Monsanto Company I

10-CT-211E-65

 

TRANSMITTAL DOCUMENT

SUBMITTED BY

Monsanto Company

800 North Lindbergh Blvd.

St. Louis, MO 63167

REGULATORY ACTION IN SUPPORT OF WHICH

THIS PACKAGE IS SUBMITTED

Application for an Experimental Use Permit

524-EUP-XXX

TRANSMITTAL DATE

July 1, 2010

Registrant Document No

10-CT-211E-65

Monsanto Company 10-CT-211E-65 Page 1 of 87

Title

 Administrative Materials in Support of an Application for an Experimental Use 

Permit to Test Combined Trait Cotton COT102 × MON 15985 and Trait Cotton 

COT102 and MON 15985, Necessary for its Production 

(EPA File Symbol 524-EUP-XXX).

Data Requirement

40 CFR § 172

Authors

Cheryl A. Florida

Kara S. Giddings

Patricia G. Keller

Bryan Savoy

Ty K. Witten

Contributors

J. Austin Burns

Tracey A. Cavato

John T. Greenplate

Melinda C. McCann

Registrant Submission Date

July 1, 2010

Registrant Submitting

Monsanto Company

800 North Lindbergh Blvd.

St. Louis, MO 63167

Registrant Document No.

10-CT-211E-65

Volume 1 of 1


Monsanto Company 10-CT-211E-65 Page 4 of 87

Volume 1

Table of Contents

Title.......................................................................................................................................1

STATEMENT OF DATA CONFIDENTIALITY CLAIM.............................................2

GLP COMPLIANCE STATEMENT ...............................................................................3

Table of Contents................................................................................................................4

List of Tables.......................................................................................................................5

EPA Form 8570-17..............................................................................................................6

Confidential Statement of Formula............................................................................... 7-9

EPA Form 8540-4 Confidential Statement of Formula.................................................10

EPA Form 8570-35 Data Matrices ............................................................................ 11-16

Section A. Product Description .........................................................................................17

COT102 Right to Cite Letter.................................................................18-19

Section B. Proposed Experimental Use Permit Label.......................................................20

Master Label Subparts.....................................................................................................21

COT102 Label........................................................................................................22-23

MON 15985 Label..................................................................................................24-25

COT102 × MON 15985 Label ...............................................................................26-27

Section C. Toxicity Data ............................................................................................. 28-29

Section D. Residue Data....................................................................................................30

Section E. Effectiveness Data ...........................................................................................31

Section F. Tolerance..........................................................................................................32

Section G.1. Proposed Experimental Program..................................................................33

G.1.a Participants in the Experimental Program.......................................................33-34

G.1.b Cooperators in the Experimental Program...........................................................35

G.1.c Counties where experimental trials will not be conducted...................................36

Section G.2. Acreage, Seed and Active Ingredient Quantities.................................... 37-50

Section G.3. Program Details, Including Containment.....................................................51

G.3.a. Crop.....................................................................................................................51

G.3.b. Target Pests.........................................................................................................51

G.3.c. Major Geographic Areas .....................................................................................51

G.3.d. Sites.....................................................................................................................51

G.3.e. Season of Use ......................................................................................................51

G.3.f. Trial Types...........................................................................................................51

Protocol 1 – Breeding and Observation Nursery...................................................52-53

Protocol 2 – Seed Increase.....................................................................................54-55

Protocol 3 – Yield and Herbicide Tolerance Trials...............................................56-57

Protocol 4 – Insect Efficacy Trials........................................................................58-59

Protocol 5 – Product Characterization and Performance Trials ............................60-61

Protocol 6 – Insect Resistance Management Trials...............................................62-63

Protocol 7 – Benefit Trials.....................................................................................64-65

Protocol 8 – Seed Treatment Trials.......................................................................66-67

G.3.g. Containment Procedures ................................................................................68-70

Section G.4. Program Objectives......................................................................................71

Section G.5. Program Quantities.......................................................................................72

Section G.6. Proposed Duration........................................................................................73

Section G.7. Method of Disposition of Plant Materials....................................................74

References..........................................................................................................................75
Monsanto Company 10-CT-211E-65 Page 5 of 87

List of Table

Table 1. List of Participants who will supervise experimental work conducted .........33-34

Table 2. Names and addresses of Cooperators involved under this EUP......................... 35

Table 3. List of counties where experimental trials will not be conducted ...................... 36

Table 4. Proposed Total Trial Acreage by State for 2010-2012....................................... 39

Table 5. Proposed Acreage by PIP Test Material by State for 2010-2012 ....................... 40

Table 6. Proposed Total PIP Test Material Active Ingredient Quantities by State .......... 41

Table 7. Proposed Active Ingredient Quantities by State for COT102 for 2010-

2012.................................................................................................................... 42

Table 8. Proposed Active Ingredient Quantities by State for MON 15985 for 

2010 -2012.......................................................................................................... 43

Table 9. Proposed Active Ingredient Quantities by State for COT102 × MON 

15985 for 2010-2012 .......................................................................................... 44

Table 10. Requested Total Trial Acreage by Protocol for Each State for 2010-

2012.................................................................................................................... 45

Table 11. Requested Trial Acreage for COT102 by State by Protocol for 2010-

2012............................................................................................................... 46

Table 12. Requested Trial Acreage for MON 15985 by State by Protocol for 

2010-2012.......................................................................................................... 47

Table 13. Requested Trial Acreage for COT102 x MON 15985 by State by 

Protocol for 2010-2012 ...................................................................................... 48

Table 14. Requested Trial Acreage for Non-PIP by State by Protocol for 2010-

2012.................................................................................................................... 49

Table 15. Requested Trial Acreage for Borders by State by Protocol for 2010-

2012.................................................................................................................... 50



Monsanto Company 10-CT-211E-65 Page 11 to 16 of 87

Data Matrix

EPA Form 8570-35 Data Matrix

{CBI Cross Reference Number 2}
Monsanto Company 10-CT-211E-65 Page 17 of 87

Section A. Product Description

Monsanto Company has used conventional breeding techniques to develop the combined 

trait cotton product COT102 (Syngenta) × MON 15985 (Monsanto) that confers insect 

protection. Each single product contributes specific benefits to the final combined 

product. 

COT102 produces the Bacillus thuringiensis Vip3Aa protein that protects against 

lepidopteran feeding pests of cotton. COT102 was reviewed as part of COT102 ×

COT67B, which was registered by the EPA (EPA Registration No 67979-9) in June 

2008. COT102 has completed FDA consultation (July 5, 2005) and USDA deregulation 

(July 20, 2005). Currently, COT102 has no pending applications at EPA as a single 

event. A letter regarding the right to cite Syngenta Company data/documents containing 

the B.t. Vip3Aa19 protein, which are needed for this EUP submission, is found on page 

18 of this document. 

MON 15985 contains the Cry2Ab2 and the β-D-glucuronidase (GUS) insert, as well as 

the MON 531 insert containing Cry1Ac, aad, and nptII. This product protects against 

major lepidopteran pests in cotton. MON 15985 was registered by the EPA (EPA 

Registration No. 524-522) in December 23, 2002. MON 15985 has completed FDA 

consultation (July 18, 2002) and USDA deregulation (November 5, 2002). 

The safety of the Vip3Aa protein has been demonstrated and is exempt from the 

requirement of a tolerance (§174.501). The safety of the Cry proteins have been 

demonstrated and are exempted from the requirement of a tolerance (Cry1A, §174.510; 

Cry2Ab2, §174.519). 

The combination of three insecticidal proteins from COT102 and MON 15985, (e.g., 

Vip3Aa, Cry1Ac, and Cry2Ab2), provide enhanced insect control and offers an 

additional insect-resistance management tool. 
Monsanto Company 10-CT-211E-65 Pages 18 to 19 of 87

Syngenta Right to Cite Letter

{CBI Cross Reference Number 3}
Monsanto Company 10-CT-211E-65 Pages 20 of 87

Section B. Proposed Experimental Use Permit Label

The attached master label for testing under the experimental use permit contains the 

following: 

a proposed label for lepitopteran-protected cotton COT102 (vector pCOT1)

a label for lepitopteran-protected cotton MON 15985 (vectors PV-GHBK4 and 

PV-GHBK11) 

a proposed label for COT102 × MON 15985 (vectors pCOT1, PV-GHBK4 and 

PV-GHBK11) appropriate to the testing anticipated under this EUP
Monsanto Company 10-CT-211E-65 Pages 21 of 87

MASTER LABEL

524-EUP-XXX

Subparts:

1. Lepidopteran-Protected Cotton: COT102

2. Lepidopteran-Protected Cotton: MON 15985

3. Lepidopteran-Protected Cotton: COT102 × MON 15985
Monsanto Company 10-CT-211E-65 Pages 22 of 87

EPA EXPERIMENTAL USE PERMIT NUMBER 524-EUP-XXX

Lepidopteran-Protected Cotton: COT102

This package contains lepidopteran-protected cotton seed producing the Vip3Aa19 insecticidal 

protein from Bacillus thuringiensis. This protein is effective in controlling feeding damage 

caused by lepidopteran insect pests.

Active Ingredient:

Bacillus thuringiensis Vip3Aa19 insecticidal protein and the genetic material necessary for its 

production (via elements of vector pCOT1) in Event COT102 cotton

(SYN-IR102 7)........... .............................................. .........................<0.000985%*

Inert Ingredient:

Hygromycin B phosphotransferase (APH4), a marker protein, and the genetic material necessary 

for its production (via elements of pCOT1) in Event COT102 cotton (SYN-IR102-7)

...................................................................................................... <0.000189%*

*Maximum percent (wt/wt) on a dry weight basis. 

Keep Out of Reach of Children

CAUTION

Syngenta Seeds, Inc.

P.O. Box 12257

3054 E. Cornwallis Rd.

Research Triangle Park, NC 27709

EPA Establishment Number 66736-NC-1 NET CONTENTS: ____ pounds of cotton seed 

DIRECTIONS FOR USE

Use of this seed in a manner inconsistent with the terms of this Experimental Use Permit (EUP) is 

a violation of Federal law. Test plots must not be located within 1⁄4 of a mile from the habitats of 

endangered/threatened lepidopteran species as listed by the US Fish and Wildlife Service.

FOR EXPERIMENTAL USE ONLY

Not for sale to or use by any person other than a participant or cooperator of 

the EPA-approved Experimental Use Program.

For use only at a site of a participant or cooperator and in accordance with the 

terms and conditions of the Experimental Use Permit.

