (Brown et al., 1985). These value

- added corn products are produced and hand led in an isolated production stream meant to preserve the identity and integrity of the product. Any variation in either the composition of the corn (accidental pollination by other, non

- specialty corn) or in starch processing (misdirection of the wrong corn variety to a corn processing facility, for example) can cause a failure to meet production specifications and may result in poor or failed performance at a manufacturing facility (Hallauer 2001). Mixing of specialty corn varieties with other corn typ es during handling or production will diminish product value and potentially disrupt downstream processing or product supply availability, for either specialty corn products or for products derived from other corn varieties.

Product differentiation and ma rket segmentation in the specialty corn industry includes mechanisms to keep track of the grain (traceability), methods for identity preservation (IP, including closed

- loop systems), and quality assurance processes (e.g., ISO9001

- 2000 certification), as we ll as contracts between growers and buyers that specify delivery agreements. Corn processing facilities, such as starch processing facilities, have quality control processes in place to maintain the integrity of their processes and assure buyers that the products produced using specialty corn will be usable for specific end products.

E THANOL P RODUCTION Ethanol, also known as grain alcohol or ethyl alcohol , is the type of alcohol produced by fermenting and distilling simple sugars from biological source s. It is the same kind of alcohol found in all alcoholic beverages, although commercial ethanol plants add a poison (two to five percent) to make it unfit for human consumption (Morris and Hill 2006)

. Corn ethanol refineries account for more than 98 % of U.S. ethanol production ( http:/ /www.ethanolrfa.org , Accessed February 2011).

Approximately 4 0% of the 2010 corn production was used for ethanol production (USDA ERS

2011 ) . Commercial ethanol production uses conventional or GE corn; organic corn production for ethanol is typic ally small scale production for herbal medicinal use. The amount of GE corn currently used for ethanol production is not tracked by the USDA National Agricultural Statistics Survey . However, biotechnology

- derived varieties make up 8 6 % of corn acreage in the U.S. (USDA

- NASS 2010b ) , suggesting that GE corn is currently used in ethanol production.

The Federal Energy Act of 2005 includes a nationwide renewable fuels standard (RFS) that will result in the use of more than 7.5 billion gallons of ethanol and biodiesel by 2012 (42 USC 15801, page 1069). The Energy Independe nce and Security Act of 2007 passed in December 2007, includes a provision to expand consumption of alternative fuels, including but not limited to ethanol, to 36 billion gallons in 2022. As of February 3

, 2011 , there were currently 194 operational corn e thanol plants with a capacity of approximately 14 billion gallons per year ( www.ethanolrfa.org )

. Biofuels have been championed and pilloried as an alternative, renewable energy source. Proponents believe that corn

- p roduced ethanol can increase energy security, reduce vehicle emissions and provide a new income stream for farmers. Critics assert that corn

based ethanol will increase energy

- price volatility, food prices, and even increase life

- cycle emissions of greenho use gases and decrease water table levels.15 There are two types of ethanol processing plants in the U

. S

. , dry

- grind and wet

- milling plants. Dry mill facilities account for 82% of ethanol production and wet mills 18% (RFA 2007) . As Event 3272 corn will be produced for dry

- grind ethanol production, this assessment will include only those related to dry

- grind ethanol pr ocessing using corn as the feedstock.

The American Coalition for Ethanol, a pro

- ethanol website, provides an interactive tour of an ethanol processing plant at http://www.ethanol.org/index.php?id=73&parentid=73 (accessed July

20 08). Below is a review of the dry

- grind ethanol process from information from the Renewable Fuels Association, also a proponent of ethanol, and Mosier and Ileleji (Mosier and Ileleji 2006) :

There are 5 major steps in the dry

- grind method of ethanol production:

1. Milling. In dry milling, the entire corn kernel is first ground into flour, which is referred to in the industry as "meal" and processed without separating out the various component parts of the grain. Water is added to the meal to create a slurry. Microbial enzymes (alpha

- amylase) are adde d to the slurry to start the conversion of starch to dextrose.

2. Liquefaction. The slurry is processed in a high

- temperature cooker to reduce bacteria levels ahead of fermentation. Jet cookers inject steam into the corn flour slurry and cook it at temperat ures above 100°C. The cooked slurry, now called mash, is allowed to cool, additional microbial alpha

- amylase is added, and liquefying continues. Sulfuric acid is typically added to maintain pH.

3. Saccharification. The mash is further cooled after liquefact ion and a second microbial enzyme (glucoamylase) is added. This enzyme completes the breakdown of the starch into glucose, a simple sugar. This step often occurs as the mash is transferred to fermenters and continues throughout the next step.

4. Fermentatio n. Yeast is added and the conversion of sugar to ethanol and carbon dioxide (CO 2 ) begins. Ammonia is added for pH control and as a nutrient to the yeast. The fermentation process generally takes about 40 to 50 hours. During this part of the process,

the ma sh is agitated and kept cool to facilitate the activity of the yeast. The CO 2 released during fermentation is either released into the atmosphere or captured and sold for use in carbonating soft drinks and beverages and the manufacture of dry ice.

5. Distill ation and recovery. After fermentation, the resulting "beer" is transferred to distillation columns where the ethanol is separated from the remaining "stillage." The ethanol is concentrated to 190 proof using conventional distillation and then is dehydrate d to approximately 200 proof in a molecular sieve system. The ethanol product is then blended with about 5% denaturant (such as natural gasoline) to render it undrinkable and thus not subject to beverage alcohol tax. It is then ready for shipment to gasoli ne terminals or retailers. The stillage is centrifuged into liquid (thin stillage) and solid (distillers’ grains) fragments. Some of the thin stillage is recycled to the beginning of the dry

- grind process to conserve the water used by the facility. The r est of the thin stillage passes through evaporators to remove a significant portion of the water to produce thickened syrup. Usually, the syrup is blended with the distillers’ grains to produce distillers’ grains with solubles (DDGS), a high quality and n utritious livestock16 feed. When markets for the feed product are close the plant, the byproduct may be sold without drying as distillers’ grains or wet distillers’ grains. DDGS in wet form is prone to deterioration, especially in warmer weather; consequen tly, the use of wet DDGS is limited to producers located close to dry grind plants (Rausch and Belyea 2006)

.

P UBLIC H EALTH Public health concerns surrounding field corn, like Event 3272 corn and the resultant ethanol co

product of DDGS , focus primarily on human and animal consumption. Non

- GE corn varieties, both those developed for conventional use and for use in organic production systems, are not routinely required to be evaluated by any regulatory agency in the U.S. for food or feed safety prior to release in the market. Under the F ederal F ood, D rug, and C osmetic A ct (FFDCA) , it is the responsibility of food and feed manufacturers to ensure that the products they market are safe and properly labeled. Food and feed derived from Event 3272 corn 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. Although a voluntary process, applicants who wish to

comm ercialize a GE variety that will be included in the food supply 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, nutrition al, or other regulatory issues regarding the bioengineered 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. For a list of completed consul tations on GE organisms, see http://www.fda.gov/Food/Biotechnology/Submissions/default.htm (access ed February 7, 2011 ). FDA completed a consultation of Event 3272 on August 7, 2007 (see Appendix H)

.

Another potential concern is worker safety, both on the farm and at the ethanol plant. Farmers use an array of chemicals, including insecticides and herbicides, which may have toxic properties during application. Ethanol plant workers deal with hazardous chemicals such as sulfuric acid and liquid ammonia dur ing the processing of corn grain to ethanol and DDGS.

G ENE M OVEMENT Gene movement to other corn plants 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 reproduction in the laboratory ) (USDA

- APHIS 200 9 )

. P ublic concern surrounding gene m ovement for GE corn is between GE and non

- GE corn plants. Currently, 8 6 % of the conventionally grown corn in the U.S. is GE corn (USDA

- NASS 2010b )

. Specialty corn, those with traits of particular interest to various markets such as blue corn, waxy corn or organic corn, are typically grown with various management pract ices that intend to limit corn pollen from reaching the specialty corn crop during the period of time that the specialty corn crop is receptive to pollen. For example, the NOP has requirements for organic plans to address pollen flow from GE crops (Kuepper 2002, Krueger 2007, Kuepper et al. 2007) . The Association of Official Seed Certifying Agencies (AOSCA) also has information for specialty corn crops, and a protocol for growing non

- GE corn (AOSCA 2008) . There is a price premium associated with growing these types of specialty crops in conjunction with the extra regimens in pl ace to maximize the purity of these specialty crops. For example, in 2008, conventional field17 corn averaged $3.90/bushel (USDA

- NASS, 2009b) , whereas organic corn averaged $7.08/bushel (USDA

- NASS 2010 a )

.

A recent paper reviewed studies investigating gene flow and cross

- fertilization studies in corn grain production fields, and using the data fro m these studies recommended 50m (approx. 164ft)

as the distance needed to isolate GE corn and non

- GE corn (Sanvido 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 et al. 1972, Di Giovanni et al. 1995, Aylor et al. 2003) that only measured pollen flow

, because pollen flow does not necessarily result in

fertiliz ation. Successful cross

- fertilization requires many different biological and physical factors, such as identical timing of flowering between corn fields, viability of pollen, and presence of physical barriers; accordingly pollen dispersal is not equivalen t to cross fertilization.

Sanv id o et al. (2008) analysis of the existing studies found that the cross

- fertilization rate in non

GE corn typically remained below 0.5% at this distance, and this result was validated when analyzing cross

- fertilization events in large scale studies (e.g. Henry et al. 2003, Weber et al. 2007)

.

There is one scientific study typically used by critics of GE crops to refute the use of distance as an isolation strategy. The study found cross

- fertilization rates higher at comparable distances

than other studies (Jones and Brooks 1950) . For example, Jones and Brooks (1950) found cross

fertilization to be as high as 2.5 % at 660ft , which is the isolation distance used by AOSCA to isolate corn fields for seed production (AOSCA 2004) . One potential reason for the discrepancy between this study and almost all other gen e 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 for hybrid varieties.

Due the biol ogy 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 et al. 2008) , allowing for a

great er chance of pollination events. Consequently, the results from Jones and Brooks (1950) may be an overestimation of cross

- fertilization potential for hybrid corn plants.

W ATER U SE Corn plants use a significant amount of water during growth. About 4,000 gallons of water are needed to produce one bushel of corn, and in 20 10

, 81 million acres, or approximately 1 2.4

b illion bushels

, of corn were harvested in the U.S (USDA

- NASS 2010b )

. However only about

15 % of U.S. corn acres are irrigated, receiving on average 750 gallons of water per day, which is the equivalent of abo ut 3% of the average rainfall on each acre grown (McLaren, 2009). Most

corn acreage only receive s the ambient rainfall in a given area and corn plants typically transpire more water than what is retained by the plant, meaning that most of the water used by a corn plant is released to the atmosphere.

Water is also used during ethanol production ; about 4.7 gallons of water are needed to produce a gallon of ethanol (Shapouri and Gallagher 2005) . Water is used during the production of

microbial alpha

- amylase, as well as during its transport by tanker truck to ethanol plants (Appendix C )

.18 A NIMAL AND P LANT C OMMUNITIES
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 many birds visit row

- crop fields such as corn,

numbers are low and few nest ther e (Patterson and Best 1996) . The red

- winged blackbird ( Agelaius phoeniceus ) is the most abundant bird in North America; they are often initially attracted to corn fields to feed on insect pests but then feed on the corn. A nnually, this bird destroys o ver 360,000 tons of field corn and substantial amounts of sweet corn (Dolbeer 1990) ; other abundant species of birds that forage and/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

- tai led ( Odocoileus virginianus ), find field corn attractive because it functions both as food and cover throughout the latter half of the growing season (Vercauteren and Hygnstrom 1993)

. Deer can significantly damage or completely destroy small corn fields that are surrounded by woody or brushy areas; however, deer damage to large corn fields is often limited to a few rows closest to th e 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 et al. 2003) . The deer

mouse feeds on a wide variety of plant and animal matter depending on availability, but primarily feeds on seeds and insects. The deer mouse has been considered beneficia l in agroecosystems because it consumes both weed and pest insect species. The meadow vole ( Microtus pennsylvanicus ) feeds primarily on fresh grass, sedges, and herbs, but also on seeds and grains. The meadow vole may also be considered beneficial for its role in the consumption of weeds, but can be a significant agricultural pest where abundant as they rely on cover absent from tilled agriculture. The lined ground squirrel ( Spermophilus tridecemlineatus ) feeds primarily on seeds of weeds and available crop s, such as corn and wheat. This species has the potential to damage agricultural crops, although it can also be considered beneficial when eating pest insects, such as grasshoppers and cutworms.

Although many of the invertebrate organisms found in corn

- p roducing areas are considered pests, such as the European corn borer ( Ostrinia nubilalis ) and the corn rootworm ( Diabrotica spp.), many others are considered beneficial. Numerous insects and related arthropods perform valuable functions; they pollinate pla nts, contribute to the decay of organic matter, cycle soil nutrients, and attack other insects and mites that are considered to be pests.

Plants The landscape surrounding a corn field varies depending on the region. In certain areas, corn fields may be b ordered by other corn (or any other crop); fields may also be surrounded by wooden and/or pasture/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 dwe ll in and around corn fields; those species will also vary depending on the region where the corn is planted. Weeds compete with crops for water, nutrients, light, and other growth factors. Each year in the U.S., corn yields are threatened by more than 200 weed species (Heap 2008) . Weed species such as giant foxtail and barnyardgrass have been shown to reduce corn yields by up to 13 and 35%, respectively (Bosnic and Swanton 1997, Fausay et al. 1 997) . Common weeds that cause problems in corn fields include velvetleaf, common cocklebur, common lambsquarters (annuals) and quackgrass and Johnsongrass (perennials).19 Biological Diversity Species diversity and abundance in corn agro

- ecosystems may di ffer between the three corn production methods; GE, conventional

, or organic. Many studies over the last 10 years have investigated the differences in biological diversity and abundance between GE and non

- GE fields, particularly those GE crops that are re sistant to insects (e.g., Bt crops) or herbicides (e.g., glyphosate

- toleran t or glufosinate

- tolerant crops )

. Each side of the GE debate has a multitude of studies to pick from to support their case; opponents of GE will point to studies that indicate

pote ntial decreases in biological diversity and/or abundance due to GE crops, particularly due to the presence of a pesticidal protein in some GE crops (Bt) (e.g., (Hansen Jesse and Obrycki 2000, Ponsard et al. 2002, Pilcher et al. 2005) . GE

- detractors will also use studies investigating how decreases in weed populations due to the u se of herbicides and herbicide

- tolerant crops results in decreases in animal populations that use weeds as a food or refuge source, which may reduce overall biological diversity in farm fields (Marshall et al. 2003) . On the other side, supporters of GE technology will use studies that compare GE crops, such as Bt corn, to non

- GE crops sprayed with insecticides to demonstrate that GE crops do not cause any changes in arthropod abundance or diversity (e.g., (Bitzer et al. 2005, Torres and Ruberson 2005, Romeis e t al. 2006, Marvier et al. 2007, Chen et al. 2008, Wolfenbarger et al. 2008) or may even increase biological diversity (e.g., (R omeis et al. 2006, Marvier et al. 2007, Wolfenbarger et al. 2008) in

agro

- ecosystems. GE proponents may also showcase studies that demonstrate herbicide

- tolerant corn, when compared to conventional corn production, does not result in changes in arthropod abundance and may even increase speci es diversity during different times of the year (e.g., (Brooks et al. 2003, Haughton et al. 2003, Hawes et al. 2003, Roy et al. 2003)

.

S OIL B IOLOGY The soil environment in and around corn fields is complex, rich in microorganisms and arthropods. The corn root system acts as a soil modifier due to its association with several microbial groups such as bacteria, fungi, protozoa, and mites. Bacteria typically represent the most abundant microbes in the soil followed by fungi. These microbial groups play an important and particular role in the ecology of the soil, including nutrimental cycling and the availability of nutrients for plant growth. In addition, certain microbial organisms may contribute to the protection of the root system against soil pathogens (OECD 2003)

.

Research shows that crop soils are prone to degradation due to the disturbance and exposure of the top surface layer by certain agronomic practices. Two environmental impacts of soil degradation are the decline in water quality and the contribution to the greenhouse effect (Lal and Bruce 1999) . It has been shown that a decline in soil quality and soil resilience 11 enhances the greenhouse effect through emissions of radi atively

- active gases 12 (CO 2 , N 2 O) and depletion of the soil carbon pool (Lal 200 3, US

- EPA 2008) . In turn, a decrease in carbon aggregation and sequestration in the soil leads to increase runoff and soil erosion.

C ONSERVATION A CREAGE The Conservation Reserve Program (CRP) is America's largest and most effective private

- lands

cons ervation program, with more than 36 million acres enrolled. Under CRP, farmers and ranchers plant grasses and trees in crop fields and along streams. The plantings stop soil and

11 The ability of a soil to restore itself.

12 Gases that absorb incoming solar radiation or outgoing infrared in turn, affecting the temperature of the atmosphere.20 nutrients from washing into regional waterways and provide habitat for wildlif e (USDA

- FSA 2007)

.

CRP had 33.7 million enrolled acres as of in 2009 (USDA

- FSA 20 09 )

. CRP parti cipants enroll

i n contracts for 10 to 15 years every three years. With the increased demand for cropland due to market demands for agricultural crops over the last few years, CRP enrollment acreage is expected to decrease. For example, of 15.7 million ac res of CRP contracts that expired in 2007, only 85% we re re

- enrolled or extended , resulting in 2.3 million acres of cropland removed from the CRP program (USDA

- FSA 2007)

. That trend has continued as another 3 million acres have been removed from the program since 2007 (USDA

- FSA 200 9 ) . The change s of some of conservation program land to agricultural s ystems may alter the biodiversity on these acres.

A NIMAL F EED : D ISTILLERS G RAINS WITH S OLUBLES A bushel of corn going into an ethanol plant yields about 2. 8 gallons of ethanol and 16

- 18 pounds of DDGS. As a result of increased ethanol production, the q uantity of distillers grains

available in the U.S. increased from 1.9 million metric tons in 1999 to 8.3 million metric tons in 2005 (a 340 percent increase)

, and then more than tripled again to over 34 million metric tons in 2010 (Hoffman and Baker, 2010)

.

Approximately 90% of the distillers grains produced in U.S. facilities are used in domestic

animal feed. In North America, over 80% of DDGS are used in ruminant diets, but DDGS are also used for hog, swine, broiler, and turkey feed. Currently, ethano l plants do not discriminate

between GE and non

- GE corn. Given that 80 % of corn production is GE, the DDGS produced and used as animal feed are likely produced with some GE corn varieties.

Because of the near complete fermentation of starch d uring the ethanol process, the remaining amino acids, fat, minerals and vitamins in DDGS increase approximately three

- fold in concentration compared to levels found in cor n (Rausch and Belyea 2006) . This can result in concerns regarding phosphorous and animal waste dispos al , sulfur diet conten t, and mycotoxin

concentrations in DDGS.

Phosphorous concentrations in DDGS can significantly increase both phosphorus diet content as well as the resulting animal waste products when used as feed (Morse et al. 1992) . Increased phosphorus in animal waste leads to waste disposal concerns (Rausch and Belyea 2006) and

concerns about increased levels of phosphorus pollution of local and national bodies of water through agricultural runoff.

High dietary sulfur concentration in DDGS may also be a concern; it can lead to excessive sulfide concentrations in the rumen, and may cause a shift in the ruminal microbial population to include bacteria that produce high levels of thiaminases (Rausch and Belyea 2006) . This reduces

the thiamine available to be absorbed from the rumen and results in an effective thiamine deficiency which may cause brain lesions (Rausch and Belyea 2006) . The bacteria also produce an analog that inhibits certain enzymes involved in energy metabolism (Kung et al. 1998)

.

Mycotoxins may also be concentrated in DDGS. Mycotoxins are poisons produced by some fung i (e.g., Aspergillus

, Fusarium

, Penicillium ). The growth of fungi and the production of mycotoxins can be associated with stressed plants in the field (e.g., drought, pest damage) or21 poor storage after harvest (Whitlow and Hagler 2005) . During ethanol processing, mycotoxins are not destroyed; the toxins are concentrated three

- fold during ethanol processing, resulting in DDGS that may contain significantly incr eased levels of mycotoxins compared to corn grain for

feed (Garcia et al. 2008)

.

One of the mycotoxin metabolites present in corn, and potentially present in the resultant DDGS, aflatoxin M1, may be a concern to human health. If M1 is present in corn feed or DDGS and fed to dairy cattle, M1 can be passed in milk for human consumption (Garcia et al. 2008) . M1, a metabolite Aflatoxin B1, is considered a potential carcinogenic compound for humans. Because of the human health concerns surrounding aflatoxins, aflatoxin is regulated by the FDA at the

action level. Action levels for poisonous or deleterious substances are established by the FDA to control levels of contaminants in human food and animal feed. These action levels represent limits at or above which FDA will take legal action to remove products from the market (FDA

200 0)

.22 

## III. Alternatives

This EA analyzes the potential environmental consequences of a proposal to grant nonregulated

status to Event 3272 corn. In order for Event 3272 corn to be granted nonregulated status, it must be found to be unlikely to pose a plant pest risk. The analysis provided in the plant pe st
risk assessment (USDA

- APHIS 200 9 ) demonstrates that Event 3272 corn is unlikely to pose a plant pest risk and therefore should no longer be subject to the regulatory requirements of 7 CFR part 340.

The regulations at 7 CFR 340.6(d)(3)(i) state that APHIS may "approve the petition in whole or in part." Because APHIS has found that Event 3272 corn is unlikely to pose a plant pest risk, there is no regulatory requirement for considering whether to approve the petition in part. The petitioner has not request ed APHIS to consider approving a partial deregulation, and the petition for Event 3272 corn only requested APHIS to grant nonregulated status to one corn line, so this

type of “in part” approval will not be considered. Therefore , there are two alternatives that will

be consid ered in this EA: (1) no action and (2) grant nonregulated status to Event 3272 corn, “ in

whole. ”



### A. No Action: Continuation as a Regulated Article

Under the "no action" alternative, APHIS would deny the petition to grant nonregulated status to

Event 3272 corn

. Event 3272 corn would continue to be subject to regulation pursuant to 7 CFR part 340. The company would have to continue to request permits and notifications for new introductions of Event 3272 corn plants. Permit conditions would be specified by APHIS

. These conditions would be designed to confine Event 3272 corn. The size of planting would be limited to help maintain confinement. In addition, the number of permits granted would be limited by agency resources, both in terms of the number of permits which could be reviewed by APHIS, and in APHIS’ ability to inspect the field trials and enforce compliance with regulations.

As such, it would be difficult for the company to commercialize Event 3272 corn under the permit or notification process. This al ternative is not the preferred alternative because APHIS has concluded through a plant pest risk assessment (USDA

- APHIS 2009) that Event 3272 corn does not pose a plant pest risk. Choosing this alternative would not be consistent with the purpose and need of APHIS to allow for the safe development and use of GE organisms given that Event 3272 corn does not pose a plant pest risk.



### B. Preferred Alternative: Determi nation that Event 3272 Corn Plants are No Longer Regulated Articles, in Whole

Under this alternative, Event 3272 corn would no longer be a regulated article under the regulations at 7 CFR part 340. Event 3272 corn should be granted nonregulated status bec ause APHIS has concluded that this GE organism does not pose a plant pest risk (USDA

- APHIS 2009)

. Permits issued or notifications ac knowledged by APHIS would no longer be required for

introductions of Event 3272 corn, or progeny derived from these events. This alternative best meets the purpose and need for agency action. The agency’s need is to make a decision on the petition that is consistent with the regula tory requirements in 7 CFR part 340. Granting nonregulated status to Event 3272 corn is consistent with the plant pest provisions of the PPA,23 the regulations codified in 7 CFR part 340, and the biotechnology regulatory policies in the

Coordinated Framewor k.



### C . Alternatives Considered but Rejected from Further Consideration
APHIS assembled a comprehensive list of alternatives that might be implemented in the decision process for Event 3272 corn. The agency individually evaluated each alternative on th e basis of consistency with the current regulations , environmental safety, efficacy, and practicality to identify which alternatives would be further considered during the decision process. Based on this evaluation, APHIS rejected several alternatives. I n the interest of transparency, these alternatives are discussed briefly below along with the specific reasons for rejecting each.

Prohibit any Event 3272 corn from being released. In response to public comments that stated a preference that no GE organis ms enter the marketplace, APHIS considered prohibiting the release Event 3272 corn, including denying any permits associated with the field testing. APHIS determined that this alternative is not appropriate given that APHIS has concluded that Event 3272 corn is unlikely to pose a plant pest risk (USDA

- APHIS 200 9 )

.

The Secretary of Agriculture is directed, through APHIS, to facilitate

— “... the smooth movement of enterable plants, plant products, biological control organisms, or other articles into, out of, or within the United States... (and to facilitate) exports, imports, and interstate commerce in agricultural products and other commodities that pose

a risk of h arboring plant pests or noxious weeds in ways that will reduce, to the extent practicable, as determined by the Secretary, the risk of dissemination of plant pests or noxious weeds... § 402(3)(5).”

The question as to how to balance this facilitation with th e protection of U.S. agriculture is unequivocally answered by the Plant Protection Act , which states that

—

[D] ecisions affecting imports, exports, and interstate movement of products regulated under (the Plant Protection Act) shall be based on sound science... § 402(4).

A risk

- management process based on sound science must, therefore, consider a growing body of scientific evidence documenting the safe use of GE organisms in U.S. agriculture, and in the rest of the world, to determine whether their use poses any unacceptable risks. Because Congress has

mandated a science

- based approach in APHIS regulations and because there is no basis in science for banning the release of Event 3272 corn, a blanket prohibition of the release of Event

3272 corn would co ntravene congressional intent and must be rejected.

Isolation distance between Event 3272 corn and non

- GE corn production and g eographic

restrictions In response to public concerns regarding possible gene movement between GE and non

- GE plants, APHIS consi dered requiring an isolation distance greater than 660 f ee t separating Event

3272 corn from non

- GE corn producti on. However, because Event 3272 corn does not pose a plant pest risk (USDA

- APHIS 2009), a n alternative based on requiring isolation distances was viewed as inconsistent with the current regulations in 7 CFR part 340

.24 APHIS also considered geographically restricting the production of Event 3272 corn based on production of non

- GE corn in organic production systems or production systems for GE

sensitive markets i n response to public concerns regarding possible gene movement between

GE and non

- GE plants

. However, a s presented in APHIS’ plant pest risk assessment for Event 3272 corn, there are no geographic differences associated with any iden tifiable plant pest risks

for Event 3272 corn (USDA

- APHIS 2009) . This alternative was rejected and not analyzed in

detail because APHIS has concluded that Event 3272 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’ regulatory authority and the biotechnology regulatory polic ies embodied in the Coordinated Framework.

Based on the foregoing, the imposition of isolation distances or geographic restrictions would not meet APHIS’ purpose and need to act on the petition in accordance with its regulatory authorities.

Nevertheless

, APHIS is not expecting significant effects. However, i ndividuals might choose on

their own to geographically isolate their non

- GE corn productions systems from Event 3272

corn, or to use isolation distances and other management practices to minimize gen e movement
between corn fields. Information to assist growers in making informed management decisions for Event 3272 is available from Association of Official Seed Certifying Agencies (AOSCA 2004) and Syngenta ( see Appendix D and Appendix G in the EA).

Requirement of testing for Event 3272 corn During the comment periods for other petitions for granting nonregulated status, some commenters requested USDA to require and provide testing for GE products in non

- GE production systems. However, there are no nationally

- established regulations involving testing

or limits of GE material in non

- GE systems. Such a requirement would be both novel and extremely difficult to implement and maintain. Additionally, because Event 3272 corn does not pose a plant pest risk (USDA

- APHIS 2009), the imposition of any type of testing requirements is

inconsistent with APHIS’ regulatory policies or the biotechnology regulatory policies embodied in the Coordinated Framework. Therefore, the requirement of tes ting for Event 3272 corn would not meet APHIS’ purpose and need to act on the petition in accordance with its regulatory

authorities

.



### D. Comparison of Alternatives
Table 2, below, briefly summarizes the results for each of the issues raised in the Environ mental Consequences (Section IV) by each of the alternatives described in the Alternatives section (Section III).

Table 2 . Comparison of Alternatives

Attribute/Measure Outcome if Alternative A

No Action is Chose Outcome if Alternative B Deregulation is Chosen Meets Purpose and Need and Objectives No Yes

Is there a plant pest risk? No (USDA

- APHIS 2009) No (USDA

- APHIS 2009) Management Practices

Corn Pro duction No change in U.S. corn

acreage . No change in use of No change in U.S. corn

acreage . No change in use of25 Attribute/Measure Outcome if Alternative A

No Action is Chose Outcome if Alternative B Deregulation is Chosen

GE or non

- GE corn varieties during corn production. No change in corn acreage for ethanol production. GE or non

- GE corn varieties during corn p roduction. U.S. corn acreage for ethanol production may decrease due to increased ethanol efficiency

of Event 3272 corn

. Cropping Practices No change in cropping practices in GE and non

- GE farming systems. No change in cropping practices for in GE and non

GE farming systems.

Pesticide Use No change in pesticide use in

GE and non

- GE farming

systems. No change in pesticide use in

GE and non

- GE farming

systems. Specialty Corn Uses and Processing Economic effects of misdirected specialty corn varieties may cause effects in food processing, but effects would not be caused by Event

3272 corn. Economic effects of misdirected specialty corn varieties may cause effects in food processing, and effects may be caused by Event 3272 corn. The magnitude of the

conseq uences of the effects

has not been studied.

Ethanol Production 2. 8 gallons/bushel if no ethanol plants are converted

for Event 3272 corn use

. 2.8 gallons/bushel + 2% or greater increase when ethanol plants are converted for Event

3272 corn use.

Human and Animal Health

Public Health: Risk to Human

Health Minimal risk due to corn consumption. Minimal risk due to Event 3272 corn consumption.

Public Health: Risk to Worker Safety Minimal risk due to corn production; Higher risk in ethanol production. Minimal risk due to Event

3272 corn production;

Decreased risk in Event 3272

converted ethanol facilities.

Animal Feed : DDGS Minimal risk due to the use of

corn in DDGS. Minimal risk due to the use of

Event 3272 corn in DDGS.

Environment

Gene Movement Low risk of gene flow from

GE corn to non

- GE corn. No risk of gene flow from GE corn to native plants. Low risk of gene flow from

GE corn to non

- GE corn. No risk of gene flow from Event 3272 corn to native plants.

Water use W ater is required for prod uction of GE and non

- GE

corn and for ethanol production

. The same amount of water used to grow corn under the

No Action alternative is needed to grow Event 3272 corn. Less water is required

for ethanol facilities converted

for Event 3272 corn use

.26 Attribute/Measure Outcome if Alternative A

No Action is Chose Outcome if Alternative B Deregulation is Chosen

Animals Minimal risk due to corn consumption. Minimal risk due to Event 3272 corn consumption.

Plants Weeds and other plants inhabit agricultural areas surrounding corn production and will be managed. The same amount of weed and plant management used to

g row corn under the No

Action alternative will occur

in Event 3272 corn production. Biological Diversity Animals will inhabit agricultural areas surrounding corn production and will be managed. The same amount of animal management used to grow

corn under the No Action

alternative will occur in Event 3272 corn production. Soil biology Soil modification may occur during corn production. The same amount of soil modification used to grow

corn under the No Action alternative may occur during Event 3272 corn p roduction. CRP Acreage Corn acres may be enrolled or

un

- enrolled in CRP. The same amount of corn

acres enrolled in CRP under

the No Action alternative will

be enrolled under this

alternative. Fewer corn acres may be un

- enrolled in CRP. Other Regulatory A pprovals for Event 3272 Corn

United States FDA consultation complete. No requirement for EPA review

Other Countries Canada, Philippines, Australia, New Zealand, Mexico, Japan, Taiwan, and Russia (Table 1)

Compliance of Granting Nonregulated Status to Event 3272 Corn with Other Laws Endangered Species Act, Clean Water Act, Clean Air Act, Executive Orders Fully compliant27 

## IV. Environmental Consequences

Potential environmental impacts f rom the “no action” alternative and the “preferred” alternative for Event 3272 corn 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 cultiv ation would be no different between the alternatives; those are described below.



### Methodology and Assumptions

The environmental effects analysis is greatly dependent on assumptions used for estimating effects. The following are key underlying assumptio ns use d to estimate effects for each

alternative.

S COPE OF THE E NVIRONMENTAL A NALYSIS
Event 3272 corn is genetically engineered to produce a microbial enzyme that facilitates ethanol production and, based upon inf ormation provided by Syngenta, ethanol production is the

intended use of Event 3272 corn

. Therefore, APHIS will primarily focus its environmental analysis on those geographic areas that produce corn and are near corn ethanol plants either currently in production or under construction.

D ue to the properties of Event 3272 corn, and the contractual obligations between growers and ethanol facilities, ethanol plant managers must make a manufacturing decision to use Event 3272 corn in their facility. Event 3272 corn cannot be used routinely in any and all ethanol plants. The inputs used for ethanol production are specific to each ethanol plant and margins of efficiency and efficacy are tied directly to characteristics of the locally grown corn (e.g., moisture content) and the specific parameters of other inputs used in a particular facility. Although the environmental analysis will include areas of corn production surrounding any corn ethanol plant, the scope of the affected environment is substantially smaller in reality and limited

to only thos e corn production areas that surround an Event 3272

- specific ethanol plant. Currently there is only one ethanol plant in operation prepared to accept Event 3272 corn, and two other ethanol plants may become operational in 2011 if nonregulated status is gr anted, out of a total of 194 corn ethanol plants.

To determine areas of corn production, APHIS used data from the National Agricultural Statistics Service (NASS) 2002 Census of Agriculture to determine where corn is produced in the United States ( www.nass.usda.gov , accessed 6/5/2008). NASS has since published the 2007 Census of Agriculture ( www.nass.usda.gov , accessed 2/16/2010) but this information has not changed the findings that were determined with the 2002 data.

The list of 49 states that produce corn grain is found in Table 3, according to the 2002 and 2007

Census es of Agriculture. As of February 20 1 1 , there are at least 194 operational corn ethanol plants with 7 plants under construction ( www.ethanolrfa.org ). The states that have operational ethanol plants or are constructing plants that will use corn as the input are also listed in Table 3.

Table 3. States that grow corn according to the 2002 and 2007 Census es of Agriculture, and whether the state also has an active corn ethanol facility or one under construction (according to28 February 201 1 data from www.e thanolrfa.org) . The states that grow corn and have an existing corn ethanol facility or one under construction will be included in the analysis for the

environmental effects for Event 3272 corn.

Corn Growing State Corn Ethanol Facility? Corn Growing State Corn Ethanol Facility?

Alabama No Nebraska Yes

Arizona Yes Nevada No

Arkansas No New Hampshire No

California Yes New Jersey No

Colorado Yes New Mexico Yes

Connecticut No New York Yes

Delaware No North Carolina Yes

Florida No North Dakota Yes Georgia Yes Ohio Yes

Hawaii No Oklahoma No

Idaho Yes Oregon Yes

Illinois Yes Pennsylvania Yes

Indiana Yes Rhode Island No

Iowa Yes South Carolina No

Kansas Yes South Dakota Yes Kentucky Yes Tennessee Yes

Louisiana No Texas Yes

Maine No Utah No Maryland No Vermont No

Massachusetts No Virginia Yes Michigan Yes Washington No

Minnesota Yes West Virginia No
Mississippi Yes Wisconsin Yes

Missouri Yes Wyoming Yes

Montana No

The corn

- growing counties within the 2 8 states that have a corn ethanol plant or one under

construct ion are listed in Appendix B. These 236 0 counties in 2 8 states are be included in the environmental effects analysis for the alternatives , even though the entirety of this area will not

include Event 3272 corn or Event 3272 corn

- specific ethanol facilities

.

G ENERAL M ANAGEMENT P RACTICES One of APHIS’ missions is to improve American agricultural productivity. Best management practices, such as planting dates, seeding rates, and harvest times are commonly accepted, practical ways to grow corn, regardless if the corn farmer is using convent ional systems, organic practices, or using genetically engineered varieties. These well

- established, widely

- practiced means to produce corn can be obtained through local Cooperative Extension Service offices and their respective websites. (A summary websi te can be found at www.ipmcenters.org/cropprofiles/index.cfm , accessed 4/24/2008).

S PECIALTY C ORN M ANAGEMENT P RACTICES It is Syngenta’s intention that Event 3272 corn will be marketed and p roduced in a manner

similar to other value

- added, specialty corn products, such as popcorn, waxy corn, high oil corn,29 high protein and modified protein corn, sweet corn, higher fermentable corn, and high amylase

corn (USGC 2006)

. As discussed above in the Affected Environment section

, these value

- added products are t ypically produced with identity preservation systems that include contracts between all handlers of the grain (seed suppliers, farmers, handlers, and processors), traceability, product tracking, process verification

, and separate channeling to minimize com mingling.

Similar to other specialty corn varieties, Event 3272 corn will require a level of control to preserve identity and avoid inadvertent mixing with other corn products. This identity preservation will include the use of a ‘closed

- loop’ system. The closed

- loop system provides more control over the specialty crop to better protect its value and to not interfere with other corn

varieties. Closed

- loop production systems involve rigorous controls to maintain the identity of the crop, tight chain of custody of the crop as it moves through the stages of production (seed, planting, harvesting, and transportation to end user) and contractual requirements of each party in the system.

