Bayer CropScience Cry1Ab Cotton EUP Amendment Application 264-E UP-140, Page 25 of 36



### Section G Proposed experimental program (Revised)

1. Summary of participants

The Participants and Cooperators who will supervise the experimental work conducted under this EUP are listed in the Confidential Attachment A.

{CBI Cross Reference Number 1}



### 2. Total Acreage by Protocol by State

State Insect Efficacy Regulatory Sample Production Total Acreage by State

AL 26.25 - 26.25
AR 26.25 15.25 41.5
AZ 94 82 176
CA 53 31 84
FL 8.75 3.75 12.5
GA 41.5 10 51.5
LA 64 17.5 81.5
MS 141.25 113.5 254.75
NC 101.75 20 121.75
PR 51 10 61
SC 184.25 - 184.25
TN 55.25 10 65.25
TX 267 174.75 441.75
TOTAL 1114.25 487.75 1602Bayer CropScience Cry1Ab Cotton EUP Amendment Application 264-E UP-140, Page 26 of 36



### 3. Acreage, seed and AI quantities by state

State Protocol* Cry1Ab Cotton Acres Non-PIP Acres Total Acreage by State Max. lb of Cry1Ab Cotton

Seed Max. Amt. of Cry1Ab Protein in Seed Planted (g)

AL E 3.75 22.5 26.25 45 0.35
AR E, R 8.5 33 41.5 102 0.79
AZ E, R 18 158 176 216 1.67
CA E, R 10 74 84 120 0.93
FL E, R 1.5 11 12.5 18 0.14
GA E, R 10.5 41 51.5 126 0.97
LA E, R 8.5 73 81.5 102 0.79
MS E, R 17.25 237.5 254.75 207 1.60
NC E, R 12.25 109.5 121.75 147 1.13
PR E, R 5 56 61 60 0.46
SC E 9.75 174.5 184.25 117 0.90
TN E, R 6.75 58.5 65.25 81 0.63
TX E, R 40.25 401.5 441.75 483 3.73
Total 152 1450 1602 1824 14.08

1 Protocols are abbreviated as follows: (E) Inse ct Efficacy, (R) Regula tory Sample Production
2 Calculated as: Cry1Ab acres x 12 lb of seed/acre
3 Calculated as: max. lb of Cry1Ab seed x 0.007718g/lb of Cry1Ab seed (based on the label

maximum concentration of 0.0017%)Bayer CropScience Cry1Ab Cotton EUP Amendment Application 264-E UP-140, Page 27 of 36



### 4. Program overview

Bayer CropScience (BCS) has developed cotton [ Gossypium hirsutum ] plants that express an insecticidal protein, Cry1Ab, from a common soil bacterium, Bacillus thuringiensis subsp. berliner ( B.t. berliner ). The Cry1Ab protein is effective in co ntrolling lepidopteran larvae, such as cotton bollworm (CBW, Helicoverpa zea ) and tobacco budworm (TBW, Heliothis virenscens ) larvae, which are common pests of cotton. These pest s cause severe economic damage to the cotton crop if not controlled. If controlled by chemic al pesticides, there is the need for large input annually to control these pests. Small scale fiel d trial experiments of cotton expressing Cry1Ab protein, conducted under notifications granted by th e U.S. Department of Agriculture's Animal and Plant Health Inspection Service (APHIS), have shown the plant’s ab ility to protect itself against these pests.

Transgenic cotton plants expressing Cry1Ab protei n provide an excellent addition to growers' options for insect control that reduces or elimin ates the need for other insecticide inputs and fits well within an integrated pest management program. Cry1Ab is a protein familiar to the Agency, but is has not been used in commercial cotton as a Plant Incorporated Protectant (PIP).

BCS is applying for an extension of this EUP (264- EUP-140) to allow further evaluation of these cotton plant lines in a larger range of environm ental conditions. All cotton plants to be evaluated under the EUP have been derived from transforma tion event T303-3 and T304-40. Several different experiments are planned, and they can be grouped into: insect efficacy trials (including a tobacco budworm high-dose verification study ), herbicide efficacy evaluation, agronomic performance evaluation, breeding (line evaluations).

In addition to these experimental plans, seed blocks may be grown as part of this program to evaluate effects on seed production and dorm ancy, and seed may be harvested for later plantings of experimental and regulatory field trials.

The primary goal of this experimental research program is to evaluate the Cry1Ab proteincontaining cotton plants for their efficacy against in sect pests of cotton, as well as to provide material for agronomic performance evaluations. These plants also contain an herbicide tolerant ingredient as a selectable marker, the phosphinot hricin acetyltransferase (PAT) protein that confers tolerance to glufosinate-ammonium herbicides.