This labeling must be in possession of the user at the time of planting.
Monsanto Company 10-CT-211E-65 Pages 23 of 87

The contents may only be used according to the approved EUP program. Cooperators must have 

at least one copy of the protocols in Section G prior to initiating any research with the contents.

DISPOSAL

Any seed not used in these experiments must be returned to Monsanto Company or seed company 

partners/affiliates, or disposed of as specified in Section G.
Monsanto Company 10-CT-211E-65 Pages 24 of 87

EPA EXPERIMENTAL USE PERMIT NUMBER 524-EUP-XXX

Lepidopteran-Protected Cotton: MON 15985

This package contains lepidopteran-protected cotton seed producing the Cry1Ac and Cry2Ab2 

insecticidal proteins from Bacillus thuringiensis. These proteins are effective in controlling 

feeding damage caused by lepidopteran insect pests.

Active Ingredients:

Bacillus thuringiensis Cry1Ac protein and the genetic material necessary for its production 

(vector PV-GHBK04) in event MON 15985 cotton......................................0.00004 – 0.00052%*

 

Bacillus thuringiensis Cry2Ab2 protein and the genetic material necessary for its production 

(vector PV-GHBK11) in event MON 15985 cotton............................................. 0.003 – 0.009%*

Inert Ingredients: 

Substance produced by the marker gene and the genetic material necessary for its production 

(vector PV-GHBK04 and PV-GHBK11) in event MON 15985 cotton ..........0.0022 – 0.0304%*

 

*Maximum percent (wt/wt) on a dry weight basis. 

Keep Out Of Reach Of Children.

CAUTION

Monsanto Company 

800 North Lindbergh Blvd. 

St. Louis, MO 63167

 

 

EPA Establishment No. 524-MO-002 NET CONTENTS____pounds of cotton seed

 

DIRECTIONS FOR USE

Use of this seed in a manner inconsistent with the terms of this Experimental Use Permit (EUP) is 

a violation of Federal law. Test plots must not be located within 1⁄4 of a mile from the habitats of 

endangered/threatened lepidopteran species as listed by the US Fish and Wildlife Service.

FOR EXPERIMENTAL USE ONLY

Not for sale to or use by any person other than a participant or cooperator of 

the EPA-approved Experimental Use Program.

For use only at a site of a participant or cooperator and in accordance with the

terms and conditions of the Experimental Use Permit.

This labeling must be in possession of the user at the time of planting.
Monsanto Company 10-CT-211E-65 Pages 25 of 87

The contents may only be used according to the approved EUP program. Cooperators must have 

at least one copy of the protocols in Section G prior to initiating any research with the contents.

DISPOSAL

Any seed not used in these experiments must be returned to Monsanto Company or seed company 

partners/affiliates, or disposed of as specified in Section G.
Monsanto Company 10-CT-211E-65 Pages 26 of 87

EPA EXPERIMENTAL USE PERMIT NUMBER 524-EUP-XXX

Insect-Protected Cotton: COT102 × MON 15985

This package contains lepidopteran-protected cotton seed producing the Vip3Aa, Cry1Ac, and 

Cry2Ab2 insecticidal proteins from Bacillus thuringiensis. These proteins are effective in 

controlling feeding damage caused by lepidopteran insect pests.

Active Ingredients:

Bacillus thuringiensis Vip3Aa protein and the genetic material necessary for its production 

(vector pCOT1) in event COT102 cotton..........................................................0.00027-.00031%*

Bacillus thuringiensis Cry1Ac protein and the genetic material necessary for its production 

(vector PV-GHBK04) in event MON 15985 cotton........................................0.000092-0.0001%**

 

Bacillus thuringiensis Cry2Ab2 protein and the genetic material necessary for its production 

(vector PV-GHBK11) in event MON 15985 cotton...............................................0.041-0.048 %**

Inert Ingredients: 

Hygromycin B phosphotranferase (APH4), a marker protein and the genetic material necessary 

for its production (by elements of vector pCOT1) in event COT102 cotton 

........................................................................................0.000022 – 0.000025%*

NPTII protein and the genetic material necessary for its production (vector PV-GHBK04) in 

event MON 15985 cotton...........................................................0.00015 – 0.00017%**

GUS protein and the genetic material necessary for its production (vector PV-GHBK11) in event 

MON 15985 cotton..........................................................................0.01 – 0.013%**

* Maximum percent (wt/wt) on a dry weight basis

** Maximum percent (wt/wt) on a fresh weight basis

Keep Out Of Reach Of Children.

CAUTION

Monsanto Company 

800 North Lindbergh Blvd. 

St. Louis, MO 63167

 

EPA Establishment No. 524-MO-002 NET CONTENTS______pounds of cotton seed

FOR EXPERIMENTAL USE ONLY

Not for sale to or use by any person other than a participant or cooperator of 

the EPA-approved Experimental Use Program.

For use only at a site of a participant or cooperator and in accordance with the 

terms and conditions of the Experimental Use Permit.

This labeling must be in possession of the user at the time of planting.
Monsanto Company 10-CT-211E-65 Pages 27 of 87

 

DIRECTIONS FOR USE

Use of this seed in a manner inconsistent with the terms of this Experimental Use Permit (EUP) is 

a violation of Federal law. Test plots must not be located within 1⁄4 of a mile from the habitats of 

endangered/threatened lepidopteran species as listed by the US Fish and Wildlife Service.

The contents may only be used according to the approved EUP program. Cooperators must have 

at least one copy of the protocols in Section G prior to initiating any research with the contents.

DISPOSAL

Any seed not used in these experiments must be returned to Monsanto Company or seed company 

partners/affiliates, or disposed of as specified in Section G.
Monsanto Company 10-CT-211E-65 Pages 28 of 87

Section C. Toxicity Data 

Safety of VIP and Cry Proteins

The environmental, human and animal safety of vegetative insecticidal proteins (VIP) 

and insecticidal crystal (Cry) Bt proteins has been demonstrated by extensive scientific 

research (Peng et al., 2006; Betz et al., 2000; Siegel, 2001; and references cited therein). 

Based on numerous toxicology studies, the EPA has determined that products containing 

VIP and Cry proteins show no adverse effects, and these products are not toxic or 

pathogenic to humans (Peng et al., 2006, McClintock et al., 1995; EPA, 1998; EPA, 

2001a, b). These findings are relevant to the safety assessment of plant-incorporated 

protectants because microbial preparations generally contain the same classes of Cry 

proteins (Cry1 and Cry2) that have been introduced into cotton plants. 

VIP and Cry proteins have been safely used in biotechnology-derived crops for over a 

decade. EPA has registered the Vip3Aa, Cry1Ac, and Cry2Ab2 proteins as produced in 

cotton. The safety of these proteins has been demonstrated and these proteins are 

exempted from the requirement of a tolerance: 

Vip3Aa Protein

The Vip3Aa protein encoded by the gene inserted into COT102 has 789 amino acids. The 

amino acid sequence is identical to the native Bt protein except for one amino acid. At 

position 284 of the Vip3Aa protein there is a glutamine (compared with lysine for the 

native protein). The Vip3Aa protein as produced in COT102 was reviewed by EPA and 

granted an exemption from the requirement of a tolerance in cotton on August 6, 2008 

(40 CFR § 174.501). 

Cry1Ac Protein

Cry1Ac protein produced in MON 15985 is identical in length (1178 amino acids) and 

99.4% identical in amino acid sequence to the protein encoded by the naturally occurring 

cry1A(c) gene (Adang, et al., 1985). EPA reviewed the safety assessment of the Cry1Ac 

protein as produced in MON 531 and granted exemption from the requirement of a 

tolerance on April 25, 2007 (40 CFR § 174.510). Recombinant DNA techniques have 

Protein

Tolerance Exemption Information

40 CFR Date Crop(s)

Vip3Aa §174.501 Aug. 6, 2008 cotton

Cry1Ac §174.510 April 25, 2007 all

Cry2Ab2 §174.519 April 25, 2007 cotton
Monsanto Company 10-CT-211E-65 Pages 29 of 87

been used to generate Bt strains with enhanced insecticidal activity for use as 

biopesticides (Baum, 1998; and Baum et al., 1999; EPA 1995a). 

Cry2Ab2 Protein

The Cry2Ab2 protein produced in MON 15985 is 88% amino acid sequence identical to 

the Cry2Aa protein produced by the Bt bacterium (Widner and Whiteley, 1990; 

Dankocsik et al., 1990) and has 97% amino acid similarity (amino acid identities and 

conservative amino acid substitutions). EPA reviewed the safety assessment of the 

Cry2Ab2 protein as produced in MON 15985 and granted permanent exemption from the 

requirement of a tolerance on April 25, 2007 (40 CFR § 174.519). 
Monsanto Company 10-CT-211E-65 Pages 30 of 87

Section D. Residue Data

Residue data are not applicable because permanent tolerance exemptions exist for the 

following proteins: Vip3Aa, Cry1Ac, and Cry2Ab2. 
Monsanto Company 10-CT-211E-65 Pages 31 of 87

Section E. Effectiveness Data 

Not required under 40 CFR § 158.202(i).
Monsanto Company 10-CT-211E-65 Pages 32 of 87

Section F. Tolerance if Testing Involves Use in Food or Feed

Permanent tolerance exemptions exist for proteins Vip3Aa, Cry1Ac, and Cry2Ab2.

Residues of Bacillus thuringiensis Vip3Aa protein (40 CFR § 174.501) in or on corn or 

cotton are exempt from the requirement of a tolerance when used as plant-incorporated 

protectants in or on the food and feed commodities of corn; corn, field; corn, sweet; corn, 

pop; and cotton; cotton, undelinted seed; cotton, refined oil; cotton, meal; cotton, hay; 

cotton, hulls; cotton, forage; and cotton, gin byproducts.

Residues of Bacillus thuringiensis Cry1Ac protein (40 CFR § 174.510) in all plants are 

exempt from the requirement of a tolerance when used as plant-incorporated protectants 

in all food commodities.

Residues of Bacillus thuringiensis Cry2Ab2 protein (40 CFR § 174.519) in or on corn or 

cotton are exempt from the requirement of a tolerance when used as a plant-incorporated 

protectant in the food and feed commodities of corn: corn, field; corn, sweet; corn, pop; 

and cotton seed, cotton oil, cotton meal, cotton hay, cotton hulls, cotton forage, and 

cotton gin byproducts.
Monsanto Company 10-CT-211E-65 Pages 33 to 34 of 87

Section G.1. Proposed Experimental Program

G.1.a. Participants in the Experimental Program 

Table 1. List of Participants who will supervise experimental work conducted under 

this EUP. 