If Event 3272 corn will be commercialized, Syngenta will have contract s and other control mechanisms in place to establish a ‘closed

- loop’ system for Event 3272 corn. This closed

- loop system includes three principle points of contact; the producer of Event 3272 corn (Syngenta), the grower of Event 3272 corn, and the end

- use r of Event 3272 corn, the ethanol plant. According to Syngenta (see Appendix G), each point of contact has specific roles and responsibilities:

“Syngenta [will] (stewardship roles and responsibilities):

**•** license the use of Event 3272 corn product to growers;

**•** sell Event 3272 corn hybrids only to licensed growers with a valid contract with an ethanol plant or an approved third party grain company that supplies corn amylase to the ethanol plant;

● ensure that grain contract includes stewardship agreement;

**•** provide incentive to grower for producing and delivery of Event 3272 corn product;

**•** provide stewardship guide to producers and handlers on the cultivation and handling of the Event 3272 corn product;

**•** prov ide specific procedures for the handling of any excess grain;

**•** ensure the domestic consumption of DDGS prior to export market approvals;

**•** make available appropriate detection methods; and,

**•** develop and implement a communication program.

Grower [will] (stewardship roles and responsibilities):

**•** execute delivery contract with ethanol plant;

**•** execute stewardship contract with Syngenta;

**•** follow Syngenta stewardship guide on cultivation; and,

**•** follow Syngenta requirement to divert excess grain to ap propriate use.30 Ethanol Plant (stewardship roles and responsibilities):

**•** contract with growers to supply Event 3272 corn product; and,

**•** ensure domestic consumption of DDGS prior to export market approvals;

These stewardship roles and responsibilities in concert with the contract relationships

between the entities in the closed

- loop system and the backstop mechanisms minimize the risk of inadvertent delivery of Event 3272 corn and commingling.”

According to Syngenta (see Appendix G), the system for Event 3272 corn will include contracts

for Event 3272 corn grown in surrounding areas of an Event 3272 corn

- contracted ethanol facility. Due to economic considerations, trucks dominate the local transportation of corn to ethanol facilities, limiting the geographic area for growing Event 3272 corn to approximately a

50

- mile radius (Denicoff 2007). The farther corn is from the ethanol plant, the greater the transportation costs to move Event 3272 corn to the ethanol facility. As a result, there is a limit to the area around the ethanol facility that is economically feasible to effectively use Event 3272

corn.

The ethanol plant will contract either directly with Event 3272 corn growers in their geographic region or indir ectly through grain suppliers. The contracts for growers will dictate the delivery location (ethanol plant or storage site within a specified radius of the field), a delivery date, and amount to be delivered. Syngenta has stated that hybrids with E vent 3272 will only be available to growers with a valid contract, and who agree to the Syngenta Stewardship Agreement (see Appendix G).

Part of the Stewardship Agreement is the requirement that growers include 12 rows of non

- Event

3272 corn as a pollen t rap to reduce the amount of Event 3272 pollen t hat may leave the corn

field. Syngenta has provided an analysis (Appendix D ) that analyzes the effect of using border rows on the minimizing corn pollen leaving an Event 3272 corn field. Syngenta’s analysis suggests that more than 99.9% of all corn pollen is captured when 12 border rows of non

- Event

3272 co rn plants are used and flowers are in synchrony with the Event 3272 corn field. This analysis is consistent with other gene flow and pollen flow studies i n the scientific literature (Jarosz et al. 2003, Jemison and Veyda 2001, Ma 2005, Ma et al. 2004, and Westgate et al. 2003).

P RODUCTION P RACTICES APHIS also recognizes that producers of non

- GE corn, particularly producers who sell their products to markets sensitive to genetically engineered traits (e.g. organic or some export markets) can be reasonably assumed to be using practices on their farm to protect their crop from

unwanted substances and maintain their price premium. For example, the National Organic Program (NOP) has recognized the practicality of protecting organically

- produced crops, and the investment farmers put into their production p ractices, by requiring that organic production plans include methods to protect organically

- produced crops.

Organic crops must be protected from contamination by prohibited substances used on adjoining lands (for example, drifting pesticides,31 fertilizer

- l aden runoff water, and pollen drift from genetically engineered...) (NCAT 2003)

.

Typically, more than one method is used under organic practices to prevent unwanted material from entering their fields including : isolation of the farm, physical barriers or buffer zones between organic production and non

- organic produc tion, as well as formal communications between neighboring farms (NCAT 2003) . The organic plan used as the basis for organic certification should include 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 transporting the crop (Riddle 2004, Krueger 2007, Kuepper et al. 2007) . Organic plans should also i nclude provisions describing how the risk of GE pollen or

seed co

- mingling will be monitored (Kuepper 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 a lso help the organic farmer reduce contamination events (NCAT 2003, Krueger 2007)

. Therefore , commonly used production practices for corn

, and the practical methods typically used by corn farmers using organic methods to protect their crop and maximize their profits and price premiums granted to corn u nder organic production, currently provide many measures that greatly reduce the likelihood of accidental gene flow between Event

3272 corn and non

- GE corn fields. APHIS will use the assumption that farmers are already using, or have the ability to use, t hese common, reasonable practices as its baseline for the analyses of the following alternatives below. Recommended organic production practices for field corn are also readily available (Kuepper 2002)

.

O THER A SSUMPTIONS
T he environmental consequences of the different alternatives described above have been analyzed under the assumption that farmers who produce conventional corn, Event 3272 corn, or produce corn using organic methods, are using reasonable, commonly accepted best management practices for their chosen system and varieties during agricultural corn production. However, APHIS recognizes that not all farmers follow these best management practices for corn. T he analyses of the environmental e ffects also include the assumption that some farmers do not follow these best management practices.

O THER I NFORMATION Syngenta provided APHIS a serie s of reports detail ing the potential benefits and impacts of Event 3272 corn in terms of ethanol production and byproducts, corn processing, and pollen

movement in corn. These are referenced in the EA and the CBI

- deleted versions are found as Appendices t o this EA.



### Corn Production

GE and non

- GE corn varieties are continually under development. Currently, 8 6 % of the total corn acres planted in 2008 are GE corn varieties (USDA

- NASS 2010b ) , and about 4 0% of the total corn production in 20 10 was used for ethanol production (USDA ERS 2011 )

. In 20 10

, growers harves t ed 81 million acres for grain, which is a slight increase from the 79.6 million

acres harvested in 2009

.32 F ARMING WITH GE AND NON

- GE V ARIETIES

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. Of the total corn acres planted in 2010 , 8 6 % were GE corn varieties (USDA

- NASS 2010b ) up from 73% in

2007 (USDA

- NASS 2007a) , and 61% in 2006 (USDA

- NASS 2006)

. Conventional production practices that use GE varieties will likely continue to increase without granting nonregulated

status to Event 3272 under the “no action” alternative , based on current acreage tren ds

. Cu rrently available seed for conventional and GE varieties will remain the same under the “no

action” alternative , except Event 3272 corn variety will be unavailable

. Corn is currently produced in 49 states (all states but Alaska according to the 200 7 Census of Agriculture) , and under the “no action” alternative, this range of production will be unchanged.

GE corn has been in production s ince the mid

- 1990’s, and in 2010 accounted for 86 % of the acres of corn planted (USDA

- NASS 2010 ) . During this time, corn production for GE

- sensitive markets has increased; for ex ample organic corn production increased 35% between 2001 and

2005 ( USGC 2006) , at the same time that corn acreage using GE varieties increased from 26% of corn acreage to 52% of corn acreage (USDA

- NASS 2001, 2005) . The amoun t of GE corn planted for conventional corn production and amount of corn planted for organic production systems in the U.S. are increasing, and both production practices will likely continue to increase without granting nonregulated status to Event 3272 un der the “no action” alternative. Management practices currently in used for GE and non

- GE corn will remain in place and used by farmers of GE varieties and those producing corn for GE

- sensitive markets. Currently

available seed for conventional and GE cor n varieties, and those corn varieties that are developed for organic production, will remain the same under the “no action” alternative.

Preferred Alternative: Acreage and Areas of Corn Production In 2008, GE corn production was planted on 8 0 % of all co rn acres currently in production in the US, and the use of GE corn has been increasing over the last 3 years (USDA

- NASS 2006, 2007a,

2008a , 2010b )

. Conventional and GE corn production occurs on land that is dedicated to crop production. Most corn is planted in f ields that have been in crop production for years. Granting nonregulated status of Event 3272 corn under the “preferred” alternative is not expected to significantly alter the range of corn cultivation as the new GE trait (alpha

- amylase) does not change th e growth habits compared to con ventional varieties (USDA

- APHIS 200 9 )

. Event 3272 corn will be marketed for use in ethanol production, and this corn variety will be limited to production area s that surround ethanol production facilities (see Appendix G)

. Currently, there

are 28 states that have corn

- based ethanol production facilities, including states with facilities under construction (see Table 3). This corn v ariety will likely be introduced to areas where corn is currently grown for ethanol production as a replacement product to other varieties (conventional and GE) already available in the market today and used for ethanol production. Further, the acreage fo r Event 3272 corn is limited to areas surrounding ethanol facilities that have been contracted to accept Event 3272 corn. As a result, under the preferred alternative, granting nonregulated status to Event 3272 corn would not increase the demand for corn

p roduction, or alone cause an increase in overall GE corn acreage.33 Event 3272 corn has the potential to decrease overall corn conventional (non

- organic) acre age needed to meet the biofuel amounts specified in the Energy Policy Act of 2005 and the Energy

In dependence and Security Act of 2007. APHIS has analyzed the projections made for ethanol production efficiency submitted by Syngenta (Appendix C). Due to the changes in ethanol processing when using Event 3272 corn, Syngenta projects a 2% or greater incr ease in efficiency, resulting in more ethanol generated per unit grain than current ethanol processes using microbial alpha

- amylase. If Event 3272 corn does result in greater efficiency during ethanol production, fewer acres of corn production may be requ ired to meet the biofuel levels mandated by Congress.

Granting nonregulated status to Event 3272 corn has the potential to result in a decrease in acreage devoted to corn produced for ethanol production due to a projected 2% or greater

increase in ethano l efficiency

– fewer bushels of corn may be needed to meet the requirements of the Energy Policy Act of 2005 and the Energy Independence and Security Act of 2007.

Corn produced using organic methods has been increasing at approximately 30% a year (USGC 2006) , even though there are no national requirements that GE corn varieties currently in production use mandated techniques for isolation between fields containing GE corn and those fields using organic production practices. In the Methodology and Assumptions section above,

the discussion contains information related t o the requirements for farmers using organic production methods under the National Organic Program. Conventional farmers growing GE varieties typically use traditional production practices, and have no requirements related to isolating pollen movement bet ween their fields and their neighbor’s field. Without any requirements in place, the production of corn using organic methods is continuing to increase.

In agricultural systems, growers may choose to grow GE or non

- GE corn, and obtain price premiums for growing 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 2008, conventional field corn averaged $3.90/bushel (USDA

- NASS, 2009b)

, whereas organic corn averaged $7.08/bushel (USDA

- NASS 2010) . USDA asserts that agricultura l practices that use conventional means, organic production systems, or genetically engineered varieties can all provide benefits to the environment, consumers, and farm income. As discussed in Section II and below under the section for “Gene Movement,” g ene flow into and out of these specialized corn production systems has been managed using various types of buffer zones or isolation practices, such as differences in tim ing of planting (which results in differences in flowering and removes the ability to pollinate neighboring crops) or making sure fields are distance from other compatible crops (such as using isolation distances).

As discussed in the Methodology and Assumption methods, Syngenta has included some measures in their Stewardship Agreement th at minimize pollen movement outside corn fields containing Event 3272 corn (see Gene Movement section below for further discussion and Appendix D). If corn growers using Event 3272 corn follow the measures, APHIS has

determined that these additional measu res to minimize pollen flow would decrease the likelihood of GE pollen movement into those corn fields that are growing non

- GE corn (see Gene Movement section below for further discussion and Appendix D). However, even if growers of

Event 3272 corn failed to use the mandated isolation techniques in the Stewardship Agreement, there is no indication that any current pollen flow from GE corn to non

- GE corn has dampened34 the organic production of corn. Currently, the use of GE corn varieties and the use of orga nic corn production systems are both increasing due to market demands, and these markets will likely continue to increase under the “preferred” alternative.

Under the “no action” alternative and und er the “preferred” alternative, granting nonregulated status to Event 3272 corn will not change the market demands for GE corn or corn grown for

GE

- sensitive markets. Granting nonregulated status to Event 3272 corn will add another GE corn variety to the market. However, adding GE varieties to the market is not related to the ability of organic production systems and other GE

- sensitive markets to maintain their market share. Between 2001 and 2005, although 5 GE corn varieties were granted nonregulated st atus, the acreage associated with the organic production of corn rose 35% (USGC 2006)

. Cumulative Effects: Acreage and Areas of Corn Production

Cumulative effects on U.S. acreage and U.S. areas of corn production due to granting nonregulated status to Event 3272 corn are unlikely. Neither the no action alternative nor granting nonregulated status to Event 3272 corn will directly cause an increase in agricultural acreage devoted to corn production, or those corn acres devoted to GE or non

- GE corn cultivation. Event 3272 corn will also not change cultivation areas for corn production in the

U.S. Currently 86% of all corn acreages are GE corn (USDA NASS 2010), and Event 3272 corn is not expected to increase the acreage of GE corn varieties in the U.S. There are no foreseeable changes to the availability of GE and non

- GE corn varieties on the market under either

alternative.

Cropping practices

Crop Rotation

No Action: Crop Rotation The current economics of corn production in GE and non

- GE systems are driving the change or perceived change in crop rotation practices. Growers make choices to plant certain corn varieties and use certain crop rotation practices based on factors such as yield, weed and disease pressures, cost of seed and other inputs, technology fees, human safety, potential for crop injury, and ease and flexibility of the produ ction system (Olson and Sander 1988, Giannessi 2005)

. Therefore, when taking into account these factors, growers will ultimately base their choice on

individual wants and needs.

As the demand for ethanol production and corn prices have increased, the frequency o f corn

- to

corn crop rotations conventional production systems has also increased in areas that support ethanol production (Hart 2006, Stockton et al. 2007)

. The average field corn prices have

increased from $2.00/bushel in 2005 (USDA

- NASS 2007b) , to $3.04/bushel in 2006, and $4.00/bushel in 2007 (USDA

- NASS 2008b)

. As of the end of 2010, the price for a bushel of corn is calculated at between $4.82 and $5.37 (USDA

- NASS 2011). These prices are driven by demand for corn products in ethanol and for feed. Under the “no action” alternative, the demand and price increase s in GE and non

- GE corn will continue to increase depending on the market for field corn, and corn

- to

- corn rotations will continue to be used by farmers if this cropping practice meets the economic and marketing strategy for the particular farmer.35 Pref erred Alternative : Crop Rotation As stated above, t he current economics of corn production are driving the change or perceived change in crop rotation practices. Granting nonregulated status to Event 3272 corn is unlikely to change the entire pricing sche me 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. Event 3272 corn is unlikely to affect a farmer’s decision to either stop using a corn

- to

c orn rotation, or to increase the overall use of corn

- to

- corn rotation as a cropping strategy with the U.S. farming community.

Cumulative Effects : Crop Rotation Under the “no action” alternative, the cumulative effects of using crop

- to

- crop rotation in G E

and non

- GE corn production systems could be a cumulative increase level in corn pests found in

corn fields or a cumulative increase in fertilizer use due to continuing use of corn crops (instead of rotating in a nitrogen

- fixing crop such as alfalfa or so ybean) (Sawyer 2007)

. (Synthetic

fertilizers are not used in corn production systems using organic methods.) Granting nonregulated status to Event 3272 corn will result in similar cumulative effects as those found under the “no action” alternative, because the use of corn

- to

- corn rotation is based on economic

de ci sions by the farmer and is not dependent on the corn varieties (GE or conventional) available

on the market.

Tillage No Action: Tillage The use of tillage and the removal of soil residue are considered agriculture practices that accentuate loss of soil organic carbon (Lal and Bruce 1999) . As described in Section II, this loss has negative impacts on the atmosphere and increas es soil erosion, among others. Under the “no action” alternative, the use of tillage methods in U.S. agricultural production of corn (in GE, conventional or organic production systems) will remain unchanged.

Preferred Alternative: Tillage Event 3272 corn does not change cultivation pr actices, including tillage, for corn production. Agronomic practices used during the production of corn for ethanol use is the same as the production of corn for grain. Granting nonregulated status to Event 3272 corn will not change the loss of soil organ ic carbon due to tillage in corn production systems with overall impacts

similar to the No Action alternative

.

Cumulative Effects: Tillage The cumulative effects of tillage include removal of soil residues over time (Lal and Bruce 1999, Triplett and Dick 2008) , and will continue independent of the status of Event 3272 corn. Event 3272 corn will not change the amount of corn acreage tilled, because it will not change acreage devoted to corn production in the U.S. Event 3272 corn will not change the management of corn fields because it will require the same agronomic practices as other conventional corn varieties.

Pes ticide Use

No Action: Pesticide Use

U nder the “no action” alternative, corn production , and pesticide use in corn, will remain as it is practiced today by the farming community. Growers make choices to use certain pesticides based on weed and disease pres sures, cost of seed and other inputs, technology fees, human36 safety, potential for crop injury, and ease and flexibility of the system (Olson and Sander 1988, Giannessi 2005) . Therefore, when taking into account these factors, growers will ultimately base

their ch oice on individual wants and needs. As an example of the pesticides used during the production of field corn, the Pesticide Action Network has an online database, including a detailed description of all the pesticides used in corn agriculture in Californi a (Kegley et al. 2008) . It lists the top 50 pesticides (e.g., herbicides, insecticides, fungicides) used in California corn production. Any effects due to pesticide use in the agricultural production of corn will

remain the same under the “no action” alternative. The no action alternative will also not change the types of pesticides used in organic production of corn (see http://ecfr.gpoaccess.gov/cgi/t/te xt/text

idx?c=ecfr&tpl=/ecfrbrowse/Title07/7cfr205_main_02.tpl for a list of approved synthetic pesticides for use in organic production).

Preferred Alternative : Pesticide Use Event 3272 corn production uses the same agricultural inputs ( e.g., pesticides , fertilizers) as corn currently grown for the ethanol production market. Event 3272 corn, and the alpha

- amylase produced by Event 3272 corn has no effect on the types of pesticides that will be used in corn production. Granting nonregulated status to Eve nt 3272 corn will not have any effect on the pesticides use d in the production of corn in the U.S., compared to the “no action” alternative.

Cumulative Effects: Pesticide Use The baseline effects of pesticides, and levels applied during the agricultural p roduction of corn in GE, conventional, and organic systems will not change due to granting nonregulated status of Event 3272 corn. Granting nonregulated status to Event 3272 corn does not cause additional or synergistic effects of pesticide use because th ere is no change in pesticide use or effects compared to the current conventional production of corn, and Event 3272 corn will not change the amount of pesticides used in organic production of corn.

Specialty Corn Uses and Processing Specialty corn, such as waxy corn, white corn, blue corn, high amylose corn, and organic corn, comprises 8% of the U.S. market (USGC 2006) . Event 32 72 corn is considered a specialty,

value

- added corn that will be grown for use in ethanol facilities. Because of the compositional differences of Event 3272 corn that increase the efficiency of ethanol production, and because

Event 3272 corn is different in amylase content than other corn varieties, Event 3272 corn will be grown in a closed loop system, using contractual arrangement s between growers and ethanol facilities to ensure Event 3272 corn only enters the ethanol production stream.

No Action: Specialty Corn Uses and Processing Under the no action alternative, Event 3272 corn will remain a regulated article, and will not be grown without a permit or notification from APHIS. Compositional differences between Event 3272 corn and other specialty c orn will remain, and the chance for misdirection of Event 3272 corn into other corn processing facilities will remain low.

Preferred Alternative: Specialty Corn Uses and Processing Under the preferred alternative, Event 3272 will be granted nonregulated s tatus and allowed to be grown without restrictions. However, as Event 3272 corn i s a specialty corn variety, the

closed

- loop system will be used extensively to maintain the identity and value of the product for the grower, who is obtaining a price premium from the ethanol producer, and the ethanol37 producer, who is paying the premium. The closed

- loop system will also help to prevent the misdirection of Event 3272 to unintended downstream processing facilities. There is no benefit to either party if Event 3272 corn is mistakenly routed.

However, there is still the potential for misdirection of Event 3272 corn in the transportation stream. Because Event 3272 corn has successfully completed the food and feed consultation process with FDA (Appendix H), there are no human health concerns if Event 3272 corn enters the food supply (see section on Public Health below for more discussion). Nonetheless, because of the thermostable properties of the alpha

- amylase present in Event 3272 corn, there may be undesirable effects in certain ty pes of proc essed products, similar to what may happen if other types of specialty corn products are misdirected into production streams for which they were not

intended.

For example, food processes that use alkaline cooking, such as processes that produce masa

tort illas and corn

- based snack foods, may be affected through changes in dough

- handling or darkened chip color, if Event 3272 corn is mistakenly included in the process (Appendices F

- H). The masa industry

, like other corn grain

- handling industries that depend on the specific properties of a corn variety, uses strict grain sourcing programs

. O nly an estimated 4% of the corn grain used for masa production is from the open market rather than through direct contracts with a grower (Appendix G). Corn hybrids used for masa production are specifically developed with improved alkaline

- cooking properties, and premiums are paid for growing, delivering, and meeting and maintain ing the purity and quality standards of corn for masa production (Appendix G), similar to how o ther specialty corn grain are handled. Both Event 3272 corn and corn hybrids for masa production are grown under strong stewardship programs, similar to other corn production processes that depend on specific and consistent grain qualities.

In the unlike ly event that Event 3272 corn were to enter a masa production facility (or another corn grain processing facility), the mixing of a truck

- load of Event 3272 corn into holding bin of corn for masa production would result in significant dilution (see Appendi x F and Appendix G for scenario development). Further dilution is also possible if the truck

- load of Event 3272 corn enters a local grain elevator, which would reduce the effects that Event 3272 corn would have on masa production (or the production of oth er corn grain derived products). Additionally, according to Syngenta, less than 10% of corn fields that currently support ethanol plants (areas where Event 3272 corn may be grown), are in the production area for masa

- contracted corn fields (Appendix G), f urther reducing the opportunity for misdirection of Event 3272 corn. Similar geographic separations of production area are expected for the grain inputs to other industrial facilities that process corn grain with restrictions similar to those in place for masa production.

Geographic limitations for specialty corn varieties destined for other specific facilities are also the result of transportation costs. For example, farmers under contract to grow Event 3272 would be in violation of the grower contract s if locations are too distant from an ethanol production plant that is ready to receive Event 3272

. (See discussion of Production Practices

above

. ) Furthermore, if those contracts are not in place, growers would be less likely to grow

Event 3272 corn at a great distance from an ethanol production plant, as the value of Event 3272 corn would diminish with the cost of shipping to the plant.38 As stated above, many types of specialty corn such as high amylose corn and waxy corn have different compositions t hat require the varieties to be grown, produced, transported, and processed separately from other corn varieties. Restrictive conditions for specialty corn are in place to maintain the integrity of both the varieties during production, and to maintain the integrity of the various processing facilities that produce specific end

- products for buyers. Event 3272 corn has been successfully grown, harvested, transported , and delivered to an ethanol facility while under permit. Syngenta did not report and APHIS is not aware of any misdirection to other corn proces sing facilities or any harm to those facilities.

Although a closed

- loop system and other identity preservation systems are standard operating procedures and meet current best management practices for the production of specialty corn, there is a chance that growers will fail to follow the contractual obligations and instead produce Event 3272 corn such that the identity of the grain is not preserved. The potential economic consequences of producing and misdirecting Event 3272 corn to a food or feed corn processing facility

, as opposed to an ethanol facility, are similar to when other specialty corn is not produced and shipped to the appropriate facility, such as if waxy corn were to be misdirected to a high amylose corn processing facility. There may be economic consequences of those actions. However, the magnitude of the economic effects, and the amount of corn needed to present detrimental effects, may be different for Event 3272 corn compared to oth er specialty corn

varieties.

Cumulative Effects: Specialty Corn and Processing Under the no action and preferred alternative, specialty corn will be grown, harvested

, and transported to specialized processing facilities. The mechanisms currently in place to protect the identity of specialty corn varieties, including Event 3272 corn, will not be affected by granting nonregulated status to Event 3272 corn. There are no cumulative effects to the natural and physical environment through the use of another specialty corn crop such as Event 3272 corn. Planting, harvesting, and transporting of Event 3272 corn will occur under rigid closed loop systems currently used for oth er specialty corn varieties. The potential economic consequences of producing and misdirecting Event 3272 corn to a food or feed corn processing facility

, as opposed to an intended corn ethanol facility, are similar to when other specialty corn is shipped to the incorrect facility . Specialty corn production, and corn production in the U.S. i n general, is currently successful and thrives, even with the understanding of economic risks of sending the wrong corn variety to the wrong place. Event 3272 corn is another specialty corn variety that will be planted, grown, harvested, and transported off the farm using a closed

- loop system, and any cumulative effects to specialty corn and processing of corn is minimal.



### Ethanol Production

Ethanol production is an i ntegral part of meeting the renewable fuels standard (RFS) in the U.S. The Federal Energy Policy Act of 2005, signed on August 8, 2005, includes a Renewable Fuels Standard that directs the doubling of the use of ethanol and biodiesel in the U.S. fuel sup ply by 2012 to 7.5 billion gallons (42 USC 15801, page 1069). The Energy Independence and Security Act of 2007 passed in December 2007, includes a provision to expand consumption of alternative fuels, including but not limited to ethanol, to 36 billion gal lons in 2022. With at least

1 94 corn ethanol plants in operation in 2 8 different states, corn

- based ethanol production may be a feasible way to meet the ethanol consumption benchmark for 2012 set in the Energy Policy Act39 of 2005 .Based on the production ou tput of ethanol production refineries operating at the end of 2010, more than 98% of ethanol produced in the U.S. is made from corn ( http://www.ethanolrfa.org , Accessed February 7, 2011). According to the U.S. Energ y Information Administration, ethanol production is projected to increase by 50,000 barrels per day (bbl/d) to 910,000 bbl/d in 2011 and then grow by an additional 10,000 bbl/d in 2012. ( htt p://www.eia.doe.gov/emeu/steo/pub/contents.html , Accessed February 9, 2011).

No Action: Ethanol Production Under the “no action” alternative, the process used for ethanol production would remain the

same. Efficiency of ethanol production would remain at 2.8 gallons/bushel of corn. Corn

- based ethanol production would still be used as one method to meet the benchmarks set by Congress in terms of biofuel consumption in the U.S

.

Preferred Alternative: Ethanol Production

The use of Event 3272 corn in ethan ol facilities that agree to receive Event 3272 corn could result in changes in inputs, methods, and capital costs during ethanol production. Each of the potential changes is due to the substitution of Event 3272 corn grain, containing a thermostable alpha

- amylase, for microbial alpha

- amylase in the liquefaction stage of ethanol production. Appendix C contains a report submitted on behalf of Syngenta, and reviewed by APHIS, that outlines these changes in detail , and what is required of ethanol plants to ef ficiently and effectively use Event 3272 corn, including potential re

- design of ethanol facilities. A brief summary of the potential changes discussed in the report are identified as issues associated with Public Health, A nimal Feed and DDGS, and Water Us e , which are discussed below.

In particular, cooking temperatures may be lowered during the liquefaction stage with Event 3272 corn ethanol production, which potentially may result in decreases in energy usage, and has the added benefit of increasing etha nol yields. Ultimately, Syngenta believes that the changes associated with using Event 3272 corn in ethanol processing, instead of adding microbial alpha

amylase during the process, will result in a 2% or greater increase in ethanol efficiency per bushel of corn. However, if the efficiency projected by Syngenta is not met, then ethanol produ ction would likely remain at 2.8 gallons/bushel of corn, as in the “no action” alternative.

Cumulative Effects: Ethanol Production

Under both the “no action” alternati ve and the “preferred” alternative, expansion of ethanol production is expected to occur to meet renewable fuel standards. The increase in production is expected to be similar under both alternatives. There are no other petitions submitted for nonregulated status for GE organisms related to ethanol production. If granted nonregulated
status, Event 3272 w ould be the only GE corn variety available specifically for ethanol production , and Event 3272 corn is likely limited to those ethanol plants that are contracted to

receive Event 3272 corn.



### Public Hea l th

Human Health Under FFDCA, it is the responsibility of food and feed manufacturers to ensure that the products they market are safe and properly labeled. Food and feed derived from Event 3272 corn mus t be40 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.

Syngenta provided the FDA with information on identity, f unction, and characterization of the genes, as well the expression levels of the gene products. They also provided information on the potential allergenicity and toxicity of the expressed proteins. The FDA considers Syngenta's consultation on alpha

- amylas e Event 3272 corn to be complete (Appendix H). Syngenta also submitted information on identity, function, characterization of genes, expression levels of gene products, as well as information on the potential allergenicity and toxicity of the expressed

pr oteins to APHIS. APHIS’ assessment of the safety of this product for humans and animals focuses on plant pest risk (USDA

- APHIS 2009) and the effects on wildlife and threatened and endangered species (see section on Animals and Threatened and Endangered Species, pages 50

- 52), and those analyses are based on the comparison of the GE

- corn to its non

- GE counterpart.

No Action: Human Health Syngenta has successfully completed the consultation process with the FDA for Event 3272 corn (Appendix H ). The status of the FDA consultation will not change under the “no action”

alternative. Other countries have also found Event 3272 corn safe for food and feed, including

Aus tralia

, Canada, Mexico

, Russia, and the Philippines (see http://cera

gmc.org/index.php?action=gm_crop_database&mode=Submit&evidcode=Event%203272 , Date Accessed: February 7 , 20 11 )

.

Preferred Alternative: Human Health

APHIS evaluated risks to human and other animal health that may result from the consumption and exposure of Event 3272 corn. Risk is a combination of hazard and exposure. APHIS first

conducted hazard identification for Event 3272 corn. APHIS assessed th e composition and nutritional quality of Event 3272 corn, and compared the composition of Event 3272 to the composition of a non

- genetically engineered control corn li ne and the natural variation found in commercial corn varieties. If the composition of E vent 3272 corn is similar to other commercial corn plants, it is unlikely that Event 3272 poses an additional health hazard. If no hazards are identified, then the risk of Event 3272 corn causing harm is unlikely, regardless of exposure

route. APHIS also assessed the exposure data presented by Syngenta to further evaluate any possible risk posed by Event 3272 corn to human or animal health.

Event 3272 corn is genetically engineered to produce a thermostable alpha

- amylase. Additionally, alpha

- amylases ar e ubiquitous enzymes found in microorganisms, plants, and

animals (Janeček et al. 1999) to which humans and other animals are frequently exposed. These enzymes play a key role in ma ny cellular metabolic processes by catalyzing the breakdown of starch, such as in germinating seed of cereal plants (Yu et al. 1996) or in the microbial decomposition of organic matter to provide carbon and energy for microbial growth

processes (Rothstein et al. 1986)

. Many types of commercial food processing, feed ingredient applications, and industrial applications also utilize alpha

- amylase enzymes, including the production of fuel and potable alcohol (brewing, distillation processes), and corn syru ps (Janeček et al. 1999, Lévêque et al. 2000, Pariza and Johnson 2001, Olempska

- Beer et al. 2006)

.41 The data presented in the petition suggests there is no difference in compositional and nutritional quality of Event 3272 corn compared to conventional corn, ap art from the presence of AMY797E and PMI. Although some of the variables measured by the applicant showed statistically significant differences between Event 3272 corn and the nontransgenic hybrid controls (Tables 6

- 1 to 6

- 6, pages 70

- 76, of the petition) , none of the values for the forage and grain composition characteristics were outside the range of natural variability of conventional corn as found in the International Life Sciences Institute Crop Composition Database (OECD 2003, Ridley et al. 2004, ILSI 2006) or in t he OECD consensus document on corn (OECD 2003) composition. Event 3272 corn does not express additional proteins, natural toxicants, allelochemicals, pheromones, hormones, etc. that could directly or indirectly affect humans or other animals. Thus, the composition of Event 3272 is not biologically d ifferent than

conventional corn.

Given that the composition of Event 3272 corn was found to be consistent with the natural variation found in conventional corn varieties, the applicant conducted studies to confirm the

absence of deleterious effects for an imals when feeding on Event 3272 corn. AMY797E is predominantly found in the corn kernels of Event 3272 plants (1627 μg/g fresh weight in the dough stage), with minute amounts found in the roots during the whole stage (<0.1 μg/g fresh weight) and in the l eaves during senescence (<1 μg/g fresh weight) (Table 3

- 3 of petition). Animals that feed primarily on corn kernels are seed

- feeding insects and rodents found in agricultural fields. During field trials, the applicant found no changes in insect feeding d amage (Table 5

- 7 of petition), indicating similar insect susceptibility for Event 3272 corn compared to

conventional corn.

Rodents, such as mice or squirrels, may seasonally feed exclusively on corn kernels. Accordingly , these animals are most likely to have a diet containing large amounts of corn kernels. Using dietary calculations (pages 93

- 94 of petition) along with the results of a mouse toxicity study (page 50 of petition and page 6 of the response letter from Syngenta), the applicant

determined tha t the No Observed Effect D ose (NOED) of AMY797E was greater than 2 times the maximum amount of alpha

- amylase corn a rodent might consume daily (page 94 of petition). Additionally, the applicant also evaluated the effects of AMY797E on birds by feeding bro iler chickens diets that contained up to 65% of Event 3272 grain for 49 days (page 94 of petition and page 6 of response letter). The applicant reported no harmful effects to chickens from a diet extremely high in AMY797E. Humans and feed animals would ha ve a significantly lower exposure to AMY797E because of feeding habits and diverse food sources.

AMY797E also does not have any significant sequence identity to any known toxins (page 7 of response letter from Syngenta). AMY797E is functionally similar t o thermostable alpha

amylases already safely used in food and feed processing (Janeček et al. 1999, Lév êque et al

. 2000, Pariza and Johnson 2001, Olempska

- Beer et al. 2006) , indicating that Event 3272 corn is unlikely to produce toxins that would negatively a ff ect humans or other animals that may eat corn kernels or plants containing AMY797E.

Concerns regarding the allergenicity of Event 3272 corn were raised by opponents of genetic engineering (Freese and Mayet 2006) centered around two components: 1) allergenicity of fungal

- derived alpha

- amylases and 2) thermostability of alpha

- amylases.42 Fungal

- derived alpha

- amylases are one of the occupational allergens in the bakery industry (Houba et al. 1996, Houba et al. 1997) . Syngenta assessed the potential allergenicity of the AMY797E alpha

- amylase and phosphomannose isomera se ( PMI ) proteins by searching for amino acid homology between these proteins and known allergen protein sequences. These searches were conducted using a database comprised of identified or putative allergen sequences from publicly available databases (Ge nPept, PIR, SWISS

- PROT, FAARP and IUIS) and additional putative allergen sequences from the scientific literature. Syngenta reported that for alpha

- amylase in Event 3272 corn, there were no significant similarities between the alpha

amylase found in Event 3272 corn and other allergens. There was one small section of the enzyme that corresponded with an allergen from an insect (American cockroach), but Syngenta maintains that this sequence identity is not biologically relevant and has no implication for th e allergenic potential of the AMY797E alpha

- amylase. AMY797E is a thermostable protein, which may be a consideration of what compounds are allergenic (Sampson 1999, Taylor and Hefle 2001) . However Syngenta conducted digestibility experiments and found that AMY797E is not stable to digestion (Appendix H), and concluded that the AMY797E is un likely to become allergenic. The FDA did not question the conclusion that AMY797E and PMI are not likely to become allergenic (Appendix H). APHIS has concluded that these studies are sufficient and agrees with the findings of the FDA.

Organisms exposed t o AMY797E will also be exposed to PMI. The pmi ( manA ) gene comes

from E. coli and encodes the PMI enzym e

. Pmi serves as a marker gene that enables selection of lines that are genetically modified, providing the plant with the ability to utilize mannose a s a sole carbon source. Table 3

- 4 of the petition gives the PMI expression data for Event 3272 corn. PMI expression was examined during 5 developmental stages of Event 3272 corn (whorl, anthesis, kernel dough, kernel maturity, and senescence). In Event 3272 corn, PMI is expressed in leaves during all stages except senescence, with maximum expression during the kernel dough stage (5.7 μg/g fresh weight). Expression of PMI in the roots occurs during all developmental stages with the highest expression of 1.0 μg/g fresh weight during the whorl stage. The highest PMI expression for kernels occurred during the kernel dough stage (0.8 μg/g fresh weight), and 8.5 μg/g fresh weight of PMI was expressed in the pollen. The expression of PMI protein in corn plant s is not expected to have deleterious effects or significant impacts on humans or other animals, based on the data provided in the petition. Dietary calculations to determine the daily dietary dose of PMI (page 93 of petition) and data from the mouse and bird toxicity studies (page 94 of petition) indicate that PMI levels in Event 3272 corn do not cause harm.