The entire program will be done on a “Crop Destruct ” basis; no cottonseed will enter commerce. Some of the plant material will be retained for sc ientific research and/or planting purposes. All other plant materials will be destroyed. There shall be no unintentional exposure to humans or domestic animals since the program will be conduc ted using containment precautions and in a crop destruct fashion. Isolation will be maintain ed in order to prevent any inadvertent outcrossing (pollination) from transgenic plants to non-tran sgenic cotton plants. No environmental impact issues related to the testing of these transgenic cotton plants have been identified. The Cry1Ab protein has a specific range of toxicity to the ta rget lepidopteran pests without having effects on the non-target beneficial insects.

In total, the program will be carried out across 13 states/territories on a total of 1602 acres, of which 152 acres will be planted to Cry1Ab cotton. The Cry1Ab-containing cotton will be planted at a maximum rate of 12 lbs/acre. The level of Cry1Ab protein is approximately 0.908 – 7.718 mg/lb of seed based on the label range of 0.0002 – 0.0 017 %; therefore the planting of these Cry1Abcontaining seeds represents an application rate of approximately 10.8 – 92.6 mg of Cry1Ab protein per acre. Our proposed experimental research program would total 1.64 - 14.08 g of Cry1Ab protein (or 0.004 - 0.031 pounds of Cry1Ab protein) for 152 acres planted to Cry1Ab-Bayer CropScience Cry1Ab Cotton EUP Amendment Application 264-E UP-140, Page 28 of 36

containing cotton. The level of Cry1Ab protei n in the different plant materials is only an estimation based on our current level of information.

Competent Bayer CropScience employees will superv ise the program which will be conducted by them, in addition to public and private cooper ators. All responsible researchers listed are professionally qualified to accomplish their stated duties.Bayer CropScience Cry1Ab Cotton EUP Amendment Application 264-E UP-140, Page 29 of 36



### 5. Program details

Experimental Protocol: EFFICACY TESTING OF INSECT RESISTANT TRANSGENIC COTTON

Objectives The purpose of these trials is to compare plant growth, morphology, and agronomic performance among insect resistant transgenic cotton lines in different genetic backgrounds and their respective non-transgenic counterparts when infest ed with (either naturally or artificially) or devoid of the target insect. Another objective of these trials is to evaluate insect resistance transgene efficacy and subsequently determine if transgene expression and resulting insect resistance affect plant growth, morphology, or any facet of agronomic performance including fiber characteristics.

Specific Objectives

 Breeding Line Evaluations - This is primarily for the purpose of transferring the Cry1Ab event into elite cotton lines, in order to evaluat e performance of the gene in elite varieties. This will include backcrossing programs for initial evaluation of lines, and for production of hand-pollinated seed to conduct preliminary yield evaluations.

 Insect Efficacy, Natural Infest ations/ Herbicide Efficacy - Th ese trials will be conducted to evaluate effectiveness of event s T303-3 and T304-40 in controlling the primary target pests CBW and TBW. The tria ls will include sites where t he infestations occur only as natural infestations. We expect large differences in the level of severity of the infestation and damage that can be seen across locations and environments; and for this reason, the evaluation of efficacy across a broad spectrum of environmental and geographical conditions is needed to accurately evaluate efficacy of the Cry1Ab events against the targeted pests under field conditions. A dditionally, evaluation by independent experts, such as in trials conducted by university researchers and other co operators, provides independently generated performance data. In addition, the tolerance to the herbicide glufosinate-ammonium provided by the marker gene will be evaluated.

 Insect Efficacy TBW, Artificial Infestations – These trials will be conducted to provide high-dose verification as part of BCS IRM strategy and in response to a deficiency identified in a letter from the Agency dated May 27, 2010. The individual component lines (Cry1Ab and Cry2Ae), a control line and TwinLink cotton will be planted and cages (8 ft X 20 ft X 6 ft) will be placed over two 20 ft row sections (one cage per line). Beginning at early squaring each cage will be infested with approximately 50 male and female tobacco budworms from a lab colony supplied by USDA-ARS in Stoneville, MS. Plants will be insp ected for sufficient oviposition to insure a heavy population of neonate TBWs in eac h line. Approximately 7-10 days after infestation at least 20 plants per line will be in spected for surviving larvae. Larvae will be collected, categorized according to size and id entified to insure that surviving larvae are TBW. Any larvae reaching third instar stage will be considered a "survivor". Second instar larvae will be collected from the plants and placed on artificial diet to determine if they survive to third instar. Cages/Lines will be infested at least three times during the season and mortality/survivorship will be deter mined, thus infestation times will serve as replications. The objective of the study will be to determine if each component line (Cry1Ab and Cry2Ae) will control the TBW populat ion at a high level to justify each Cry gene component of TwinLink as a "high dose". The trials may include sites where the infestations occur as natural infestations, as well as artificially infested to enhance the opport unity to observe extreme CBW pressure. We expect large differences in the level of severity of the infestation and damage that can beBayer CropScience Cry1Ab Cotton EUP Amendment Application 264-E UP-140, Page 30 of 36

seen across locations and environments; and fo r this reason, the evaluation of efficacy across a broad spectrum of environmental and geographical conditions is needed to accurately evaluate efficacy of the Cry1Ab events against the targeted pests under field conditions. Additionally, evaluation by independ ent experts, such as in trials conducted by university researchers and other coo perators, provides independently generated performance data.