All Participants are research scientists employed by Monsanto Company.

{CBI Cross Reference Number 4}
Monsanto Company 10-CT-211E-65 Pages 35 of 87

G.1.b. Cooperators in the Experimental Program

Table 2. Names and addresses of the Cooperators involved in the experimental work conducted 

under this EUP.

The names, addresses and telephone numbers of the Cooperators involved in the experimental 

work conducted under this EUP will be provided to the Agency prior to planting.
Monsanto Company 10-CT-211E-65 Pages 36 of 87

G.1.c. Counties containing threatened or endangered Lepidoptera habitats 

Experimental trials will only be conducted in EUP approved states. Experimental trials will not 

be conducted within 1⁄4 mile buffer of identified threatened or endangered Lepidoptera habitats. 

The most accurate information available to Monsanto Company regarding the location of 

threatened or endangered Lepidoptera habitats was considered when creating the list of counties 

that contain locations where EUP trials will not be conducted. The sources used to create the list 

of counties are stated below. The counties are listed in Table 3. 

County-Level species presence information in Table 3 was obtained from FIFRA Endangered 

species Task Force (FESTF) Information Management System (IMS). For this analysis, countylevel species location data from two sources was considered:

1. U.S. Fish and Wildlife Service (USFWS)

2. FESTF Multi-Jurisdictional Database (also known as “NatureServe”)a

Prior to the time of planting, if any location is removed from FIFRA Endangered Species Task 

Force (FESTF) Information Management System (IMS), experimental trials may be conducted 

in those areas under this EUP.

Table 3: Counties containing threatened or endangered Lepidoptera habitats

State County

AL Bibb, Greene, Tuscaloosa, Hale, Perry, Colbert, Fayette, Franklin, Lauderdale, 

Marengo, Pickens, Sumter

CA Alameda , Contra Costa , Del Norte , Humboldt , Kern , Lassen , Los Angeles , 

Marin , Mendocino , Monterey , Napa , Orange , Riverside , Sacramento, San 

Bernardino , San Diego , San Francisco , San Luis Obispo , San Mateo , Santa 

Barbara , Santa Clara , Santa Cruz , Solano , Sonoma , Yolo, Ventura

FL Miami-Dade, Monroe

HI Hawaii, Honolulu, Kauai, Maui

MS Alcorn, Itawamba, Lee, Prentiss, Tippah, Tishomingo, Union

NC Cumberland, Hoke

NV Carson City (city)b, Washoeb

TN Hardin

VA Carroll , Floyd , Franklin , Montgomery , Patrick , Pulaski

a

Sub-county location data are available in the counties where NatureServe is identified as a data 

source.

b

These Nevada counties are adjacent to California.
Monsanto Company 10-CT-211E-65 Pages 37 of 87

Section G.2 Acreage, Seed and Active Ingredient Quantities by State or Possession

A total of 13663 acres are requested for this EUP over an 18 month period, which includes 1897

acres of PIP test materials, 909 acres of non-PIP material, and 10857 acres of border materials 

(Table 4). The total amount of active ingredient for each test material of the following: Vip3Aa, 

Cry1Ac, and Cry2Ab2, were calculated as a sum of the planting seed of all events that produce 

this protein per state over the 18 month period of the EUP (Table 5). This is based on the 

amount of active ingredient calculated per PIP test material product per state (Tables 6-9). 

Calculation of active ingredient for each protein below is based on a maximum level of that 

protein in cotton seed on a dry weight basis. The calculations to determine the amount of 

active ingredient (lbs) in each PIP test material in Tables 6-9 were performed using an 

estimated planting density of 35 pounds of planting seed per acre equivalent to approximately 

161000 seeds per acre. Expression levels for calculation of each protein for PIP test materials 

are as follows: 

Protein Expression level (μg/g)

Vip3Aa 9.85

Cry1Ac 5.2

Cry2Ab2 88.7

Note: Total values may differ slightly from the sum of individual values due to rounding.

Tables 4 and 5 list the proposed acreage by PIP by state for 2010 through 2012. The PIP test 

materials included in this EUP, referred to as “PIP test materials” are as follows (inclusive):

Primary Test Material

COT102 MON 15985

Other Test Materials

COT102 

MON 15985 

The acreage for PIP test materials includes acreage for each individual PIP test material. The 

acreage for non-PIP materials includes acreage for any non-PIP breeding materials and any 

non-intentional recipients of PIP test materials within the experimental trial. The acreage for 

border materials includes acreage intentionally planted as border adjacent to trial (if 

applicable).
Monsanto Company 10-CT-211E-65 Pages 38 of 87

Table 10 lists the requested total trial acreage by protocol for each state for 2010-2012. The 

protocols are listed by number code as follows:

Protocol 1 - Breeding and Observation Nursery 

Protocol 2 - Seed Increase

Protocol 3 - Yield and Herbicide Tolerance Trials 

Protocol 4 - Insect Efficacy Trials 

Protocol 5 - Product Characterization and Performance Trials 

Protocol 6 - Insect Resistance Management Trials 

Protocol 7 - Benefit Trials

Protocol 8 - Seed Treatment Trials

Tables 11-15 list the requested trial acreage for each PIP test material, by state and by protocol 

for 2010-2012. 
Monsanto Company 10-CT-211E-65 Pages 39 of 87

Table 4. Proposed Total Trial Acreage by State for 2010-2012

STATE

Acres

PIP Non-PIP Border Total

AL 117 55 626 798

AR 177 81 775 1033

AZ 159 83 797 1039

CA 48 30 546 624

FL 48 16 63 127

GA 117 55 644 816

HI 13 3 20 36

KS 51 29 548 628

KY 42 26 536 604

LA 129 59 644 832

MD 42 26 536 604

MO 102 46 605 753

MS 135 59 665 859

NM 92 44 586 722

NC 120 56 635 811

OK 42 26 536 604

PR 64 34 135 233

SC 129 59 644 832

TN 111 51 626 788

TX 132 62 658 852

VA 27 9 32 68

Total 1897 909 10857 13663
Monsanto Company 10-CT-211E-65 Pages 40 of 87

Table 5. Proposed Acreage by PIP Test Material by State for 2010-2012 

States

Acres × PIP Test Materials

COT102 MON 15985

COT102

X

MON 15985

AL 37 37 43

AR 57 57 63

AZ 51 51 57

CA 14 14 20

FL 16 16 16

GA 37 37 43

HI 3 4 6

KS 15 15 21

KY 12 12 18

LA 41 41 47

MD 12 12 18

MO 32 32 38

MS 43 43 49

NM 29 28 35

NC 38 38 44

OK 12 12 18

PR 20 21 23

SC 41 41 47

TN 35 35 41

TX 42 42 48

VA 9 9 9

Total 596 597 704
Monsanto Company 10-CT-211E-65 Pages 41 of 87

Table 6. Proposed Total PIP Test Material Active Ingredient Quantities by state for 2010-2012

States Planting Seed (lb) Acres × PIP Test Materials (grams)

Vip3Aa Cry1Ac Cry2Ab2

AL 4095 12.51 6.60 112.65

AR 6195 18.76 9.91 168.99

AZ 5565 16.89 8.92 152.09

CA 1680 5.32 2.81 47.87

FL 1680 5.00 2.64 45.06

GA 4095 12.51 6.60 112.65

HI 455 1.41 0.83 14.08

KS 1785 5.63 2.97 50.69

KY 1470 4.69 2.48 42.25

LA 4515 13.76 7.26 123.92

MD 1470 4.69 2.48 42.25

MO 3570 10.94 5.78 98.57

MS 4725 14.38 7.60 129.55

NM 3220 10.00 5.20 88.72

NC 4200 12.82 6.77 115.47

OK 1470 4.69 2.48 42.25

PR 2240 6.73 3.63 61.96

SC 4515 13.76 7.26 123.92

TN 3885 11.88 6.27 107.03

TX 4620 14.08 7.43 126.73

VA 945 2.82 1.48 25.34

Total 66395 203.27 107.40 1832.04

Active Ingredient –

Pounds (lbs) 0.45* 0.24* 4.04*

* Activity Total is the sum of a given active ingredient from all PIPs in the program: single event COT102 or MON 15985, 

and the combined trait product COT102 × MON 15986. 
Monsanto Company 10-CT-211E-65 Pages 42 of 87

Table 7. Proposed Active Ingredient Quantities by State for COT102 for 2010-2012 

States Planting seed (lb) Active Ingredient in Planting Seed (g)

Vip3Aa Cry1Ac Cry2Ab2

AL 1295 5.79 - -

AR 1995 8.91 - -

AZ 1785 7.98 - -

CA 490 2.19 - -

FL 560 2.50 - -

GA 1295 5.79 - -

HI 105 0.47 - -

KS 525 2.35 - -

KY 420 1.88 - -

LA 1435 6.41 - -

MD 420 1.88 - -

MO 1120 5.00 - -

MS 1505 6.72 - -

NM 1015 4.53 - -

NC 1330 5.94 - -

OK 420 1.88 - -

PR 700 3.13 - -

SC 1435 6.41 - -

TN 1225 5.47 - -

TX 1470 6.57 - -

VA 315 1.41 - -

Total 20860 93.21 - -

Active Ingredient -

Pounds (lbs) 0.21 - -

 
Monsanto Company 10-CT-211E-65 Pages 43 of 87

Table 8. Proposed Active Ingredient Quantities by State for MON 15985 for 2010-2012 

States Planting seed (lb)

Active Ingredient in Planting Seed (g)

Vip3Aa Cry1Ac Cry2Ab2

AL 1295 - 3.05 52.10

AR 1995 - 4.71 80.27

AZ 1785 - 4.21 71.82

CA 490 - 1.16 19.71

FL 560 - 1.32 22.53

GA 1295 - 3.05 52.10

HI 140 - 0.33 5.63

KS 525 - 1.24 21.12

KY 420 - 0.99 16.90

LA 1435 - 3.38 57.74

MD 420 - 0.99 16.90

MO 1120 - 2.64 45.06

MS 1505 - 3.55 60.55

NM 980 - 2.31 39.43

NC 1330 - 3.14 53.51

OK 420 - 0.99 16.90

PR 735 - 1.73 29.57

SC 1435 - 3.38 57.74

TN 1225 - 2.89 49.29

TX 1470 - 3.47 59.14

VA 315 - 0.74 12.67

Total 20895 - 49.27 840.68

Active Ingredient -

Pounds (lbs) - 0.11 1.85
Monsanto Company 10-CT-211E-65 Pages 44 of 87

Table 9. Proposed Active Ingredient Quantities by State for COT102 × MON 15985 for 2010-2012 