Additionally, the EPA has granted an exemption from the requirement of a tolerance for the PMI protein as an inert ingredient in all plants (69 FR 26770

- 26775). The DNA encoding the PMI protein is not toxic. At the 80

- amino acid peptide level, the PMI protein shares no significant homology with proteins known to be toxic or allergenic. Within one of the 80

- amino acid windows, there was one region of sequence homology of eight contiguous amino

- acids between PMI and a known allergen, α

- parvalbumin from a Rana (frog) species. Further testing found no

cross

- reactivity between PMI and the human serum immunoglobulin E (IgE) and Bovine Serum Albumin (BSA ), indicating that the low degree of sequence identity between the PMI used in

Event 3272 corn and α

- parvalbumin from Rana sp. is not biologically relevant.

Based on the assessment of laboratory data provided by Syng enta in the submitted petition and an analysis of the scientific literature (USDA

- APHIS 200 9 ) , along with the completion of the43 consultation process with FDA regar ding Event 3272 corn (Appendix H ), and consideration that

other countries have also found Event 3272 safe for food and feed use (Table 1)

, APHIS has concluded that under this alternative, the proposed action to grant nonregulated status to Event 3272 corn would have no significant impacts on human or animal health.

Cumulative Effects: Human Health There are no significant impacts on human or animal health related to the no action alternative or granting nonregulated status to Event 3272 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 “n o action” alternative, exposure to these agricultural chemicals during corn production would remain the same.

Ethanol processing of corn grain into ethanol involves the use of hazardous chemicals. In standard ethanol production, pH is adjusted at sever al stages to maintain efficient processing using sulfuric acid and liquid ammonia during different ethanol processing steps for pH adjustment. Under the “no action” alternative, exposure to these ethanol processing chemicals

would remain the same.

Prefer red Alternative: Worker Safety Worker safety issues related to the use of pesticides during agricultural production of Event 3272

corn would remain the same as the “no action” alternative. As discussed under the issue of “Pesticide Use”, Event 3272 corn d oes not change the agronomic practices , or use of chemicals such as pesticides, associated with corn production.

The use of Event 3272 corn in ethanol production cou ld result in changes in inputs and methods during ethanol production in those ethanol facilities that receive Event 3272 corn . Each of the potential changes is due to the substitution of Event 3272 corn grain, containing a thermostable alpha

- amylase, for microbial alpha

- amylase in the liquefaction stage of ethanol product ion. Appendix C contains a report submitted on behalf of Syngenta , and reviewed by APHIS, that outlines these changes in detail. A brief summary of the potential changes suggested to worker safety is provided below.

Using Event 3272 corn in the ethanol production process, instead of standard field corn with microbial alpha

- amylase added, may eliminate constant adjustment of pH during processing. In standard ethanol production, pH is adjusted at several stages to maintain efficient processing.

With the use of Event 3272 corn, and the potential consistency in pH during ethanol production

, the use of sulfuric acid may be reduced by half because less sulfuric acid is needed to maintain pH. Urea may also be used as a substitute for liquid ammonia for additi onal pH balancing, because of the pH properties of ethanol processing due to the use of Event 3272 corn. Thus, the possible replacement and reduction of these hazardous chemicals (sulfuric acid and liquid ammonia) would reduce both environmental and workpl ace safety exposure

.44 Cumulative Effects: Worker Safety Worker safety issues related to the use of pesticides during agricultural production of corn would

continue and remain the same under both alternative s . As discussed under the issue of “Pesticide Use”, Event 3272 corn does not change the agronomic practices, or use of chemicals such as pesticides, associated with corn production. Worker safety issues related to ethanol production

would continue to occur under both alternatives. The use of Event 3272 would reduce worker exposure to some hazardous chemicals used in ethanol production. There are no cumulative

effects identified for this issue.

Animal Feed

- DDGS

Dried d istillers grains with solubles (DDGS) is a co

- prod uct of ethanol production, and is mainly used domestically as feed for dairy cattle, but also for hogs, swines, broilers and turkeys. As a result of increased ethanol production, the quantity of distillers grains marketed for use in animal

feed has increa sed from 1.89 million metric tons in 1999 to 8.35 million metric tons in 2005 (a 340 percent increase) and then more than tripled again to over 34 million metric tons in 2010 (Hoffman and Baker, 2010).

Approximately 90% of the distillers grains produced in U.S. facilities are used in domestic

animal feed. Currently, ethanol plants do not discriminate between GE and non

- GE corn. Given that 80% of corn production is GE, the DDGS produced and used as anim al feed are likely produced with some GE corn varieties.

Because of the near complete fermentation of starch during the ethanol process, the remaining amino acids, fat, minerals and vitamins in DDGS increase approximately three

- fold in

concent ration compared to levels found in corn (R ausch and Belyea 2006) . This can result in concerns regarding phosphorous and animal waste dispose, sulfur diet content, and mycotoxin

concentrations in DDGS.

No Action: Animal Feed

- DDGS Under the “no action” alternative, DDGS will still be produced during ethanol production.

Concerns will still surround DDGS nutritional content in terms of increased phosphorus

concentration and the disposal of the resulting a nimal waste, high sulfur diet content, and

increased mycotoxin concentrations as discussed i n Section II

. The successful completion of the

consultation w ith FDA for Event 3272 corn (Appendix H ) will not change under the “no action”

alternative.

Preferred Alternative: Animal Feed

- DDGS DDGS are currently made with GE corn, so the use of a new G E corn variety in ethanol production will not change the availability of GE DDGS as an animal feed. Syngenta has completed the consultation process on Even t 3272 corn with FDA (Appendix H ), and has

concluded that Event 3272 corn is considered safe for use in food and feed. Based on this FDA consultation, APHIS supports this conclusion.

Overall, the use of Event 3272 corn is not anticipated to change the composition of DDGS. Ethanol production e xperiments at a laboratory scale conducted with Event 327 2 corn found no difference in DDGS produced with conventional corn and with Event 3272 corn (Singh et al. 2006) . However, the experiments were run using only 3% of Event 3272 corn during the ethanol45 processing, and no comparisons of DDGS c omposition were conducted using 100% Event 3272

corn.

The use of Event 3272 corn in ethanol production is not anticipated to change the phosphorous or mycotoxin concerns surrounding the use of DDGS as animal feed , as discussed in Section II

. However, if the use of Event 3272 corn in ethanol production decreased the use of sulfuric acid, the resultant DDGS may have a reduced sulfur content. This reduction in sulfur content in DDGS is a potential benefit as DDGS produced using the current ethanol produ ction process could have a sulfur content that results in toxicity to animals (Rausch and Belyea 2006)

.

Under the “ preferred ” alternative, Event 3272 corn would be available for ethanol processing, and has the potential to alleviate some of the concerns surrounding sulfur content in DDGS, an ethanol by

- product used for animal feed. If the reductions in sulfuric acid during ethanol processing due to Event 3272 corn are not realized impacts regarding sulfur content would be

similar to the no action alternative. Overall, the use of Event 3272 corn would still be considered safe for food and feed use, including DDGS (Appendix H )

.

Cumulative Effects: Animal Feed

- DDGS DDGS are currently made with GE corn and this production is expected to continue. T he use of a new GE corn variety in ethanol p roduction will not change the availability of GE DDGS as an

animal feed. Syngenta has completed the consultation process on Even t 3272 corn with FDA (Appendix H ), and Event 3272 corn is considered safe for use in food and feed. There are no

cumulative effects identified for this issue.



### Gene Movement

G ene flow is the transfer of genetic information between different individuals and/or populations. Pollen flow, or the movement of genes from one plant to another, occurs between plants that are sexually

- c ompatible, or able to receive pollen at the appropriate time during the appropriate plant stage . Corn does not have sexually

- compatible relatives found in ‘natural’ area; corn is only able to reproduce with other corn plants in the U.S. (USDA

- APHIS 200 9 )

.

Corn pollen moves by the wind to other corn fields that are nearby. Successful gene movement from one plant to another requires many different biological an d physical factors, such as timing of flowering between corn fields, viability of pollen, an d presence of physical barriers; consequently pollen movement is not equivalent to gene movement

. A recent paper (Sanvido 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. 164 f ee t), and this result was validated when analyzing cross

- fertilization events in large scale studies (Henry et al. 2003, Weber 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 almost all other gene flow studies in corn may be due to the type of corn used in this study. Jones and Brooks (1950) investigated the appropriate isolation distance for seed production in open

- pollinated varieties, and not for hybrid varieties. Due the biology of open

- pollinated varieties, these types46 of plants may be more receptive to pollen over a longer period of time than hybrid corn plants (Sanvido et al. 2008) , allowing for a greater chance of pollination events. Therefore, the results from Jones and Brooks (1950) may be an overestimation of cross

- fertilization potential for hybrid corn plants.

No Action: Gene Movement Under the “no action” alternative, Event 3272 corn would remain a regulated article and would require a pe rmit or notification for release into the environment. Under regulated releases, GE corn is typically separated from non

- regulated corn by a distance of 660ft , based on distances 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 limit gene movement from occurring outside the release site.

Preferred Alternative: Gene Movement In 2008, GE corn production was planted on 80% of all corn acres currently in production in the US, and the use of GE corn has been increasing over the last 3 years (USDA

- NASS 2006, 2007a, 2008a, 2010b)

. Concurrently, organic corn acreage is also increasing at approximately 30% a

year (USDA

- ERS 2008a) , even though there are no requirements that GE corn currently in production use mandated techniques for separation between the two types of corn varieties. There are also no mandated separation distances betwe en other corn types. Syngenta has included some mandatory measures in their Stewardship Agreement, including measures to minimize pollen movement outside corn fields containing Event 3272 corn.

Best management guidelines for Event 3272 corn (Appendix D and Appendix G) require the use

of 12 border rows of non

- Event 3272 corn to reduce the likelihood of gene movement between

Event 3272 corn and other corn fields. The border rows of non

- Event 3272 corn are used as a ‘pollen trap.’ Corn pollen is heavy and most pollen does not travel far from the corn field (Sanvido et al. 2008) . When pollen does move from Event 3272 corn, the border rows of corn will ‘catch’ the pollen in a ‘trap,’ an d prevent the movement of pollen, and subsequently genes, outside of the field of Event 3272 corn. Additionally, Event 3272 corn is being grown as a specialty corn crop; maintaining the purity of this corn v ariety is also important. Hence, t o a lesser de gree, the border rows also act as a ‘pollen trap’ to prevent other corn pollen from entering the field of Event 3272 corn. The use of border rows may result in a reduction of up to more than 99.9% of Event 3272 corn pollen leaving the corn field (Appendix D).

In agricultural systems, growers may choose to grow GE or non

- GE corn, and obtain price premiums for growing particular varieties of corn for particular markets. In 2008, conventional field corn averaged $3.90/bushel (USDA

- NASS, 2009b) , whereas organic corn averaged $7.08/bushel (USDA

- NASS 2010) . USDA asserts that agricultural practices th at use conventional means, organic production systems, or genetically engineered varieties can all provide benefits to the environment, consumers, and farm income. Gene movement into and out of these specialized corn production systems have been managed u sing various types of buffer zones or isolation practices, such as differences in planting (which results in differences in flowering) or making sure fields are distance from other compatible crops (such as using isolation distances).47 For example, beside s the typical identity preservation and closed loop systems in place for specialty crops, those farmers using organic production also put in place measures to maintain purity for their crops. Typically, more than one method is used by farmers using organi c methods to prevent unwanted material from entering their fields including; isolation of the farm, physical barriers or buffer zones between organic production and non

- organic production, as well as formal communications between neighboring farms (NCAT 2003) . The plan used as the basis for organic certification sh ould include 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 transporting the crop (Riddle 2004, Krueger 2007, Kuepper et al. 2007) . Plans for organic systems also should include of how the risk of

GMO contamination will be monitored (Kuepper et al. 2007) . F armers 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 farmer reduce contamination events (NCAT 2003, Krueger 2007) . Commonly used production practices for corn and the practical methods

ty pically used by corn farmers who use organic methods, or those that grown specialty crops, to protect their crop and maximize their profits and premiums granted to corn produced using approved plans currently provide many measures that will greatly reduce the likelihood of accidental gene flow between Event 3272 corn and other corn fields.

As more and more corn acreage is using GE varieties, corn production for GE

- sensitive markets has increased; for example corn produced using organic methods increased 35% between 2001

and 2005 (USGC 2006) , at the same time that cor n acreage using GE varieties increased by 50% (USDA

- NASS 2001, 2005) . During this time, there was no mandated use of separation distance

or other measures to mi nimize gene movement between corn fields, except for those measures taken by farmers who use organic production practices. No acreage data for corn production under organic methods is available since 2005, thus no further trend comparisons were

conducted.

Based on the 2005 information, if growers of Event 3272 corn fail to use the separation techniques mandated in their contract with Syngenta, there is no indication that any pollen flow

from GE corn to non

- GE corn would dampen the organic production of corn. Event 3272 corn

wil l be one of many GE varieties already in the marketplace, and GE corn already accounts for 86% of corn planted in the U.S . Event 3272 corn will not increase the acreage of GE corn planted in the U.S. There are no new procedures or requirements that growers of non

- GE

varieties may need to implement as baseline market and economic conditions are already in place to account for GE varieties in U.S. corn production. Acreage using GE varieties has increased by 40% since 2005, and the re is no corresponding data for acres of corn production under organic methods. Overall impacts of gene movement would be similar to the no action

alternative. The pollen flow prevention measures in place for Event 3272 corn, along with geographic isolat ion and other factors that will be part of the closed

- loop produc tion system for

Event 3272 corn will also minimize gene flow to other conventional and specialty corn varieties (See Production Practices above).

Cumulative Effect: Gene Movement

Event 3272 corn would be an additional GE and specialty corn variety that may be available to the farming community. Curren tly, GE corn is approximately 86 % of all acreage of corn production in the U.S. The production area for Event 3272 corn will likely be limite d to areas in48 states that currently have ethanol production facilities because this GE corn variety has been specifically developed for that purpose. Because GE corn is currently used in ethanol production, and Event 3272 corn will not change the amount o f GE corn produced in the U.S., the availability of Event 3272 corn will not result in cumulative effects on gene movement. Event 3272 corn does not change the amount of GE pollen in the environment.



### Water Use in Ethanol Production
Ethanol production is an integral part of meeting the renewable fuels standard (RFS) in the U.S. Almost all (more than 98%) of U.S. ethanol is made from corn ( http://www.ethanolrfa.org

, Accessed February 7, 2011). Ethanol production t ypically uses 4.7 gallons of water to produce 1 gallon of ethanol (Shapouri and Gallagher 2005) . Water is also used during the manufacturing and transport of microbial alpha

- amylase for conventional ethanol production.

No Action: Water Use in Ethanol Production Under the “no action” alternative, there is no change in the conventional processes used for ethanol production. Water would still be used during the manufacturing and transport of microbial alpha

- amylase for conventional ethanol production, and approxima tely 4.7 gallons of

water would be used to produce 1 gallon of ethanol.

Preferred Alternative: Water Use in Ethanol Production

The use of Event 3272 corn could result in the water cons ervation in two ways. There is a potential for a decrease in w ater usa ge during ethanol production during the saccharification phase due to the properties of Event 3272 corn (Appendix C ) . However, large

- scale ethanol plant research and development have not verified this potential savings. T he use of Event 3272

corn may res ult in the same water us e in ethanol production as now [ 4.7 gallons of water for 1 gallon of ethanol, (Shapouri and Gallagher 2005) ]

.

Currently, water is intensively used during the production of microbial alpha

- amylase and microbial alpha

- amylase is also transported to the ethanol facility in a water solution. If Event

3272 corn was used in ethanol production, water usage could be reduced because microbial alpha

- amylase would not be needed in ethanol production due to the replacement by the alpha

amylase in Event 3272 corn.

Cumulative Effect: Water Use in Ethanol Production

Water is cur rently used in the production of ethanol and this practice will continue with the availability of Event 3272 corn. Ethanol production typically uses 4.7 gallons of water to produce 1 gallon of ethanol (Shapouri and Gallagher 2005) . Water is also used during the manufacturing and transport of microbial alpha

- amylase for conventional ethanol production. Granting nonregulated status to Event 3272 corn may potentially decrease water use. However,

no cumulative effects have been identified for this issue.



### Animal and Plant Communities

Animals Corn production systems in agriculture are host many animal species. Mammals and birds may seasonally use grain, and invertebrates can feed on the plant during the entire growing season.49 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 releases of Event 3272 corn would be under regulation. Animal incursions are limited during regulated field trials, but may occur. However,

t he consultation with FDA has been succ essfully completed for Event 3272 corn (Appendix H ), which addressed any concerns of composition, as well as demonstrated a lack of toxicity and allergenicity, of Event 3272 corn for human and animal consumption. Based upon the FDA consultation, APHIS sup ports Syngenta’s conclusions that Event 3272 corn is considered safe for animal consumption.

Preferred Alternative: Animals

APHIS has reviewed and accepts the data submitted by the applicant that confirmed the absence

of deleterious effects for wildlife when feeding on Event 3272 corn. The data are similar to what was submitted during the FDA consultation process for Event 3272 corn (Appendix H )

. Agronomic practices used to produce Event 3272 corn will be the same as those used to produc e conventionally grown corn, and the effects discussion for Event 3272 corn on animals will focus solely on the effects of the introduced proteins in Event 3272 corn, the alpha

- amylase enzyme and the phosphomannose enzyme (used as a selectable marker for Event 3272 corn).

Event 3272 corn contains the alpha

- amylase protein, AMY797E, which is found predominantly

in the corn kernels of Event 3272 corn plants (1627 μg/g fresh weight in the dough stage), with minute amounts found in the roots during the whole stage (<0.1 μg/g fresh weight) and in the leaves during senescence (<1 μg/g fresh weight) (Table 3

- 3 of petition). Animals that feed primarily on corn kernels are seed

- feeding insects and rodents found in agricultural fields. During field trials, the applicant found no c hanges in insect feeding damage (Table 5

- 7 of petition), indicating similar insect susceptibility for Event 3272 corn compared to conventional

corn. Event 3272 corn has not been genetically engineered to produce any pesticides.

Rodents, such as mice or s quirrels, may seasonally feed exclusively on corn kernels. Accordingly , these animals are most likely to have a diet containing large amounts of corn kernels. Using the dietary calculations (pages 93

- 94 of petition) along with the results of a

mouse toxi city study (page 50 of petition and page 6 of response letter), the applicant determined that the no observed effect dose (NOED) of AMY797E was greater than 2 times the maximum amount of alpha

- amylase corn a rodent might consume daily (page 94 of petition) . Other mammals, such as deer, would have even lower exposure to AMY797E because of feeding habits; for example, deer nibble on tips of corn ears as opposed to kernels (Steffey et al. 1999)

. APHIS has reviewed this information and has determined that there would be no negative effects to mammals that forage in Event 3272 corn.

The applicant also evaluated the effects of AMY797E on birds by feeding broiler chickens diets that contained up to 65% of Event 3272 grain for 49 days (page 94 of petition and page 6 of response letter). The applicant reported no harmful effects to chickens from a diet extremely high in AMY797E. Moreover, given that diets high in AMY797E do not result in harmful effects, the diets of wild birds that occasionally forage in corn fields ar e unlikely to contain high amounts of AMY797E for 49 consecutive days as corn availability is limited by seed50 germination and harvest. APHIS has reviewed this information and has determined that there would be no negative effects to birds that forage in E vent 3272 corn.

Additionally, animals have been previously exposed to alpha

- amylases in the environment, as they are ubiquitous enzymes found in microorganisms, plants, and animals (Jan eček et al. 1999)

. AMY797E also does not have any significant sequence identity to any known toxins (page 7 of response letter). As stated earlier, AMY797E is functionally similar to thermostable alpha

amylases already safely used in food and feed pro cessing (Janeček et al. 1999, Lév êque et al. 2000, Pariza and Johnson 2001, Olempska

- Beer et al. 2006) , indicating that Event 3272 corn is unlikely to produce toxins that would negatively affect animals that may eat corn kernels or plants containing AMY797E.

Organisms exposed to the AMY797E protein in Event 3272 corn will also be exposed to PMI , a protein used by the applicant to select genetically engineered plants during breeding . Additional species, such as leaf

- feeding animals, butterflies and bees, may not be exposed to AMY797E but will be exposed to PMI because of its expression in vegetative tissue and pollen (Table 3

- 4 of petition). The pmi ( manA ) gene comes from E. coli and encodes the enzyme phosphomannose isomerase (PMI). Pmi serves as a marker g ene that enables selection of lines that are genetically engineered , providing the plant with the ability to utilize mannose as a sole carbon source. Table

3

- 4 of the petition gives the PMI expression data for Event 3272 corn. PMI expression was

examined during 5 developmental stages of Event 3272 corn (whorl, anthesis, kernel dough, kernel maturity, and senescence). In Event 3272 corn, PMI is expressed in leaves during all stages except senescence, with maximum expression during the kernel dough stage ( 5.7 μg/g fresh weight). Expression of PMI in the roots occurs during all developmental stages with the highest expression of 1.0 μg/g fresh weight during the whorl stage. The highest PMI expression for kernels occurred during the kernel dough stage (0.8 μg/g fresh weight), and 8.5 μg/g fresh weight of PMI was expressed in the pollen. The expression of PMI protein in corn plants is not expected to have deleterious effects or significant impacts on non

- target organisms or TES organisms, based on the data p rovided in the petition. Dietary calculations to determine the daily dietary dose of PMI (page 93 of petition) and data from the mouse and bird toxicity studies (page 94 of petition) indicate that PMI levels in Event 3272 corn do not cause harm in wildlif e populations, including threatened and endangered species.

Additionally, the EPA has granted an exemption from the requirement of a tolerance for the PMI protein as an inert ingredient in all plants (69 FR 26770

- 26775). The DNA encoding the PMI protein is not toxic. At the 80

- amino acid peptide level, the PMI protein shares no significant homology with proteins known to be toxic or allergenic. Within one of the 80

- amino acid windows, there was one region of sequence homology of eight contiguous amino

- a cids between PMI and a known allergen, α

- parvalbumin from a Rana (frog) species. Further testing found no

cross

- reactivity between PMI and the human serum immunoglobulin E (IgE) and Bovine Serum Albumin (BSA), indicating that the low degree of sequence id entity between the PMI used in

Event 3272 corn and α

- parvalbumin from Rana sp. is not biologically relevant.

Plants Corn production systems in agriculture are host many plant species as well. The landscape surrounding a corn field varies depending on the region. In certain areas, corn fields may be bordered by other corn (or any other crop); fields may also be surrounded by wooden and/or51 pasture/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 dwell in and around corn fields; those species will also vary depending on the region where the corn is planted. Cor n itself is not sexually compatible with any other plant species found in the U.S. (USDA

- APHIS 200 9 )

. The cumulative effects analysis for this issue is found bel ow at “Cumulative Effects: Plants, Animals, Biodiversity.”

No Action: Plants Under the “no action” alternative, environmental releases of Event 3272 corn would be under regulation. Plant species that typically inhabit corn production systems will be m anaged as in conventional corn production, including the use of mechanical, cultural, and chemical control

methods.

Preferred Alternative: Plants If Event 3272 corn was granted nonregulated status, agricultural practices used for conventional

corn would be used for plant management during the cultivation of Event 3272 corn. Impacts

would be similar to the no action alternative. Event 3272 corn does not exhibit characteristics associated with weedy growth and will not compete with plants found outside of agricultural production. Weeds within fields of Event 3272 corn will be managed using mechanical, cultural, and chemical control, as weeds are now managed in conventional corn systems. As there are no toxic effects on animals (see Animals discussion abo ve), there are no toxic effects on animals that could be pollinators of other plants in or around fields cultivated with Event 3272 corn. Event 3272 corn 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 may have even been bred fo r a particular purpose. In the case of corn agriculture, corn varieties have been developed for food processing needs (e.g., waxy corn), varietal development (e.g., blue corn or white corn), or for use as a vegetable (e.g. sweet corn). In conventional ag riculture, farmers want to encourage 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, particularly insect and other pest species will also be managed through chemical and cultural controls to protect the crop from destruction by animals

. Therefore, the biological diversity in agricultural systems (the agro

- ecosystem) is highly managed and may be lower than i n the surrounding habita ts. 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, environmental releases of Event 3272 corn would be under regulation. Animal and plant species that typically inhabit corn production systems will be managed as in conventional corn production, including the use of mechan ical, cultural, and

chemical control methods.

Preferred Alternative: Biological Diversity Under the “preferred” alternative c ultivation of Event 3272 corn requires the same agronomic practices as conventional corn production. Animal and plant species t hat typically inhabit corn52 production systems will be managed as in conventional corn production, likely with the use of mechanical, cultural, and chemical control methods. Overall impacts would be similar to the no

action alternative.

Cumulative Effect s: Animals, Plants, Biodiversity

Event 3272 corn 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 et al. 2002, Brooks et al. 2003, Haughton et al. 2003, Hawes et al. 2003, Marshall et al. 2003, Roy et al. 2003, Romeis et al. 2004, Sisterson et al. 2004, Bitzer et al. 2005, Pilcher et al. 2005, Torres and Ruberson 2005, Romeis et al. 2006, Marvier et al. 2007, Chen et al. 2008, Wolfenbarger et al. 20 08) , Event 3272 corn is unlikely to affect the animal or plant communities found in conventional corn production systems because of the lack of toxicity and allergen i city, and because there is no change to agronomic pra ctices due to the cultivation of Event 3272 corn.



### Soil Biology

The soil environment in and around corn fields is complex, rich in microorganisms and arthropods. The corn root system acts as a soil modifier due to its association with several microbial gr oups such as bacteria, fungi, protozoa, and mites. Bacteria typically represent the most abundant microbes in the soil followed by fungi. These microbial groups play an important and particular role in the ecology of the soil, including nutrimental cycli ng and the availability of nutrients for plant growth.

Research shows that crop soils are prone to degradation due to the disturbance and exposure of the top surface layer by certain agronomic practices. Two environmental impacts of soil degradation are the decline in water quality and the contribution to the greenhouse effect (Lal and Bruce 1999)

.

No Action: Soil Biology Under the “no action” alternative, environmental releases of Event 3272 corn would be under regulation. Interactions with the soil would be limited t o the areas that were approved for regulated releases. Cultivation practices associated with regulated releases of Event 3272 corn would be the same as conventional corn production. The soil environment would be modified by corn roots and crop soils woul d still be affected by agronomic practices associated with

conventional corn cultivation.

Preferred Alternative: Soil Biology Under the “preferred” alternative, soil interactions with Event 3272 corn would occur at a large scale. Cultivation practices as sociated with regulated releases of Event 3272 corn would be the same as conventional corn production. The soil environment would be modified by corn roots and crop soils would still be affected by agronomic practices associated with conventional corn

cul tivation. Overall impacts would be similar to the no action alternative.

Event 3272 corn was engineered in such a way that the alpha

- amylase protein is produced nearly exclusively in the grain. This is important in the case of roots, because they are i n contact with53 the soil and could possibly affect soil biology. The protein was detected in the roots of four out of ten plants tested at the whorl stage, but not in any plants at any later stage s of development.

The level was extremely low, less than 0.1 μg/g fresh weight, which is more than two thousand

fold lower than in kernels. For this reason, roots are not likely to contribute biologically significant amounts of alpha

- amylase to the soil.

Because roots are not likely to contribute alpha

- amylase to the soil, and because larger amounts are found in grain, the degradation of grain is the main avenue for alpha amylase to find its way

into the soil

. Event 3272 grain contains 1627 μg/g fresh we ight of alpha

- amylase in the grain in the dough stage (Table 3

- 3, page 44 of Petition). It has be e n estimated (Wolt and Karaman, 2007) that approximately 1% of grain is lost during harvest, which falls to the ground in the

field. Wolt and Karaman (2007) estimated potential amounts of alpha

- amyla se that might accumulate in agricultural soils over time and space due to the use of Event 3272 corn. The study assumed levels of alpha

- amylase in grain, leaves, and roots to extrapolate and estimate environmental amounts of alpha

- amylase that might accum ulate in corn fields in Iowa. According to their calculations, they projected an order of magnitude increase in the amount of alpha

- amylase, due to increased amounts of alpha

- amylase, in soils in Iowa corn fields. However, the authors did not present any empirical evidence to suggest that alpha

- amylase would persist, and more importantly, did not consider the potential for natural degradation of alpha

- amylase. An abstract was cited (Kosaki et al. 2006) tha t found soil persistence of another thermostable protein, however

, the comparison between the two proteins was based solely on the thermostability of each of the proteins, and not the degradability of the proteins. Thermostability in and of itself is not a characteristic related to the ability of an enzyme to degrade, or persist for that matter, in agricultural soils.

Amylases, including alpha

- amylases, are enzymes that are ubiquitous in nature and naturally

occur in soils. In so il, a high diversity of alpha

- amylases are commonly found (Rondon et a l. 2000) . The microbial soil community and activity is complex, with a multitude of chemical and enzymatic interactions. Soils that have healthy microbial activity produce sufficient levels of proteases to degrade most proteins (Marx et al. 2005)

. Digestibility data presented to APHIS and FDA (Appendix H) by Syngenta show degradation of alpha

- amylase from Event 3272 corn by a single protease, pepsin. Although pepsi n is not normally found in soils, the data do suggest that alpha

- amylase is not inherently indigestible or non

- degradable.

Degradation of grain would have to occur before alpha

- amylase would be released into soil.

Grain left in the field does not immedi ately result in the increased amounts of alpha

- amylase in soil. Even if present, amylase would have no effect unless specific conditions for enzyme activity are met including substrate availability, inducers, availability of other nutrients, physical

and chemical parameters such as moisture, temperature, and pH. These characteristics vary in time and space and from one microenvironment to another. Additionally, according to Syngenta, alpha

- amylase from Event 3272 is constructed for maximum activity at 1 76oF (80oC), and only has 10% of its maximal activity under 86oF (30oC). None of these limiting factors were considered in the estimations and conjecture of potential contributes of Event 3272 grain to alpha

- amylase levels in the soil by Wolt and Karaman (2007).54 Cumulative Effects: Soil Biology The impacts to soil biology under the preferred alternative are unlikely to differ significantly

from the No Action Alternative. Soil environments where Event 3272 corn is grown will not be significantly change d by the increased exposure to amylase

. Alpha

- amylase from Event 3272

corn will not have significant activity within soils, and is expected to be readily degraded by microorganisms. No cumulative effects are identified for this issue.



### Conservation Re serve Program
The Conservation Reserve Program (CRP), administered by the USDA, is the largest private

lands conservation program in the U.S. According to the Farm Service Agency : CRP plantings and practices offer our nation vast environmental benefits, including reducing soil erosion, improving surface and ground water quality, creating wildlife habitat, restoring wetlands, sequestering carbon, preserving soil productivity, and reducing offsite wind erosion damages. Some of CRP’s substantial 2007 accompl ishments include: • Reducing runoff of sediment (207 million tons), nitrogen (480 million pounds), and phosphorus (108 million pounds) from agricultural soils. • Restoring and protecting 2.1 million acres of wetlands and adjacent buffers. • Establishing 1. 9 million acres of grass and forested buffers along the nation’s

rivers and streams. • Improving populations of Prairie Pothole ducks, ring

- necked pheasants, sage grouse, bobwhite quail, and other grassland birds. • Sequestering over 50 million tons of car bon. (USDA

- FSA 2007) There is public concern that the demand for ethanol production, and subsequent corn prices, are a catalyst resulting in removing acreage from the Conservat ion Reserve Program to place into corn production (Hart 2006)

.

No Action: Conservation Reserve Program Acres removed or maintained in the Conservation Reserve Program will not change under the “no action” alternative. Demands for ethanol production and corn prices may result in growers removing acres from the CRP and placing them into corn production, unde r the “no action”

alternative

. Market demands and corn prices are the drivers behind decisions made by growers

to remove acres from CRP (Hart 2006)

.

Preferred Alternative: Conservation Reserve Program

As stated in Section II , the demands for ethanol production and increased corn prices may result in growers removing acres from the CRP and place them into corn produ ction, independent of granting Event 3272 nonregulated status. Overall impacts on the Conservation Reserve Program

are similar to the no action alternative.

Cumulative Effects: Conservation Reserve Program The demands for ethanol production and increas ed corn prices are likely greater factors in determining if a farmer would remove acres from the CRP and place them into corn production,

than granting Event 3272 nonregulated status. No cumulative effects identified for this issue.55 

### Role of Assumptions in Environmental Consequences Analyses
Event 3272 corn will be grown under strict contractual agreements and a closed loop system to protect the value of the corn for ethanol production , similar to how other specialty corn varieties

are grown, harvested, transported, and marketed in the U.S . However, APHIS recognizes that there is a small chance that some members of the system may not live up to the contractual obligations. Each of the above issues was analyzed with the consideratio n that the assumptions made by APHIS were not met. Failure to follow contractual agreements will not result in significant environmental impacts in terms of corn production, ethanol production, public health, water use in ethanol production, animal and plant communities, soil biology, and Conservation Reserve Program acreage. Event 3272 corn is planted, grown, and harvested similar to other conventional corn. Event 3272 corn does not change the acreage devoted to corn production in the U.S., does not change where corn can be produced in the U.S., and will not change the acreage of corn planted to GE varieties (86% of corn acreage in the U.S. is planted to GE varieties, which is also used in corn ethanol facilities).

Planting, harvesting, and transporting of Event 3272 corn wi ll occur under rigid closed loop systems currently used for other specialty corn varieties. The potential economic consequences

of lapses of the close

- loop system have been analyzed. As stated above, there is no harm to the natural or physical environment of planting, growing, or harvesting Event 3272 corn. However, once the truckload of corn has left the farm, it is possible that Event 3272 corn may be del ivered

to an incorrect facil ity. The economic and marketing effects of m isdirecting Event 3272 corn to

a food or feed corn processing facility

, as opposed to an intended corn ethanol facility, are similar to the economic and marketing effects that occur when other specialty corn is shipped to the incorrect facility , or when manufacturing ingredients end up at an incorrect processing plant. There is the potential for economic and marketing harms for food processing facilities when material ends up at an unintended facility, whether i t is high amylose corn, waxy corn, or Event

3272 corn.



### Other Cumulative Effects
All potential cumulative effects regarding specific issues have been analyzed and addressed above. No further potential cumulative effects have been identified. Stacked va rieties, those crop varieties that may contain more than one trait, are currently found in the marketplace and in agricultural production. If granted nonregulated status, Event 3272 corn may be combined with other GE corn varieties by traditional breeding techniques, resulting in amylase corn that, for example, may also be resistant to herbicides or insects. Some GE corn varieties used for any commercial breeding program have already been found not to pose a plant pest risk, and have been granted nonregul ated status by APHIS. Syngenta currently has 4 GE corn varieties that may be stacked with Event 3272 corn: three varieties that have an insect

- resistance trait (Bt11 and Mir604) and one variety that has an herbicide

- tolerance trait (GA21). These corn lin es (Bt11, MIR604, GA21, MIR 162) have all been granted non

- regulated status, and the environmental assessments and FONSI determinations conducted by APHIS for each of these products can be found at h ttp://www.aphis.usda.gov/brs/not_reg.html . There is no guarantee that Event 3272 corn will be stacked with any particular deregulated GE variety, as company plans and market demands play a significant role in those business decisions. APHIS current regul ations at 7 CFR part 340 do not provide for Agency oversight of these GE corn varieties previously granted nonregulated status, nor over stacked varieties combining deregulated GE varieties unless it can be positively shown that such stacked varieties were somehow likely to56 pose a plant pest risk. Further, there is no guarantee that Event 3272 corn will be stacked with any particular deregulated GE variety, as company plans and market demands play a significant role in those business decisions. Moreover, E vent 3272 corn could even be combined with non

GE corn varieties. Thus, postulating and predicting any and all potential combinations of stacked varieties that could be created using both deregulated GE corn varieties and also non

- GE

corn varieties is too hypothetical and purely speculative.57 

### Threatened and Endangered Species

APHIS evaluated the potential for negative effects on Federal Threatened and Endangered Species (TES) as listed by the U.S. Fish and Wildlife Service (FWS) ( http://ecos.fws.gov/tess_public/pub/SpeciesReport.do?dsource=animals,
http://ecos.fws.gov/tess_public/pub/SpeciesReport.do?dsource=plants

, accessed 8/12/08 and 2/7/11 ), from cultivation of Event 3272 corn and its progeny. To identify negative effects or significant impacts on TES animal species, APHIS evaluated the risks to TES animals from consuming Event 3272 corn. Risk is a combination of hazard and exposure. APHIS first conducted hazard identification for Event 3272 corn. APHIS assessed the composition and nutritional quality of Event 3272 corn, and compared the composition of Event 3272 to the composition of a non

genetically engineered control corn line and the natura l variation found in commercial corn varieties. Corn is a feed common for many livestock and wildlife (e.g., birds, deer, and rodents). If the composition of Event 3272 corn is similar to other commercial corn plants, it is unlikely that Event 3272 poses a hazard to TES animal species. If no hazards are identified, then the risk

of Event 3272 corn harming TES animal species is also unlikely, regardless of exposure. However, APHIS also assessed the exposure data presented by Syngenta to further elucidate the exposure posed by Event 3272 corn to TES animals.