States Planting seed (lb) Active Ingredient in Planting Seed (g)

Vip3Aa Cry1Ac Cry2Ab2

AL 1505 6.72 3.55 60.55

AR 2205 9.85 5.20 88.72

AZ 1995 8.91 4.71 80.27

CA 700 3.13 1.65 28.16

FL 560 2.50 1.32 22.53

GA 1505 6.72 3.55 60.55

HI 210 0.94 0.50 8.45

KS 735 3.28 1.73 29.57

KY 630 2.81 1.49 25.35

LA 1645 7.35 3.88 66.18

MD 630 2.81 1.49 25.35

MO 1330 5.94 3.14 53.51

MS 1715 7.66 4.05 69.00

NM 1225 5.47 2.89 49.29

NC 1540 6.88 3.63 61.96

OK 630 2.81 1.49 25.35

PR 805 3.60 1.90 32.39

SC 1645 7.35 3.88 66.18

TN 1435 6.41 3.38 57.74

TX 1680 7.51 3.96 67.59

VA 315 1.41 0.74 12.67

Total 24640 110.06 58.13 991.36

Active Ingredient -

Pounds (lbs) 0.24 0.13 2.19
Monsanto Company 10-CT-211E-65 Pages 45 of 87

Table 10. Requested Total Trial Acreage by Protocol for Each State for 2010-2012

Protocol Number

STATE 1 2 3 4 5 6 7 8 Total

AL 0 0 26 20 632 43 68 9 798

AR 0 85 26 14 762 53 84 9 1033

AZ 0 163 40 0 746 31 50 9 1039

CA 0 0 20 0 604 0 0 0 624

FL 0 0 8 0 24 33 62 0 127

GA 0 0 26 18 638 49 76 9 816

HI 36 0 0 0 0 0 0 0 36

KS 0 0 0 0 612 8 8 0 628

KY 0 0 0 0 604 0 0 0 604

LA 0 0 26 18 642 53 84 9 832

MD 0 0 0 0 604 0 0 0 604

MO 0 0 22 0 632 37 62 0 753

MS 10 28 28 14 642 53 84 0 859

NM 0 0 26 2 624 27 43 0 722

NC 0 0 26 18 642 47 78 0 811

OK 0 0 0 0 604 0 0 0 604

PR 36 177 0 20 0 0 0 0 233

SC 0 0 26 18 642 53 84 9 832

TN 0 0 22 8 642 45 71 0 788

TX 0 0 58 6 642 53 84 9 852

VA 0 0 0 0 20 20 28 0 68

Total 82 453 380 156 10958 605 966 63 13663
Monsanto Company 10-CT-211E-65 Pages 46 of 87

Table 11. Requested Trial Acreage for COT102 by State by Protocol for 2010-2012

Protocol Number

STATE 1 2 3 4 5 6 7 8 Total

AL 0 0 3 2 16 6 9 1 37

AR 0 8 3 2 25 7 11 1 57

AZ 0 14 3 0 23 4 6 1 51

CA 0 0 2 0 12 0 0 0 14

FL 0 0 1 0 3 4 8 0 16

GA 0 0 3 2 16 6 9 1 37

HI 3 0 0 0 0 0 0 0 3

KS 0 0 0 0 13 1 1 0 15

KY 0 0 0 0 12 0 0 0 12

LA 0 0 3 2 17 7 11 1 41

MD 0 0 0 0 12 0 0 0 12

MO 0 0 3 0 16 5 8 0 32

MS 1 2 3 2 17 7 11 0 43

NM 0 0 3 1 15 4 6 0 29

NC 0 0 3 2 17 6 10 0 38

OK 0 0 0 0 12 0 0 0 12

PR 3 15 0 2 0 0 0 0 20

SC 0 0 3 2 17 7 11 1 41

TN 0 0 3 0 17 6 9 0 35

TX 0 0 4 2 17 7 11 1 42

VA 0 0 0 0 3 3 3 0 9

Total 7 39 40 19 280 80 124 7 596
Monsanto Company 10-CT-211E-65 Pages 47 of 87

Table 12. Requested Trial Acreage for MON 15985 by State by Protocol for 2010-2012

Protocol Number

STATE 1 2 3 4 5 6 7 8 Total

AL 0 0 3 2 16 6 9 1 37

AR 0 8 3 2 25 7 11 1 57

AZ 0 14 3 0 23 4 6 1 51

CA 0 0 2 0 12 0 0 0 14

FL 0 0 1 0 3 4 8 0 16

GA 0 0 3 2 16 6 9 1 37

HI 4 0 0 0 0 0 0 0 4

KS 0 0 0 0 13 1 1 0 15

KY 0 0 0 0 12 0 0 0 12

LA 0 0 3 2 17 7 11 1 41

MD 0 0 0 0 12 0 0 0 12

MO 0 0 3 0 16 5 8 0 32

MS 1 2 3 2 17 7 11 0 43

NM 0 0 3 0 15 4 6 0 28

NC 0 0 3 2 17 6 10 0 38

OK 0 0 0 0 12 0 0 0 12

PR 4 15 0 2 0 0 0 0 21

SC 0 0 3 2 17 7 11 1 41

TN 0 0 3 0 17 6 9 0 35

TX 0 0 4 2 17 7 11 1 42

VA 0 0 0 0 3 3 3 0 9

Total 9 39 40 18 280 80 124 7 597
Monsanto Company 10-CT-211E-65 Pages 48 of 87

Table 13. Requested Trial Acreage for COT102 x MON 15985 by State by Protocol for 2010-2012

Protocol Number

STATE 1 2 3 4 5 6 7 8 Total

AL 0 0 3 2 22 6 9 1 43

AR 0 8 3 2 31 7 11 1 63

AZ 0 14 3 0 29 4 6 1 57

CA 0 0 2 0 18 0 0 0 20

FL 0 0 1 0 3 4 8 0 16

GA 0 0 3 2 22 6 9 1 43

HI 6 0 0 0 0 0 0 0 6

KS 0 0 0 0 19 1 1 0 21

KY 0 0 0 0 18 0 0 0 18

LA 0 0 3 2 23 7 11 1 47

MD 0 0 0 0 18 0 0 0 18

MO 0 0 3 0 22 5 8 0 38

MS 1 2 3 2 23 7 11 0 49

NM 0 0 3 1 21 4 6 0 35

NC 0 0 3 2 23 6 10 0 44

OK 0 0 0 0 18 0 0 0 18

PR 6 15 0 2 0 0 0 0 23

SC 0 0 3 2 23 7 11 1 47

TN 0 0 3 0 23 6 9 0 41

TX 0 0 4 2 23 7 11 1 48

VA 0 0 0 0 3 3 3 0 9

Total 13 39 40 19 382 80 124 7 704
Monsanto Company 10-CT-211E-65 Pages 49 of 87

Table 14. Requested Trial Acreage for Non-PIP by State by Protocol for 2010-2012

Protocol Number

STATE 1 2 3 4 5 6 7 8 Total

AL 0 0 5 4 30 6 9 1 55

AR 0 16 5 2 39 7 11 1 81

AZ 0 26 9 0 37 4 6 1 83

CA 0 0 4 0 26 0 0 0 30

FL 0 0 1 0 3 4 8 0 16

GA 0 0 5 4 30 6 9 1 55

HI 3 0 0 0 0 0 0 0 3

KS 0 0 0 0 27 1 1 0 29

KY 0 0 0 0 26 0 0 0 26

LA 0 0 5 4 31 7 11 1 59

MD 0 0 0 0 26 0 0 0 26

MO 0 0 3 0 30 5 8 0 46

MS 1 2 5 2 31 7 11 0 59

NM 0 0 5 0 29 4 6 0 44

NC 0 0 5 4 31 6 10 0 56

OK 0 0 0 0 26 0 0 0 26

PR 3 27 0 4 0 0 0 0 34

SC 0 0 5 4 31 7 11 1 59

TN 0 0 3 2 31 6 9 0 51

TX 0 0 12 0 31 7 11 1 62

VA 0 0 0 0 3 3 3 0 9

Total 7 71 72 30 518 80 124 7 909
Monsanto Company 10-CT-211E-65 Pages 50 of 87

Table 15. Requested Trial Acreage for Borders by State by Protocol for 2010-2012

Protocol Number

STATE 1 2 3 4 5 6 7 8 Total

AL 0 0 12 10 548 19 32 5 626

AR 0 45 12 6 642 25 40 5 775

AZ 0 95 22 0 634 15 26 5 797

CA 0 0 10 0 536 0 0 0 546

FL 0 0 4 0 12 17 30 0 63

GA 0 0 12 8 554 25 40 5 644

HI 20 0 0 0 0 0 0 0 20

KS 0 0 0 0 540 4 4 0 548

KY 0 0 0 0 536 0 0 0 536

LA 0 0 12 8 554 25 40 5 644

MD 0 0 0 0 536 0 0 0 536

MO 0 0 10 0 548 17 30 0 605

MS 6 20 14 6 554 25 40 0 665

NM 0 0 12 0 544 11 19 0 586

NC 0 0 12 8 554 23 38 0 635

OK 0 0 0 0 536 0 0 0 536

PR 20 105 0 10 0 0 0 0 135

SC 0 0 12 8 554 25 40 5 644

TN 0 0 10 6 554 21 35 0 626

TX 0 0 34 0 554 25 40 5 658

VA 0 0 0 0 8 8 16 0 32

Total 46 265 188 70 9498 285 470 35 10857
Monsanto Company 10-CT-211E-65 Pages 51 of 87

Section G.3 Program Details, Including Containment

G.3.a. Crop

Cotton (Gossypium hirsutum L.)

This program involves testing of the combined insecticidal cotton product COT102 MON 

15985 (vectors pCOT1, PV-GHBK4 and PV-GHBK11), and two single products, COT102

(vector pCOT1) and MON 15985 (PV-GHBK04 and PV-GHBK11). Other registered PIP and/or 

non-PIP test materials may be planted in experimental trials.

Monsanto Company has used conventional breeding techniques to develop the combined trait 

cotton product COT102 (Syngenta) × MON 15985 (Monsanto), that confers insect protection. 

Each biotechnology-derived trait contributes specific benefits to the final combined product. 