Event 3272 corn is genetically engineered to produce a thermostable alpha

- amylase. Alpha

amylases are ubiquitous in the environment, being naturally present in microorganisms, plants

and animals (Janeček et al. 1999) . These enzymes play a key role in many cellular metabolic processes by catalyzing the breakdown of starch, such as in germinating seed of cereal plants (Yu et al. 1996) or in the microbial decomposition of organic matter to provide carbon and energy for microbial growth processes (Rothstein et al. 1986) . Many types of commercial food processing, feed ingredient applications, and industrial applications also util ize alpha

- amylase enzymes, including the production of fuel and potable alcohol (brewing, distillation processes), and corn syrups (Janeček et al. 1999, Lév êque et al. 2000, Pariza and Johnson 2001, Olempska

Beer et al. 2006)

.

The data presented in the petiti on suggests there is no difference in compositional and nutritional quality of Event 3272 corn compared to conventional corn, apart from the presence of AMY797E and PMI. Although some of the variables measured by the applicant showed statistically signifi cant differences between Event 3272 corn and the nontransgenic hybrid controls (Tables 6

- 1 to 6

- 6, pages 70

- 76 , of the petition ), none of the values for the forage and grain composition characteristics were outside the range of natural variability of conve ntional corn as found in the International Life Sciences Institute Crop Composition Database (OECD 2003, Ridley et al. 2004, ILSI 2006) or in the OECD consensus do cument on corn (OECD 2003) composition. Event 3272 corn does not express additional proteins, natural toxicants, allelochemicals, pheromones, hormones, etc. that could directly or indirectly affect a listed TES or species proposed for listing. Thus, the composition of Event 3272 is not biologically different

than conventional corn.

Given that the composition of Event 3272 corn was found to be consistent with the natural variation found in conventional corn varieties, the applicant conducted studies to confirm the

absence of deleterious effects for animals when feeding on Event 3272 corn. AMY797E is predominantly found in the corn kernels of Event 3272 plants (1627 μg/g fresh weight in the58 dough stage), with minute amounts found in the roots during the whole stage (<0.1 μg/g fresh weight) and in the leaves during senescence (<1 μg/g fresh weight) (Table 3

- 3 of petition).

Animals th at feed primarily on corn kernels are seed

- feeding insects and rodents found in agricultural fields. During field trials, the applicant found no changes in insect feeding damage (Table 5

- 7 of petition), indicating similar insect susceptibility for Event 3 272 corn compared to

conventional corn.

Rodents, such as mice or squirrels, may seasonally feed exclusively on corn kernels. Accordingly , these animals are most likely to have a diet containing large amounts of corn kernels. Using dietary calculations ( pages 93

- 94 of petition) along with the results of a mouse toxicity study (page 50 of petition and page 6 of response letter from Syngenta ), the applicant

determined that the N o O bserved E ffect D ose (NOED) of AMY797E was greater than 2 times

the maximum am ount of alpha

- amylase corn a rodent might consume daily (page 94 of petition). Other mammals, such as deer, would have even lower exposure to AMY797E because of feeding habits; for example, deer nibble on tips of corn ears as opposed to kernels (Steffey et al. 1999)

. Therefore , there would be no negative impact to T ES mammals that forage in Event 3272 corn

.

The applicant also evaluated the effects of AMY797E on birds by feeding broiler chickens diets that contained up to 65% of Event 3272 grain for 49 days (page 94 of petition and page 6 of response letter). The ap plicant reported no harmful effects to chickens from a diet extremely high in AMY797E. Moreover, given that diets high in AMY797E do not result in harmful effects, the diets of wild birds that occasionally forage in corn fields, including threatened and

e ndangered species such as the whooping crane, are unlikely to contain high amounts of AMY797E for 49 consecutive days as corn availability is limited by seed germination and

harvest.

Additionally, th reatened and endangered species have been previously exp osed to alpha

amylases in the environment, as they are ubiquitous enzymes found in microorganisms, plants,

and animals (Janeček et al. 1999) . AMY797E also does not have any significan t sequence identity to any known toxins (page 7 of response letter from Syngenta ). As stated earlier, AMY797E is functionally similar to thermostable alpha

- amylases already safely used in food and feed processing (Janeček et al. 1999, Lév êque et al. 2000, Pariza and Johnson 2001, Olempska

- Beer et al. 2006) , indicating that Event 3272 corn is unlikely to produce toxins that would negatively a ffect animals that may eat corn kernels or plants containing AMY797E.

Organisms exposed to AMY797E will also be exposed to PMI. Additional species, such as leaf

feeding animals, butterflies and bees, may not be exposed to AMY797E but will be exposed to PMI because of its expression in vegetative tissue and pollen (Table 3

- 4 of petition). The pmi ( manA ) gene comes f rom E. coli and encodes the enzyme PMI

. Pmi serves as a marker gene that enables selection of lines that are genetically modified, providing the plant with the ability to

utilize mannose as a sole carbon source. Table 3

- 4 of the petition gives the PMI ex pression data for Event 3272 corn. PMI expression was examined during 5 developmental stages of Event 3272 corn (whorl, anthesis, kernel dough, kernel maturity, and senescence). In Event 3272 corn, PMI is expressed in leaves during all stages except sene scence, with maximum expression during the kernel dough stage (5.7 μg/g fresh weight). Expression of PMI in the roots occurs during all developmental stages with the highest expression of 1.0 μg/g fresh weight during the whorl stage. The highest PMI expr ession for kernels occurred during the kernel dough stage (0.8 μg/g59 fresh weight), and 8.5 μg/g fresh weight of PMI was expressed in the pollen. The expression of PMI protein in corn plants is not expected to have deleterious effects or significant impact s on TES organisms, based on the data provided in the petition. Dietary calculations to determine the daily dietary dose of PMI (page 93 of petition) and data from the mouse and bird toxicity studies (page 94 of petition) indicate that PMI levels in Event 3272 corn do not cause harm for threatened and endangered species.

Additionally, the EPA has granted an exemption from the requirement of a tolerance for the PMI protein as an inert ingredient in all plants (69 FR 26770

- 26775). The DNA encoding the PMI protein is not toxic. At the 80

- amino acid peptide level, the PMI protein shares no significant homology with proteins known to be toxic or allergenic. Within one of the 80

- amino acid windows, there was one region of sequence homology of eight contiguous amino

- acids between PMI and a known allergen, α

- parvalbumin from a Rana (frog) species. Further testing found no

cross

- reactivity between PMI and the human serum immunoglobulin E (IgE) and Bovine Serum Albumin (BSA), indicating that the low degree of seq uence identity between the PMI used in

Event 3272 corn and α

- parvalbumin from Rana sp. is not biologically relevant.

Corn itself is not sexually compatible with any TES plant species; therefore there is no potential

for a direct effect of Event 3272 corn on TES plants . Indirect effects of Event 3272 corn on TES plant species were also evaluated. As stated above, Event 3272 corn has no negative effect on animals, including animals such as insects or bats, which may be pollinators for TES plants. As

a res ult, there are no indirect effects of Event 3272 corn on TES plant species.

Cultivation of Event 3272 corn is not expected to differ from typical corn cultivation. Event 3272

corn is not genetically engineered to produce a toxin or pesticide, and is not genetically engineered to be tolerant to an herbicide. Although the extent to which Event 3272 corn will be grown is ultimately unknown, this product is expected to replace other GE and non

- GE corn varieties currently grown for the ethanol market. After reviewing the possible effects of granting nonregulated status to Event 3272 corn, APHIS has not identified any stressor caused directly by this product that could affect the reproduction, numbers, or distribution of a listed TES or species proposed for li sting. The potential environmental impacts on TES of this product are those associated with typical corn agriculture. Growers planting Event 3272 corn, as with any other corn variety, genetically engineered or not, should consider the environmental impact s of agronomic practices on those TES found in and around their corn field.

After reviewing possible effects of granting nonregulated status to Event 3272 corn, APHIS has not identified any stressor that could affect the reproduction, numbers, or distrib ution of a listed TES or species proposed for listing. Consequently, an exposure analysis for individual species is not necessary. APHIS has considered the effect of Event 3272 corn production on designated

critical habitat or habitat proposed for design ation and could identify no difference from affects that would occur from the production of other corn varieties. APHIS has reached a conclusion that the release of Event 3272 corn, following a determination of nonregulated status, would

have no effect on federally listed threatened or endangered species or species proposed for listing, nor is it expected to adversely modify designated critical habitat or habitat proposed for designation, compared to current agricultural practices. Consequently, a written concurrence or formal consultation with the USFWS is not required for this action. Based on this analysis, there60 is no apparent potential for significant impact on threatened or endangered species if APHIS were to grant the petition for nonregulated statu s to Event 3272 corn.



### Compliance with Statutes, Executive Orders and Regulations

Executive Order (EO) 12898 (US

- NARA 2008) , “Federal Actions to Address Environmental Justice in Minority Populations and Low

- Income Populations”, requires Federal agencies to conduct their programs, policies, and activities that substantially affect human health or the environment in a manner so as not to exclude pers ons and populations from participation in or benefiting from such programs. It also enforces existing statutes to prevent minority or low

income communities from being subjected to disproportionately high and adverse human health

or environmental effects. EO 13045 (US

- NARA 2008) , “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 metabolic activity levels, and behavior patterns, as compared to adults. The EO (to the extent permitted by law and consistent with the agency’s mission) required eac h Federal agency to identify, assess, and address environmental health risks and safety risks that may disproportionately 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, Event 3272 corn is not significantly different than conventional corn and has successfully completed FDA consultation for food and feed use. Therefore, Event 3272 corn is not expected to have a disproportionate adverse effect on mi norities, low

- income populations, or children.

EO 13112 (US

- NARA 2008) , “Invasive Species”, states that Federal agencies take action to prevent the introd uction of invasive species, to provide for their control, and to minimize the economic, ecological, and human health impacts that invasive species cause. Both non

- GE and GE corn varieties that have been granted nonregulated status are widely grown in the U .S. Based on historical experience with corn and the data submitted by the applicant and assessed by APHIS, Event 3272 corn plants are very similar in fitness characteristics to other corn varieties currently grown and are not expect ed to become weedy or invasive

. (USDA

- APHIS 200 9 )

.

EO 13186 (US

- NARA 2008) , “Resp onsibilities 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 Fish 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 quality of Event 3272 corn comp ared to conventional corn, apart from the presence of AMY797E and PMI. Syngenta also conducted feeding experiments on broiler chickens to evaluate the effects of AMY797E on birds (page 94 of petition and page 6 of response letter). The applicant reported no harmful effects to chickens from a diet extremely high in AMY797E. Moreover, given that diets high in AMY797E do not result in harmful effects, the diets of migratory birds that occasionally forage in corn fields are unlikely to contain high amounts o f AMY797E as corn availability is limited by seed germination and harvest. Based on APHIS’ assessment of Event 3272 corn it is unlikely that granting nonregulated status to this corn variety will have a negative effect on migratory bird populations.61 I N TERNATIONAL I MPLICATIONS
EO 12114 (US

- NARA 2008) , “Environmental Effects Abroad of Major Federal Actions”, requires Federal officials to take into considera tion any potential 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. should nonregul ated status be granted to Event 3272 corn. It should be noted that all the considerable, existing national and international regulatory authorities and phytosanitary regimes that currently apply to introductions of new corn cultivars internationally, apply equally to those covered by an APHIS determination of nonregulated status under 7 CFR part 340. Any

international traffic of Event 3272 corn subsequent to a determination of nonregulated status for the product would be fully subject to national phytosani tary requirements and be in accordance with phytosanitary standards developed under the International Plant Protection Convention (IPPC).

The purpose of the IPPC “is to secure a common and effective action to prevent the spread and introduction of pests of plants and plant products and to promote appropriate measures for their

control” (IPP 2008) ; the protection it affords extends 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 phytosanitary certifi cation among the nations that have signed or acceded to the Convention ( 177 countries as of February 2011 )

. In April 2004, a standard for pest risk analysis (PRA) of living modified organisms (LMOs) was adopted at a meeting of the governing body of the IPPC as a supplement to an existing standard, International Standard for Phytosanitary Measure No. 11 (ISPM

- 11, Pest Risk Analysis for Quarantine 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 pest risk assessment procedures for genetically engineered organisms are consistent with the guidance developed under the IPPC. In addition, issues that may relate to commercialization and transboundary movement of particular

a gricultural commodities produced through biotechnology are being addressed in other international forums and through national regulations.

The Cartagena Protocol on Biosafety is a treaty under the United Nations Convention on Biological Diversity (CBD) t hat established a framework for the safe transboundary movement, with respect to the environment and biodiversity, of LMOs, which includes those modified through biotechnology. The Protocol came into force on September 11, 2003, and 160 countries

are Parti es to it as of February 2011 (CBD 2011 ) . Although the U.S. is not a party to the CBD,

and ther efore not a party to the Cartagena Protocol on Biosafety, U.S. exporters will still need to comply with domestic regulations that importing countries that are Parties to the Protocol have put in place to comply with their obligations. The first intentiona l transboundary movement of LMOs intended for environmental release (field trials or commercial planting) will require consent from the importing country under an advanced informed agreement (AIA) provision, which includes a requirement for a risk assessme nt consistent with Annex III of the Protocol, and the required documentation.

LMOs imported for food, feed, or processing (FFP) are exempt from the AIA procedure, and are

covered under Article 11 and Annex II of the Protocol. Under Article 11 Parties mu st post62 decisions to the Biosafety Clearinghouse database on domestic use of LMOs for FFP that may be subject to transboundary movement. To facilitate compliance with obligations to this protocol,

the U.S. Government has developed a website that provides the status of all regulatory reviews completed for different uses of bioengineered products (NBII 2008) . These data will be available to the Biosafety Clearinghouse. APHIS continues to work toward harmonization of biosafety and biotechnolo gy consensus documents, guidelines, and regulations, including within the North American Plant Protection Organization (NAPPO), which includes Mexico, Canada, and the U.S., and within the Organization for Economic Cooperation and Development. NAPPO

has com pleted three modules of a standard for the Importation and Release into the Environment of Transgenic Plants in NAPPO Member Countries (NAPPO 2008) . APHIS also participates in the North American Biotechnology Initiative (NABI), a forum for information exchange and cooperation on agricultural biotechnology issues for the U.S., Mexico and Canada. In addition, bilateral discussions on biotechnology regulatory issues are held regularly with other countries including: Argentina, Brazil, Japan, China, and Korea.

C OMPLIANCE WITH C LEAN W ATER A CT AND C LEAN A IR A CT This Environmental Assessment evaluated the changes in corn production due to the unrestricted use of Event 3272 corn. Event 3272 corn will not lead to the increased production of corn in U.S. agriculture. There is no expected change in water use due to the production of Event 3272 corn, nor is it expected that air quality will change to do the production of Event 3272 corn. If APHIS grants nonregulated status to Event 3272 corn, APHIS will be fully compliant with the

Clean Water Act and the Clean Air Ac t.



## V. Listing of Agencies and Persons Consulted

Dr. Levis Handley Biotechnology Regulatory Services

APHIS

Rhey Solomon E&T Solutions, LLP

Rhonda Solomon

Environmental Services

APHIS

Dr. Michael Watson Biotechnology Regulatory Services

APHIS

Office of General Counsel

USDA63 

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- 929. Stallman, H. R. and L. B. Best. 1996. Small

- mammal use of an experimental strip intercropping system in northeastern Iowa. American Midland Naturalist. 135 :226

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mammals and birds on low

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Resources. Taylor, S. L. and S. L. Hefl e. 2001. Will genetically engineered food be allergenic? Current Reviews of Allergy and Clinical Immunology. 103 :765

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world. February 2007 :26

- 27. Thomison, P. no date. Specialty Corns: waxy, high

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D.C.

US

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– crop production practices., United States Department of Ag riculture, Economic Research Service. http://www.ers.usda.gov/Data/ARMS/app/Crop.aspx . Access date: August 22, 2008.

USDA

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Research Service. http://www.ers.usda.gov/data/feedgrains/ . Access date: August 25,

2008.

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Research Service. http://www.ers.usda.gov/data/feedgrains/ . Access date: February 2011

. USDA

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Service.69 http://usda.mannlib.cornell.edu/MannUsda/viewDocume ntInfo.do?documentID=1000

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. Date Accessed: September 1, 2008.

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Statistics Service. http://usda.mannlib.cornell.edu/MannUsda/viewDocumentInfo.do?documentID=1000

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- 1289.71 

## Appendix A . Biotech Seed Products Available for the

2011 Planting Season 1,2,3

PRODUCT REGISTRANT

TRADE NAME CHARACTERISTIC EVENT JAPAN

APPROVED EU FOOD

APPROVAL EU

PROCESSED

FEED

APPROVAL

Syngenta AgrisureTM CB/LL Cry1Ab, Corn Borer

Glufosinate herbicide

Tolerance Bt11 Yes Yes Yes

Agrisure VipteraTM 3110 Vip3A, Cry1Ab,

European and

Southwestern

Corn Borers, Southern

Cornstalk Borer,

Fall and Beet

Armyworm,

Black and Western

Bean Cutworm,

Sugarcane Borer,

Common Stalk borer

and

Dingy Cutworm

protection

Glyphosate tolerance

Glufosinate tolerance MIR162+Bt11+GA21 Yes No XVII. No

Agrisure VipteraTM 3111 Vip3A, Cry1Ab,

European and

Southwestern

Corn Borers, Southern

cornstalk borer,

Fall and Beet

armyworm,

Black and Western

Bean Cutworm,

Sugarcane borer,

Western, Northern and

Mexican

corn rootworm,

Common stalk borer

and Dingy cutworm

protection MIR162+Bt11+GA21+MIR604 Yes No No72 Glyphosate tolerance

Glufosinate tolerance

DowAgrosciences Pioneer Hi

Bred

Herculex® I Cry1F, Western Bean

Cutworm, Corn Borer,

Black Cutworm and

Fall Armyworm

resistance Glufosinate

herbicide tolerance. TC1507 Yes Yes Yes

Monsanto YieldGard® Corn

Borer Cry1Ab, European and

Southwestern

Corn Borers,

Sugarcane Borer and

Southern Cornstalk

Borer protection. Mon810 Yes Yes Yes

Monsanto YieldGard® Corn

Borer with

Roundup Ready® Corn 2 Cry1Ab, European and

Southwestern

Southwestern Corn

Borers, Sugarcane

Borer

and Southern Cornstalk

Borer protection.

Glyphosate herbicide

tolerance. Mon810+NK603 Yes Yes Yes

Monsanto YieldGard®

Rootworm

with Roundup Ready® Corn 2 Cry3Bb1, Western,

Northern and Mexican

Corn Rootworm

Protection.

Glyphosate herbicide

tolerance. Mon863+NK603 Yes Yes Yes

Monsanto YieldGard®

Rootworm Cry3Bb1, Western,

Northern and Mexican,

Corn Rootworm

protection. Glyphosate

herbicide tolerance. Mon863 Yes Yes Yes

Monsanto Roundup Ready®

Corn 2 Glyphosate herbicide

tolerance. NK603 Yes Yes Yes73 Bayer CropScience

LibertyLink® Glufosinate herbicide

tolerance. XVIII. T25 Yes Yes Yes

Monsanto YieldGard® Plus Cry1Ab, Cry3Bb1,

European and

Southwestern, Corn

Borers, Sugarcane

Borer, Southern

Cornstalk Borer, , and

Western, Northern and

Mexican Corn

Rootworm protection. Mon810+Mon863 Yes Yes Yes

Monsanto YieldGard® Plus

with Roundup Ready® Corn 2 Cry1Ab, Cry3Bb1,

European a nd

Southwestern Corn

Borers, Sugarcane

Borer, Southern

Cornstalk Borer, and

Western, Northern and

Mexican Corn

Rootworm protection.

Glyphosate herbicide

tolerance. XIX. Mon810+Mon863+NK603 XX. Yes XXI. Yes XXII. Yes

DowAgrosciences Pioneer Hi

Bred Herculex® I

Monsanto Roundup Ready®

Corn 2 Cry1F, Western Bean

Cutworm, Corn Borer,

Black Cutworm and Fall

Armyworm resistance.

Glyphosate herbicide

tolerance

Glufosinate herbicide

tolerance. TC1507+NK603 Yes Yes Yes

Syngenta Agrisure® GT Glyph osate herbicide

tolerance. SYTGA21 Yes Yes Yes

Syngenta Agrisure®

GT/CB/LL Cry1AB, European and

Southwestern Corn

borer protection

Glyphosate herbicide

tolerance

Glufosinate herbicide

tolerance. Bt11+GA21 Yes Yes Yes74 Dow AgroSciences Pioneer Hi

Bred Herculex® RW Cry34/35Ab1, Western

Corn Rootworm,

Northern Corn

Rootworm protection.

Glufosinate herbicide

tolerance. DAS59122

- 7 Yes Yes Yes

Dow AgroSciences Pioneer Hi

Bred Herculex® Xtra Cry1 F, Western Bean

Cutworm, Corn Borer,

Black Cutworm and Fall

Armyworm resistance

Northern Corn

Rootworm. Western

Corn Rootworm

protection.

Glufosinate herbicide

tolerance. TC1507+DAS59122

- 7 Yes Yes Yes

Dow AgroSciences Pioneer Hi

Bred

Herculex® RW

Monsanto Roundup Ready®

Corn 2 Cry34/35Ab1, Western

Corn Rootworm,

Northern Corn

Rootworm protection.

Glufosinate herbicide

tolerance.

Glyphosate herbicide

tolerance.

. DAS59122

- 7+NK603 Yes Yes Yes

Dow AgroSciences Pioneer Hi

Bred Herculex® Xtra

Monsanto Roundup® Corn 2 Cry1F, Western Bean

Cutworm, Corn Borer,

Black Cutworm and Fall

Armyworm resistance.

Glufosinate herbicide

tolerance.

Cry34/35Ab1, Western

Corn Rootworm

Northern Corn

Rootworm Protection.

Glyphosate herbicide

toleran ce. TC1507+DAS59122

- 7+NK603 Yes Yes Yes

Monsanto YieldGard VTTM

Rootworm/RR2® Cry3Bb1, Western,

Northern, and Mexican

Corn Rootworm

protection. Mon88017 Yes Yes Yes75 Glyphosate Herbicide

Tolerance.

Monsanto YieldGard VTTM

Triple Cry1Ab, Cry3Bb1,

European and

Southwestern Corn

Borer, Sugarcane Borer

and South ern Cornstalk

Borer and Western,

Northern, and Mexican

Corn Rootworm

protection.

Glyphosate herbicide

tolerance. Mon810+Mon88017 Yes Yes Yes

XXIII. Syngenta Agrisure®

RW XXIV. Modified

Cry3A,

Protection of

Western,

Northern and

Mexican Corn

Rootworm. XXV. MIR604 Yes Yes Yes

Syngenta® GT/RW Modified Cry3A,

Protection of Western,

Northern and Mexican

Corn Rootworm

Glyphosate herbicide

tolerance MIR604+SYTGA21 Yes No No

Syngenta Agrisure®

CB/LL/RW Cry1Ab, Corn Borer

protection

Modified Cry3A,

Protection of Western,

Northern and Mexican

Corn Rootworm

Glufosinate herbicide

tolerance. Bt11+MIR604 Yes No No

Syngenta Agris ure® 3000GT XXVI. Cry1Ab, Corn

Borer

protection.

Modified

Cry3A,

Protection of

Western, XXVII. SYTGA21+Bt11+MIR604 Yes No No76 Northern and

Mexican Corn

Rootworm.

Glufosinate

herbicide

tolerance.

Glyphosate

tolerance.

Monsanto GenuityTM VT

Double PROTM Cry1A.105, Cry2Ab2,

European and

Southwestern Corn

Borers, Sugarcane

Borer, Southern

Cornstalk Borer, Corn

Earworm, and Fall

Armyworm protection.

Glyphosate herbicide

tolerance. Mon89034+NK603 Yes No No

Monsanto GenuityTM VT Triple

PROTM Cry1A.105, Cry2Ab2,

Cry3Bb1, European

and Southwestern Corn

Borers, Sugarcane

Borer, Southern

Cornstalk Borer, Corn

Earworm, Fall

Armyworm, Western

Corn Rootworm,

Northern Corn

Rootworm, and

Mexican Corn

Rootworm protection.

Glyphosate herbicide

tolerance. Mon88017+Mon89034 Yes XXVIII. No XXIX. No

Monsanto

GenuityTMSmartStaxTM

DowAgrosciences

SmartStaxTM Cry1A.105, Cry2Ab2,

Cry1F, Cry3Bb1,

Cry34/35Ab1 Western,

Northern, and Mexican

Corn Rootworms,

European and

Southwestern Corn

Borers, Sugarcane

Borer, Southern

Cornstalk Borer,

Western Bean and

Black Cutworms, Corn

Earworm, Fall77 Armyworm protection.

Glyph osate herbicide

tolerance.

Glufosinate herbicide

tolerance.

1 This list is representative of available products but may not include all corn biotechnology hybrids currently

available.

2 All of the hybrids listed have full food and feed approval in the United States.

3 Not all varieties are approved for all export market uses.



### References

NCGA. 2008. Approval Status of Biotech Corn Hybrids: Know Before you Grow SM

. National Corn Growers Association, St. Louis, Missouri ( http://www.ncga.com/know

- you

grow
, ). Accessed on Feb ruary , 20 10

.78 

## Appendix B

. Corn

- producing counties in the 2 8
states that have corn ethanol facilities

State/County State/County State/County State/County

Arizona \ Apache Iowa \ Marion Missouri \ Perry South Dakota \ Grant

Arizona \ Cochise Iowa \ Marshall Missouri \ Pettis South Dakota \ Gregory

Arizona \ Coconino Iowa \ Mills Missouri \ Phelps South Dakota \ Haakon

Arizona \ Gila Iowa \ Mitchell Missouri \ Pike South Dakota \ Hamlin

Arizona \ Graham Iowa \ Monona Missouri \ Platte South Dakota \ Hand

Arizona \ Greenlee Iowa \ Monroe Missouri \ Polk South Dakota \ Hanson

Arizona \ La Paz Iowa \ Montgomery Missouri \ Pulaski South Dakota \ Harding

Arizona \ Maricopa Iowa \ Muscatine Missouri \ Putnam South Dakota\ Hughes

Arizona \ Mohave Iowa \ O'Brien Missouri \ Ralls South Dakota \ Hutchinson

Arizona \ Navajo Iowa \ Osceola Missouri \ Randolph South Dakota \ Hyde

Arizona \ Pima Iowa \ Page Missouri \ Ray South Dakota \ Jackson

Arizona \ Pinal Iowa \ Palo Alto Missouri \ Reynolds South Dakota \ Jerauld

Arizona \ Santa Cruz Iowa \ Plymouth Missouri \ Ripley South Dakota \ Jones

Arizona \ Yavapai Iowa \ Pocahontas Missouri \ St. Charles South Dakota \ Kingsbury

Arizona \ Yuma Iowa \ Polk Missouri \ St. Clair South Dakota \ Lake

California \ Alameda Iowa \ Pottawattamie Missouri \ Ste. Genevieve South Dakota \ Lawrence

California \ Alpine Iowa \ Poweshiek Missouri \ St. Francois South Dakota \ Lincoln

California \ Amador Iowa \ Ringgold Missouri \ St. Louis South Dakota \ Lyman

California \ Butte Iowa \ Sac Missouri \ Saline South Dakota \ McCook

California \ Calaveras Iowa \ Scott Missouri \ Schuyler South Dakota \ McPherson

California \ Colusa Iowa \ Shelby Missouri \ Scotland South Dakota \ Marshall

California \ Contra Costa Iowa \ Sioux Missouri \ Scott South Dakota \ Meade

California \ Del Norte Iowa \ Story Missouri \ Shannon South Dakota \ Mellette

California \ El Dorado Iowa \ Tama Missouri \ Shelby South Dakota \ Miner

California \ Fresno Iowa \ Taylor Missouri \ Stoddard South Dakota \ Minnehaha

California \ Glenn Iowa \ Union Missouri \ Stone South Dakota \ Moody

California \ Humboldt Iowa \ Van Buren Missouri \ Sullivan South Dakota \ Pennington

California \ Imperial Iowa \ Wapello Missouri \ Taney South Dakota \ Perkins

California \ Inyo Iowa \ Warren Missouri \ Texas South Dakota \ Potter

California \ Kern Iowa \ Washington Missouri \ Vernon South Dakota \ Roberts

California \ Kings Iowa \ Wayne Missouri \ Warren South Dakota \ Sanborn

California \ Lake Iowa \ Webster Missouri \ Washington South Dakota \ Shannon

California \ Lassen Iowa \ Winnebago Missouri \ Wayne South Dakota \ Spink

California \ Los Angeles Iowa \ Winneshiek Missouri \ Webster South Dakota \ Stanley

California \ Madera Iowa \ Woodbury Missouri \ Worth South Dakota \ Sully

California \ Mariposa Iowa \ Worth Missouri \ Wright South Dakota \ Todd

California \ Merced Iowa \ Wright Nebraska \ Adams South Dakota \ Tripp

California \ Modoc Kansas \ Allen Nebraska \ Antelope South Dakota \ Turner

California \ Mono Kansas \ Anderson Nebraska \ Arthur South Dakota \ Union

California \ Monterey Kansas \ Atchison Nebraska \ Banner South Dakota \ Walworth

California \ Napa Kansas \ Barber Nebraska \ Blaine South Dakota \ Yankton

California \ Nevada Kansas \ Barton Nebraska \ Boone South Dakota \ Ziebach

California \ Orange Kansas \ Bourbon Nebraska \ Box Butte Tennessee \ Anderson79 State/County State/County State/County State/County

California \ Placer Kansas \ Brown Nebraska \ Boyd Tennessee \ Bedford

California \ Plumas Kansas \ Butler Nebraska \ Brown Tennessee \ Benton

California \ Riverside Kansas \ Chase Nebraska \ Buffalo Tennessee \ Bledsoe

California \ Sacramento Kansas \ Chautauqua Nebraska \ Burt Tennessee \ Blount

California \ San Benito Kansas \ Cherokee Nebraska \ Butler Tennessee \ Bradley

California \ San

Bernardino Kansas \ Cheyenne Nebraska \ Cass Tennessee \ Campbell

California \ San Diego Kansas \ Clark Nebraska \ Cedar Tennessee \ Cannon

California \ San

Francisco Kansas \ Clay Nebraska \ Chase Tennessee \ Carroll

California \ San Joaquin Kansas \ Cloud Nebraska \ Cherry Tennessee \ Carter

California \ San Luis Obispo Kansas \ Coffey Nebraska \ Cheyenne Tennessee \ Cheatham

California \ San Mateo Kansas \ Comanche Nebraska \ Clay Tennessee \ Chester

California \ Santa

Barbara Kansas \ Cowley Nebraska \ Colfax Tennessee \ Claiborne

California \ Santa Clara Kansas \ Crawford Nebraska \ Cuming Tennessee \ Clay

California \ Shasta Kansas \ Decatur Nebraska \ Custer Tennessee \ Cocke

California \ Sierra Kansas \ Dickinson Nebraska \ Dakota Tennessee \ Coffee

California \ Siskiyou Kansas \ Doniphan Nebraska \ Dawes Tennessee \ Crockett

California \ Solano Kansas \ Douglas Nebraska \ Dawson Tennessee \ Cumberland

California \ Sonoma Kansas \ Edwards Nebraska \ Deuel Tennessee \ Davidson

California \ Stanislaus Kansas \ Elk Nebraska \ Dixon Tennessee \ Decatur

California \ Sutter Kansas \ Ellis Nebraska \ Dodge Tennessee \ DeKalb

California \ Tehama Kansas \ Ellsworth Nebraska \ Douglas Tennessee \ Dickson

California \ Tulare Kansas \ Finney Nebraska \ Dundy Tennessee \ Dyer

California \ Tuolumne Kansas \ Ford Nebraska \ Fillmore Tennessee \ Fayette

California \ Ventura Kansas \ Franklin Nebraska \ Franklin Tennessee \ Fentress

California \ Yolo Kansas \ Geary Nebraska \ Frontier Tennessee \ Franklin

California \ Yuba Kansas \ Gove Nebraska \ Furnas Tennessee \ Gibson

Colorado \ Adams Kansas \ Graham Nebraska \ Gage Tennessee \ Giles

Colorado \ Alamosa Kansas \ Grant Nebraska \ Garden Tennessee \ Grainger

Colorado \ Arapahoe Kansas \ Gray Nebraska \ Garfield Tennessee \ Greene

Colorado \ Archuleta Kansas \ Greeley Nebraska \ Gosper Tennessee \ Grundy

Colorado \ Baca Kansas \ Greenwood Nebraska \ Grant Tennessee \ Hamblen

Colorado \ Bent Kansas \ Hamilton Nebraska \ Greeley Tennessee \ Hamilton

Colorado \ Boulder Kansas \ Harper Nebraska \ Hall Tennessee \ Hancock

Colorado \ Broomfield Kansas \ Harvey Nebraska \ Hamilton Tennessee \ Hardeman

Colorado \ Chaffee Kansas \ Haskell Nebraska \ Harlan Tennessee \ Hardin

Colorado \ Cheyenne Kansas \ Hodgeman Nebraska \ Hayes Tennessee \ Hawkins

Colorado \ Clear Creek Kansas \ Jackson Nebraska \ Hitchcock Tennessee \ Haywood

Colorado \ Conejos Kansas \ Jefferson Nebraska \ Holt Tennessee \ Henderson

Colorado \ Costilla Kansas \ Jewell Nebraska \ Hooker Tennessee \ Henry

Colorado \ Crowley Kansas \ Johnson Nebraska \ Howard Tennessee \ Hickman

Colorado \ Custer Kansas \ Kearny Nebraska \ Jefferson Tennessee \ Houston

Colorado \ Delta Kansas \ Kingman Nebraska \ Johnson Tennessee \ Humphreys

Colorado \ Denver Kansas \ Kiowa Nebraska \ Kearney Tennessee \ Jackson80 State/County State/County State/County State/County

Colorado \ Dolores Kansas \ Labette Nebraska \ Keith Tennessee \ Jefferson

Colorado \ Douglas Kansas \ Lane Nebraska \ Keya Paha Tennessee \ Johnson

Colorado \ Eagle Kansas \ Leavenworth Nebraska \ Kimball Tennessee \ Knox

Colorado \ Elbert Kansas \ Lincoln Nebraska \ Knox Tennessee \ Lake

Colorado \ El Paso Kansas \ Linn Nebraska \ Lancaster Tennessee \ Lauderdale

Colorado \ Fremont Kansas \ Logan Nebraska \ Lincoln Tennessee \ Lawrence

Colorado \ Garfield Kansas \ Lyon Nebraska \ Logan Tennessee \ Lewis

Colorado \ Gilpin Kansas \ McPherson Nebraska \ Loup Tennessee \ Lincoln

Colorado \ Grand Kansas \ Marion Nebraska \ McPherson Tennessee \ Loudon

Colorado \ Gunnison Kansas \ Marshall Nebraska \ Madison Tennessee \ McMinn

Colorado \ Hinsdale Kansas \ Meade Nebraska \ Merrick Tennessee \ McNairy

Colorado \ Huerfano Kansas \ Miami Nebraska \ Morrill Tennessee \ Macon

Colorado \ Jackson Kansas \ Mitchell Nebraska \ Nance Tennessee \ Madison

Colorado \ Jefferson Kansas \ Montgomery Nebraska \ Nemaha Tennessee \ Marion

Colorado \ Kiowa Kansas \ Morris Nebraska \ Nuckolls Tennessee \ Marshall

Colorado \ Kit Carson Kansas \ Morton Nebraska \ Otoe Tennessee \ Maury

Colorado \ Lake Kansas \ Nemaha Nebraska \ Pawnee Tennessee \ Meigs

Colorado \ La Plata Kansas \ Neosho Nebraska \ Perkins Tennessee \ Monroe

Colorado \ Larimer Kansas \ Ness Nebraska \ Phelps Tennessee \ Montgomery

Colorado \ Las Animas Kansas \ Norton Nebraska \ Pierce Tennessee \ Moore

Colorado \ Lincoln Kansas \ Osage Nebraska \ Platte Tennessee \ Morgan

Colorado \ Logan Kansas \ Osborne Nebraska \ Polk Tennessee \ Obion

Colorado \ Mesa Kansas \ Ottawa Nebraska \ Red Willow Tennessee \ Overton

Colorado \ Mineral Kansas \ Pawnee Nebraska \ Richardson Tennessee \ Perry

Colorado \ Moffat Kansas \ Phillips Nebraska \ Rock Tennessee \ Pickett

Colorado \ Montezuma Kansas \ Pottawatomie Nebraska \ Saline Tennessee \ Polk

Colorado \ Montrose Kansas \ Pratt Nebraska \ Sarpy Tennessee \ Putnam

Colorado \ Morgan Kansas \ Rawlins Nebraska \ Saunders Tennessee \ Rhea

Colorado \ Otero Kansas \ Reno Nebraska \ Scotts Bluff Tennessee \ Roane

Colorado \ Ouray Kansas \ Republic Nebraska \ Seward Tennessee \ Robertson

Colorado \ Park Kansas \ Rice Nebraska \ Sheridan Tennessee \ Rutherford

Colorado \ Phillips Kansas \ Riley Nebraska \ Sherman Tennessee \ Scott

Colorado \ Pitkin Kansas \ Rooks Nebraska \ Sioux Tennessee \ Sequatchie

Colorado \ Prowers Kansas \ Rush Nebraska \ Stanton Tennessee \ Sevier

Colorado \ Pueblo Kansas \ Russell Nebraska \ Thayer Tennessee \ Shelby

Colorado \ Rio Blanco Kansas \ Saline Nebraska \ Thomas Tennessee \ Smith

Colorado \ Rio Grande Kansas \ Scott Nebraska \ Thurston Tennessee \ Stewart

Colorado \ Routt Kansas \ Sedgwick Nebraska \ Valley Tennessee \ Sullivan

Colorado \ Saguache Kansas \ Seward Nebraska \ Washington Tennessee \ Sumner

Colorado \ San Juan Kansas \ Shawnee Nebraska \ Wayne Tennessee \ Tipton

Colorado \ San Miguel Kansas \ Sheridan Nebraska \ Webster Tennessee \ Trousdale

Colorado \ Sedgwick Kansas \ Sherman Nebraska \ Wheeler Tennessee \ Unicoi

Colorado \ Summit Kansas \ Smith Nebraska \ York Tennessee \ Union

Colorado \ Teller Kansas \ Stafford New Mexico \ Bernalillo Tennessee \ Van Buren

Colorado \ Washington Kansas \ Stanton New Mexico \ Catron Tennessee \ Warren

Colorado \ Weld Kansas \ Stevens New Mexico \ Chaves Tennessee \ Washington81 State/County State/County State/County State/County

Colorado \ Yuma Kansas \ Sumner New Mexico \ Cibola Tennessee \ Wayne

Georgia \ Appling Kansas \ Thomas New Mexico \ Colfax Tennessee \ Weakley

Georgia \ Atkinson Kansas \ Trego New Mexico \ Curry Tennessee \ White

Georgia \ Bacon Kansas \ Wabaunsee New Mexico \ De Baca Tennessee \ Williamson

Georgia \ Baker Kansas \ Wallace New Mexico \ Dona Ana Tennessee \ Wilson

Georgia \ Baldwin Kansas \ Washington New Mexico \ Eddy Texas \ Anderson

Georgia \ Banks Kansas \ Wichita New Mexico \ Grant Texas \ Andrews

Georgia \ Barrow Kansas \ Wilson New Mexico \ Guadalupe Texas \ Angelina

Georgia \ Bartow Kansas \ Woodson New Mexico \ Harding Texas \ Aransas

Georgia \ Ben Hill Kansas \ Wyandotte New Mexico \ Hidalgo Texas \ Archer

Georgia \ Berrien Kentucky \ Adair New Mexico \ Lea Texas \ Armstrong

Georgia \ Bibb Kentucky \ Allen New Mexico \ Lincoln Texas \ Atascosa

Georgia \ Bleckley Kentucky \ Anderson New Mexico \ Los Alamos Texas \ Austin

Georgia \ Brantley Kentucky \ Ballard New Mexico \ Luna Texas \ Bailey

Georgia \ Brooks Kentucky \ Barren New Mexico \ McKinley Texas \ Bandera

Georgia \ Bryan Kentucky \ Bath New Mexico \ Mora Texas \ Bastrop

Georgia \ Bulloch Kentucky \ Bell New Mexico \ Otero Texas \ Baylor

Georgia \ Burke Kentucky \ Boone New Mexico \ Quay Texas \ Bee

Georgia \ Butts Kentucky \ Bourbon New Mexico \ Rio Arriba Texas \ Bell

Georgia \ Calhoun Kentucky \ Boyd New Mexico \ Roosevelt Texas \ Bexar

Georgia \ Camden Kentucky \ Boyle New Mexico \ Sandoval Texas \ Blanco

Georgia \ Candler Kentucky \ Bracken New Mexico \ San Juan Texas \ Borden

Georgia \ Carroll Kentucky \ Breathitt New Mexico \ San Miguel Texas \ Bosque

Georgia \ Catoosa Kentucky \ Breckinridge New Mexico \ Santa Fe Texas \ Bowie

Georgia \ Charlton Kentucky \ Bullitt New Mexico \ Sierra Texas \ Brazoria

Georgia \ Chatham Kentucky \ Butler New Mexico \ Socorro Texas \ Brazos

Georgia \ Chattahoochee Kentucky \ Caldwell New Mexico \ Taos Texas \ Brewster

Georgia \ Chattooga Kentucky \ Calloway New Mexico \ Torrance Texas \ Briscoe

Georgia \ Cherokee Kentucky \ Campbell New Mexico \ Union Texas \ Brooks

Georgia \ Clarke Kentucky \ Carlisle New Mexico \ Valencia Texas \ Brown

Georgia \ Clay Kentucky \ Carroll New York \ Albany Texas \ Burleson

Georgia \ Clayton Kentucky \ Carter New York \ Allegany Texas \ Burnet

Georgia \ Clinch Kentucky \ Casey New York \ Bronx Texas \ Caldwell

Georgia \ Cobb Kentucky \ Christian New York \ Broome Texas \ Calhoun

Georgia \ Coffee Kentucky \ Clark New York \ Cattaraugus Texas \ Callahan

Georgia \ Colquitt Kentucky \ Clay New York \ Cayuga Texas \ Cameron

Georgia \ Columbia Kentucky \ Clinton New York \ Chautauqua Texas \ Camp

Georgia \ Cook Kentucky \ Crittenden New York \ Chemung Texas \ Carson

Georgia \ Coweta Kentucky \ Cumberland New York \ Chenango Texas \ Cass

Georgia \ Crawford Kentucky \ Daviess New York \ Clinton Texas \ Castro

Georgia \ Crisp Kentucky \ Edmonson New York \ Columbia Texas \ Chambers

Georgia \ Dade Kentucky \ Elliott New York \ Cortland Texas \ Cherokee

Georgia \ Dawson Kentucky \ Estill New York \ Delaware Texas \ Childress

Georgia \ Decatur Kentucky \ Fayette New York \ Dutchess Texas \ Clay

Georgia \ DeKalb Kentucky \ Fleming New York \ Erie Texas \ Cochran

Georgia \ Dodge Kentucky \ Floyd New York \ Essex Texas \ Coke82 State/County State/County State/County State/County

Georgia \ Dooly Kentucky \ Franklin New York \ Franklin Texas \ Coleman

Georgia \ Dougherty Kentucky \ Fulton New York \ Fulton Texas \ Collin

Georgia \ Douglas Kentucky \ Gallatin New York \ Genesee Texas \ Collingsworth

Georgia \ Early Kentucky \ Garrard New York \ Greene Texas \ Colorado

Georgia \ Echols Kentucky \ Grant New York \ Hamilton Texas \ Comal

Georgia \ Effingham Kentucky \ Graves New York \ Herkimer Texas \ Comanche

Georgia \ Elbert Kentucky \ Grayson New York \ Jefferson Texas \ Concho

Georgia \ Emanuel Kentucky \ Green New York \ Kings Texas \ Cooke

Georgia \ Evans Kentucky \ Greenup New York \ Lewis Texas \ Coryell

Georgia \ Fannin Kentucky \ Hancock New York \ Livingston Texas \ Cottle

Georgia \ Fayette Kentucky \ Hardin New York \ Madison Texas \ Crane

Georgia \ Floyd Kentucky \ Harlan New York \ Monroe Texas \ Crockett

Georgia \ Forsyth Kentucky \ Harrison New York \ Montgomery Texas \ Crosby

Georgia \ Franklin Kentucky \ Hart New York \ Nassau Texas \ Culberson

Georgia \ Fulton Kentucky \ Henderson New York \ New York Texas \ Dallam

Georgia \ Gilmer Kentucky \ Henry New York \ Niagara Texas \ Dallas

Georgia \ Glascock Kentucky \ Hickman New York \ Oneida Texas \ Dawson

Georgia \ Glynn Kentucky \ Hopkins New York \ Onondaga Texas \ Deaf Smith

Georgia \ Gordon Kentucky \ Jackson New York \ Ontario Texas \ Delta

Georgia \ Grady Kentucky \ Jefferson New York \ Orange Texas \ Denton

Georgia \ Greene Kentucky \ Jessamine New York \ Orleans Texas \ DeWitt

Georgia \ Gwinnett Kentucky \ Johnson New York \ Oswego Texas \ Dickens

Georgia \ Habersham Kentucky \ Kenton New York \ Otsego Texas \ Dimmit

Georgia \ Hall Kentucky \ Knott New York \ Putnam Texas \ Donley

Georgia \ Hancock Kentucky \ Knox New York \ Queens Texas \ Duval

Georgia \ Haralson Kentucky \ Larue New York \ Rensselaer Texas \ Eastland

Georgia \ Harris Kentucky \ Laurel New York \ Richmond Texas \ Ector

Georgia \ Hart Kentucky \ Lawrence New York \ Rockland Texas \ Edwards

Georgia \ Heard Kentucky \ Lee New York \ St. Lawrence Texas \ Ellis

Georgia \ Henry Kentucky \ Leslie New York \ Saratoga Texas \ El Paso

Georgia \ Houston Kentucky \ Letcher New York \ Schenectady Texas \ Erath

Georgia \ Irwin Kentucky \ Lewis New York \ Schoharie Texas \ Falls

Georgia \ Jackson Kentucky \ Lincoln New York \ Schuyler Texas \ Fannin

Georgia \ Jasper Kentucky \ Livingston New York \ Seneca Texas \ Fayette

Georgia \ Jeff Davis Kentucky \ Logan New York \ Steuben Texas \ Fisher

Georgia \ Jefferson Kentucky \ Lyon New York \ Suffolk Texas \ Floyd

Georgia \ Jenkins Kentucky \ McCracken New York \ Sullivan Texas \ Foard

Georgia \ Johnson Kentucky \ McCreary New York \ Tioga Texas \ Fort Bend

Georgia \ Jones Kentucky \ McLean New York \ Tompkins Texas \ Franklin

Georgia \ Lamar Kentucky \ Madison New York \ Ulster Texas \ Freestone

Georgia \ Lanier Kentucky \ Magoffin New York \ Warren Texas \ Frio

Georgia \ Laurens Kentucky \ Marion New York \ Washington Texas \ Gaines

Georgia \ Lee Kentucky \ Marshall New York \ Wayne Texas \ Galveston

Georgia \ Liberty Kentucky \ Martin New York \ Westchester Texas \ Garza

Georgia \ Lincoln Kentucky \ Mason New York \ Wyoming Texas \ Gillespie

Georgia \ Long Kentucky \ Meade New York \ Yates Texas \ Glasscock83 State/County State/County State/County State/County

Georgia \ Lowndes Kentucky \ Menifee North Carolina \ Alamance Texas \ Goliad

Georgia \ Lumpkin Kentucky \ Mercer North Carolina \ Alexander Texas \ Gonzales

Georgia \ McDuffie Kentucky \ Metcalfe North Carolina \ Alleghany Texas \ Gray

Georgia \ McIntosh Kentucky \ Monroe North Carolina \ Anson Texas \ Grayson

Georgia \ Macon Kentucky \ Montgomery North Carolina \ Ashe Texas \ Gregg

Georgia \ Madison Kentucky \ Morgan North Carolina \ Avery Texas \ Grimes

Georgia \ Marion Kentucky \ Muhlenberg North Carolina \ Beaufort Texas \ Guadalupe

Georgia \ Meriwether Kentucky \ Nelson North Carolina \ Bertie Texas \ Hale

Georgia \ Miller Kentucky \ Nicholas North Carolina \ Bladen Texas \ Hall

Georgia \ Mitchell Kentucky \ Ohio North Carolina \ Brunswick Texas \ Hamilton

Georgia \ Monroe Kentucky \ Oldham North Carolina \ Buncombe Texas \ Hansford

Georgia \ Montgomery Kentucky \ Owen North Carolina \ Burke Texas \ Hardeman

Georgia \ Morgan Kentucky \ Owsley North Carolina \ Cabarrus Texas \ Hardin

Georgia \ Murray Kentucky \ Pendleton North Carolina \ Caldwell Texas \ Harris

Georgia \ Muscogee Kentucky \ Perry North Carolina \ Camden Texas \ Harrison

Georgia \ Newton Kentucky \ Pike North Carolina \ Carteret Texas \ Hartley

Georgia \ Oconee Kentucky \ Powell North Carolina \ Caswell Texas \ Haskell

Georgia \ Oglethorpe Kentucky \ Pulaski North Carolina \ Catawba Texas \ Hays

Georgia \ Paulding Kentucky \ Robertson North Carolina \ Chatham Texas \ Hemphill

Georgia \ Peach Kentucky \ Rockcastle North Carolina \ Cherokee Texas \ Henderson

Georgia \ Pickens Kentucky \ Rowan North Carolina \ Chowan Texas \ Hidalgo

Georgia \ Pierce Kentucky \ Russell North Carolina \ Clay Texas \ Hill

Georgia \ Pike Kentucky \ Scott North Carolina \ Cleveland Texas \ Hockley

Georgia \ Polk Kentucky \ Shelby North Carolina \ Columbus Texas \ Hood

Georgia \ Pulaski Kentucky \ Simpson North Carolina \ Craven Texas \ Hopkins

Georgia \ Putnam Kentucky \ Spencer North Carolina \ Cumberland Texas \ Houston

Georgia \ Quitman Kentucky \ Taylor North Carolina \ Currituck Texas \ Howard

Georgia \ Rabun Kentucky \ Todd North Carolina \ Dare Texas \ Hudspeth

Georgia \ Randolph Kentucky \ Trigg North Carolina \ Davidson Texas \ Hunt

Georgia \ Richmond Kentucky \ Trimble North Carolina \ Davie Texas \ Hutchinson

Georgia \ Rockdale Kentucky \ Union North Carolina \ Duplin Texas \ Irion

Georgia \ Schley Kentucky \ Warren North Carolina \ Durham Texas \ Jack

Georgia \ Screven Kentucky \ Washington North Carolina \ Edgecombe Texas \ Jackson

Georgia \ Seminole Kentucky \ Wayne North Carolina \ Forsyth Texas \ Jasper

Georgia \ Spalding Kentucky \ Webster North Carolina \ Franklin Texas \ Jeff Davis

Georgia \ Stephens Kentucky \ Whitley North Carolina \ Gaston Texas \ Jefferson

Georgia \ Stewart Kentucky \ Wolfe North Carolina \ Gates Texas \ Jim Hogg

Georgia \ Sumter Kentucky \ Woodford North Carolina \ Graham Texas \ Jim Wells

Georgia \ Talbot Michigan \ Alcona North Carolina \ Granville Texas \ Johnson

Georgia \ Taliaferro Michigan \ Alger North Carolina \ Greene Texas \ Jones

Georgia \ Tattnall Michigan \ Allegan North Carolina \ Guilford Texas \ Karnes

Georgia \ Taylor Michigan \ Alpena North Carolina \ Halifax Texas \ Kaufman

Georgia \ Telfair Michigan \ Antrim North Carolina \ Harnett Texas \ Kendall

Georgia \ Terrell Michigan \ Arenac North Carolina \ Haywood Texas \ Kenedy

Georgia \ Thomas Michigan \ Baraga North Carolina \ Henderson Texas \ Kent

Georgia \ Tift Michigan \ Barry North Carolina \ Hertford Texas \ Kerr84 State/County State/County State/County State/County

Georgia \ Toombs Michigan \ Bay North Carolina \ Hoke Texas \ Kimble

Georgia \ Towns Michigan \ Benzie North Carolina \ Hyde Texas \ King

Georgia \ Treutlen Michigan \ Berrien North Carolina \ Iredell Texas \ Kinney

Georgia \ Troup Michigan \ Branch North Carolina \ Jackson Texas \ Kleberg

Georgia \ Turner Michigan \ Calhoun North Carolina \ Johnston Texas \ Knox

Georgia \ Twiggs Michigan \ Cass North Carolina \ Jones Texas \ Lamar

Georgia \ Union Michigan \ Charlevoix North Carolina \ Lee Texas \ Lamb

Georgia \ Upson Michigan \ Cheboygan North Carolina \ Lenoir Texas \ Lampasas

Georgia \ Walker Michigan \ Chippewa North Carolina \ Lincoln Texas \ La Salle

Georgia \ Walton Michigan \ Clare North Carolina \ McDowell Texas \ Lavaca

Georgia \ Ware Michigan \ Clinton North Carolina \ Macon Texas \ Lee

Georgia \ Warren Michigan \ Crawford North Carolina \ Madison Texas \ Leon

Georgia \ Washington Michigan \ Delta North Carolina \ Martin Texas \ Liberty

Georgia \ Wayne Michigan \ Dickinson North Carolina \ Mecklenburg Texas \ Limestone

Georgia \ Webster Michigan \ Eaton North Carolina \ Mitchell Texas \ Lipscomb

Georgia \ Wheeler Michigan \ Emmet North Carolina \ Montgomery Texas \ Live Oak

Georgia \ White Michigan \ Genesee North Carolina \ Moore Texas \ Llano

Georgia \ Whitfield Michigan \ Gladwin North Carolina \ Nash Texas \ Loving

Georgia \ Wilcox Michigan \ Gogebic North Carolina \ New Hanover Texas \ Lubbock

Georgia \ Wilkes Michigan \ Grand Traverse North Carolina \ Northampton Texas \ Lynn

Georgia \ Wilkinson Michigan \ Gratiot North Carolina \ Onslow Texas \ McCulloch

Georgia \ Worth Michigan \ Hillsdale North Carolina \ Orange Texas \ McLennan

Idaho \ Ada Michigan \ Houghton North Carolina \ Pamlico Texas \ McMullen

Idaho \ Adams Michigan \ Huron North Carolina \ Pasquotank Texas \ Madison

Idaho \ Bannock Michigan \ Ingham North Carolina \ Pender Texas \ Marion

Idaho \ Bear Lake Michigan \ Ionia North Carolina \ Perquimans Texas \ Martin

Idaho \ Benewah Michigan \ Iosco North Carolina \ Person Texas \ Mason

Idaho \ Bingham Michigan \ Iron North Carolina \ Pitt Texas \ Matagorda

Idaho \ Blaine Michigan \ Isabella North Carolina \ Polk Texas \ Maverick

Idaho \ Boise Michigan \ Jackson North Carolina \ Randolph Texas \ Medina

Idaho \ Bonner Michigan \ Kalamazoo North Carolina \ Richmond Texas \ Menard

Idaho \ Bonneville Michigan \ Kalkaska North Carolina \ Robeson Texas \ Midland

Idaho \ Boundary Michigan \ Kent North Carolina \ Rockingham Texas \ Milam

Idaho \ Butte Michigan \ Keweenaw North Carolina \ Rowan Texas \ Mills

Idaho \ Camas Michigan \ Lake North Carolina \ Rutherford Texas \ Mitchell

Idaho \ Canyon Michigan \ Lapeer North Carolina \ Sampson Texas \ Montague

Idaho \ Caribou Michigan \ Leelanau North Carolina \ Scotland Texas \ Montgomery

Idaho \ Cassia Michigan \ Lenawee North Carolina \ Stanly Texas \ Moore

Idaho \ Clark Michigan \ Livingston North Carolina \ Stokes Texas \ Morris

Idaho \ Clearwater Michigan \ Luce North Carolina \ Surry Texas \ Motley

Idaho \ Custer Michigan \ Mackinac North Carolina \ Swain Texas \ Nacogdoches

Idaho \ Elmore Michigan \ Macomb North Carolina \ Transylvania Texas \ Navarro

Idaho \ Franklin Michigan \ Manistee North Carolina \ Tyrrell Texas \ Newton

Idaho \ Fremont Michigan \ Marquette North Carolina \ Union Texas \ Nolan

Idaho \ Gem Michigan \ Mason North Carolina \ Vance Texas \ Nueces

Idaho \ Gooding Michigan \ Mecosta North Carolina \ Wake Texas \ Ochiltree85 State/County State/County State/County State/County

Idaho \ Idaho Michigan \ Menominee North Carolina \ Warren Texas \ Oldham

Idaho \ Jefferson Michigan \ Midland North Carolina \ Washington Texas \ Orange

Idaho \ Jerome Michigan \ Missaukee North Carolina \ Watauga Texas \ Palo Pinto

Idaho \ Kootenai Michigan \ Monroe North Carolina \ Wayne Texas \ Panola

Idaho \ Latah Michigan \ Montcalm North Carolina \ Wilkes Texas \ Parker

Idaho \ Lemhi Michigan \ Montmorency North Carolina \ Wilson Texas \ Parmer

Idaho \ Lewis Michigan \ Muskegon North Carolina \ Yadkin Texas \ Pecos

Idaho \ Lincoln Michigan \ Newaygo North Carolina \ Yancey Texas \ Polk

Idaho \ Madison Michigan \ Oakland North Dakota \ Adams Texas \ Potter

Idaho \ Minidoka Michigan \ Oceana North Dakota \ Barnes Texas \ Presidio

Idaho \ Nez Perce Michigan \ Ogemaw North Dakota \ Benson Texas \ Rains

Idaho \ Oneida Michigan \ Ontonagon North Dakota \ Billings Texas \ Randall

Idaho \ Owyhee Michigan \ Osceola North Dakota \ Bottineau Texas \ Reagan

Idaho \ Payette Michigan \ Oscoda North Dakota \ Bowman Texas \ Real

Idaho \ Power Michigan \ Otsego North Dakota \ Burke Texas \ Red River

Idaho \ Shoshone Michigan \ Ottawa North Dakota \ Burleigh Texas \ Reeves

Idaho \ Teton Michigan \ Presque Isle North Dakota \ Cass Texas \ Refugio

Idaho \ Twin Falls Michigan \ Roscommon North Dakota \ Cavalier Texas \ Roberts

Idaho \ Valley Michigan \ Saginaw North Dakota \ Dickey Texas \ Robertson

Idaho \ Washington Michigan \ St. Clair North Dakota \ Divide Texas \ Rockwall

Illinois \ Adams Michigan \ St. Joseph North Dakota \ Dunn Texas \ Runnels

Illinois \ Alexander Michigan \ Sanilac North Dakota \ Eddy Texas \ Rusk

Illinois \ Bond Michigan \ Schoolcraft North Dakota \ Emmons Texas \ Sabine

Illinois \ Boone Michigan \ Shiawassee North Dakota \ Foster Texas \ San Augustine

Illinois \ Brown Michigan \ Tuscola North Dakota \ Golden Valley Texas \ San Jacinto

Illinois \ Bureau Michigan \ Van Buren North Dakota \ Grand Forks Texas \ San Patricio

Illinois \ Calhoun Michigan \ Washtenaw North Dakota \ Grant Texas \ San Saba

Illinois \ Carroll Michigan \ Wayne North Dakota \ Griggs Texas \ Schleicher

Illinois \ Cass Michigan \ Wexford North Dakota \ Hettinger Texas \ Scurry

Illinois \ Champaign Minnesota \ Aitkin North Dakota \ Kidder Texas \ Shackelford

Illinois \ Christian Minnesota \ Anoka North Dakota \ LaMoure Texas \ Shelby

Illinois \ Clark Minnesota \ Becker North Dakota \ Logan Texas \ Sherman

Illinois \ Clay Minnesota \ Beltrami North Dakota \ McHenry Texas \ Smith

Illinois \ Clinton Minnesota \ Benton North Dakota \ McIntosh Texas \ Somervell

Illinois \ Coles Minnesota \ Big Stone North Dakota \ McKenzie Texas \ Starr

Illinois \ Cook Minnesota \ Blue Earth North Dakota \ McLean Texas \ Stephens

Illinois \ Crawford Minnesota \ Brown North Dakota \ Mercer Texas \ Sterling

Illinois \ Cumberland Minnesota \ Carlton North Dakota \ Morton Texas \ Stonewall

Illinois \ De Kalb Minnesota \ Carver North Dakota \ Mountrail Texas \ Sutton

Illinois \ De Witt Minnesota \ Cass North Dakota \ Nelson Texas \ Swisher

Illinois \ Douglas Minnesota \ Chippewa North Dakota \ Oliver Texas \ Tarrant

Illinois \ Du Page Minnesota \ Chisago North Dakota \ Pembina Texas \ Taylor

Illinois \ Edgar Minnesota \ Clay North Dakota \ Pierce Texas \ Terrell

Illinois \ Edwards Minnesota \ Clearwater North Dakota \ Ramsey Texas \ Terry

Illinois \ Effingham Minnesota \ Cook North Dakota \ Ransom Texas \ Throckmorton

Illinois \ Fayette Minnesota \ Cottonwood North Dakota \ Renville Texas \ Titus86 State/County State/County State/County State/County

Illinois \ Ford Minnesota \ Crow Wing North Dakota \ Richland Texas \ Tom Green

Illinois \ Franklin Minnesota \ Dakota North Dakota \ Rolette Texas \ Travis

Illinois \ Fulton Minnesota \ Dodge North Dakota \ Sargent Texas \ Trinity

Illinois \ Gallatin Minnesota \ Douglas North Dakota \ Sheridan Texas \ Tyler

Illinois \ Greene Minnesota \ Faribault North Dakota \ Sioux Texas \ Upshur

Illinois \ Grundy Minnesota \ Fillmore North Dakota \ Slope Texas \ Upton

Illinois \ Hamilton Minnesota \ Freeborn North Dakota \ Stark Texas \ Uvalde

Illinois \ Hancock Minnesota \ Goodhue North Dakota \ Steele Texas \ Val Verde

Illinois \ Hardin Minnesota \ Grant North Dakota \ Stutsman Texas \ Van Zandt

Illinois \ Henderson Minnesota \ Hennepin North Dakota \ Towner Texas \ Victoria

Illinois \ Henry Minnesota \ Houston North Dakota \ Traill Texas \ Walker

Illinois \ Iroquois Minnesota \ Hubbard North Dakota \ Walsh Texas \ Waller

Illinois \ Jackson Minnesota \ Isanti North Dakota \ Ward Texas \ Ward

Illinois \ Jasper Minnesota \ Itasca North Dakota \ Wells Texas \ Washington

Illinois \ Jefferson Minnesota \ Jackson North Dakota \ Williams Texas \ Webb

Illinois \ Jersey Minnesota \ Kanabec Ohio \ Adams Texas \ Wharton

Illinois \ Jo Daviess Minnesota \ Kandiyohi Ohio \ Allen Texas \ Wheeler

Illinois \ Johnson Minnesota \ Kittson Ohio \ Ashland Texas \ Wichita

Illinois \ Kane Minnesota \ Koochiching Ohio \ Ashtabula Texas \ Wilbarger

Illinois \ Kankakee Minnesota \ Lac qui Parle Ohio \ Athens Texas \ Willacy

Illinois \ Kendall Minnesota \ Lake Ohio \ Auglaize Texas \ Williamson

Illinois \ Knox Minnesota \ Lake of the

Woods Ohio \ Belmont Texas \ Wilson

Illinois \ Lake Minnesota \ Le Sueur Ohio \ Brown Texas \ Winkler

Illinois \ La Salle Minnesota \ Lincoln Ohio \ Butler Texas \ Wise

Illinois \ Lawrence Minnesota \ Lyon Ohio \ Carroll Texas \ Wood

Illinois \ Lee Minnesota \ McLeod Ohio \ Champaign Texas \ Yoakum

Illinois \ Livingston Minnesota \ Mahnomen Ohio \ Clark Texas \ Young

Illinois \ Logan Minnesota \ Marshall Ohio \ Clermont Texas \ Zapata

Illinois \ McDonough Minnesota \ Martin Ohio \ Clinton Texas \ Zavala

Illinois \ McHenry Minnesota \ Meeker Ohio \ Columbiana Virginia \ Accomack

Illinois \ McLean Minnesota \ Mille Lacs Ohio \ Coshocton Virginia \ Albemarle

Illinois \ Macon Minnesota \ Morrison Ohio \ Crawford Virginia \ Alleghany

Illinois \ Macoupin Minnesota \ Mower Ohio \ Cuyahoga Virginia \ Amelia

Illinois \ Madison Minnesota \ Murray Ohio \ Darke Virginia \ Amherst

Illinois \ Marion Minnesota \ Nicollet Ohio \ Defiance Virginia \ Appomattox

Illinois \ Marshall Minnesota \ Nobles Ohio \ Delaware Virginia \ Arlington

Illinois \ Mason Minnesota \ Norman Ohio \ Erie Virginia \ Augusta

Illinois \ Massac Minnesota \ Olmsted Ohio \ Fairfield Virginia \ Bath

Illinois \ Menard Minnesota \ Otter Tail Ohio \ Fayette Virginia \ Bedford

Illinois \ Mercer Minnesota \ Pennington Ohio \ Franklin Virginia \ Bland

Illinois \ Monroe Minnesota \ Pine Ohio \ Fulton Virginia \ Botetourt

Illinois \ Montgomery Minnesota \ Pipestone Ohio \ Gallia Virginia \ Brunswick

Illinois \ Morgan Minnesota \ Polk Ohio \ Geauga Virginia \ Buchanan

Illinois \ Moultrie Minnesota \ Pope Ohio \ Greene Virginia \ Buckingham

Illinois \ Ogle Minnesota \ Ramsey Ohio \ Guernsey Virginia \ Campbell87 State/County State/County State/County State/County

Illinois \ Peoria Minnesota \ Red Lake Ohio \ Hamilton Virginia \ Caroline

Illinois \ Perry Minnesota \ Redwood Ohio \ Hancock Virginia \ Carroll

Illinois \ Piatt Minnesota \ Renville Ohio \ Hardin Virginia \ Charles City

Illinois \ Pike Minnesota \ Rice Ohio \ Harrison Virginia \ Charlotte

Illinois \ Pope Minnesota \ Rock Ohio \ Henry Virginia \ Chesterfield

Illinois \ Pulaski Minnesota \ Roseau Ohio \ Highland Virginia \ Clarke

Illinois \ Putnam Minnesota \ St. Louis Ohio \ Hocking Virginia \ Craig

Illinois \ Randolph Minnesota \ Scott Ohio \ Holmes Virginia \ Culpeper

Illinois \ Richland Minnesota \ Sherburne Ohio \ Huron Virginia \ Cumberland

Illinois \ Rock Island Minnesota \ Sibley Ohio \ Jackson Virginia \ Dickenson

Illinois \ St. Clair Minnesota \ Stearns Ohio \ Jefferson Virginia \ Dinwiddie

Illinois \ Saline Minnesota \ Steele Ohio \ Knox Virginia \ Essex

Illinois \ Sangamon Minnesota \ Stevens Ohio \ Lake Virginia \ Fairfax

Illinois \ Schuyler Minnesota \ Swift Ohio \ Lawrence Virginia \ Fauquier

Illinois \ Scott Minnesota \ Todd Ohio \ Licking Virginia \ Floyd

Illinois \ Shelby Minnesota \ Traverse Ohio \ Logan Virginia \ Fluvanna

Illinois \ Stark Minnesota \ Wabasha Ohio \ Lorain Virginia \ Franklin

Illinois \ Stephenson Minnesota \ Wadena Ohio \ Lucas Virginia \ Frederick

Illinois \ Tazewell Minnesota \ Waseca Ohio \ Madison Virginia \ Giles

Illinois \ Union Minnesota \ Washington Ohio \ Mahoning Virginia \ Gloucester

Illinois \ Vermilion Minnesota \ Watonwan Ohio \ Marion Virginia \ Goochland

Illinois \ Wabash Minnesota \ Wilkin Ohio \ Medina Virginia \ Grayson

Illinois \ Warren Minnesota \ Winona Ohio \ Meigs Virginia \ Greene

Illinois \ Washington Minnesota \ Wright Ohio \ Mercer Virginia \ Greensville

Illinois \ Wayne Minnesota \ Yellow Medicine Ohio \ Miami Virginia \ Halifax

Illinois \ White Mississippi \ Adams Ohio \ Monroe Virginia \ Hanover

Illinois \ Whiteside Mississippi \ Alcorn Ohio \ Montgomery Virginia \ Henrico

Illinois \ Will Mississippi \ Amite Ohio \ Morgan Virginia \ Henry

Illinois \ Williamson Mississippi \ Attala Ohio \ Morrow Virginia \ Highland

Illinois \ Winnebago Mississippi \ Benton Ohio \ Muskingum Virginia \ Isle of Wight

Illinois \ Woodford Mississippi \ Bolivar Ohio \ Noble Virginia \ James City

Indiana \ Adams Mississippi \ Calhoun Ohio \ Ottawa Virginia \ King and Queen

Indiana \ Allen Mississippi \ Carroll Ohio \ Paulding Virginia \ King George

Indiana \ Bartholomew Mississippi \ Chickasaw Ohio \ Perry Virginia \ King William

Indiana \ Benton Mississippi \ Choctaw Ohio \ Pickaway Virginia \ Lancaster

Indiana \ Blackford Mississippi \ Claiborne Ohio \ Pike Virginia \ Lee

Indiana \ Boone Mississippi \ Clarke Ohio \ Portage Virginia \ Loudoun

Indiana \ Brown Mississippi \ Clay Ohio \ Preble Virginia \ Louisa

Indiana \ Carroll Mississippi \ Coahoma Ohio \ Putnam Virginia \ Lunenburg

Indiana \ Cass Mississippi \ Copiah Ohio \ Richland Virginia \ Madison

Indiana \ Clark Mississippi \ Covington Ohio \ Ross Virginia \ Mathews

Indiana \ Clay Mississippi \ DeSoto Ohio \ Sandusky Virginia \ Mecklenburg

Indiana \ Clinton Mississippi \ Forrest Ohio \ Scioto Virginia \ Middlesex

Indiana \ Crawford Mississippi \ Franklin Ohio \ Seneca Virginia \ Montgomery

Indiana \ Daviess Mississippi \ George Ohio \ Shelby Virginia \ Nelson

Indiana \ Dearborn Mississippi \ Greene Ohio \ Stark Virginia \ New Kent88 State/County State/County State/County State/County

Indiana \ Decatur Mississippi \ Grenada Ohio \ Summit Virginia \ Northampton

Indiana \ DeKalb Mississippi \ Hancock Ohio \ Trumbull Virginia \ Northumberland

Indiana \ Delaware Mississippi \ Harrison Ohio \ Tuscarawas Virginia \ Nottoway

Indiana \ Dubois Mississippi \ Hinds Ohio \ Union Virginia \ Orange

Indiana \ Elkhart Mississippi \ Holmes Ohio \ Van Wert Virginia \ Page

Indiana \ Fayette Mississippi \ Humphreys Ohio \ Vinton Virginia \ Patrick

Indiana \ Floyd Mississippi \ Issaquena Ohio \ Warren Virginia \ Pittsylvania

Indiana \ Fountain Mississippi \ Itawamba Ohio \ Washington Virginia \ Powhatan

Indiana \ Franklin Mississippi \ Jackson Ohio \ Wayne Virginia \ Prince Edward

Indiana \ Fulton Mississippi \ Jasper Ohio \ Williams Virginia \ Prince George

Indiana \ Gibson Mississippi \ Jefferson Ohio \ Wood Virginia \ Prince William

Indiana \ Grant Mississippi \ Jefferson Davis Ohio \ Wyandot Virginia \ Pulaski

Indiana \ Greene Mississippi \ Jones Oregon \ Baker Virginia \ Rappahannock

Indiana \ Hamilton Mississippi \ Kemper Oregon \ Benton Virginia \ Richmond

Indiana \ Hancock Mississippi \ Lafayette Oregon \ Clackamas Virginia \ Roanoke

Indiana \ Harrison Mississippi \ Lamar Oregon \ Clatsop Virginia \ Rockbridge

Indiana \ Hendricks Mississippi \ Lauderdale Oregon \ Columbia Virginia \ Rockingham

Indiana \ Henry Mississippi \ Lawrence Oregon \ Coos Virginia \ Russell

Indiana \ Howard Mississippi \ Leake Oregon \ Crook Virginia \ Scott

Indiana \ Huntington Mississippi \ Lee Oregon \ Curry Virginia \ Shenandoah

Indiana \ Jackson Mississippi \ Leflore Oregon \ Deschutes Virginia \ Smyth

Indiana \ Jasper Mississippi \ Lincoln Oregon \ Douglas Virginia \ Southampton

Indiana \ Jay Mississippi \ Lowndes Oregon \ Gilliam Virginia \ Spotsylvania

Indiana \ Jefferson Mississippi \ Madison Oregon \ Grant Virginia \ Stafford

Indiana \ Jennings Mississippi \ Marion Oregon \ Harney Virginia \ Surry

Indiana \ Johnson Mississippi \ Marshall Oregon \ Hood River Virginia \ Sussex

Indiana \ Knox Mississippi \ Monroe Oregon \ Jackson Virginia \ Tazewell

Indiana \ Kosciusko Mississippi \ Montgomery Oregon \ Jefferson Virginia \ Warren

Indiana \ LaGrange Mississippi \ Neshoba Oregon \ Josephine Virginia \ Washington

Indiana \ Lake Mississippi \ Newton Oregon \ Klamath Virginia \ Westmoreland

Indiana \ LaPorte Mississippi \ Noxubee Oregon \ Lake Virginia \ Wise

Indiana \ Lawrence Mississippi \ Oktibbeha Oregon \ Lane Virginia \ Wythe

Indiana \ Madison Mississippi \ Panola Oregon \ Lincoln Virginia \ York

Indiana \ Marion Mississippi \ Pearl River Oregon \ Linn Virginia \ Chesapeake City

Indiana \ Marshall Mississippi \ Perry Oregon \ Malheur Virginia \ Suffolk

Indiana \ Martin Mississippi \ Pike Oregon \ Marion Virginia \ Virginia Beach City