COT102 produces an insecticidal protein that protects against feeding damage caused by several 

lepidopteran species including, but not limited to Helicoverpa zea (cotton bollworm), Heliothis 

virescens (tobacco budworm, Pectinophora gossypiella (pink bollworm), Spodoptera frugiperda

(fall armyworm), Spodoptera exigua (beet armyworm), Pseudoplusia includens (soybean 

looper), Trichoplusia ni (cabbage looper), and Bucculatrix thurberiella (cotton leaf perforator).

MON 15985 produces two Bacillus thuringiensis insecticidal proteins, Cry1Ac and Cry2Ab2

that protect against damage by major lepidopteran pests including cotton bollworm, tobacco 

budworm, pink bollworm, and armyworm.

G.3.b. Target Pests 

Lepidopteran insect pests.

G.3.c. Major Geographic Areas

Twenty-one states and the Commonwealth of Puerto Rico, representing diverse agronomic 

zones.

G.3.d. Sites 

Research stations, seed production fields, privately owned farms, and university farms.

G.3.e. Season of Use 

Plantings from December 1, 2010 through June 30, 2012

G.3.f. Trial Types

Protocol 1 - Breeding and Observation Nursery 

Protocol 2 - Seed Increase

Protocol 3 - Yield and Herbicide Tolerance Trials 

Protocol 4 - Insect Efficacy Trials 

Protocol 5 - Product Characterization and Performance Trials 

Protocol 6 - Insect Resistance Management Trials 

Protocol 7 - Benefit Trials

Protocol 8 - Seed Treatment Trials
Monsanto Company 10-CT-211E-65 Pages 52 of 87

G.3.f.1. Protocol 1 - Breeding and Observation Nursery

In the instance that USDA regulated plant materials are planted in an EUP plot, Monsanto will 

manage the EUP plot consistent with USDA Performance Standards in 7 CFR § 340.3 or as 

directed in USDA permits. 

Objective

The objectives of these activities are to: 1) develop breeding material intended for future 

experimental testing and observation, 2) maintain these biotechnology-derived products through 

seed production, and 3) conduct small plot field trials to evaluate phenotype and agronomic 

performance. 

Description

The cotton plants may be open-pollinated or hand-pollinated. The cotton plants may be selfpollinated, backcrossed, or crossed to conventional or other biotechnology-derived cotton plants.

Progeny derived from crosses may be increased to provide adequate seed quantities for future 

experimental testing. Samples may be collected for various types of analyses. Other registered 

PIP or Non-PIP material may be used in these trials.

Genotypes and Vectors

The PIP test materials will consist of insect-protected cotton in different genetic backgrounds as 

previously described.

Sampling

Plant tissue (including seed) and/or whole plant samples may be collected during the growing 

season and returned to Monsanto or other laboratories for analysis.

Isolation

PIP test material will be isolated to prevent persistence in the environment. Isolation methods 

will be implemented consistent with USDA Performance Standards at 7 CFR § 340.3(c).

Schedule

Hawaii and Puerto Rico: Number of plantings per site: continuous, but includes a minimum 30-

day irrigated and monitored fallow period after each harvest. Rain may substitute for irrigation 

during that period in Puerto Rico. 

Continental U.S.: Number of plantings per site: up to eight.

 Approx. Planting Dates: February through July

 Approx. Harvest Dates: August through January

Harvest Procedures

Plots may be harvested by hand or mechanically. Harvested seed cotton will be placed in labeled 

containers of such construction to prevent loss of seed. Containers may include but are not 

limited to mesh or cloth bags, boxes, bins, etc. Harvested seed cotton may be used for further 

research or future planting seed. Border rows, or plots that will not be used for further research 

or planting seed purposes, may be mechanically or hand harvested in bulk and disposed of as 

described below (see Final Disposition). Seed cotton or seed not to be used in future research, or 
Monsanto Company 10-CT-211E-65 Pages 53 of 87

as future planting seed will be disposed of as described below (see Final Disposition). All 

equipment will be thoroughly cleaned prior to exiting the plot area.

Final Disposition

Unused planting seed either 1) will be used for other research purposes, 2) will be returned to 

Monsanto or seed company partners and affiliates according to approved methods, 3) will be 

destroyed by grinding, burning, incineration, autoclaving, composting, or burial in the test field, 

or 4) returned to a secure on-site storage location. Seed and other plant materials may be 

harvested and used in further research or as future planting seed. Following completion of data 

collection and/or harvest, all cotton plants in the trial will be destroyed (see G.7).

Volunteer Plants

If plants in the trial plot are allowed to flower, the plot area and border rows will be monitored 

for volunteers and all volunteers will be destroyed. The occurrence of volunteer plants will be 

minimized by growing regulated material in defined areas in the field and by utilizing adequate 

termination protocols as outlined in the Final Disposition Procedures in Section G.7. In the 

continental U.S., the plot area will be monitored for a minimum period of twelve months after 

harvest and/or crop destruct. If volunteers still are present after this period, the Cooperator will 

continue to monitor the test plots until no volunteers are present. Plots in Hawaii and Puerto 

Rico will utilize a minimum 30-day irrigated fallow period, or any additional time period 

mandated by the State Department of Agriculture, during which plots will be monitored for 

germinating cotton plants. Rain may substitute for irrigation during that period in Puerto Rico. 

All volunteers will be removed prior to flowering. Volunteer monitoring will not be required for 

plots that are destroyed prior to flowering. Volunteer monitoring requirements are not applicable 

if all materials under this EUP are deregulated by the USDA.
Monsanto Company 10-CT-211E-65 Pages 54 of 87

G.3.f.2. Protocol 2 - Seed Increase 

In the instance that USDA regulated plant materials are planted in an EUP plot, Monsanto will 

manage the EUP plot consistent with USDA Performance Standards in 7 CFR § 340.3 or as 

directed in USDA permits. 

Objective

Produce seed and plant material for further testing and product development in various genetic 

backgrounds.

Description

Plants may be hand-pollinated or allowed to open pollinate in isolation to generate seed. Other 

registered PIP or Non-PIP material may be used in these trials.

Genotypes and Vectors

The PIP test materials will consist of insect-protected cotton in different genetic backgrounds as 

previously described.

Sampling

Plant tissue (including seed) and/or whole plant samples may be collected during the growing 

season and returned to Monsanto or other laboratories for analysis.

Isolation

PIP test material will be isolated to prevent persistence in the environment. Isolation methods 

will be implemented consistent with USDA Performance Standards at 7 CFR § 340.3(c).

Schedule

Hawaii and Puerto Rico: Number of plantings per site: continuous, but includes a minimum 30-

day irrigated and monitored fallow period after each harvest. Rain may substitute for irrigation 

during that period in Puerto Rico. 

Continental U.S.: Number of plantings per site: up to eight.

 Approx. Planting Dates: February through July

 Approx. Harvest Dates: August through January

Harvest Procedures

Plots may be harvested by hand or mechanically. Harvested seed cotton will be placed in labeled 

containers of such construction to prevent loss of seed. Containers may include but are not 

limited to mesh or cloth bags, boxes, bins etc. Harvested seed cotton may be used for further 

research or future planting seed. Border rows, or plots that will not be used for further research 

or planting seed purposes, may be mechanically or hand harvested in bulk and disposed of as 

described below (see Final Disposition). Seed cotton or seed not to be used in future research, or 

as future planting seed will be disposed of as described below (see Final Disposition). All 

equipment will be thoroughly cleaned prior to exiting the plot area.
Monsanto Company 10-CT-211E-65 Pages 55 of 87

Final Disposition

Unused planting seed either 1) will be used for other research purposes, 2) will be returned to 

Monsanto or seed company partners and affiliates according to approved methods, 3) will be 

destroyed by grinding, burning, incineration, autoclaving, composting, or burial in the test field, 

or 4) returned to a secure on-site storage location. Seed and other plant materials may be 

harvested and used in further research or as future planting seed. Following completion of data 

collection and/or harvest, all cotton plants in the trial will be destroyed (see G.7).

Volunteer Plants

If plants in the trial plot are allowed to flower, the plot area and border rows will be monitored 

for volunteers and all volunteers will be destroyed. The occurrence of volunteer plants will be 

minimized by growing regulated material in defined areas in the field and by utilizing adequate 

termination protocols as outlined in the Final Disposition Procedures in Section G.7. In the 

continental U.S., the plot area will be monitored for a minimum period of twelve months after 

harvest and/or crop destruct. If volunteers still are present after this period, the Cooperator will 

continue to monitor the test plots until no volunteers are present. Plots in Hawaii and Puerto 

Rico will utilize a minimum 30-day irrigated fallow period, or any additional time period 

mandated by the State Department of Agriculture, during which plots will be monitored for 

germinating cotton plants. Rain may substitute for irrigation during that period in Puerto Rico. 

All volunteers will be removed prior to flowering. Volunteer monitoring will not be required for 

plots that are destroyed prior to flowering. Volunteer monitoring requirements are not applicable 

if all materials under this EUP are deregulated by the USDA.
Monsanto Company 10-CT-211E-65 Pages 56 of 87

G.3.f.3. Protocol 3 - Yield and Herbicide Tolerance Trials 

In the instance that USDA regulated plant materials are planted in an EUP plot, Monsanto will 

manage the EUP plot consistent with USDA Performance Standards in 7 CFR § 340.3 or as 

directed in USDA permits. 

Objective

Assess any potential agronomic effects of the genetic insertion in PIP test materials and any 

effects of combining these in genetic backgrounds. In addition, crops may be sprayed with 

herbicides to assess crop tolerance.

Description

Cotton lines will be evaluated for agronomic characteristics, such as yield, fiber quality, 

maturity, and plant growth characteristics. Lines may also be sprayed with herbicides to assess 

crop tolerance, or may be sampled for various tests. Other registered PIP or Non-PIP material 

may be used in these trials.

Genotypes and Vectors

The PIP test materials will consist of insect-protected cotton in different genetic backgrounds as 

previously described.

Sampling

Plant tissue (including seed) and/or whole plant samples may be collected during the growing 

season and returned to Monsanto or other laboratories for analysis.

Isolation

PIP test material will be isolated to prevent persistence in the environment. Isolation methods 

will be implemented consistent with USDA Performance Standards at 7 CFR § 340.3(c).

Schedule

Hawaii and Puerto Rico: Number of plantings per site: continuous, but includes a minimum 30-

day irrigated and monitored fallow period after each harvest. Rain may substitute for irrigation 

during that period in Puerto Rico. 

Continental U.S.: Number of plantings per site: up to eight.