Indiana \ Miami Mississippi \ Pontotoc Oregon \ Morrow Wisconsin \ Adams

Indiana \ Monroe Mississippi \ Prentiss Oregon \ Multnomah Wisconsin \ Ashland

Indiana \ Montgomery Mississippi \ Quitman Oregon \ Polk Wisconsin \ Barron

Indiana \ Morgan Mississippi \ Rankin Oregon \ Sherman Wisconsin \ Bayfield

Indiana \ Newton Mississippi \ Scott Oregon \ Tillamook Wisconsin \ Brown

Indiana \ Noble Mississippi \ Sharkey Oregon \ Umatilla Wisconsin \ Buffalo

Indiana \ Ohio Mississippi \ Simpson Oregon \ Union Wisconsin \ Burnett

Indiana \ Orange Mississippi \ Smith Oregon \ Wallowa Wisconsin \ Calumet

Indiana \ Owen Mississippi \ Stone Oregon \ Wasco Wisconsin \ Chippewa

Indiana \ Parke Mississippi \ Sunflower Oregon \ Washington Wisconsin \ Clark89 State/County State/County State/County State/County

Indiana \ Perry Mississippi \ Tallahatchie Oregon \ Wheeler Wisconsin \ Columbia

Indiana \ Pike Mississippi \ Tate Oregon \ Yamhill Wisconsin \ Crawford

Indiana \ Porter Mississippi \ Tippah Pennsylvania \ Adams Wisconsin \ Dane

Indiana \ Posey Mississippi \ Tishomingo Pennsylvania \ Allegheny Wisconsin \ Dodge

Indiana \ Pulaski Mississippi \ Tunica Pennsylvania \ Armstrong Wisconsin \ Door

Indiana \ Putnam Mississippi \ Union Pennsylvania \ Beaver Wisconsin \ Douglas

Indiana \ Randolph Mississippi \ Walthall Pennsylvania \ Bedford Wisconsin \ Dunn

Indiana \ Ripley Mississippi \ Warren Pennsylvania \ Berks Wisconsin \ Eau Claire

Indiana \ Rush Mississippi \ Washington Pennsylvania \ Blair Wisconsin \ Florence

Indiana \ St. Joseph Mississippi \ Wayne Pennsylvania \ Bradford Wisconsin \ Fond du Lac

Indiana \ Scott Mississippi \ Webster Pennsylvania \ Bucks Wisconsin \ Forest

Indiana \ Shelby Mississippi \ Wilkinson Pennsylvania \ Butler Wisconsin \ Grant

Indiana \ Spencer Mississippi \ Winston Pennsylvania \ Cambria Wisconsin \ Green

Indiana \ Starke Mississippi \ Yalobusha Pennsylvania \ Cameron Wisconsin \ Green Lake

Indiana \ Steuben Mississippi \ Yazoo Pennsylvania \ Carbon Wisconsin \ Iowa

Indiana \ Sullivan Missouri \ Adair Pennsylvania \ Centre Wisconsin \ Iron

Indiana \ Switzerland Missouri \ Andrew Pennsylvania \ Chester Wisconsin \ Jackson

Indiana \ Tippecanoe Missouri \ Atchison Pennsylvania \ Clarion Wisconsin \ Jefferson

Indiana \ Tipton Missouri \ Audrain Pennsylvania \ Clearfield Wisconsin \ Juneau

Indiana \ Union Missouri \ Barry Pennsylvania \ Clinton Wisconsin \ Kenosha

Indiana \ Vanderburgh Missouri \ Barton Pennsylvania \ Columbia Wisconsin \ Kewaunee

Indiana \ Vermillion Missouri \ Bates Pennsylvania \ Crawford Wisconsin \ La Crosse

Indiana \ Vigo Missouri \ Benton Pennsylvania \ Cumberland Wisconsin \ Lafayette

Indiana \ Wabash Missouri \ Bollinger Pennsylvania \ Dauphin Wisconsin \ Langlade

Indiana \ Warren Missouri \ Boone Pennsylvania \ Delaware Wisconsin \ Lincoln

Indiana \ Warrick Missouri \ Buchanan Pennsylvania \ Elk Wisconsin \ Manitowoc

Indiana \ Washington Missouri \ Butler Pennsylvania \ Erie Wisconsin \ Marathon

Indiana \ Wayne Missouri \ Caldwell Pennsylvania \ Fayette Wisconsin \ Marinette

Indiana \ Wells Missouri \ Callaway Pennsylvania \ Forest Wisconsin \ Marquette

Indiana \ White Missouri \ Camden Pennsylvania \ Franklin Wisconsin \ Menominee

Indiana \ Whitley Missouri \ Cape Girardeau Pennsylvania \ Fulton Wisconsin \ Milwaukee

Iowa \ Adair Missouri \ Carroll Pennsylvania \ Greene Wisconsin \ Monroe

Iowa \ Adams Missouri \ Carter Pennsylvania \ Huntingdon Wisconsin \ Oconto

Iowa \ Allamakee Missouri \ Cass Pennsylvania \ Indiana Wisconsin \ Oneida

Iowa \ Appanoose Missouri \ Cedar Pennsylvania \ Jefferson Wisconsin \ Outagamie

Iowa \ Audubon Missouri \ Chariton Pennsylvania \ Juniata Wisconsin \ Ozaukee

Iowa \ Benton Missouri \ Christian Pennsylvania \ Lackawanna Wisconsin \ Pepin

Iowa \ Black Hawk Missouri \ Clark Pennsylvania \ Lancaster Wisconsin \ Pierce

Iowa \ Boone Missouri \ Clay Pennsylvania \ Lawrence Wisconsin \ Polk

Iowa \ Bremer Missouri \ Clinton Pennsylvania \ Lebanon Wisconsin \ Portage

Iowa \ Buchanan Missouri \ Cole Pennsylvania \ Lehigh Wisconsin \ Price

Iowa \ Buena Vista Missouri \ Cooper Pennsylvania \ Luzerne Wisconsin \ Racine

Iowa \ Butler Missouri \ Crawford Pennsylvania \ Lycoming Wisconsin \ Richland

Iowa \ Calhoun Missouri \ Dade Pennsylvania \ McKean Wisconsin \ Rock

Iowa \ Carroll Missouri \ Dallas Pennsylvania \ Mercer Wisconsin \ Rusk

Iowa \ Cass Missouri \ Daviess Pennsylvania \ Mifflin Wisconsin \ St. Croix90 State/County State/County State/County State/County

Iowa \ Cedar Missouri \ DeKalb Pennsylvania \ Monroe Wisconsin \ Sauk

Iowa \ Cerro Gordo Missouri \ Dent Pennsylvania \ Montgomery Wisconsin \ Sawyer

Iowa \ Cherokee Missouri \ Douglas Pennsylvania \ Montour Wisconsin \ Shawano

Iowa \ Chickasaw Missouri \ Dunklin Pennsylvania \ Northampton Wisconsin \ Sheboygan

Iowa \ Clarke Missouri \ Franklin Pennsylvania \ Northumberland Wisconsin \ Taylor

Iowa \ Clay Missouri \ Gasconade Pennsylvania \ Perry Wisconsin \ Trempealeau

Iowa \ Clayton Missouri \ Gentry Pennsylvania \ Philadelphia Wisconsin \ Vernon

Iowa \ Clinton Missouri \ Greene Pennsylvania \ Pike Wisconsin \ Vilas

Iowa \ Crawford Missouri \ Grundy Pennsylvania \ Potter Wisconsin \ Walworth

Iowa \ Dallas Missouri \ Harrison Pennsylvania \ Schuylkill Wisconsin \ Washburn

Iowa \ Davis Missouri \ Henry Pennsylvania \ Snyder Wisconsin \ Washington

Iowa \ Decatur Missouri \ Hickory Pennsylvania \ Somerset Wisconsin \ Waukesha

Iowa \ Delaware Missouri \ Holt Pennsylvania \ Sullivan Wisconsin \ Waupaca

Iowa \ Des Moines Missouri \ Howard Pennsylvania \ Susquehanna Wisconsin \ Waushara

Iowa \ Dickinson Missouri \ Howell Pennsylvania \ Tioga Wisconsin \ Winnebago

Iowa \ Dubuque Missouri \ Iron Pennsylvania \ Union Wisconsin \ Wood

Iowa \ Emmet Missouri \ Jackson Pennsylvania \ Venango Wyoming \ Albany

Iowa \ Fayette Missouri \ Jasper Pennsylvania \ Warren Wyoming \ Big Horn

Iowa \ Floyd Missouri \ Jefferson Pennsylvania \ Washington Wyoming \ Campbell

Iowa \ Franklin Missouri \ Johnson Pennsylvania \ Wayne Wyoming \ Carbon

Iowa \ Fremont Missouri \ Knox Pennsylvania \ Westmoreland Wyoming \ Converse

Iowa \ Greene Missouri \ Laclede Pennsylvania \ Wyoming Wyoming \ Crook

Iowa \ Grundy Missouri \ Lafayette Pennsylvania \ York Wyoming \ Fremont

Iowa \ Guthrie Missouri \ Lawrence South Dakota\ Aurora Wyoming \ Goshen

Iowa \ Hamilton Missouri \ Lewis South Dakota \ Beadle Wyoming \ Hot Springs

Iowa \ Hancock Missouri \ Lincoln South Dakota \ Bennett Wyoming \ Johnson

Iowa \ Hardin Missouri \ Linn South Dakota \ Bon Homme Wyoming \ Laramie

Iowa \ Harrison Missouri \ Livingston South Dakota \ Brookings Wyoming \ Lincoln

Iowa \ Henry Missouri \ McDonald South Dakota \ Brown Wyoming \ Natrona

Iowa \ Howard Missouri \ Macon South Dakota \ Brule Wyoming \ Niobrara

Iowa \ Humboldt Missouri \ Madison South Dakota\ Buffalo Wyoming \ Park

Iowa \ Ida Missouri \ Maries South Dakota \ Butte Wyoming \ Platte

Iowa \ Iowa Missouri \ Marion South Dakota \ Campbell Wyoming \ Sheridan

Iowa \ Jackson Missouri \ Mercer South Dakota \ Charles Mix Wyoming \ Sublette

Iowa \ Jasper Missouri \ Miller South Dakota \ Clark Wyoming \ Sweetwater

Iowa \ Jefferson Missouri \ Mississippi South Dakota \ Clay Wyoming \ Teton

Iowa \ Johnson Missouri \ Moniteau South Dakota \ Codington Wyoming \ Uinta

Iowa \ Jones Missouri \ Monroe South Dakota \ Corson Wyoming \ Washakie

Iowa \ Keokuk Missouri \ Montgomery South Dakota \ Custer Wyoming \ Weston

Iowa \ Kossuth Missouri \ Morgan South Dakota \ Davison

Iowa \ Lee Missouri \ New Madrid South Dakota \ Day

Iowa \ Linn Missouri \ Newton South Dakota \ Deuel

Iowa \ Louisa Missouri \ Nodaway South Dakota\ Dewey

Iowa \ Lucas Missouri \ Oregon South Dakota \ Douglas

Iowa \ Lyon Missouri \ Osage South Dakota \ Edmunds

Iowa \ Madison Missouri \ Ozark South Dakota \ Fall River91 State/County State/County State/County State/County

Iowa \ Mahaska Missouri \ Pemiscot South Dakota \ Faulk92 

## Appendix C

. Economic Impact Report submitted by Sygenta93949596979899100101102103104105106107108109110111112113114115116117118119120121122123124125126127128129130131132133 

## Appendix D

. Pollen

- mediated gene flow report submitted by Syngenta

Minimization of Pollen

- Mediated Gene Flow from Corn Amylase Corn through Planting Border Rows

Over the past several years a number of studies have been published on the distribution of corn pollen and the potential transgenes to fertilize other corn. The USDA APHIS BRS reviewed this topic in their Draft Programmatic Environmental Impact Statement on the Introduction of Genetically Engineered Organisms (USDA 2007). Syngenta’s analysis of pollen

- mediated gene flow (PMGF) from corn amylase corn (CA) to non

- corn amylase corn (NCA) is built upon several published studies, i.e., Jarosz et al. (2003), Je mison and Veyda (2001), Ma (2005), Ma et al. (2004) and Westgate et al. (2003). However, emphasis was placed upon the work of Dr. Ma, Eastern Cereal and Oilseed Research Center, Agriculture and Agri

- Food Canada ( http://www.isb.vt.edu/articles/feb0502.htm ). Dr. Ma’s findings were consistent with or more conservative than the other studies, i.e., the potential for pollen movement over a longer distance and a g reater probability for PMGF at the same distance. Furthermore, Dr. Ma’s exponential decline model indicates essentially a zero probability of detecting PMGF beyond six hundred and sixty

- six feet (200 meters) (Table 1).

Using Dr. Ma’s model, Syngenta cal culated the probability of PMGF from a 75

- acre field of 100% CA corn to NCA corn planted on all four sides. The analysis was performed with and without 12 NCA corn border rows surrounding the CA field. The NCA corn was planted

immediately adjacent to the CA corn (Scenario 1, Figure 1) or the 12 th border row (Scenario 2, Figure 2). As previously communicated to APHIS BRS on September 6, 2007, Syngenta,

post

- commercialization, will instruct growers to plant 12 border rows around each CA field.

The followi ng assumptions were made in calculating PMGF in these two scenarios:

1. CA corn pollen will flow to the NCA corn bordering all four sides and corners of

the CA field.

2. The probability of CA corn PMGF beyond the edge of the CA field or 12 th border to 200 meters in the NCA corn fields was calculated using Dr. Ma’s exponential decline model (Ma 2005).

3. 100% of the corn in the CA field contains the corn amylase trait.

Based on these assumptions, and the Ma exponential decline model, und er Scenario 1, Syngenta calculates that 0.19% of the NCA corn planted adjacent to the CA corn to 200 meters will contain the CA gene (Table 1, Row 1). Under Scenario 2, 0.005% of the NCA

corn planted adjacent to the 12 th border row to 200 meters will cont ain the CA gene (Table 1, Row 12). Consequently, the 12 border rows surrounding the CA corn field capture 97.2% of all CA pollen (0.190%

- 0.005%) / (0.190%) = 97.2%.134 By blending the border rows with the harvested CA corn, >99.9% of all CA is captured . For example, assume 100 acres of CA is planted and 170 acres of NCA is planted adjacent to the CA corn to achieve the 200 meters distance cited by Ma (Figure 1).

With yields of 180 bushels per acre (bpa), the total production of CA would be:

10 0 CA acres x 180 bpa = 18,000 CA bushels

170 NCA acres x 180 bpa = 30,600 NCA bushels x 0.19% adventitious CA = 58 bushels

Total CA bushels from 100 acre CA field and 170 acre NCA field = 18,058 bushels

With 12 border rows the CA production bey ond the 12 border rows is 1.5 bushels (170 acres x 180 bpa x 0.005% adventitious CA). Therefore, the total CA capture is 18,058 bushels less the 1.5 bushels divided by 18,058 = 99.99%.

Literature cited

Jarosz, N., B. Loubet, ., B. Durand, A. McCartney

, X. Foueillassar and L. Huber. 2003. Field measurements of airborne concentration and deposition rate of maize pollen. Agricultural and Forest Meteorology 119, 37

– 51.

Jemison, J. and M.E. Vayda. 2001. Cross pollination from genetically e ngineered corn: wind transport and seed source. AgBioForum 4, 87

- 92.

Ma B.L. 2005. Frequency of pollen drift in genetically engineered corn. http://www.isb.vt.edu/articles/feb0502.htm

. Ma B.L., K.D. Subedi and L.M. Reid. 2004. Extent of cross

- ferti lization in maize by pollens from neighboring transgenic hybrid. Crop Sci. 44, 1273

- 1282.

Westgate ME, J. Lizaso and W. Batchelor. 2003. Quantitative relationship between pollen

shed density and grain yield in maize. Crop Sci. 43, 934

- 942.135 Figure 1. Planting configuration of corn amylase (CA) and non

- corn amylase (NCA) corn

without 12 separating border rows (Scenario 1).

200 meters136 Figure 2. Planting configuration of corn amylase (CA) and non

- corn amylase (NCA) corn with 12 separating border rows (Scenario 2).

200 meters

12 NCA

border rows137 Table 1. Percent pollen

- mediated gene flow downwind and upwind of source corn pollen as

extrapolated from Ma (2005) 1

.

Row # 2 Distance (meters) Downwind Upwind Downwind

Mean Upwind

Mean Overall

Mean Beyond Mean 3

0 0.00 0.00% 0.00% 0.00% 0.00% 0.00% 0.19%

1 0.75 20.35% 9.47% 20.35% 9.47% 14.91% 0.19%

2 1.50 14.96% 5.83% 17.66% 7.65% 12.65% 0.13%

3 2.25 11.00% 3.59% 15.44% 6.30% 10.87% 0.10%

4 3.00 8.09% 2.21% 13.60% 5.27% 9.44% 0.07%

5 3.75 5.95% 1.36% 12.07% 4.49% 8.28% 0.05%

6 4.50 4.38% 0.84% 10.79% 3.88% 7.34% 0.04%

7 5.25 3.22% 0.52% 9.71% 3.40% 6.55% 0.03%

8 6.00 2.37% 0.32% 8.79% 3.02% 5.90% 0.02%

9 6.75 1.74% 0.20% 8.01% 2.70% 5.35% 0.01%

10 7.50 1.28% 0.12% 7.33% 2.44% 4.89% 0.01%

11 8.25 0.94% 0.07% 6.75% 2.23% 4.49% 0.01%

12 9.00 0.69% 0.05% 6.25% 2.05% 4.15% 0.005%

13 9.75 0.51% 0.03% 5.81% 1.89% 3.85% 0.00%

14 10.50 0.37% 0.02% 5.42% 1.76% 3.59% 0.00%

15 11.25 0.28% 0.01% 5.08% 1.64% 3.36% 0.00%

16 12.00 0.20% 0.01% 4.77% 1.54% 3.16% 0.00%

17 12.75 0.15% 0.00% 4.50% 1.45% 2.97% 0.00%

18 13.50 0.11% 0.00% 4.26% 1.37% 2.81% 0.00%

19 14.25 0.08% 0.00% 4.04% 1.30% 2.67% 0.00%

20 15.00 0.06% 0.00% 3.84% 1.23% 2.53% 0.00%

30 22.50 0.00% 0.00% 2.56% 0.82% 1.69% 0.00%

40 30.00 0.00% 0.00% 1.92% 0.62% 1.27% 0.00%

50 37.50 0.00% 0.00% 1.54% 0.49% 1.02% 0.00%

60 45.00 0.00% 0.00% 1.28% 0.41% 0.85% 0.00%

70 52.50 0.00% 0.00% 1.10% 0.35% 0.73% 0.00%

80 60.00 0.00% 0.00% 0.96% 0.31% 0.63% 0.00%

90 67.50 0.00% 0.00% 0.85% 0.27% 0.56% 0.00%

100 75.00 0.00% 0.00% 0.77% 0.25% 0.51% 0.00%

120 90.00 0.00% 0.00% 0.64% 0.21% 0.42% 0.00%

140 105.00 0.00% 0.00% 0.55% 0.18% 0.36% 0.00%

160 120.00 0.00% 0.00% 0.48% 0.15% 0.32% 0.00%

180 135.00 0.00% 0.00% 0.43% 0.14% 0.28% 0.00%

200 150.00 0.00% 0.00% 0.38% 0.12% 0.25% 0.00%

220 165.00 0.00% 0.00% 0.35% 0.11% 0.23% 0.00%

240 180.00 0.00% 0.00% 0.32% 0.10% 0.21% 0.00%

260 195.00 0.00% 0.00% 0.30% 0.09% 0.20% 0.00%

267 200.25 0.00% 0.00% 0.29% 0.09% 0.19% 0.00%

1 Data presented in this table was calculated using the exponential decline model described in Ma (2005, http://www.isb.vt.edu/articles/feb0502.htm ).

2 Number of rows from the edge of the Corn Amylase (CA) field into a field of non

- CA corn.

3 The “Beyond Mean” refers to the percent of pollen mediated gene flow in non

- CA corn from the distance indicated t o 200 meters from the edge of the CA field.138 

## Appendix E

. Food processing report submitted by Syngenta

Event 3272 Corn and Food Processing

Event 3272 corn expresses the thermostable AMY797E alpha

- amylase in the endosperm of the grain. This enzyme hydrolyzes starch to sugar during the liquefaction phase of dry grind ethanol production. As presented in Syngenta’s petition 05

- 280

- 01p , the compositional analysis of Event 3272 corn demo nstrated it is not materially different from commercial corn hybrids. Although the presence of this alpha

- amylase may have an impact on certain processed corn products due to the conversion of starch to dextrins and sugars during processing, neither the s tarch itself nor any other nutritional component of Event 3272 corn has been altered by the genetic modification. Furthermore, as demonstrated by Syngenta’s concluded consultation with the FDA (BNF 0095), there is no food safety issue associated with the p ossible presence of the AMY797E alpha

- amylase enzyme in foods.

While beneficial for ethanol production, the activity of AMY797E alpha

- amylase may either

have no effect or potentially may have de sirable or undesirable effects o n certain types of processed corn food products

. AMY797E alpha

- amylase is not able to act on the starch in

the intact kernel. However, during or after processing with the addition of water and heat, enzyme activity could result in the conversion of some starch to dextrin s and sugars. It is this potential for alpha

- amylase starch hydrolyzing activity, during or after processing, that may impact properties of the corn or its components used in food.

However, the quality and properties of processed corn (e.g. dough handlin g characteristics) and finished processed food products (e.g. corn chips) are dependent on many factors. These include the characteristics and condition of the raw corn as well as specific process variables such as, cooking time, temperature, type of equi pment, degree of grinding (particle size), moisture content and baking time. Furthermore, commercial hybrid seed companies have specifically developed corn hybrids with improved properties desirable by the food industry,

e.g. (white, waxy, hard endosperm, high oil, low temperature dried, high extractable starch, etc.) that have good yield and are adapted across regions of the U.S. corn belt. These hybrids are specified by buyers and end

- users of corn for production and premiums are paid for growing, deliv ering, and meeting and maintaining the purity and quality standards of the corn. In addition to grain sourcing, a number of additional measures are employed throughout the food process including inspection and grading of the corn, cooking the corn,

formul ation and testing of the final food product. These commonly employed quality control measures ensure that corn food products are acceptable to consumers.

References :139 Corn: Chemistry and Technology, Second Edition, White, P.J. and Johnson, L.A. eds.,

Amer ican Association of Cereal Chemists, Inc., 2003.

U.S. Grains Council, Value Enhanced Corns Report 2005/2006

, available at http://www.grains.org/galleries/technical_publications/USGC%20Value%20Enhanced%20C orn%20Report%202006%20%20(English).pdf

Hallauer, A.R., Specialty Corn , Second Edition CRC Press, 2000.140 

## Appendix F

. Food processing report specific to masa submitted by Syngenta

Dilution of Corn in Masa Supply Chain

To estimate the dilution effect of Corn amylase (CA) corn in non

- CA (NCA) corn that could potentially reach a masa production facility, Syngenta conducted interviews with grain merchandisers to understand the movement of corn and the commingling with other corn that occurs during transport and storage. This information was further supplemented with Syngenta internal analysis of grain dilution pat terns.

Masa facilities typically source their grains from within 50 miles of the plant (C. Morley, Global Risk Management, Eden Prairie, MN,). White corn comprises approximately 80% of the grain used for masa and is typically contracted because it i s higher in density, protein and oil and lower in moisture, thins and starch basis in comparison to yellow corn (Sparks 2003; U.S. Grains Council 2007). Yellow corn may also be contracted and is used for color and texture (Sparks 2003). While both white and yellow corn varieties are often contracted (up to 80%; C. Morley), if bought on the open market , the grain would either be obtained directly from a grower, local elevator or other local third party supplier. These same

distribution channels would als o apply to CA grain that is mistakenly delivered to a masa plant. Considering these likely distribution paths, Syngenta analyzed the grain dilution that

occurs for each.

The first case assumes a CA grower delivers a truck load of CA grain directly from a production field or on

- farm silo to a masa plant. In this case the truck contains only CA corn was and is unloaded into a holding bin of non

- CA corn at the plant. Drawing corn through a bin will commingle the grain 3x to 16x depending on bin size and fullness at the time of delivery (Syngenta unpubl. data). To be conservative, Syngenta estimates a dilution rate of 5x for direct delivery. If, however, the grain first went to a local elevator and then later delivered to a masa facility it would have be drawn through two bins and one truck before being used by the facility. This dilution factor is estimated at approximately 50x, i.e., approximately 10x at the local elevator (larger bins) and 5x at the masa storage bin (10x time 5x = 50x).

It is impor tant to note that there is no food safety issue associated with the possible presence of the alpha

- amylase enzyme in foods as demonstrated by Syngenta’s concluded consultation with the FDA (BNF 0095). Furthermore, the likelihood of grain being mistakenly delivered directly to a masa plant by a grower from their farm is very remote. Growers will be contracted and paid a premium for producing CA grain and consequently, will have both contractual and financial incentives to deliver this grain to the ethanol plant. In addition, a large percentage of both white and yellow grain (up to 80%) is contracted and yellow represents just 20% of the total used to produce masa. Nonetheless, if a masa plant or other141 food processing company desires to determine whether their grain contains the alpha

amylase enzyme a test method will be available for their use.

References :

U.S. Grains Council, Value Enhanced Corns Report 2005/2006 , available at http://www.grains.org/galleries/technical_publications/USGC%20Value%20Enhanced%20C orn%20Report%202006%20%20(English).pdf

Sparks Companies, Inc., The US Corn Masa Industry: Structure and Im plications for the Great Plains Region , Prepared for Agricultural Marketing Resource Center ( http://www.agmrc.org ), Kansas State University, 2003. Available at http://www.agmrc.org/NR/rdonlyres/EC8E389D

- 7085

- 40ED

- B8B2

- 0B02B0AC4552/0/sparkswhitecornpaper.pdf142 

## Appendix G

. Bryson and Roberts report submitted by Syngenta

The Role of the Specialty Grain, Closed

- Loop Production System for Event 3272 in Supporting a Finding of No Significant Impact

Under the National Environmental Policy Act

By Nancy S. Bryson

and

Michael T. Roberts

December 19, 2007143 I.

Introduction

The deregulation of new plant biotechnology events by APHIS is a federal action that requires evaluation under the National Environmental Policy Act (“NEPA”). 13 APHIS has historically found that the deregulation of such events in corn have not been “major federal actions significantly affecting the human environment.” 14 NEPA compliance has been effectuated through an environmental assessment taking the “hard loo k” at the environmental effects of the action that has been required by reviewing courts, 15 and a finding of no significant impact.

Event 3272 fits comfortably into this body of NEPA analysis. Event 3272 corn is a novel transgenic corn variety that cont ains a genetically inserted thermostable alpha

- amylase enzyme that facilitates the production of ethanol by the dry grind process. Alpha

- amylases are ubiquitous in the environment. They have a long history of safe consumption by humans

and animals and the particular alpha

- amylase incorporated in Event 3272 has successfully completed the Food and Drug Administration (FDA) food safety consultation process.

Event 3272 presents no unique or novel issues relating to the environmental impacts

that have been identified by APHIS as relevant environmental effects for NEPA analysis. 16
Like other specialty corns in the marketplace, Event 3272 may not be suitable for use in some processing applications, but this does not change the NEPA analysis. Event 3272 will

b e grown and managed in the marketplace as a specialty grain produced in a tightly

controlled, closed loop system. This system will prevent any effect arising from a lack of suitability for uses for which it is not intended from rising to a level of “signi ficance” under the relevant NEPA factors of context and intensity. Accordingly, the deregulation of this event will not significantly affect the human environment within the meaning of NEPA.

The commercial value of Event 3272 is for dedicated ethanol pr oduction in the dry grind process. The realization of that value will be ensured by a closed

- loop system that reliably channels this corn from the production field into dry grind ethanol production and

13 42 U.S.C.§§ 4321 et seq.

14 42 U.S.C. § 4332(2)(C).

15 Citizens to Preserve Overton Park, Inc. v. Volpe , 401 U.S. 402 (1971).

16 USDA, Animal and Plant Health Inspection Service (APHIS), Introduction of Genetically Engineered Organisms, Draft Programmatic Environmental Impact State ment

– July 2007 , 67

– 90 (“DEIS”).144 away from the commercial channels intended for the pr oduction of processed corn food products such as masa. This process includes the Event 3272

- specific Syngenta Stewardship Program, the well

- established and recognized ability of the existing commercial marketplace to manage specialty grain product under c ontract and quality management systems, the enhanced legal traceability recordkeeping requirements of the Bioterrorism Act, and a reliable and widely available Event 3272 detection test.

Event 3272 will be deregulated in the context of a market replete w ith numerous other specialty grain products produced and marketed in the U.S. under well

- defined identity preservation and traceability systems. As another specialty grain product, the intensity of the impact of its deregulation will be very low. As such , a finding of no significant impact can be made on the basis of the robust controls that govern these systems.

II.

Requirements of the National Environmental Policy Act (NEPA)

NEPA requires that agencies undertaking a maj or Federal action “significantly affecting the quality of the human environment” provide a detailed statement of the environmental impact of the proposed action, any adverse environmental effects that cannot be avoided, and alternatives to the action.” 17 W here the significance of an action is uncertain, agencies may use an Environmental Assessment (“EA”) to identify, analyze, and evaluate the impacts of the proposed action. The EA will satisfy the NEPA obligation where it provides sufficient evidence and an alysis to support a “finding of no significant impact (“FONSI”).” 18

In order to support such a finding, the agency must have:

**•** accurately identified the relevant environmental concern;

**•** taken a hard look at the problem in preparing the environmental asse ssment, and

**•** made a convincing case for the FONSI.

If there is an impact of true significance, the preparation of an environmental impact statement can be avoided only if changes are made that reduce the impacts of the action to a

minimum. 19

17 Id. The test of wh ether an action “significantly” affects the environment requires

18 See e.g., Coalition on Sensible Transp., Inc. v. Dole , 826 F. 2d 60 (D.C. Cir. 1987).

19 Grand C anyon Trust v. FAA , 290 F. 3d 339, 340

- 41 (D.C. Cir. 2002).145 considerations of both context and intensity. 20 The term “context” refers to the setting within which the proposed action takes place. 21 In considering context, an agency must look

at the s ignificance of an action “analyzed in several contexts such as society as a whole (human, national), the affected region, the affected interests, and the locality.” 22 The term “intensity” refers to “the severity of the impact.” 23

20 40 C.F.R. § 1508.27.

21 40 C.F.R. § 1508.27(a).

22 Marsh v. Oregon Natural Resources Council , 490 U.S. 360, 374 (1989) (quoting 40 C.F.R. § 1508.27).

23 Id. With regard to the intensity element of the “significance” determination, the Council on Environmental Quality (CEQ) regulations provide the follow ing ten (10) factors to guide the analysis:

(1) Impacts that may be both beneficial and adverse. A significant effect may exist even if the Federal agency

believes that on balance the effect will be beneficial.

(2) The degree to which the proposed actio n affects public health or safety.

(3) Unique characteristics of the geographic area such as proximity to historic or cultural resources, park lands, prime farmlands, wetlands, wild and scenic rivers, or ecologically critical areas.

(4) The degree to whi ch the effects on the quality of the human environment are likely to be highly

controversial.

(5) The degree to which the possible effects on the human environment are highly uncertain or involve unique

or unknown risks.

(6) The degree to which the actio n may establish a precedent for future actions with significant effects or represents a decision in principle about a future consideration.

(7) Whether the action is related to other actions with individually insignificant but cumulatively significant imp acts. Significance exists if it is reasonable to anticipate a cumulatively significant impact on the environment. Significance cannot be avoided by terming an action temporary or by breaking it down into small

component parts.

(8) The degree to which the action may adversely affect districts, sites, highways, structures, or objects listed in or eligible for listing in the National Register of Historic Places or may cause loss or destruction of significant scientific, cultural, or historical resources.

(9) The degree to which the action may adversely affect an endangered or threatened species or its habitat that has been determined to be critical under the Endangered Species Act of 1973.

(10) Whether the action threatens a violation of Federal, State, or local law or requirements imposed for the protection of the environment.146 A recent unreported deci sion 24 from the Northern District of California in Geertson

Seed Farms v. Johanns held that APHIS was required to prepare an Environmental Impact Statement (EIS) to support a decision to grant deregulation to a glyphosate

- resistant (i.e.,

Round

- up Ready) al falfa product, and rejected the APHIS EA and FONSI as insufficient. 25
The court examined the factors of “context” and “intensity” in light of the NEPA goals of “attaining the widest range of beneficial uses of the environment without degradation, risk to

h ealth and safety, or other undesirable and unintended consequences” and “maintain[ing], wherever possible, an environment which supports diversity and variety of individual

choice.” 26 The court concluded that substantial and unanswered questions existed re garding gene transmission to non

- genetically engineered alfalfa, the development of alfalfa weeds resistant to herbicides, and increased use of glyphosate. 27 The court held that the possibility that the deregulation of Round

- up Ready alfalfa will degrade th e human environment by eliminating a farmer’s choice to grow non

- genetically engineered alfalfa and a consumer’s choice to consume such food is a significant impact that requires an EIS.

By contrast, the Syngenta Event 3272 presents a fundamentally different situation. Unlike the alfalfa industry, the segmentation and specialization of corn production has generated a suite of standard industry practices in the U.S. grain marketing system that has

been shown to fa cilitate coexistence among different varieties and obviate the potential gene flow consequences posed by alfalfa, as addressed in Geertson . Event 3272 corn is being released in an environment in which the diversity of corn is closely guarded, and continua lly enhanced. This is evident from the commercial channels and identity preservation programs that have been developed in this modern grain system to manage the co

- existence of a variety of corn products.

40 C.F.R. § 1508.27(b).

24 Unreported decisions issued by the courts are binding on the parties, but are not generally accepted as precedent

- even by the issuing court.

25 Geertson Seed Farms v. Johanns , No. C 06

- 01075 (N.D. Cal., Civ. Feb. 13, 2007).

26 42 U.S.C. § 4331(b)(3)

- (4).

27 Geertson, supra note 13.147 III.

Modern U.S. Grain System: Identity Preserva tion and Co

- Existence

A. Factors Enabling Identity Preservation and Co

- Existence

Th e modern U.S. grain system is characterized by identity preservation, product differentiation, and market segmentation. 28 This system has evolved in response to the development of a variety of specialty grain crops that require some form of segregation fro m conventional grain commodities in order to maintain their value to the end users. Examples

include popcorn, waxy (high amylopectin) corn, high oil corn, high protein and modified protein corn, sweet corn, white corn, blue corn, Indian corn, higher ferme ntable corn and high amylose corn

. 29 As a result, robust identity preservation systems with separate marketing channels prevent inadvertent commingling of specialty crops.

A number of factors have helped create this segmented market, including biotechnology, consumer preferences and demand, global concerns for safety and quality, and the increasing demand for food products originating from diverse sources. 30 For example, the corn wet

- milling industry produces starches and sweeteners or syrups tai lored to specific food and industrial uses. This has led to an increased demand for trait

- specific corn types such as high

- amylose corn. 31

Innovations in transportation, logistics, and information technologies and changes in the international regulatory environment have also facilitated the marketing of differentiated grains. Web

- based monitoring software can remotely assess the quantity and quality of grain

28 See generally Aziz Elbehri, The Changing Face of the U.S. Grain System: Differentiation and Identity

Preservation Trends , U SDA ERS (Feb 2007) at 1, available at www.ers. us da.gov/publications/err35/err35.pdf

.

29 U.S. Grains Council, Value Enhanced Corns Report 2005/2006, available at

w ww. grains .org/.../technical_publications/ US GC%20 Value %20 Enhanced %20Corn%20 Report %202006%20% 20(English).pdf

30 See Linus U. Opara, Traceability in agriculture and food supply: a review of basic concepts, technological implications, and future prospects , Food and Agr iculture & Environment Vol. 1 (1), 101 (2003).

31 See generally Kevin B. Hicks, et al., Potential New Uses for Corn Fiber , Corn Utilization and Technology Conference (2002), available at http://www.ars.usda.gov/research/publications/publications.htm?SEQ_NO_115=132521 ; Corn Refiners Association, Starch Products

, available at www.corn.org/starch.htm ; Corn Refiners Associa tion, Sweeteners

, available at http://www.corn.org/sweeteners.htm

.148 inventories in a supplier’s storage facilities. Communication networks and increased reliance

o n the Internet are cutting the costs of differentiation. 32 The implementation of the European Union food traceability and labeling directives 33 and the Cartagena Protocol on Biosafety 34
have compelled agribusinesses, particularly grain elevators and grain war ehouses at the front of the supply chain, to track closely shipments and supplies. 35

B. Identity Preservation Tools

Identity preservation or product differentiation relies on source verification

– trace ability, product tracking, and process verification. Traceability then is an important tool that “helps facilitate the identification of product(s) and/or batches.” 36 Agricultural traceability has been defined as the

collection, documentation, maintenanc e, and application of information related to all processes in the supply chain in a manner that provides guarantees to the consumer and other stakeholders on the origin, location, and life history of a product as well as assisting in crises management in t he event of a safety and quality breach. 37

32 Elberhri, supra note 16, at 5.

33 See generally Margaret Rosso Grossman, Traceability and Labeling of Genetically Modified Crops, Food

, and Feed in the European Union , 1 J. OF FOOD L. & POL'Y 43 (2005 ) (discussing traceability, labeling, and coexistence measures in the European Union).

34 The Cartagena Protocol on Biosafety was adopted on January 29, 2000, signed by 107 parties, and by

S eptember 2003 was ratified by 50 countries, the minimum required for the Protocol to enter into force. Countries that ratified the Protocol became Parties to the Protocol and are required to comply with and implement all of its provisions. Countries that have not signed but that export Living Modified Organisms (LMOs) to member countries are encouraged to comply with the Protocol’s provisions implemented in the importing country.

35 See Elberhri, supra note 16, at 25.

36 Opara, supra note 18, at 103.

37 Id. at 102.149 A major feature in a traceable supply chain is the ability to trace

- back or trace

- forward and track the physical location of the product in the overall supply chain. 38

Commercial contracts provide another important tool for product differentiation in the modern grain system. Contracts for identity preserved grains govern relations and establish stewardship controls between farmers and seed suppliers, handlers, intermediary firms, and processors. These cont racts are typically uniform and establish terms for specific variety, delivery time, delivery place, dedicated storage, and quality control. 39

C. Private Market Initiatives

To retain their competitive ad vantage in the global market and to address domestic food safety and quality issues, U.S. grain producers and handlers have relied on the contract and traceability tools described above to implement methods to produce, handle, and market trait specific gra ins, including documentation systems that trace raw materials back to the farm. Traceability and documentation are considered core competencies for grain operations. 40

These competencies are especially well developed for value

- added grains or specialty corn products. These value

- added products are produced in large volumes and are segregated by channeling. 41 Segregation by channeling flows from the farm to the transporting truck to the grain elevator and commingling is minimized by running equipment empt y before switching varieties or designating certain days of the week or alternate sites for receiving and shipping these specialty crops. 42 Crops handled by this method include a variety of value

added crops, such as white, waxy, yellow food

- grade, high

- oi l, non

- GM corn, and durum wheat grown in the Southwest. 43

38 See id.

39 See Elbehri, supra note 16, at 18.

40 See Tim Herrman, White Paper on Traceability in the U.S. Grain and Plant Protein Feed Ingredient

Industries (July 2, 2002), at 1, available at www.oznet.ksu.edu/grsiext/ White % 2 0 Paper % 2 0CVM.PDF

.

41 See Elbehri, supra note 16, at 6.

42 Id. at 6

- 7.

43 Id. at 7.150 Additional identity

- preservation controls are found in what is commonly referred to

as a “closed

- loop system.” This system is used to preserve the identity of value

- added grain products, such as high

- amylose corn. As noted by the Economic Research Service (ERS) for the USDA, a closed

- loop delivery system has distinct advantages over segregation by channeling:

A closed

- loop system provides more controls than mere channeling, and better protects the value of a specialty crop such as high

- sucrose soybeans, high

- oleic soybeans, or high

amylose corn. Production occurs almost exclusively under a contract between the grower and end

- user. Typically, these production contracts mandate delivery of all p roduction to a specified location, and require midseason inspections and return of all unused seed to the seed company. Third

- party auditors also verify that the system is in fact a closed loop and that all requirements have been adhered to throughout the

system. 44

This closed

- loop system for the delivery of value

- added, specialized corn or grain product clearly contrasts with the marketing of commodity corn, primarily yellow dent corn. Commodity corn is used for animal feed and is processed by wet and dr y milling industries for numerous and diverse food and industrial products, including ethanol. Specialty corn, on the other hand, especially in the closed

- loop system, is handled differently than commodity corn throughout the supply chain

– from seed prod uction to grain handling

– in order to ensure that a sufficient quantity of high quality corn is produced and delivered to the end

user for its particular use. This control also helps to ensure that the specialty corn is not used in unsuitable application s, which could result in process breakdowns or products with altered

or undesired characteristics.

D. Supplemental Mechanisms, Standards, and Recordkeeping Requirements

In addition to private initiatives, a variety of supporting mechanisms, standards, and legal requirements support product identification and the management of coexistence in the U.S. grain system today.

1. BIO Quality Management Program Guide

The Biotechnology Industry Organization (BIO) recently launched a new

44 Id. at 7.151 program, Excellence Through Stewardship: Advancing Best Practices in Agricultural Biotechnology , the first industry

- coordinated effort to address product stewardship and quality managem ent. 45 This industry stewardship program is described in the Quality Management Program Guide to Maintaining Plant Product Integrity

. 46 The guide is for member companies and others involved in agricultural biotechnology research and development to use in u nderstanding and implementing their own best practices. The Guide provides detailed guidance on how to develop and implement a quality management program that will assist product developers in maintaining plant product integrity from discovery to

commerci alization and post

- market activities. It includes a series of comprehensive and informative educational modules that can be adapted to the specific activities pertinent to the user’s own operations, including incorporation into existing quality management systems. Common to all of the modules is an emphasis on the importance of product identification and traceability. 47

Syngenta supports the new BIO stewardship program. As a company, Syngenta has adopted the following best practices and quality manageme nt principles to serve as a guide when it launches new products:

**•** Syngenta will conduct market and trade assessments to identify key import markets for all of its biotech products prior to product

commercialization.

**•** For each biotech product, at the time U .S. submissions are completed, Syngenta will begin to consult with the major, relevant trade and value chain stakeholders on detailed plans for pre

- commercial activities, and

full

- scale commercialization.

**•** Syngenta will meet all necessary regulatory requir ements in key exporting countries (where the seed will be commercialized) and importing countries that have functioning regulatory systems, which currently include the United States, Canada and Japan, prior to commercialization, unless determined otherwise in consultation with

45 See Excellence Through Stewardship website, BIO Launche s Excellence Through StewardshipSM Program Initiative Introduces Best Practices For Quality Management of Plant Biotechnology Products , July 25, 2007,

available at htt p://excellencethroughstewardship.org/press/newsitem.asp?id=2007_0725_01

.

46 See id.

47 See id.152 the value chain that a dedicated grain management system is workable for a specific product.

**•** Syngenta will make available prior to commercialization a reliable detection method or test that enables event identity in the crop.

**•** Syngen ta is committed to the principles of good stewardship, which are exemplified through the responsible management of Syngenta products across their lifecycle, from research through development and

commercialization to their discontinuation and withdrawal fro m the

market.

**•** Syngenta will continue to work at the global level with the value chain to engage in efforts to harmonize science

- based agriculture biotechnology regulatory approaches to achieve Global Adventitious Presence tolerances and synchronous author izations. 48

2. The Public Health Security and Bioterrorism Preparedness and Response Act of 2002 (Bioterrorism Act)

The Food and Drug Administration (FDA) is responsible for assuring that food is not

adulterated

– i.e. poisonous, deleterious, unclean, decomposed, exposed to unsanitary conditions, or contaminated with filth. 49 Following the September 11, 2001, terrorist attacks, the FDA’s authority to deter, prevent, and respond expeditiously to food safety emergencies was enhanced by enactment of the Bi oterrorism Act. 50 The Act adopts the central feature of a traceable supply chain by requiring “step

- back/step

- forward” recordkeeping for all grain product moving within commercial channels. 51

48 Syngenta website, Biotech Launch Policy Syngenta Implementation Principles, available at Entities in the grain industry, including grain elevators, feed mills, grain processors, feed ingredient manufacturers, retail feed stores, feed

http://www.syngenta

- us.com/scenter/index.asp?nav=biotech_policy_main

.

49 See 21 U.S.C. § 342(a)(1)

- (7), (c).

50 Public Health Security and Bioterrorism Preparedness and Response Act of 2002, Pub. L. No. 107

- 188 (codified in scattered sections of 42 U.S.C.).

51 See 21 U.S.C. § 350(c).153 dealers, and transporters (truckers, railroads, and barge lines) are required to establish and

maintain records containing information to help identify the grain product’s immediate previous sources and the immediate subsequent recipients. 52 Guidance from the FDA and grain industry provides useful, practical guidance on how to deal with commingling, lot code

or num bers, and an adequate description of product to ensure that the recordkeeping requirements are lawfully met. 53

As more fully outlined in the companion white paper detailing these recordkeeping requirements, the Bioterrorism Act further bolsters identity p reservation in the U.S. value

- added grain industry. 54

3. American Institute of Baking Quality Systems Management Program Guide (AIB)

The American Institute of Baking (AIB) is a non

- profit corporation, founded in 1919 by the North American wholesale and retail baking industries as a technology center for bakers and food processors. 55 The AIB has introduced to the grain handling industry its Quality Systems Evaluation (QSE), a comprehensive audit that thoroughly evaluates a supplier’s quality system. 56

52 See Establishment and Maintenance of Records Under the Public Health Security and Bioterrorism Preparedness and Response Act of 2002, 69 Fed. Reg. 71, 561

- 62 (Dec. 9, 2004) (codified at 21 C.F.R. pts. 1, 11). Th e QSE covers such elements as raw materials, process control, process verification, finished product acceptability, storage and shipping, training, plant programs, and quality policies. Each grain facility is expected to pass an AIB Food Safety

audit. In 2000, Farmland Grain Division first adopted QSE as its quality management system.

53 See FDA, Questions and Answers Regarding Establishment and Maintenance of Records (Edition 4) (Sept. 2006), available at http://www.cfsan.fda.gov/~dms/recguid4.html ; See National Grain an d Feed Association (NGFA), FDA’s Bioterrorism Recordkeeping Requirements . . . A Compliance Guide for Grain Elevators, Feed Manufacturers, Feed Dealers, Integrators, Grain Processors and Transporters . . . , April 2006.

54 See White Paper, The Bioterrorism Act of 2002: A Valuable Tool to Identity Preservation in the U.S. Value

Add Grain Industry (Dec. 2007).

55 See AIB website, https://www.aibonline.org/

.

56 See P.F. Stevenson, AIB’s Quality Systems Evaluation , prese nted at International Quality Grains Conference, July 2004, available at http://cobweb.ecn.purdue.edu/~grainlab/IQGC1/proc/text/stevenson.htm

.154 Since then, many other grain handlers have utilized QSE as their template to achieve AIB or ISO recognition. 57

4. USDA Initiatives

The mission of the USDA GIPSA is to fa cilitate the marketing of grains, oilseeds, and related agricultural commodities. 58 The process Verification Program offered by GIPSA ensures traceability and managing risk to settle disputes. GIPSA recently implemented a Proficiency Program to evaluate t he performance of laboratories that test cereals, oilseeds, and feed ingredients for the presence of GM events in the U.S. 59 Other USDA initiatives are documented in recent ERS

reports. 60

5. American Seed Trade Association (ASTA) Guide to Seed Quality

Ma nagement Practices

The American Seed Trade Association (ASTA) was founded in 1883 and consists of about 850 companies involved in seed production and distribution, plant breeding, and related

industries. 61 ASTA is drafting a Guide to Seed Quality Managem ent Practices that provides general guidance for the development of quality management practices for use in the development and production of seed products intended for food, feed, or fiber use. The guide is intended to establish and maintain the specific identity of a seed product and the purity of that seed product by using appropriate quality management measures. 62

57 See id. Syngenta supports the

58 See USDA GIPSA website, http://www.gipsa.usda.gov/GIPSA/webapp?area=home&subject=landing&topic=landing

59 See United States Department of Agriculture Grain Inspection, Packers and Stockyards Administration, Federal Grain Inspection Service Directive No. 9180.79, Jan. 29, 2007, available at 151.121.3.117/reference

library/directives/9180

- 79.pdf.

60 See e.g., USDA ERS Report, Traceability in the U.S. Food Supply: Economic Theory and Industry Studies

, Mar. 2004 (Grains and Oilseeds, pp. 22

- 26), available at http://www.ers.usda.gov/Publications/AER830/

.

61 See American Seed Trade Association website, About ASTA

, avai lable at http://www.amseed.com/about.asp

.

62 The Guide to Seed Quality Management Practices is still in the form of a working draft and has not yet been published.155 efforts of ASTA, as it provides another example of efforts in the food industry to preserve product identification.

IV.

Syngenta Stewardship Program for Event 3272 Corn

A. Description of Event 3272 Corn

Event 3272 corn has been genetically modified to express an optimized alpha

- amyla se enzyme specifically developed for use in dry grind ethanol production. This product will serve as a source of the amylase enzyme in the dry grind ethanol process, replacing the addition of liquid microbially produced enzyme and significantly improving the overall efficiency of dry grind ethanol production. 63 In addition, it will enable process flexibility that could generate real benefits at multiple points in the ethanol plant, such as increased ethanol yield, reduced energy costs, reduced water usage, and reduced chemical usage. Commercialization could also increase energy yield of corn per acre, ease current tight markets for corn, and contribute substantially to the advancement of next generation biofuels. 64

Alpha

- amylases are ubiquitous in the envi ronment and are safe. There is a long history of safe consumption by humans and animals and/or exposure to both naturally occurring and commercially produced alpha

- amylases. They are naturally present in microorganisms, plants (including corn), and anima ls. They are commercially produced for food processing uses such as baking bread, brewing beer, and producing corn syrup. In addition, a co

- product of ethanol production is sold as an animal feed ingredient. Event 3272 has successfully completed the FDA food safety consultation process. 65

63 Enzymes such as alpha

- amylase and glucoamyl ase are added during the ethanol production process because they provide a fast, safe and economical conversion of starch to sugars. A more efficient conversion of starch translates into process efficiencies in ethanol production. The improved alpha

- amyl ase facilitates this more

efficient starch to ethanol conversion. There is no food safety issue associated with the possible presence of the AMY797E alpha

- amylase enzyme in foods.

64 See generally John M. Urbanchuk, Economic Impact Analysis of Event 3272 Corn on Dry Mill Ethanol

Production , CBI Deleted Copy prepared for Syngenta Biotechology, Inc. (July 2007). The United States ethanol industry has grown at a tremendous rate, driven by energy security, the high price of foreign oil, the renewable energy mandates of the Energy Bill of 2005, and the President’s Biofuels Initiative. More than over 100 dry

mill ethanol plants (the majority of capacity) are currently dedicated specifically to the production of ethanol, with a minimum of an additional 50 planned or under construction. The ethanol industry used an estimated 1.5 billion bushels of corn in 2005 (14% of tota l corn demand), with ethanol currently representing the third most important market for US corn, after feed and export uses. This explosive growth in ethanol has created a significant domestic market for US corn producers. See id.

65 See Letter from Laura M. Tarantino, Ph.D., Director, Office of Food Additive Safety, Center for Food Safety and Applied Nutrition, to Ann Tuttle, Manager, Regulatory Affairs, Syngenta Seeds, Inc. (Aug. 2007) (acknowledging that based on the information availab le, Event 3272 corn did not raise safety or other issues that would require pre

- market review or approval by the FDA).156 B. Utilization of Event 3272 Corn

Notwithstanding its safety as food, Event 3272 will be commercialized for the value it brings to ethanol production. In order to capture the highest value of Event 3272 corn, it will be branded, marketed, and sold for that use

– i.e., with an indication o f its unique characteristic relating to the expression of an alpha

- amylase. This commercial approach is comparable to the marketing of many other specialty corns.

It is expected that Event 3272 corn may be unsuitable for some processing applications fo r which it will not be branded, marketed or sold. Specialty corns, such as

waxy corn, are developed for a specific purpose and generally are unsuitable for use in other applications. The AMY797E alpha

- amylase enzyme in Event 3272 corn is not able to act on the starch in the intact kernel and neither the starch itself nor any other nutritional or functional component of Event 3272 corn has been altered by the genetic modification. 66
However, during or after processing, enzyme activity could result in the c onversion of some starch to dextrins and sugars. It is th is potential for alpha

- amylase starch hydrolyzing activity , during or after processing, that may make it unsuitable for some applications

. For example, Event 3272 could have an effect in alkaline c ooking, the process used to produce

masa. M asa flour and dough are used to make tortillas and chips . The effect decreases the handling characteristics of the masa dough and the acceptability of the appearance of the finished product produced from that d ough. 67

The strict grain sourcing procedures in the masa industry make it highly unlikely that Event 3272 corn would be diverted to a masa facility. The masa industry has developed quality control measures to meet production requirements,

66 As presented in the Syngenta Petition for deregulation of Event 3272 corn, compositional analysis demonstrated it is not materially different from commercial corn hybrids. This is in contrast to the specialty corns in which the starch, oil, protein, or color content itself is intentionally modified. See Syngenta Petition for the Determination of Non

- Regulated Status: Corn Rootworm Pro tected Transformation Event MIR 604

, APHIS,

Docket No. 2006

- 0157

- 0003 (Jan. 10, 2007), available at http://www.regulations.gov/fdmspublic/component/main

.

67 The information contained in th is paragraph is documented by the September 5, 2007 Syngenta presentation

to BRS. See generally, L.W. Rooney & S.O. Serna

- Saldivar, Food Use of Whole Corn and Dry

- Milled Fractions in Corn: Chemistry and Technology , (P.J. White & L.A. Johnson eds.) (2003); U.S. Grains Council, Value Enhanced Corns Report 2005/2006

, available at

www. grains .org/.../technical_publications/ US GC%20 Value %20 Enhanced %20Corn%20 Report %202006%20% 20(English).pdf ; A.R. Hallauer, Specialty Corn, (2 nd ed. CRC Press 2000); Sparks Companies, Inc., The US Corn Masa Industry: Structure and Implications for the Great Plains Region , Prepared for Agricultural Marketing Resource Center, http://www.agmrc.org , Kansas State Univ . (2003), available at www.agmrc.org/NR/rdonlyres/EC8E389D

- 7085

- 40ED

- B8B2

- 0B02B0AC4552/0/ sparks white corn paper.pdf.157 prevent process disruptions, and ensure industry standards for finished products are maintained. The use of dedicated supply contracts limits the total supply of food

quality yellow corn from the open market used in masa production to 4%. 68
Commercial hybrid seed companies have specifically developed corn hybrids with improved alkaline

- cooking properties such as hard endosperm corn that have good yield and are adapted across regions of the U.S. corn belt. These hybrids are specified by buy ers and end

- users of corn for masa production and premiums are paid for growing, delivering, and meeting and maintaining the purity and quality standards of this corn. In addition to grain sourcing, a number of additional measures are employed throughout the process including grading, inspection of the corn, cooking the corn, handling the masa, formulation, and testing of the final food product. These commonly employed quality control measures ensure that masa products and finished food products meet indu stry standards. 69 These market controls in the masa industry make the likelihood of Event 3272 corn being mistakenly delivered directly to a masa plant by a grower from their farm extremely remote.

Even if corn is somehow diverted to a masa facility, ot her checks reduce the possibility of an effect. Syngenta will make available prior to commercialization a reliable detection method or test that enables crop identity verification for intended use by corn food processors capable of detecting Event 3272 in any yellow corn purchased from the open

- market, the only possible source of Event 3272 in the corn supply. Finally, dilution rates would further minimize the possible effect. In the unlikely event that corn were directly delivered from the farm to a mas a facility, it

would be co

- mingled in a masa storage bin, and depending on the actual size of the bin and fullness at the time of delivery, the dilution rate would be a conservative estimate of 5x. Corn entering a masa facility through a local elevator wo uld be drawn through two bins and one truck before being used by the facility. The dilution rate would be approximately 50x, 10x at the local elevator (due to larger bins) and 5x at the masa storage bin (10x times 5x = 50x). 70

68 Approximately 80% of the corn used in masa production is food

- quality white corn, while food

- quality yellow corn constitutes only 20% of t he total used to produce masa. Both types of corn are typically grown by producers under contract with major buyers or end

- users. In fact, of the 20% drawn from the yellow corn market, 80% is grown under dedicated supply contracts with the masa plant, le aving only 20% as sourced from the open market. This means that only 4% of the total supply of corn used in masa production is food

- quality yellow corn from the open market.

69 See generally L.W. Rooney, supra note 55; U.S. Grains Council, supra note 55; Sparks, supra note 55.

70 These dilution factors were calculated based on Syngenta’s interviews with grain merchandisers to understand the movement of corn and the commingling with other corn that occurs during transport and storage.

This information was further supplemented with Syngenta’s internal analysis of grain dilution patterns.158 C. Closed

- Loop System

The already low likelihood that Event 3272 corn would be diverted from the farm to the ethanol plant is reduced even further by the strict controls evident in the commercial system developed by Syngenta for this product. Event 3272 will be commercialized solely in a

traditional closed

- loop system, replete with effective control points. The system relies on concentrated acreage, contract grown product within the geographic footprint of the ethanol plant, stewardship manda tes, and grower agreements for delivery to ensure a smooth, uninterrupted, and verifiable supply of grain to end users

– dry grind ethanol plants.

These dry grain ethanol plants will contract directly with growers in their geographic area or indirectly through third party grain suppliers for the production and delivery of the Event 3272 corn. These grower grain contracts will specify a delivery location that either will be an ethanol plant or a storage site (country elevator) within a specified radius, a delivery date for which a window for delivery will be specified, and the acres or bushels per acre to be delivered. 71 The delivery will be made either by the growers delivering the corn directly to the ethanol plant or to an elevator or truck transporte r. Syngenta will sell hybrids with Event 3272 only to growers with a valid contract with an ethanol plant and who execute a Syngenta Stewardship Agreement that will ensure and facilitate appropriate cultivation, handling, detection, communication, inspecti on, and audits. The contracts in this closed

- loop system will contain legal and financial incentives for compliance.

Syngenta’s closed

- loop system diverts Event 3272 away from the production of processed corn food products such as masa (tortillas and c hips). The stewardship controls imposed in Syngenta’s closed

- loop system are designed to prevent the contracted grower from inadvertently delivering Event 3272 to a masa facility or to a local elevator that is an open

market supplier to a masa facility.

D. Stewardship Roles and Responsibilities Within Closed

- Loop System

In this closed

- loop system, the principle contract actors

– the producer, the grower, and the ethanol plant (end user)

– e ach have specific stewardship roles and responsibilities. These stewardship roles and responsibilities are consistent with Syngenta’s adoption of the best practices and quality management principles derived from the BIO stewardship program and are delinea ted below.

Syngenta (stewardship roles and responsibilities):

71 A “country grain elevator” is a grain elevator that services a limited geographic area. See John S. Seitz, Memorandum, Nov. 1995 at 3, available at www.epa.gov/ttncaaa1 /t5/memoranda/grainfnl.pdf

. It is very rare even in the commodity corn market for corn to be delivered from distances greater than 100 miles from the plants and only occurs where there is a shortage of local corn due to drought.159 **•** license the use of Event 3272 corn product to growers;

**•** sell Event 3272 corn hybrids only to licensed growers with a valid contract with an ethanol plant or approved third party grain company that supplies corn amylase to the ethanol plant;

● ensure that grain contract includes stewardship agreement;

**•** provide incentive to grower for producing and delivery of Event 3272 corn product;

**•** provide stewardship guide to producers and handl ers on the cultivation and handling of the Event 3272 corn product;

**•** provide specific procedures for the handling of any excess grain;

**•** ensure the domestic consumption of DDGS prior to export market approvals;

**•** make available appropriate detection me thods; and,

**•** develop and implement a communication program.

Grower (stewardship roles and responsibilities):

**•** execute delivery contract with ethanol plant;

**•** execute stewardship contract with Syngenta;

**•** follow Syngenta stewardship guide on cultivation; and,

**•** follow Syngenta requirement to divert excess grain to appropriate use.

Ethanol Plant (stewardship roles and responsibilities):

**•** contract with growers to supply Event 3272 corn product; and,

**•** ensure domest ic consumption of DDGS prior to export market approvals;

These stewardship roles and responsibilities in concert with the contract relationships

between the entities in the closed

- loop system and the backstop mechanisms minimize the

risk of inadvertent de livery of Event 3272 corn and commingling.

V.

The Deregulation of Event 3272 is not a Major Federal Action160 As demonstrated above, the deregulation of Event 3272 will result in the market entry of another specialty corn product that is identity preserved, safe for food and feed, produced in a closed loop system, and carefully managed consistent with the robust suite of safegua rds in place in the market for value

- added specialty grain products. NEPA analysis for this event necessarily begins with an identification of the relevant environmental concerns, a hard look at the significance of those concerns for a specialty grain pro duct, and an explanation of the conclusions resulting from that hard look. 72

A. Identification of Relevant Environmental Concerns

As described in the July 7, 2007 draft programmati c EIS, APHIS has identified three general categories of effects that are relevant to its assessment of risks to the human environment under NEPA. These include potential changes in weediness and invasiveness (including gene flow and persistence in the env ironment); potential effects of plants on the soil; and impacts on human health (including human allergenicity). 73

B. A “Hard

- Look” Analysis of the Effect of the Identity

- Preserved, Closed

Loop Specialty Grain Management System, Legal Recordkeeping Requirements and Supporting Quality Management Systems

.

The hard look required of the agency prior to issuing a FONSI is a critical evaluation of whether the impacts of the action are significant in terms of context and intensity. Context refers to the setting within which the proposed action take s place. 74 In considering context, an agency must look at the significance of an action analyzed in several contexts such as society as a whole (human, national), the affected region, the affected interests, and the locality.” 75 The term “intensity” refers to “the severity of the impact” 76

72 Gran d Canyon Trust v. FAA, supra note 3, at 340

– 41. judged on a number of factors. As demonstrated below, the deregulation of Event 3272 is not significant on either

factor.

73 DEIS, supra note 4, at 67

– 90.

74 49 C.F.R. §1508.27(a).

75 Marsh

, supra note 10, at 374 (quoting 40 C.F.R. §1508.27).

76 Id

.161 1. Potential Changes in Weediness and Invasiveness

Event 3272 contains an added alpha

- amylase en zyme that facilitates ethanol production. This creates a valuable output trait

– better suiting this specialty corn to the production of ethanol in the dry grind process

– but it does nothing to make it a better competitor vis

- à

- vis other corn. Accordin gly, it presents none of the weediness and invasiveness concerns associated with plant

- incorporated protectants that provide resistance to pests or herbicides.

Event 3272 does present the same gene flow potential between itself and other corn

varieties that is acknowledged as a scientific reality in the draft EIS. 77 This is not a concern respecting gene flow between corn and other species. As noted in the environmental assessment supporting the deregulation of MIR604 (corn rootworm), corn does not have weedy relatives with which it outcrosses in the United States. 78 With respect to the impact of gene flow between different varieties of corn, it is important to note that corn is an intensively cultivated and managed crop with which producers have a great d eal of experience. The draft EIS documents the well

- understood mechanisms for effective management of this gene flow potential in seed production, noting that “[I]solation and borders effectively limit the level of unintended off

- types in the final produc t and their use is supported by decades of experience with plant breeders and the seed industry.” 79

The record shows that with respect to the potential for gene flow between Event 3272 corn and other corn varieties, the Syngenta Stewardship Program provide s for all contracted growers to include 12 border rows in the harvest of Event 3272 corn to address adventitious pollination. The record further shows that 97.12% of adventitious pollination occurs in the first 12 rows. This preventative measure means tha t, by including the first 12 non

- corn

amylase rows on all sides of the field in the harvest of the Event 3272 corn, 99.9% of amylase containing grain will be accounted for in the dedicated grain supply system and directed to dry grind ethanol production. 80

77 See DEIS, supra note 4, at 64.

78 See Environmental Assessment for Prosposed Determination of Nonregulated Status for Corn Gentically Engineered for Insect Resistance , APHIS, Docket No. 2006

- 0157

- 0003 (Jan. 10, 2007), available at http://www.regulations.gov/fdmspub lic/component/main

.

79 DEIS, supra note 4, at 64.

80 This point was developed in the September 6, 2007 Syngenta presentation to the Biotechnology Regulatory Service (BRS) and in the November 16, 2007 Syngenta presentation to USDA.162 To the extent adventitious pollination result in corn amylase entering the yellow dent corn commodity supply, it will be limited to 0.1% of the grown corn amylase. As discussed above, these minute quantities will present no food safety issues and will not affect the suitability of yellow dent commodity corn for corn processing. This 0.1% will be diluted at least 5 times, and possibly up to 50 times, with non

- amylase yellow dent corn prior to entering masa production. This is far below the 0.1% at whic h corn amylase begins to affect masa production

. Moreover, as affirmed by APHIS, it will not become a seed production issue given the sophistication of current seed production techniques, isolation distances, and

the like.

Accordingly, this effect is no t significant in either context or intensity.

2. Potential Effects of Plants on the Soil

There is no suggestion in the scientific evidence presented in support of Syngenta’s petition to deregulate or in the literature that Event 3272 corn would have any effect on the

soil. 81 What the record does show is that alpha amylases are ubiquitous in the environment and are safe. They are naturally present in microorganisms, plants (including corn), and animals. The FDA food safety consultation process indicates that the Event 3272 corn presents no different issues for toxicity or allergenicity. 82

Accordingly, this effect is not significant in either context or intensity.

3. Impacts on Human Health

As discussed above, the successful conclusion of the FDA food safety consultation

demonstrates that there are no effects of concern for human health. 83 Accordingly, this effect is not significant in either context or intensity.

4. Functionality of Event 3272 for Production of Processed Corn

Products

The suitability of a new specialty corn biotechnology event for use in markets for which it is not intended has not been identified by APHIS as a factor considered to be a

81 See Syngenta Petition, supra note 54, at 87

- 88; 94

- 95.

82 See FDA Letter, supra note 53.

83 See id.163 relevant environmental effect in the draft EIS for purposes of NEPA analysis in supp ort of deregulation. 84 This approach is consistent with Supreme Court precedent on the types of environmental effects that trigger the NEPA procedural assessment requirements. “[T]he theme of [§ 4332(c) of NEPA] is sounded by the adjective ‘environmental’ . . . the context of the statute shows that Congress was talking about the physical environment

– the world around us, so to speak.” 85

The suitability effect is a commercial use issue, not an effect on the physical

environment. Unlike the Geerston court’s evaluation of the alfalfa deregulation decision, the deregulation of Event 3272 does not present any substantial or unanswered questions regarding gene transmission to non

- genetically engineered corn. Corn is an intensively managed annual crop into which multiple biotech events have already been introduced. Alfalfa, by contrast, is a perennial crop pollinated by bees to which herbicide tolerance is being newly introduced. Nor does deregulation of Event 3272 present any issue relating to the development of additional pesticide resistance or increased use of herbicides. Event 3272 introduces a special output trait. As noted above, it does not enhance the competitiveness or resistance of this corn plant.

Moreover, even if APHIS were to determine that commercial use issues should be included in its analysis of relevant environmental effects, the functionality effects of Event 3272 have been effectively mitigated by requiring steps to ensure that this specialty grain is directed to its economically produ ctive use and away from use as an ingredient in certain applications. The identity

- preserved, closed

- loop specialty grain management systems, grain traceability recordkeeping requirements, and supporting quality management systems in use in the grain mana gement industry today are fully capable of ensuring the separation necessary to avoid any unintended commingling and associated commercial use effect. Treated as mitigation measures, these steps are sufficient to bring the potential adverse suitability im pacts below the level of significance.

Accordingly, Syngenta’s decision to commercialize Event 3272 as a specialty grain produced in a closed

- loop system would be fully supported by an EA and FONSI even if the functionality effect were considered to be a n effect on the human environment for purposes of NEPA analysis. It is well

- established that even where an effect may be considered to be significant if unmitigated, an EA and FONSI will satisfy NEPA’s “hard look” requirements where mitigation can reduce those effects below the level of significance. 86

The masa example provides an excellent example of how the mitigation created by

the closed

- loop production system reduces the significance of the effects of Event 3272 on

84 See DEIS, supra note 4, at 69

- 90.

.

85 See Metropolitan Edison Co. v. People Against Nuclear Energy , 460 U.S. 766, 772 (1983); Baltimore Gas &

Elec. Co. v. Natural Res

. Def

. Council, Inc. , 462 U.S. 87, 96 (1983).

86 Robertson v. Methow Valley Citizens Counsel , 490 U.S. 322 (1989)164 non

- target users. There are three scenarios under which the dedicated grain supply system between the seed supplier, the contracted grower and the ethanol plant for Event 3272 could be breached, creating the possibility for Event 3272 to enter the yellow corn commodity market. First, adv entitious pollination could result in Event 3272 growing in neighboring fields. Second, a contracted grower could inadvertently deliver Event 3272 corn directly to a masa production facility. Third, the contracted grower could inadvertently deliver Event 3272 to a local elevator that is an open

- market supplier to a masa production facility.

As discussed above, the probability of the first scenario is remote. First, less than 10% of corn fields supporting ethanol plants (the geographic area in which E vent 3272 will be grown) are in the vicinity of masa plants. The twelve border row requirement for harvest assures that 99.9% of the grown corn amylase will be accounted for in the Event 3272 dedicated grain supply system. This level, even before the certa in dilution with other yellow

dent corn bound for the masa facility, is below the level which would impact masa production.

The likelihood of the second and third scenarios is effectively reduced by the series of supply

- chain control points between the fi eld and the masa facility. These control points include those in the masa industry and those in the closed

- loop system for Event 3272 corn. The controls in the masa industry make it highly unlikely that Event 3272 corn would be

diverted to a masa facilit y. Even if a diversion occurs, testing and dilution rates further

minimize the risk. The controls in the closed

- loop system build stewardship responsibilities for the farm, the transporters, and the ethanol plants that substantially reduce this risk even more. Whatever risk there is even further reduced by the mechanisms, standards, and recordkeeping requirements in the modern grain system that promote product segregation and identity preservation.

It is important to note that the context and intens ity effects as analyzed in this hard

look analysis do not differ from those associated with other specialty corns in commerce today. Many of these specialty corn products are unsuitable for some generic corn uses. For example, waxy corn starch has a uniq ue starch matrix of more than 98% amylopectin starch. It delivers a unique functionality that differs from normal yellow dent corn starch, which is a blend of 70% amylopectin and 30% amylose. Should waxy corn be introduced into a wet mill that produces s tarch products from standard yellow dent corn, the products will not meet commercial specifications.

Based on the hard

- look analysis of the context of introduction of Event 3272 as a specialty grain and the intensity of its impact, the effect is not sign ificant. The Syngenta Stewardship Program, the sophisticated ability of the U.S. grain supply system to manage and distribute differentiated specialty corn grain, and the traceability record

- keeping provisions of the Bioterrorism Act all support a determi nation that the deregulation of Event 3272 will not significantly affect the human environment.

VI.

Conclusion165 For the foregoing reasons, the deregulation of Event 3272 will not significantly affect the human environment and Syngenta’s petition should be granted based upon a finding of no significant impact.166 

## Appendix H. FDA memo on Event 3272 corn

consultation

FDA Home Page | CFSAN Home | Search/Subject Index | Q & A | CFSAN/Office of Food Additive Safety August 7, 2007 Help 

# Biotechnology Consultation

Note to the File

BNF No. 000095

Date: August 7, 2007

Subject: Biotechnology Notification File (BNF) 000095, corn transformation event

3272.

Keywords: Zea mays L., maize, corn, corn event 3272, AMY797E, alpha

amylase, amylase, Agrobacterium tumefaciens , PMI, phosphomannose isomerase

1. Introduction

In a submission dated August 31, 2005, Syngenta Seeds, Inc. (Syngenta) provided to the Food and Drug Administration (FDA) a safety and nutritional assessment of genetically engineered AMY797E alpha

- amylase corn designated as corn event 3272. Syngenta provided additional information to the FDA on May 1, 2006, August 2, 2006, November 8, 2006, January 19, 2007, March 2, 2007, March 6, 2007, March 7, 2007, May 31, 2007, July 11, 2007 and July 12, 2007. Syngenta

concluded that food and feed derived from corn event 3272 are as safe and

nutritious as food and feed derived from conventional corn varieties.

2. Intended Effect

Corn event 3272 was genetically engineered to contain a chimeric, thermostable, alpha

- amylase gene derived from alpha

- amylase genes from three hyperthermophilic microorganisms within the order Thermococcales . The resulting transformed corn is intended to be mixed with other corn for use in dry grind fuel ethanol production as a source of alpha

- amylase, replacing the need for the addition of micro bially

-167 produced amylase during processing. The enzyme catalyzes the hydrolysis of the alpha

- 1,4

- glucosidic bonds of amylase and amylopectin polymers into dextrins, maltose, and glucose. The recombinant enzyme is stable and active at high temperatures of dr y grind ethanol production.

Syngenta states that, based on its safety assessment, corn event 3272 is as safe as other corn lines, and as such may be used as food, feed or in the production of food products or ingredients. However, Syngenta states that gra in from corn event 3272 is primarily targeted for the dry grind fuel ethanol production industry in the United

States.

http://www.cfsan.fda.gov/~rdb/bnfm095.html

3. Genetic Modifications and Characterization

3.1 Transformation Plasmid and Parential Var iety

Syngenta constructed the plasmid vector pNOV7013. The plasmid contains two expression

cassettes within its T

- DNA region. The T

- DNA region contains a single copy of the amy797E alpha

- amylase expression cassette and a single copy of the phosphomannose isomerase ( pmi ) expression cassette as a selectable marker. The T

- DNA region of pNOV7013 was incorporated into immature corn embryo cells from a proprietary corn line (NP2499/NP2500) using Agrobacterium

- mediated transformation. The T

- DNA region of pNOV7013 contains the following genetic elements.