 Approx. Planting Dates: February through July

 Approx. Harvest Dates: August through January

Harvest Procedures

Plots may be harvested by hand or mechanically. Harvested seed cotton will be placed in labeled 

containers of such construction to prevent loss of seed. Containers may include but are not 

limited to mesh or cloth bags, boxes, bins etc. Harvested seed cotton may be used for further 

research or future planting seed. Border rows, or plots that will not be used for further research 

or planting seed purposes, may be mechanically or hand harvested in bulk and disposed of as 

described below (see Final Disposition). Seed cotton or seed not to be used in future research, or
Monsanto Company 10-CT-211E-65 Pages 57 of 87

as future planting seed will be disposed of as described below (see Final Disposition). All 

equipment will be thoroughly cleaned prior to exiting the plot area. 

Final Disposition

Unused planting seed either 1) will be used for other research purposes, 2) will be returned to 

Monsanto or seed company partners and affiliates according to approved methods, 3) will be 

destroyed by grinding, burning, incineration, autoclaving, composting, or burial in the test field, 

or 4) returned to a secure on-site storage location. Seed and other plant materials may be 

harvested and used in further research or as future planting seed. Following completion of data 

collection and/or harvest, all cotton plants in the trial will be destroyed (see G.7).

Volunteer Plants

If plants in the trial plot are allowed to flower, the plot area and border rows will be monitored 

for volunteers and all volunteers will be destroyed. The occurrence of volunteer plants will be 

minimized by growing regulated material in defined areas in the field and by utilizing adequate 

termination protocols as outlined in the Final Disposition Procedures in Section G.7. In the 

continental U.S., the plot area will be monitored for a minimum period of twelve months after 

harvest and/or crop destruct. If volunteers still are present after this period, the Cooperator will 

continue to monitor the test plots until no volunteers are present. Plots in Hawaii and Puerto 

Rico will utilize a minimum 30-day irrigated fallow period, or any additional time period 

mandated by the State Department of Agriculture, during which plots will be monitored for 

germinating cotton plants. Rain may substitute for irrigation during that period in Puerto Rico. 

All volunteers will be removed prior to flowering. Volunteer monitoring will not be required for 

plots that are destroyed prior to flowering. Volunteer monitoring requirements are not applicable 

if all materials under this EUP are deregulated by the USDA.
Monsanto Company 10-CT-211E-65 Pages 58 of 87

G.3.f.4. Protocol 4 - Insect Efficacy Trials

In the instance that USDA regulated plant materials are planted in an EUP plot, Monsanto will 

manage the EUP plot consistent with USDA Performance Standards in 7 CFR § 340.3 or as 

directed in USDA permits. 

Objective

Assess the efficacy in various genetic backgrounds against key cotton pests. Develop data to 

assess the benefits of PIP test plants in various combinations.

Description

Field trials will be conducted at various locations and data generated and analyzed to assess the 

benefits of PIP test plants in controlling key cotton insect pests. Other registered PIP or Non-PIP 

material may be used in these trials.

Genotypes and Vectors

The PIP test materials will consist of insect-protected cotton in different genetic backgrounds as 

previously described.

Sampling

Plant tissue (including seed) and/or whole plant samples may be collected during the growing 

season and returned to Monsanto or other laboratories for analysis.

Isolation

PIP test material will be isolated to prevent persistence in the environment. Isolation methods 

will be implemented consistent with USDA Performance Standards at 7 CFR § 340.3(c).

Schedule

Hawaii and Puerto Rico: Number of plantings per site: continuous, but includes a minimum 30-

day irrigated and monitored fallow period after each harvest. Rain may substitute for irrigation 

during that period in Puerto Rico. 

Continental U.S.: Number of plantings per site: up to eight.

 Approx. Planting Dates: February through July

 Approx. Harvest Dates: August through January

Harvest Procedures

Plots may be harvested by hand or mechanically. Harvested seed cotton will be placed in labeled 

containers of such construction to prevent loss of seed. Containers may include but are not 

limited to mesh or cloth bags, boxes, bins etc. Harvested seed cotton may be used for further 

research or future planting seed. Border rows, or plots that will not be used for further research 

or planting seed purposes, may be mechanically or hand harvested in bulk and disposed of as 

described below (see Final Disposition). Seed cotton or seed not to be used in future research, or 

as future planting seed will be disposed of as described below (see Final Disposition). All 

equipment will be thoroughly cleaned prior to exiting the plot area. 
Monsanto Company 10-CT-211E-65 Pages 59 of 87

Final Disposition

Unused planting seed either 1) will be used for other research purposes, 2) will be returned to 

Monsanto or seed company partners and affiliates according to approved methods, 3) will be 

destroyed by grinding, burning, incineration, autoclaving, composting, or burial in the test field, 

or 4) returned to a secure on-site storage location. Seed and other plant materials may be 

harvested and used in further research or as future planting seed. Following completion of data 

collection and/or harvest, all cotton plants in the trial will be destroyed (see G.7).

Volunteer Plants

If plants in the trial plot are allowed to flower, the plot area and border rows will be monitored 

for volunteers and all volunteers will be destroyed. The occurrence of volunteer plants will be 

minimized by growing regulated material in defined areas in the field and by utilizing adequate 

termination protocols as outlined in the Final Disposition Procedures in Section G.7. In the 

continental U.S., the plot area will be monitored for a minimum period of twelve months after 

harvest and/or crop destruct. If volunteers still are present after this period, the Cooperator will 

continue to monitor the test plots until no volunteers are present. Plots in Hawaii and Puerto 

Rico will utilize a minimum 30-day irrigated fallow period, or any additional time period 

mandated by the State Department of Agriculture, during which plots will be monitored for 

germinating cotton plants. Rain may substitute for irrigation during that period in Puerto Rico. 

All volunteers will be removed prior to flowering. Volunteer monitoring will not be required for 

plots that are destroyed prior to flowering. Volunteer monitoring requirements are not applicable 

if all materials under this EUP are deregulated by the USDA.
Monsanto Company 10-CT-211E-65 Pages 60 of 87

G.3.f.5. Protocol 5 – Product Characterization and Performance Trials

In the instance that USDA regulated plant materials are planted in an EUP plot, Monsanto will 

manage the EUP plot consistent with USDA Performance Standards in 7 CFR § 340.3 or as 

directed in USDA permits. 

Objective

Produce seed cotton and plant material for product characterization and performance studies.

Description

Plants will be either hand- or open-pollinated to generate seed and/or sampled for various types 

of laboratory analyses. Some or all lines may be hand pollinated or open pollinated with 

conventional or other biotechnology-derived cotton. Other registered PIP or Non-PIP material 

may be used in these trials.

Genotypes and Vectors

The PIP test materials will consist of insect-protected cotton in different genetic backgrounds as 

previously described.

Sampling

Plant tissue (including seed) and/or whole plant samples may be collected during the growing 

season and returned to Monsanto or other laboratories for analysis.

Isolation

PIP test material will be isolated to prevent persistence in the environment. Isolation methods 

will be implemented consistent with USDA Performance Standards at 7 CFR § 340.3(c).

Schedule

Hawaii and Puerto Rico: Number of plantings per site: continuous, but includes a minimum 30-

day irrigated and monitored fallow period after each harvest. Rain may substitute for irrigation 

during that period in Puerto Rico. 

Continental U.S.: Number of plantings per site: up to eight.

 Approx. Planting Dates: February through July

 Approx. Harvest Dates: August through January

Harvest Procedures

Plots may be harvested by hand or mechanically. Harvested seed cotton will be placed in labeled 

containers of such construction to prevent loss of seed. Containers may include but are not 

limited to mesh or cloth bags, boxes, bins etc. Harvested seed cotton may be used for further 

research or future planting seed. Border rows, or plots that will not be used for further research 

or planting seed purposes, may be mechanically or hand harvested in bulk and disposed of as 

described below (see Final Disposition). Seed cotton or seed not to be used in future research, or 

as future planting seed will be disposed of as described below (see Final Disposition). All 

equipment will be thoroughly cleaned prior to exiting the plot area. 
Monsanto Company 10-CT-211E-65 Pages 61 of 87

.

Final Disposition

Unused planting seed either 1) will be used for other research purposes, 2) will be returned to 

Monsanto or seed company partners and affiliates according to approved methods, 3) will be 

destroyed by grinding, burning, incineration, autoclaving, composting, or burial in the test field, 

or 4) returned to a secure on-site storage location. Seed and other plant materials may be 

harvested and used in further research or as future planting seed. Following completion of data 

collection and/or harvest, all cotton plants in the trial will be destroyed (see G.7).

Volunteer Plants

If plants in the trial plot are allowed to flower, the plot area and border rows will be monitored 

for volunteers and all volunteers will be destroyed. The occurrence of volunteer plants will be 

minimized by growing regulated material in defined areas in the field and by utilizing adequate 

termination protocols as outlined in the Final Disposition Procedures in Section G.7. In the 

continental U.S., the plot area will be monitored for a minimum period of twelve months after 

harvest and/or crop destruct. If volunteers still are present after this period, the Cooperator will 

continue to monitor the test plots until no volunteers are present. Plots in Hawaii and Puerto 

Rico will utilize a minimum 30-day irrigated fallow period, or any additional time period 

mandated by the State Department of Agriculture, during which plots will be monitored for 

germinating cotton plants. Rain may substitute for irrigation during that period in Puerto Rico. 

All volunteers will be removed prior to flowering. Volunteer monitoring will not be required for 

plots that are destroyed prior to flowering. Volunteer monitoring requirements are not applicable 

if all materials under this EUP are deregulated by the USDA.
Monsanto Company 10-CT-211E-65 Pages 62 of 87

G.3.f.6. Protocol 6 - Insect Resistance Management Trials

In the instance that USDA regulated plant materials are planted in an EUP plot, Monsanto will 

manage the EUP plot consistent with USDA Performance Standards in 7 CFR § 340.3 or as 

directed in USDA permits. 

Objective

Studies will be performed to assess the level of control that PIP test plants provide of key cotton 

pests, potentially including cotton bollworm, tobacco budworm, pink bollworm, beet and fall 

armyworms.

Description

Testing may include rating levels of damage and mortality for the relevant species in naturally or 

artificially infested field plots. These field plot data may be used to assess levels of larval 

movement among plants for these species under field conditions. These data may be used to 

support the insect resistance management (IRM) plans. Other registered PIP or Non-PIP 

material may be used in these trials.

Genotypes and Vectors

The PIP test materials will consist of insect-protected cotton in different genetic backgrounds as 

previously described.

Sampling

Plant tissue (including seed) and/or whole plant samples may be collected during the growing 

season and returned to Monsanto or other laboratories for analysis.

Isolation

PIP test material will be isolated to prevent persistence in the environment. Isolation methods 

will be implemented consistent with USDA Performance Standards at 7 CFR § 340.3(c).