Table 1. Genetic elements contained in the T

- DNA region of the plasmid vector pNOV7013.168169 Left border sequence from the nopaline Ti plasmid

. Delineates the T

- DNA region for Agrobacterium

- mediated transformation.170171 Plasmid pNOV7013 contains several genes on its plasmid backbone necessary for maintenance and selection of the plasmid. These sequences are not intended for transfer into

the plant genome. The plasmid backbone of pNOV7013 contains origins of replication that allow replication of the plasmid in both Agrobacterium tumefaciens and Escherichia coli

. The plasmid contains the spec gene encoding the Tn7 adenylyltransferase conferring resistance to erythromycin, streptomycin and spectinomycin. pNOV7013 also contains the

virG gene, which is a gene involved in regulation of virulence in A. tumefaciens , and the repA gene, which is a gene responsible for plasmid replication in Gram

- negativ e, plant

associated bacteria.

3.2 Characterization, Stability and Inheritance of the Introduced DNA

Syngenta performed polymerase chain reaction (PCR), restriction digests, and Southern blot analysis to support its conclusion that corn event 3272 contai ns one intact copy of its amy797E and pmi expression cassette. Syngenta determined the nucleotide sequence of the

T

- DNA region present in corn event 3272 to demonstrate the integrity of the insert.172 Syngenta reports that corn event 3272 contains no detectab le genetic material from the pNOV7013 backbone, including the spec , virG , and repA genes.

Syngenta examined the stability and inheritance pattern of the introduced traits through conventional breeding over five generations consisting of four rounds of bac kcrossing to conventional inbred lines. Syngenta reported no significant differences in the observed

- to

expected segregation ratios for the amy797E gene over five generations, as demonstrated by

the

chi

- square (χ 2 ) values. Syngenta reported that these segregation data indicate a single

locus, Mendelian inheritance pattern for the insert in corn event 3272.

4. Introduced Substances

- AMY797E Alpha

- Amylase and PMI Enzymes

Syngenta provided information on the identity, function, and characterization of the genes as well the expression levels of the gene products. They also provided information on the potential allergenicity and toxicity of the expressed proteins.

4.1 Identity, Function, and Characterization

4.1.1 AMY797E Alpha

- Amyla se

Syngenta compared the amino acid sequence of the AMY797E alpha

- amylase protein to other alpha

- amylases, stating that AMY797E shared a 93% homology to a microbially derived alpha

- amylase BD5088 (Innovase LLC) produced through a similar recombination

tec hnique as AMY797E. The BD5088 alpha

- amylase was the subject of a generally recognized as safe (GRAS) notice (GRN 126) for use in food. Syngenta also provided the complete amino acid sequence of AMY797E. Additionally, Syngenta provided information

about the functional activity of AMY797E to demonstrate its functionality as an alpha

amylase.

4.1.2 PMI

The pmi gene, introduced as a selectable marker into corn event 3272, encodes a phosphomannose isomerase (PMI) enzyme that catalyzes the inter

- conversion of

m annose

- 6 phosphate and fructose

- 6

- phosphate. The expression of the pmi gene in the plant allows the plant to survive and grow on media containing only mannose as the only or primary carbon source, which facilitates selection of transformed plants. Syngent a reports that PMI enzymes exist widely in nature among both prokaryotes and eukaryotes. Syngenta states that PMI enzymes have been found in plants such as tobacco, walnut, and soybean and other legumes, although Syngenta notes that sequence homology for

t he pmi gene introduced into event 3272 is highest for enteric Gram

- negative bacteria (70

- 100% homology).

4.2 Expression Levels

4.2.1 AMY797E Alpha

- Amylase173 Syngenta reports that AMY797E alpha

- amylase expression levels were measured by

enzyme

- linked immun osorbent assay (ELISA) in several tissues at various stages of development. Syngenta notes that expression of AMY797E is directed primarily to the kernel by the maize gamma

- zein promoter, and as expected, the highest concentration of AMY797E alpha

- amylase is in the kernel. Mean concentration measured in mature or senescent kernels ranged from 838 ± 268 μg/g fresh weight (fw) (1004 ± 322 μg/g dry weight (dw)) to 955 ± 225 μg/g fw (1335 ± 358 μg/g dw). Mean concentration in younger "dough" stage (R4)

kernels ranged from 874 ± 160 μg/g fw (1994 ± 228 μg/g dw) to 1627 ± 338 μg/g fw (3365 ± 780 μg/g dw). Concentration of AMY797E alpha

- amylase varied in whole plant samples, at various growth stages, from < 12 μg/g fw (< 37 μg/g dw) to 281 ± 108 μg/g fw (668 ± 248 μg/g dw).

4.2.2 PMI

PMI protein concentrations were also measured in various tissues and developmental stages. PMI was detected in most tissues tested and concentrations were similar regardless of the developmental stage tested. The highest levels were d etected in pollen, with concentrations ranging from 8.0 to 8.5 μg/g fw (17.0

- 18.2 μg/g dw). Mean concentrations of PMI in kernels over all growth stages ranged from < 0.4 μg/g fw (< 0.5 μg/g dw) to 0.8 ± 0.1 μg/g fw (1.8 ± 0.4 μg/g dw). Concentrations of PMI in whole plant samples from all developmental stages ranged from < 0.3 μg/g fw (< 0.6 μg/dw) to 1.5 ± 0.3 μg/g fw (3.6 ± 0.9 μg/g dw).

4.3. Assessment of Potential Allergenicity

Syngenta states that the potential allergenicity of the AMY797E alpha

- a mylase and PMI proteins were assessed by searching for amino acid homology between these proteins and known allergen protein sequences. These searches were conducted using a database comprised of identified or putative allergen sequences from publicly avai lable databases (GenPept, PIR, SWISS

- PROT, FAARP and IUIS) and additional putative allergen sequences from the scientific literature. Syngenta also assessed the stability of the AMY797E and PMI proteins using in vitro digestibility assays.

4.3.1 AMY797E A lpha

- Amylase

Syngenta reports that the donor organisms ( Thermococcus / Pyrococcus ) used to develop AMY797E alpha

- amylase protein are not known to be allergenic.

Syngenta reports that for AMY797E, there were no amino acid sequence identities of greater than 35% in segments of 80 amino acids with any entries in the database. Syngenta does note that there was a single segment of 8 contiguous amino acids in AMY797E that matched a known allergenic sequence derived from an insect. However, Syngenta notes

that the allergenic epitopes are known for this allergen (American cockroach, Per a 3) and there is no overlap between these binding epitopes and the eight amino acid region of sequence identity with AMY797E alpha

- amylase. Therefore, Syngenta maintains that this

s equence identity is not biologically relevant and has no implication for the allergenic174 potential of the AMY797E alpha

- amylase.

Syngenta provides data on the in vitro digestibility of AMY797E. Syngenta reports that AMY797E was susceptible to proteolytic d egradation in simulated gastric fluid (SGF) containing pepsin, indicating that AMY797E is degraded within 5 minutes. Syngenta states that AMY797E is not stable to digestion and is therefore unlikely to become allergenic.

Syngenta notes that AMY797E is a t hermostable protein. In addition, Syngenta reports that analysis of AMY797E as expressed in corn event 3272 does not reveal evidence of

post translational glycosylation.

4.3.2 PMI

Syngenta reports that the donor organism ( E. coli ) used to develop the PMI protein is not known to be allergenic.

Syngenta reports that for PMI, there were no amino acid sequence identities of greater than 35% in segments of 80 amino acids with any entries in the database. Syngenta does note that

there was a single segment of 8 contiguous amino acids in the PMI protein that matched a known allergen, α

- parvalbumin from Rana species CH2001, an edible frog. Syngenta reports that further investigation of PMI using serum from one known α

- parvalbumin

- sensitive individual demonstrated a lack of reactivity with PMI. Syngenta therefore concluded that the sequence identity between PMI and α

- parvalbumin is not biologically relevant.

Syngenta provides data on the in vitro digestibility of PMI. Syngenta reports that the PMI protein was found to be degraded when sampled immediately after time zero. Syngenta states that PMI is not stable to digestion and is therefore unlikely to become allergenic.

Syngenta notes that the PMI protein is labile to heat. In addition, Syng enta reports that PMI is unlikely to be glycosylated, given that the PMI protein does not contain consensus amino acid sequences required for N

- glycosylation and the protein is not targeted to a cellular glycosylation pathway.

4.4 Assessment of Potential Toxicity

4.4.1 AMY797E Alpha

- Amylase

Syngenta reported results from an acute oral toxicity study in mice where 1511 milligrams/kilogram body weight (mg/kg bw) AMY797E protein, or a control, was given by gavage. Syngenta indicates that the AMY797E test su bstance was prepared from event

3272 grain and was determined to be 42% AMY797E protein. Syngenta states that the animals were monitored for 14 days and were sacrificed. Syngenta reports that there were no effects of treatment on any observation, including body weight, food consumption, organ weight, or histopathology.

Syngenta reports that the potential toxicity of AMY797E was also assessed by comparing175 its amino acid sequence against all publicly available protein sequences identified as toxins

in the Na tional Center for Biotechnology Information Entrez Protein Database. Syngenta reports that no significant sequence homology to any known toxins was identified.

4.4.2 PMI

Syngenta reports results from an acute oral toxicity study where PMI protein was giv en to mice by gavage at a dose of 3030 mg/kg bw. Syngenta notes that the PMI protein used in this study was obtained by over

- expressing the protein in E. coli . Syngenta states that the animals were monitored for 14 days and were sacrificed. Syngenta report s that there were no effects of treatment on any observation, including body weight, food consumption, organ weight, or histopathology.

Syngenta reports that the potential toxicity of PMI was also assessed comparing its amino acid sequence against all pub licly available protein sequences identified as toxins in the National Center for Biotechnology Information Entrez Protein Database. Syngenta reports that no significant sequence homology to any known toxins was identified.

5. Food and Feed Uses of Corn

Syngenta notes that corn grown in the U.S. is primarily the yellow dent type, a commodity crop used primarily to feed domestic animals, either as grain or forage. The remainder of the crop being exported or processed by wet or dry milling to yield human fo od products such as high fructose corn syrup, starch or oil, grits and flour. The by

- products of wet and dry milling are commonly used in animal feed.

6. Compositional Analysis

Syngenta evaluated the composition of forage and grain from event 3272

- derive d corn hybrids relative to negative segregant (near

- isogenic) control corn hybrids of similar genetic background.

Compositional analyses were performed on forage and grain. Syngenta analyzed corn grown
at 10 different locations over a two year period, wit h three replicate plots of each genotype 1

planted at each location in randomized complete blocks. Grain was harvested from 6

locations in both years and forage was harvested from 6 locations in 2003 and 7 locations in 2004.

Three of the locations used in 2003 were again used in 2004 for growing corn grain, and four of the locations used in 2003 were again used for growing corn forage in 2004. Two

hybrid pairs 2

were grown in 2003 and one hybrid pair was grown in 2004. The data were combined for statistical analysis across locations, hybrid pairs, and growing seasons. Compositional data were statistically analyzed using a mixed model analysis of variance with locations serving as blocks. Statistical significance was assigned at p < 0.05 indicating that the difference between the treatments was statistically different at the 5% customary level. Syngenta176 compared the compositional data with published literature values for each analyte to assess whether statistically significant differences in the com position of the event 3272

- derived hybrids and the corresponding near

- isogenic control maize were biologically meaningful.

6.1 Forage Composition

Syngenta determined the levels of the following analytes in forage from corn event

3272 derived and the near

- isogenic control hybrids collected at the R4 development stage. The following groups of analytes were measured:

z Proximates

z Minerals

A list of specific analytes contained in each group is shown in Table 2. Syngenta reported

mean values for these analytes fell within published literature ranges. 3

, 4 Syngenta conducted

a combined statistical analysis of forage data from all three field trials for analytes 5 marked

with an asterisk (*) in Table 2, and found statistically significant differences betwee n the event

3272 derived hybrids and their near

- isogenic control lines for protein, carbohydrate and acid detergent fiber (ADF). Protein levels were statistically significantly higher, and carbohydrates and ADF were significantly lower in event 3272

- derive d hybrids than their respective near

- isogenic control lines. Syngenta concluded that these differences were not biologically meaningful.

Table 2. Components measured in event 3272

- derived forage and grain177 Syngenta determined the levels of the following analytes in grain from corn event 3272

- derived and the near

- isogenic control hybrids collected at the R6 growth stage (maturity). The following groups of analytes were measured:

z Proximates

z Minerals

z Amino Acids z Fatty Acids

z Anti

- nutrients z Secondary metabolites

z Vitamins

A list of specific components contained in each group is shown in Table 2. Syngenta reported mean values were within published literature ranges, with minor exceptions

noted. 6

, 7 Syngenta conducted a combined statistical analysis of grain data from all field trials
for all analytes listed in Table 2, excluding γ

- tocopherol and β

- cryptoxanthin 8 and the nine analytes below the limit of quantitation. Exclusion of these analytes did not, however, affect Syngenta's

conclusion s. Low levels of these nutrients and anti

- nutrients are consistent with values reported in the published literature and variable levels of certain nutrients (e.g. selenium) found in soil throughout the U.S.

Syngenta reported statistically significant diff erences in mean values between event 3272

-178 derived hybrids and controls for protein and manganese, which were higher in event 3272

- derived hybrids than near

- isogenic control hybrids. Syngenta also reported statistically significant differences in the mean v alues of carbohydrates, total dietary fiber, neutral detergent fiber, vitamin B

6 , β

- carotene, inositol, and ferulic acid, which were lower in event

3272

- derived

hybrids. Event 3272

- derived hybrids had significantly higher levels of almost all amino

acids compared to the near

- isogenic control lines, with the exception of arginine, cysteine, and lysine which were not different. Syngenta has compared all mean values to published literature ranges and concludes differences between event 3272

- derived and near

- i sogenic control hybrids are not biologically meaningful.

7. Wholesomeness Study

Syngenta reports results from a 42

- day broiler chicken feeding study comparing birds fed

event 3272 grain versus the near

- isogenic control and commercially

- available corn. Sy ngenta indicates that all of the diets derived from treatment and control diets supported rapid growth

with low mortality rates and excellent feed conversion ratios. 9 Syngenta also reports no evidence of biologically

- significant differences in growth or f eed conversion in chickens fed

event

- 3272 grain compared to near

- isogenic control or commercially available corn. Furthermore, Syngenta notes the absence of any adverse nutritional or toxic effects in

chickens fed these diets.

8. Conclusions

Syngenta has concluded that AMY797E alpha

- amylase corn event 3272 is not materially different in composition, safety, or any other relevant parameter from corn now grown, marketed, and consumed. At this time, based on Syngenta's data and information, the

agency consi ders Syngenta's consultation on AMY797E alpha

- amylase corn event 3272 to be complete.

Richard E. Bonnette

(1) The term genotype is used by Syngenta to refer only to the presence or absence of

Event 3272.

(2) Syngenta defines hybrid pairs as the result o f an initial cross

- breeding of event 3272 and a

non

- transgenic line followed by self

- crossing or cross

- breeding of the resulting progeny one or more times and the selection of homozygous trait positive and negative segregants.179 (3) Syngenta reported fat le vels below the 0.1% limit of quantification in forage from both

event 3272

- derived hybrids and their respective near

- isogenic control lines at a single field site. Excluding data from this site, reported mean values for fat were within published

literature ranges.

(4) Values were within the ranges found in the International Life Sciences Institute Crop Composition Database (version 3.0, released April 10, 2006, available at: www.cropcomposition.org

(5) Syngenta limited forage analytes in the combined statistical analysis to those suggested by the OECD (2002). Fat was excluded from the combined statistical analysis due to level s below the limit of detection. ). Excluding phosphorous, means for all analytes also fell within values cited by the Organisation for Economic Co

- operation and Development (OECD) "Consensus Document on Compositional Considerations for New Varieties of Maize (Zea Mays): Key Food and Feed Nutrients, Anti

- Nutrients and Secondary Plant M etabolites." (Series on the Safety of Novel Foods and Feeds, No. 6, 2002).

(6) The following 9 grain analytes had one or more observations below the limit of quantitation of the assay: sodium, selenium, raffinose, phytic acid, furfural, tocopherols (α

- , β

- and γ

- ), and vitamin C. An observation rep resents the mean of 3 replicate plots per location, with grain from 15 plants pooled from each plot.

(7) Values were within the ranges found in the International Life Sciences Institute Crop Composition Database (version 3.0, released April 10, 2006, avail able at: www.cropcomposition.org

(8) For several of the analytes, Syngenta reported that the treatment (genotype) effect differed by location. However, based on the statistical model used, the interpretation of the data was

not affected. ). Fatty acids mean values were within ranges cited by OECD (2002).

(9) Syngenta notes that the non

- transgenic control grain appeared to contain

low levels (approximately 1.6

- 2.6%) of event

- 3272 corn.

Biotechnology | Products: Completed Consultations

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## Appendix I

. Report on DDGS submitted by Syngenta

Impact of Event 3272 Corn on Distiller s Grains

John M. Urbanchuk

Director, LECG LLC

December 4, 2007

Syngenta Biotechnology Inc. has developed a novel transgenic corn variety, designated as

Event 3272 that contains a genetically inserted thermostable alpha

- amylase enzyme. Event

3272 is agronomically equivalent to No. 2 Yellow Corn and has been shown to be as safe for

human and animal consumption as conventional yellow corn; its unique properties make it

especially suited for use as a feedstock for ethanol produced by the dry grind process.

Dry Grind Ethanol Production

Ethanol is produced from corn and other grain s using one of two production processes: wet

milling and dry milling. The main difference between the two is the initial treatment of the

grain. Most new ethanol facilities built in the U.S. in recent years have been dry grind plants.

According to the Ren ewable Fuels Association dry mill facilities account for 82 percent of

ethanol production and wet mills 18 percent.

In the typical dry grind process , the entire corn kernel is ground into a meal or flour and is

processed without separating out the various component parts of the grain. Water is added to

form a mash to which enzymes are added to convert the starch to dextrose, a simple sugar.

Ammonia is added for pH control and as a nutrient for the yeast. The mash is processed in a

high

- temperature cooker to reduce bacteria levels ahead of fermentation. The mash is cooled

and transferred to fermenters where yeast is added and the conversion of sugar to ethanol and

carbon dioxide (CO 2 ) begins.181 After fermentation, the resulting mixture called “ beer ” is transferred to distillation columns

where the ethanol is separated out . The ethanol is concentrated to 190 proof using

conventional distillation and then is dehydrated to approximately 200 proof in a molecular

sieve system. The anhydrous ethanol is the n blended with a denaturant such as regular

gasoline to render it non potable and thus not subject to beverage alcohol tax. It is then ready

for shipment to gasoline terminals for blending with gasoline.

After the fermented mash is distilled to remove the alcohol, the remaining slurry contains 5

to 10 percent dry matter called whole or spent stillage. Whole stillage is processed by

various techniques to remove the large volume of water associated with the residual dry

matter. The first step involves scre ening and pressing, or centrifuging to remove the solids

composed of coarser grain particles. These solids, called Distillers’ Grains

- DG, can be

subsequently handled in several ways but are ultimately sold in various forms for animal

feed. The distille rs grains can be sold as a wet product (Wet Distillers’ Grains

– WDG) or

can be dried (Dried Distillers’ Grains

– DDG). The liquid remaining after screening or

centrifuging the whole stillage contains fine grain particles and yeast cells and is called thin

stillage. Thin stillage is generally evaporated to produce a syrup called solubles which may

be added back to the distillers grains. The mixture is then dried to produce Dried Distillers

Grains with Solubles (DDGS), a high quality, medium

- protein livest ock feed.

Alpha

- amylase is one of two enzymes (the other being glucoamylase) that convert the starch

in corn to sugar which is then fermented and distilled into ethanol. Microbial produced

alpha

- amylase is already used commercially in the starch to sugar step of the dry grind and

wet mill process of ethanol production. The Syngenta innovation prompts corn to produce its

own heat

- resistant alpha

- amylase, thus eliminating the need for externally applied microbial

alpha

- amylase.182 Impact of Event 3272 on Distillers grains

While efficiencies from Event 3272 are gained throughout the ethanol production process,

the greatest benefits are realized within the starch

- to

- sugar conversion phase. The most

apparent of these benefits is the elimination of microbial alpha

- amylase as a raw material

input. Alpha

- amylase is used to break down the starch component of the corn into short

chain dextrins, or complex sugars. Glucoamylase is then added during saccharification to

further break down dextrins into simple sugar s that can be fermented into ethanol. Because

the alpha

- amylase enzyme is already present in the corn kernel of Event 3272, this input is

eliminated from the process.

The alpha

- amylase enzyme produced by Event 3272 catalyzes the same reaction as other

microbial alpha

- amylases. The enzyme, whether exogenously introduced in microbial form

or provided by the corn via Event 3272, only directly affects the conversion of starch to

sugar, which eventually becomes ethanol. Use of Event 3272 corn is unlikely t o have any

adverse impact on the composition or quality of the coarse grain (distillers grains) or solubles

left after distillation that, when added to distillers grains, become DDGS.

However, the use of Event 3272 corn will provide important efficiencies to the ethanol

production process that may result in direct benefits to the distillers grains produced from

Event 3272 corn.

**•** The first of these is a reduction by half in sulfuric acid used to maintain pH levels.

When conventional yellow corn is used in a dry grind plant, pH must be adjusted at

several stages to create optimal conditions for chemical reactions to take place. With

Event 3272 corn, pH is maintained at a constant level of approximately 4.8

throughout the production process. This reduces th e use of sulfuric acid by half.

Sulfuric acid ultimately becomes a sulfur salt in the DDGS product. According to

research conducted at the University of Minnesota the sulfur content of distillers

grains can be both very high and highly variable. “If not m anaged properly, high S

concentrations in the diet, coupled with S from drinking water, may negatively affect183 both animal performance and animal health”. 87 The effect of reducing sulfuric acid

use in dry grind ethanol production through the use of Event 32 72 corn will enable

feedlot operators to more effectively manage sulfur concentrations and avoid

potential toxicity issues associated with conventional distillers grains.

**•** Event 3272 provides an additional benefit to the ethanol production process by

incre asing solids content in the starch

- to

- sugar phase. An increased solids/liquid ratio

decreases water use which may also reduce the amount of drying required to

evaporate the water in the stillage to produce a marketable distillers grains product.

Reduced d rying requirements may both improve consistency and quality of distillers

grains. One of the most commonly cited problems with DDGS is scorching or

burning that result in darkening of the distillers grains, creating an offensive odor, and

a potential degra dation of nutritional properties. 88

87 Grant Crawford. “Managing High Sulfur Concentrations in Beef Cattle Feedlot Rations”. University of

Minnesota Beef Center. 2007. Availab le online at www.extension.umn.edu/beef The use of Event 3272 corn may

enable ethanol producers to avoid these problems.

88 D.O. Connor. “The Impact of Changes in the Ethanol Production Process on the Nutritional Value of Distillers Grains”. (S&T) 2 Consultants. 2007.184 

## Appendix J

. Glossary

A

Abiotic Stress Stress due to non

- living, environmental factors such as cold, heat, drought, flooding, salinity, toxic substances, and ultraviolet light. **Agrobacterium**
**tumefaciens** A bacterium that causes crown gall disease in some plants. The bacterium characteristically infects a wound and incorporates a piece of its own DNA into the host plant genome, causing the host cell to grow into a tumor

- like structure. This DNA

- transfer mechanism is commonly exploited in the genetic engineering of plants.

Agrobacterium
Tumefaciens

mediated
Transformation The process of DNA transfer from Agrobacterium tumefaciens to plants, which occurs naturally during crown gall disease and can be

used as a method of transformation.

Allergen Any substance that causes an allergic reaction. Alpha

- amylase The major form of amylase found in humans and other mammals. Amylase is the name given to glycoside hydrolase enzymes that break

down starch into glucose molecules.

Anthesis Time at which the corn sheds pollen.

Antibiotic

Resistance Marker

Gene Genes (usually of bacterial origin) used as selection mar kers in transformation because their presence allows cell survival in the presence of normally toxic antibiotic agents.

Antinutritional or

Antinutritive Compound A compound in food or animal feed that has a negative impact on nutrition or the absorption o f nutrients.

APHIS Animal and Plant Health Inspection Service

. Archael order

Thermococcales In taxonomy , the Thermococcales are an order of the Thermococci , in

the Archaea domain. Archaea are a major division of microorganisms . Like bacteria , archaea are single

- celled organisms lacking nuclei and are therefore prokaryotes , classified as belonging to kingdom Monera in the traditional five

- kingdom taxonomy . Although

there is still uncertainty in the approximate phylogeny of the groups, Archaea, Eukaryota and Bacteria are the fundamental classifications in

what is called the three

- domain system . Despite being prokaryotes,

archaea are mo re closely related to eukaryotes than to bacteria.


### B
Barren plants Corn plants that do not develop an ear.

Benefical organisms Any organism directly or indirectly advantageous to commodities, including biological control agents [ISPM No. 3, 2005]. Biotechnology Making specific modifications to the genome of an organism using techniques based on molecular biology, such as gene manipulation, gene transfer, DNA typing, and cloning of plants and animals. Breeding The process of sexual reproduction and production of offspring. Plant breeding is an applied science for the development of plants suited for185 the use of humans, rather than their ability to survive in the wild.
BRS Biotechnology Regulatory Services (USDA

– APHIS). 

### C

CFR Code of Federal Regul ations (U.S.).
Conservation Tillage A broad range of soil tillage systems that leave crop residue on the soil surface, substantially reducing the effects of soil erosion from wind

and water.
Constitutive Expression Describing a gene that is expressed (i.e., “turned on”) at a relatively constant level in all cells of an organism without regard to cell

environmental conditions.

Counterpart A plant variety (or varieties) that represents the closest appropriate genotype to the transgenic plant in question and is a suitable control taking into account the breeding history of the transgenic plant. In some instances, it may be appropriate to use a transgenic progenitor plant as a counterpart in addition to, or as a substitute for, a non

- transgenic counterpart . (NAPPO, RSPM No. 14, Transgenic Materials).


### D

DNA See Deoxyribo nucleic Acid. Deoxyribonucleic

Acid A nucleic acid that carries the genetic information of a cell. The structure of DNA is two long chains, consisting of chemical building

blocks called ‘nucleotides,’ twisted into a double helix. The order of nucleotides determines hereditary characteristics. Diploid The status of having two complete sets of chromosomes, most commonly one set of paternal origin and the other of maternal origin.

S omatic tissues of higher plants and animals are ordinarily diploid in chromosome constitution, in contrast with the haploid gametes (FAO).

Donor An organism that provides a gene or gene fragment used in the genetic transformation of another organism, call ed the “recipient.” Dry

- grind ethanol

process In dry milling, the entire corn kernel or other starchy grain is first ground into flour, which is referred to in the industry as "meal" and processed without separating out the various component parts of the grain. In contrast, wet

- milling involves soaking or steeping the grain in water and dilute sulfurous acid for 24 to 48 hours. (RFA website http://www.ethanolrfa.org/resource/made/)


### E
Early root lodging Percent of plants per plot leaning greater than 30 degrees from vertical at the root prior to anthesis.

ELISA (enzyme

linked

immunosorbent serologic assay) A sensitive assay for detecting a specific protein that uses antibodies to

bind to the protein.

EPA U.S. Environmental Protection Agency.

Event See Transformation Event.186 Expression The means by which a gene’s information stored in DNA (or RNA in some viruses) is turned into biochemical information such as RNA or protein.


### F

F 1 Hybrid Abbreviation for filial generation 1. The initial hybrid generation resulting from a cross between two parents (FAO).

FDA Food and Drug Administration

. FFDCA Federal Food, Drug, and Cosmetic Act. Flanking Region The DNA sequences extending on either side of a specific sequence.

FPPA Federal Plant Pest Act

. 

### G

GE See Genetically Engineered.

Gene The basic unit of heredity transmitted from generation to generation during sexual or asexual reproduction; an ordered sequence of nucleotide bases, comprising of a segment of DNA. A gene contains the sequence of DNA that encodes an individual RNA or protein. Gene Expression The process by which a gene produces mRNA and protein and ultimately exerts its effect on the phenotype of an organism.

Gene Flow The spread o f genes from one population to another by the movement of individuals, gametes, seeds, or spores.

Gene Introgression See Introgression.

Genetic Engineering Genetic engineering refers to the process in which one or more genes and other genetic elements from one or more organism(s) are inserted into the genetic material of a second organism using recombinant DNA techniques. Genetically Engineered (GE) Modified in genotype and, hence, phenotype using recombinant DNA techniques. GE Organism Genetically engineered organisms. (See Genetically Engineered.)

GE Plant Genetically engineered plant. (See Genetically Engineered.)

Genetic Marker A gene that is a reliable indicator that a particular organism possesses a specific trait of interest. Markers may be used to select certain individual organisms , e.g., cells that have inherited resistance to an antibiotic will be the only ones in a population that survive an

antibiotic treatment.

Genetic
Transformation See T ransformation.

Genome All of th e hereditary material in a cell including DNA present in the

cell nucleus

, as well as in other locations such as plant chloroplasts and

mitochondria. Genotype The total genetic makeup that an individual receives from its parents.

Germination The initial st ages in the growth of a seed to form a seedling (FAO).
GRAS Generally recognized as safe.187 

### H

Herbicide

Resistance or

Tolerance The ability of a plant to remain relatively unaffected by the application

of what would otherwise be a highly damaging dose of a n herbicide.

HGT Horizontal gene transfer

Horizontal Gene
Transfer The transfer of genetic material from one organism (the donor) to another organism (the recipient) that is not sexually compatible with

the donor. Hybrid The offspring of two genetically dissimilar organisms. Hyperthermophilic organism that thrives in extremely hot environments (Hyperthermophile)


### I

Industrial Plant A plant genetically engineered with a gene whose effect is primarily of industrial use, as opposed to an agricultural or nutritional purpose.

Inserted Gene A piece of DNA that has been inserted into an organism using recombinant DNA technology. Introgression The introduction of genes from one species into the gene pool of another via sexual crossing. The p rocess begins with hybridization between the two species, followed by repeated backcrossing to one of the parent species.


### K
Kernel dough About 24 to 28 days after silking, the kernel's milky inner fluid is changing to a 'doughy' consistency as starch accumulation continues in the endosperm. The shelled cob is now light red or pink. By dough stage, four embryonic leaves have formed and about half of the mature kernel dry weight is now in place. Kernel abortion is much less likely

once kernels have reach ed early dough stage, but severe stress can continue to affect eventual yield by reducing kernel weight. Kernel moisture content is approximately 70 percent. http://www.ces.purdue.edu/extmedia/CL/ CL

- 10.html Kernel maturity About 55 to 65 days after silking, kernel dry weight usually reaches its maximum and kernels are said to be physiologically mature and safe from frost. Physiological maturity occurs shortly after the kernel milk

line disappears and just before the kernel black layer forms at the tip of the kernels. Severe stress after physiological maturity has little effect on grain yield, unless the integrity of the stalk or ear is compromised (e.g., ECB damage or stalk rots). Kernel moisture content at physiological maturity averages 30 percent, but can vary from 25 to 40 percent grain moisture. http://www.ces.purdue.edu/extmedia/CL/CL

- 10.html


### L

Late root lodging Perce nt of plants per plot leaning greater than 30 degrees from vertical

at the root after anthesis . Used as a measure of abiotic stress tolerance.

Late season Rating of late

- season integrity of the plant above the ear188 intactness



### M

Marker Gene A gene of known function or known location that is inherited in Mendelian fashion and facilitates the study of inheritance of a nearby

gene. Microorganism An organism that is microscopic (too small to be seen by the human eye).

Monocot A flowering plant with only one embryonic seed leaf. Examples include grasses, irise s, lilies, and onions. ( See Dicot.)

Monoecious A plant species that has separate male and female flowers on the same plant (e.g. maize) (FAO)

MOU Memorandum of Understanding.


### N

NEPA The National Environmental Policy Act of 1969 and subsequent

amentments.
NOI Notice of Intent.

Non

- target organisms An organism which is affected by a treatment (e.g. pesticide application) for which it was not the intended recipient (FAO).

Northern corn leaf blight Corn disease caused by the fungus Exserohilum turcicum

Notification An administratively

- streamlined al ternative to a permit for the introduction of a regulated GE plant. The GE plant must meet specified eligibility criteria, and the introduction must meet certain pre

- defined performance standards. 

### O

OECD Organisation for Economic Co

- operation and Develo pment

. Outcrossing The tendency of a plant species to produce offspring that result from the mating of two different individual plants. (See S elf

- pollinated. ) Overwintering Time at which plants experience ‘winter conditions’ where growth of vegetative tissues and reproductive structures becomes minimal or ceases completely. (wiki)


### P
Percent snapped plants Percent of plants per plot broken prior to anthesis due to adverse environmental conditions, such as high wind speeds. Used as a

measure of abioti c stress tolerance.

Permits An application to BRS for the introduction of GE organisms that pose a plant pest risk, including plants, insects, or microbes.

Persistence Ability of an organism to remain in a particular setting for a period of

time after it is introduced (FAO) Phenotype The appearance or other characteristics of an organism, resulting from the interaction of its genetic constitution with the environment. Phosphomannose
isomerase Phosphomannose isomerase (PMI), an enzyme not present in many plants, catalyzes the reversible interconversion of mannose 6

- phosphate and fructose 6

- phosphate. Plant cells lacking this enzyme189 are incapable of surviving on synthetic medium containing mannose

. Phytosanitary Plant health, including quarantine (FAO)

P lant Pest Any living stage (including active and dormant forms) of insects, mites, nematodes, slugs, snails, protozoa, or other invertebrate animals, bacteria, fungi, other parasitic plants or reproductive parts thereof; viruses; or any organisms similar to or allied with any of the foregoing; or any infectious agents or substances, which can directly or indirectly injure or cause disease or damage in or to any plants or parts thereof, or any processed, manufactured, or other products of plants. (7 CFR

340 .1)

Plasmid An circular self

- replicating non

- chromosomal DNA molecule found in many bacteria, capable of transfer between bacterial cells of the same species, and occasionally of different species. Antibiotic resistance genes are frequently located on pla smids. Plasmids are particularly important as vectors for genetic engineering (FAO)

PPA Plant Protection Act.
PPQ Plant Protection and Quarantine (USDA, APHIS).

Proline An amino acid. Some plant cells accumulate proline as an

osmoprotectant.

Promoter A region of DNA locat ed upstream of a gene that controls to what degree, where, and/or when a gene is expressed.
Push Test Scores The number of plants out of 10 plants tested that break at the stalk or have root failure after pushing to 45 degrees from ver tical.


### R

Recombinant DNA Technology The manipulation of DNA in which DNA, including DNA from different organisms, is cut apart and recombined using enzymes.

Recombination The physical exchange of genetic material between two genetic sequences that produces new combinations of genetic information. (See Homologous recombination and Non

- homologous

r ecombination.) Regulated Article Subject to APHIS regulation under 7 CFR p ar t 340. Regulatory

sequence A DNA sequence involved in regulating the expression of a gene, e.g. a promoter or operator region (in the DNA molecule) (FAO)

Ribonucleic Acid (RNA) A nucleic acid composed of a long, often single

- stranded chain of

chemical building blocks called ‘nucleotides.’ RNA has multiple functions in the process of translating information stored in genes (DNA) into proteins.

Ribonucleic Acid See RNA.

Rotation In crop production, the cycle of crops grown in successive years in the

same field. Rotations are instituted to limit the spread and accumulation of diseases (especially soil

- borne diseases) and pests and to manage plant nutrients.



### S190 Self

- pollinated Th e tendency of a plant species to produce offspring that result from a flower pollinating itself. (See Outcrossing.)

Senescence A late stage in the development of multicellular organisms, during which irreversible loss of function and degradation of biolo gical components occur. The physiological ageing process in which cells and tissues deteriorate and finally die (FAO).
Southern corn leaf blight Corn disease caused by the fungus Bipolaris maydis

Stratification Chilling or warming seeds, for a period of time, to improve germination. Sympatric populations Species undergoing sympatric speciation are not geographically isolated by, for example, a mountain or a river. The diverging populations generally share the same territory. (wiki)

Synergy The interacti on of two or more factors so that their combined effect is greater than the sum of their individual effects.



### T

TES Threatened and Endangered Species

. Tetraploid An organism, or a tissue whose cells contain four haploid sets of chromosomes (FAO)

Thermostable A molecule which retains its biological activity at some specified higher temperature (FAO)

Trait A characteristic of an organism that manifests itself in the phenotype. Traits may be the result of a single gene or may be polygenic, resulting from the simultaneous expression of more than one gene.

Transformant A cell or organism that has been genetically altered through the integration of a transgene(s). A “primary” transformant is the first generation following the transformation event.

Tra nsformation The uptake and integration of DNA in a cell’s genome, in which th e introduced DNA is intended to change the phenotype of the recipient organism in a predictable manner.

Transformation

Event A single successful integration of a gene or gene fra gment into a cell or a successful deletion of a gene or gene fragment from a cell. Transgene A foreign gene that is inserted into the genome of a cell via recombinant DNA techniques. Transgenic Organism An organism whose genome has been modified via the stable incorporation of a piece of foreign DNA (a transgene).


### U

V

Vector The agent, such as a plasmid, used by researchers to carry new genes

into cells.

Vigor A measure of plant growth, health and robustness during the vegetative growth stage191 Volunteer Plants resulting from crop seed that escapes harvest and remains in the field until subsequent seasons, where it germinates along with the succeeding crop.


### W

Weediness The ability of a plant to colonize a disturbed habitat and compete with cultivated species.

Whorl Vegetative stage of the corn plant