Schedule

Hawaii and Puerto Rico: Number of plantings per site: continuous, but includes a minimum 30-

day irrigated and monitored fallow period after each harvest. Rain may substitute for irrigation 

during that period in Puerto Rico. 

Continental U.S.: Number of plantings per site: up to eight.

 Approx. Planting Dates: February through July

 Approx. Harvest Dates: August through January

Harvest Procedures

Plots may be harvested by hand or mechanically. Harvested seed cotton will be placed in labeled 

containers of such construction to prevent loss of seed. Containers may include but are not 

limited to mesh or cloth bags, boxes, bins etc. Harvested seed cotton may be used for further 

research or future planting seed. Border rows, or plots that will not be used for further research 

or planting seed purposes, may be mechanically or hand harvested in bulk and disposed of as 

described below (see Final Disposition). Seed cotton or seed not to be used in future research, or 
Monsanto Company 10-CT-211E-65 Pages 63 of 87

as future planting seed will be disposed of as described below (see Final Disposition). All 

equipment will be thoroughly cleaned prior to exiting the plot area. 

Final Disposition

Unused planting seed either 1) will be used for other research purposes, 2) will be returned to 

Monsanto or seed company partners and affiliates according to approved methods, 3) will be 

destroyed by grinding, burning, incineration, autoclaving, composting, or burial in the test field, 

or 4) returned to a secure on-site storage location. Seed and other plant materials may be 

harvested and used in further research or as future planting seed. Following completion of data 

collection and/or harvest, all cotton plants in the trial will be destroyed (see G.7).

Volunteer Plants

If plants in the trial plot are allowed to flower, the plot area and border rows will be monitored 

for volunteers and all volunteers will be destroyed. The occurrence of volunteer plants will be 

minimized by growing regulated material in defined areas in the field and by utilizing adequate 

termination protocols as outlined in the Final Disposition Procedures in Section G.7. In the 

continental U.S., the plot area will be monitored for a minimum period of twelve months after 

harvest and/or crop destruct. If volunteers still are present after this period, the Cooperator will 

continue to monitor the test plots until no volunteers are present. Plots in Hawaii and Puerto 

Rico will utilize a minimum 30-day irrigated fallow period, or any additional time period 

mandated by the State Department of Agriculture, during which plots will be monitored for 

germinating cotton plants. Rain may substitute for irrigation during that period in Puerto Rico. 

All volunteers will be removed prior to flowering. Volunteer monitoring will not be required for 

plots that are destroyed prior to flowering. Volunteer monitoring requirements are not applicable 

if all materials under this EUP are deregulated by the USDA.
Monsanto Company 10-CT-211E-65 Pages 64 of 87

G.3.f.7. Protocol 7 - Benefit Trials 

In the instance that USDA regulated plant materials are planted in an EUP plot, Monsanto will 

manage the EUP plot consistent with USDA Performance Standards in 7 CFR § 340.3 or as 

directed in USDA permits. 

Objective

Develop data to assess the benefits of COT102 × MON 15985 alone and/or in combination with 

other test materials.

Description

Plants with various treatments will be evaluated for efficacy against key cotton insect pests, and 

any effects on agronomic characteristics, such as yield, fiber quality, maturity, plant growth 

characteristics, germination and seedling vigor compared to controls. Tests may be either 

naturally or artificially infested. Other registered PIP or Non-PIP material may be used in these 

trials.

Genotypes and Vectors

The PIP test materials will consist of insect-protected cotton in different genetic backgrounds as 

previously described.

Sampling

Plant tissue (including seed) and/or whole plant samples may be collected during the growing 

season and returned to Monsanto or other laboratories for analysis.

Isolation

PIP test material will be isolated to prevent persistence in the environment. Isolation methods 

will be implemented consistent with USDA Performance Standards at 7 CFR § 340.3(c).

Schedule

Hawaii and Puerto Rico: Number of plantings per site: continuous, but includes a minimum 30-

day irrigated and monitored fallow period after each harvest. Rain may substitute for irrigation 

during that period in Puerto Rico. 

Continental U.S.: Number of plantings per site: up to eight.

 Approx. Planting Dates: February through July

 Approx. Harvest Dates: August through January

Harvest Procedures

Plots may be harvested by hand or mechanically. Harvested seed cotton will be placed in labeled 

containers of such construction to prevent loss of seed. Containers may include but are not 

limited to mesh or cloth bags, boxes, bins etc. Harvested seed cotton may be used for further 

research or future planting seed. Border rows, or plots that will not be used for further research 

or planting seed purposes, may be mechanically or hand harvested in bulk and disposed of as 

described below (see Final Disposition). Seed cotton or seed not to be used in future research, or 
Monsanto Company 10-CT-211E-65 Pages 65 of 87

as future planting seed will be disposed of as described below (see Final Disposition). All 

equipment will be thoroughly cleaned prior to exiting the plot area. 

Final Disposition

Unused planting seed either 1) will be used for other research purposes, 2) will be returned to 

Monsanto or seed company partners and affiliates according to approved methods, 3) will be 

destroyed by grinding, burning, incineration, autoclaving, composting, or burial in the test field, 

or 4) returned to a secure on-site storage location. Seed and other plant materials may be 

harvested and used in further research or as future planting seed. Following completion of data 

collection and/or harvest, all cotton plants in the trial will be destroyed (see G.7).

Volunteer Plants

If plants in the trial plot are allowed to flower, the plot area and border rows will be monitored 

for volunteers and all volunteers will be destroyed. The occurrence of volunteer plants will be 

minimized by growing regulated material in defined areas in the field and by utilizing adequate 

termination protocols as outlined in the Final Disposition Procedures in Section G.7. In the 

continental U.S., the plot area will be monitored for a minimum period of twelve months after 

harvest and/or crop destruct. If volunteers still are present after this period, the Cooperator will

continue to monitor the test plots until no volunteers are present. Plots in Hawaii and Puerto 

Rico will utilize a minimum 30-day irrigated fallow period, or any additional time period 

mandated by the State Department of Agriculture, during which plots will be monitored for 

germinating cotton plants. Rain may substitute for irrigation during that period in Puerto Rico. 

All volunteers will be removed prior to flowering. Volunteer monitoring will not be required for 

plots that are destroyed prior to flowering. Volunteer monitoring requirements are not applicable 

if all materials under this EUP are deregulated by the USDA.
Monsanto Company 10-CT-211E-65 Pages 66 of 87

G.3.f.7. Protocol 8 - Seed Treatment Trials 

In the instance that USDA regulated plant materials are planted in an EUP plot, Monsanto will 

manage the EUP plot consistent with USDA Performance Standards in 7 CFR § 340.3 or as 

directed in USDA permits. 

Objective

Evaluate effects of diverse chemical seed treatments, in combination with PIP test materials and 

cotton pests.

Description

Seed may be coated with a variety of seed treatments. Plants with the various treatments will be 

evaluated for efficacy against key cotton insect pests, and any effects of the seed treatment on 

agronomic characteristics, such as germination, emergence and seedling vigor. Other registered 

PIP or Non-PIP material may be used in these trials.

Genotypes and Vectors

The PIP test materials will consist of insect-protected cotton in different genetic backgrounds as 

previously described.

Sampling

Plant tissue (including seed) and/or whole plant samples may be collected during the growing 

season and returned to Monsanto or other laboratories for analysis.

Isolation

PIP test material will be isolated to prevent persistence in the environment. Isolation methods 

will be implemented consistent with USDA Performance Standards at 7 CFR § 340.3(c).

Schedule

Hawaii and Puerto Rico: Number of plantings per site: continuous, but includes a minimum 30-

day irrigated and monitored fallow period after each harvest. Rain may substitute for irrigation 

during that period in Puerto Rico. 

Continental U.S.: Number of plantings per site: up to eight.

 Approx. Planting Dates: February through July

 Approx. Harvest Dates: August through January

Harvest Procedures

Plots may be harvested by hand or mechanically. Harvested seed cotton will be placed in labeled 

containers of such construction to prevent loss of seed. Containers may include but are not 

limited to mesh or cloth bags, boxes, bins etc. Harvested seed cotton may be used for further 

research or future planting seed. Border rows, or plots that will not be used for further research 

or planting seed purposes, may be mechanically or hand harvested in bulk and disposed of as 

described below (see Final Disposition). Seed cotton or seed not to be used in future research, or 

as future planting seed will be disposed of as described below (see Final Disposition). All 

equipment will be thoroughly cleaned prior to exiting the plot area. 
Monsanto Company 10-CT-211E-65 Pages 67 of 87

Final Disposition

Unused planting seed either 1) will be used for other research purposes, 2) will be returned to 

Monsanto or seed company partners and affiliates according to approved methods, 3) will be 

destroyed by grinding, burning, incineration, autoclaving, composting, or burial in the test field, 

or 4) returned to a secure on-site storage location. Seed and other plant materials may be 

harvested and used in further research or as future planting seed. Following completion of data 

collection and/or harvest, all cotton plants in the trial will be destroyed (see G.7).

Volunteer Plants

If plants in the trial plot are allowed to flower, the plot area and border rows will be monitored 

for volunteers and all volunteers will be destroyed. The occurrence of volunteer plants will be 

minimized by growing regulated material in defined areas in the field and by utilizing adequate 

termination protocols as outlined in the Final Disposition Procedures in Section G.7. In the 

continental U.S., the plot area will be monitored for a minimum period of twelve months after 

harvest and/or crop destruct. If volunteers still are present after this period, the Cooperator will 

continue to monitor the test plots until no volunteers are present. Plots in Hawaii and Puerto 

Rico will utilize a minimum 30-day irrigated fallow period, or any additional time period 

mandated by the State Department of Agriculture, during which plots will be monitored for 

germinating cotton plants. Rain may substitute for irrigation during that period in Puerto Rico. 

All volunteers will be removed prior to flowering. Volunteer monitoring will not be required for 

plots that are destroyed prior to flowering. Volunteer monitoring requirements are not applicable 

if all materials under this EUP are deregulated by the USDA.
Monsanto Company 10-CT-211E-65 Pages 68 of 87

G.3.g. Containment Procedures

Once test materials have been registered, all containment procedures are no longer in effect.

Maintenance of Test Plant Materials

1. Storage of Test Seed

Test seed will be stored in a secure location and handled in a manner to prevent unintended 

release into the environment. All test seed bags will be color coded, marked, tagged, or 

otherwise distinguished to allow easy recognition.

2. Disposal of Leftover Planting Seed

Seed unused for planting either will be:

1) used for other research purposes or

2) returned to a secure on-site storage location or

3) returned to Monsanto Company or seed company partners/affiliates, using approved 

shipment procedures or

4) destroyed by grinding, burning, incineration, autoclave, composting or buried in the test 

field.

3. Transporting of Test Plant Materials

PIP test seed will be transported in secured, closed containers of such construction to avoid loss 

of seed outside the container during transport. 

Field Operations

1. Personnel

Workers in the trial site area will be informed that no plant material is to be removed from the 

marked trial site without direct authorization. Where trials are located on privately owned farms, 

the responsible researcher will inform the owner of the farm as to the nature of the experiment 

and the need to maintain the plants and seed separate from other plant material not part of the 

trial.

2. Planting

2a. Site

These trials will be planted at locations specified in USDA notifications or permits (if applicable) 

and in states approved by the EPA Experimental Use Permit. 
Monsanto Company 10-CT-211E-65 Pages 69 of 87

2b. Equipment

After planting is completed and before the planter leaves the field, the planter will be thoroughly 

cleaned out to prevent mixing with other cotton seed.

 1) Small amounts of seed cleaned out of the planter may be allowed to fall into the field and 

to germinate along with the test crop.

2) Significant quantities of seed cleaned out from the planter can be securely packaged and 

returned to the secure storage location, Monsanto Company or seed company 

partners/affiliates.

2.c. Pollen Containment

One or more of the following methods may be used: 

 Test plants will be destroyed prior to anthesis. 

 Test plants will be located at least 660 feet from open-pollinating cotton that is not a part of 

this research. 

 If there will be any cotton grown within 660 feet of the regulated cotton, a perimeter (border 

rows) of a non-regulated cotton should completely surround the regulated cotton test area to 

act as a pollen sink for insect pollinators. This border must be continuous around the entire 

field trial area. These border rows must be destroyed, as outlined in the Destruction section 

G.7., and volunteer monitored if monitoring of the EUP plot area is required. 

3. Equipment

The field equipment will be examined before leaving the trial site and any viable plant material

will be removed.

4. Trial harvest

The test and border seed cotton may be harvested for further research purposes. Equipment will 

be thoroughly cleaned after harvesting operations.

5. Trial completion

5.a. Destruction

After harvest or completion of data collection, all cotton plants in the trial, including test, 

controls, and border plants will be destroyed. Methods standard for the area will be employed 

(bush-hog, shred and disk, etc.). Waste plant materials may be plowed under, buried in the 

regulated field plot, burned in the regulated field plot or otherwise devitalized.

5.b. Crop rotation

Options for replanting the trial site next growing season:

1. Leave the site fallow and control any volunteer plants.
Monsanto Company 10-CT-211E-65 Pages 70 of 87

2. Plant to another appropriate rotational crop that would allow the clear identification and 

control of volunteer cotton plants. Replanting to cotton not regulated by EPA or USDA is 

NOT allowed. 

3. Plant to the same or another regulated cotton material and follow the requirements for that 

regulated cotton material. 

In Hawaii and Puerto Rico, continuous cropping of cotton may occur, but must include at least a 

30-day irrigated fallow period after each harvest, during which time the plots will be monitored 

for germinating cotton plants.

5.c. Monitoring and Control of Volunteer Cotton

If plants in the trial plot are allowed to flower, the plot area and border rows will be monitored 

for volunteers and all volunteers will be destroyed. The occurrence of volunteer plants will be 

minimized by growing regulated material in defined areas in the field and by performing the 

Final Disposition Procedures outlined in Section G.7. Plots in Hawaii and Puerto Rico will 

utilize a minimum 30-day irrigated fallow period, or any additional time period mandated by the 

State Department of Agriculture, during which plots will be monitored for germinating cotton 

plants. All volunteers will be removed prior to seed set or flowering. Volunteer monitoring will 

not be required for plots that are destroyed prior to flowering. Volunteer monitoring 

requirements are not applicable if all materials under this EUP are deregulated by the USDA.
Monsanto Company 10-CT-211E-65 Pages 71 of 87

Section G.4. Program Objectives

The research program is multi-faceted and will include product advancement and agronomic 

tests to evaluate how cotton with the insect-protected and herbicide-protected traits can be 

utilized by growers. Testing is separated into several key areas, as described below.

Protocol 1 - Breeding and Observation Nursery

The objectives of these activities are to: 1) develop breeding material intended for future 

experimental testing and observation, 2) maintain these biotechnology-derived products through 

seed production, and 3) conduct small plot field trials to evaluate phenotype and agronomic 

performance. 

Protocol 2 - Seed Increase 

Produce seed and plant material for further testing and product development in various genetic 

backgrounds.

Protocol 3 - Yield and Herbicide Tolerance Trials

Assess any agronomic effects of the genetic insertion in PIP test materials and any effects of 

combining these in genetic backgrounds. In addition, crops may be sprayed with herbicides to 

assess crop tolerance.

Protocol 4 - Insect Efficacy Trials

Assess the efficacy in various genetic backgrounds against key cotton pests. Develop data to 

assess the benefits of PIP test plants in various combinations.

Protocol 5 - Product Characterization and Performance Trials

Produce seed cotton and plant material for product characterization and performance studies. 

Protocol 6 - Insect Resistance Management Trials

Studies will be performed to assess the level of control that PIP test plants provide of key cotton 

pests, potentially including cotton bollworm, tobacco budworm, pink bollworm, beet and fall 

armyworms. 

Protocol 7 - Benefit Trials

Develop data to assess the benefits of COT102 × MON 15985 alone and/or in combination with 

other test materials. 

Protocol 8 - Seed Treatment Trials

Evaluate effects of diverse chemical seed treatments, in combination with PIP test materials and 

cotton pests. 
Monsanto Company 10-CT-211E-65 Pages 72 of 87

Section G.5. Program Quantities

Total plantings of 13663 acres are requested for PIP, Non-PIP, and border acres in 2010-

2012:

PIP materials are estimated to be on 1897 acres, 

Non-PIP materials are estimated to be on 909 acres, 

Border materials are estimated to be on 10857 acres, 

This acreage is necessary to adequately address the extensive program objectives, varied 

insect species, infestation levels, and weather conditions found in U.S. cotton growing 

regions. 

In the development of insect-protected test plants, it is necessary to properly characterize the 

performance of the crop on a relevant scale and to produce sufficient seed to continue similar 

research and development in subsequent seasons.
Monsanto Company 10-CT-211E-65 Pages 73 of 87

Section G.6. Proposed Duration

Plantings from December 1, 2010 through June 30, 2012.
Monsanto Company 10-CT-211E-65 Pages 74 of 87

Section G.7. Method of Disposition of Plant Materials

Disposition of plant materials at the conclusion of the testing program will be as follows:

Unused seed for planting will be either:

1. used for other research purposes or

2. returned to a secure on-site storage location or

3. returned to Monsanto Company or seed company partners/affiliates, using approved 

shipment procedures or

4. destroyed by grinding, burning, incineration, autoclaving or burial in the test field.

Harvested seed and other plant materials may be used:

1. in further research or

2. as future planting seed.

After harvest or completion of data collection, all cotton plants in the trial, including test plants, 

control plants, and buffer row plants will be destroyed. Standard methods for the area will be 

employed and conducted in compliance with local plow down regulations. Waste plant materials 

may be plowed under, burned in the field (subject to local burning ordinances), or otherwise 

devitalized.
Monsanto Company 10-CT-211E-65 Pages 75 of 87

References

Adang, M.J., M.J. Staver, T.A. Rocheleau, J. Leighton, R.F. Barker, and D.V. Thompson. 1985. 

Characterized full-length and truncated plasmid clones of the crystal protein of Bacillus 

thuringiensis subsp. kurstaki HD-73 and their toxicity to Manduca sexta. Gene 36(3):289-300.s

Baum, J.A. 1998. Transgenic Bacillus thuringiensis. Phytoprotection 79 (suppl.): 1270130.

Baum, J.A., T.B. Johnson, and B.C. Carlton. 1999. Bacillus thuringiensis. Natural and 

recombinant biopesticide products. In Methods in Biotechnology: Biopesticides: Use and 

Delivery. Vol.5. p. 1890209. Hall, F.R. and J.J. Menn (eds). Humana Press Inc., Totowa, NJ.

Betz, F.S., B.G. Hammond, and R.L. Fuchs. 2000. Safety and advantages of Bacillus 

thuringiensis protected plants to control insect pests. Regulatory Tox. Pharm. 32:1560173.

Dankocsik, C., W.P. Donovan, and C.S. Jany. 1990. Activation of a cryptic crystal protein gene 

of Bacillus thuringiensis subspecies kurstaki by gene fusion and determination of the crystal 

protein insecticidal specificity. Molecular Microbiology 4:2087-2094.

EPA. 1995a. Cry1A(c) and Cry1C derived delta endotoxins of Bacillus thuringiensis

encapsulated in killed Pseudomonas fluorescens; Exemption from the requirement of a tolerance; 

Final rule. U.S. Environmental Protection Agency. Fed. Reg. 60(177): 47487047489.

EPA. 1998. Reregistration Eligibility Decision (R.E.D.) Bacillus thuringiensis. U.S. 

Environmental Protection Agency. EPA 7380R0980004.

EPA. 2001a. Bacillus thuringiensis Cry3Bb1 and Cry2Ab2 Protein and the Genetic Material 

Necessary for its Production in Corn and Cotton; Exemption from the Requirement of a 

Tolerance; Final rule. U.S. Environmental Protection Agency. Fed. Reg. 66(92): 24061024066.

EPA. 2001b. Biopesticides Registration Action Document: Bacillus thuringiensis

plant-incorporated protectants. U.S. Environmental Protection Agency. 

McClintock, J.T., C.R. Schaffer, and R.D. Sjoblad. 1995. A comparative review of the 

mammalian toxicity of Bacillus thuringiensis-based pesticides. Pestic. Sci. 45:950105.

Peng, D., S. Chen, L. Ruan, L. Li, Z. Yu, M. Sun. 2006. Safety assessment of transgenic 

Bacillus thuringiensis with VIP insecticidal protein gene by feeding studies.

Siegel, J.P. 2001. The mammalian safety of Bacillus thuringiensis based insecticides. 

Minireview. J. Invert. Pathol. 77:13.

Widner, W.R., and H.R. Whiteley. 1990. Location of the dipteran specificity region in a

lepidopteran-dipteran crystal protein from Bacillus thuringiensis. J. Bacteriol. 172, 2826-2832.