 Insect Efficacy CBW, Artificial Infestatio ns - These trials will be conducted to evaluate effectiveness of events T303-3 and T304-40 in controlling the primary target pest CBW. The trials may include sites where the infestations occur as natural infestations, as well as artificially infested to enhance the opport unity to observe extreme CBW pressure. We expect large differences in the level of severity of the infestation and damage that can be seen across locations and environments; and fo r this reason, the evaluation of efficacy across a broad spectrum of environmental and geographical conditions is needed to accurately evaluate efficacy of the Cry1Ab events against the targeted pests under field conditions. Additionally, evaluation by independ ent experts, such as in trials conducted by university researchers and other coo perators, provides independently generated performance data.

 Insect Efficacy, other Leps - Though the pr imary target pest for the Cry1Ab events are TBW and CBW, the Cry1Ab protein has been shown to have activity against other lepidopteran pests of cotton, including Fall Armyworm and Pink Bollworm. These pests can be significant in certain seasons and in certain geographical regions, though the overall incidence and damage attributable to these pests is probably less significant than for the two main pests. Since these and ot her lepidopteran pests can cause significant economic damage in some environments, the oppor tunity for Cry1Ab to provide control of additional lepidopteran pests will be evaluated. The acreage available under this EUP will allow the flexibility to test for these addi tional insect pests in planned experiments.

 Agronomic Evaluation - These trials will be used to document yield and agronomic performance of the even ts containing Cry1Ab. Some tria ls will be small plot evaluations to look at effects on flowering, maturity, s eed size, herbicide sensitivity, and yield, for example. Other trials will use larger fields to evaluate whether having Cry1Ab provides agronomic and yield benefits for production fields that affect the economics of producing cotton seed. One could reasonably expect that use of Cry1Ab in cotton lines will significantly reduce the amount of insecticid es that will need to be applied to production fields, and will also substantially increase seed yields in production fields affected by the targeted pests. Evaluation of these questions is not possible under small plot situations, and requires the acreage provided by this EUP.

Cooperators and Participants The Participants and Cooperators who will supervise the experimental work conducted under this EUP are listed in the Confidential Attachment A.

{CBI Cross Reference Number 1}

Locations The total maximum acreage by state planted with seed containing Cry1Ab protein for efficacy testing is listed below.Bayer CropScience Cry1Ab Cotton EUP Amendment Application 264-E UP-140, Page 31 of 36

Efficacy Testing Locations and Acreage



### State Non-PIP Acres Cry1Ab Cotton Acres Total Acreage by State Max. lb of Cry1Ab Cotton Seed

AL 22.5 3.75 26.25 45
AR 22.5 3.75 26.25 45
AZ 83 11 94 132
CA 46 7 53 84
FL 7.5 1.25 8.75 15
GA 33 8.5 41.5 102
LA 58 6 64 72
MS 132.5 8.75 141.25 105
NC 93.5 8.25 101.75 99
PR 48 3 51 36
SC 174.5 9.75 184.25 117
TN 50.5 4.75 55.25 57
TX 243 24 267 288
Total 1014.5 99.75 1114.25 1197

1 Calculated as: Cry1Ab acres x 12 lb of seed/acre

Genotypes and vectors Transgenic cotton plants expressing the cry1ab gene (event T304-40 and T303-3) Experimental lines with different backgrounds Non transgenic plants Coker 315, Coker 312 or equivalents Transgenic commercial control Non transgenic commercial control

Trial DesignThe preferred statistical design is a split-plot where level of insect infestation (sprayed vs. nonsprayed) is the main plot and line/transgene is t he subplot. A split-strip design may also be used which facilitates insecticide tr eatments. A randomized comple te block design (RCBD) would suffice, but would not provide maximal precision.

Agronomic Treatments Typical agronomic inputs for conventionally grown cotton for the area, including, but not limited to:

 Conventional herbicide treatments, both pre- and post-planting

 Granular insecticide and/or fungicide application at planting

 Fertilizer applications

 Necessary in-season insecticide applications for non-target and/or target insects only (see test treatments below)

 Growth regulator application (this should be done sparingly if at all)

 Additional hand weeding as necessary

 Chemical defoliation without boll-opening desiccants

Test TreatmentsBayer CropScience Cry1Ab Cotton EUP Amendment Application 264-E UP-140, Page 32 of 36

Test treatments involve natural infestations of tar get insects, chemical control of target insects, and chemical control of non-target insects. Treatments include but are not limited to:

 Complete insect control, both target and non-target in sprayed plots

 Insect control of non-target insects that allo ws or encourages natural infestation of target insect, in non sprayed plots.

Border rows The EUP test plants or the trial will be surrounded by one or more border rows of cotton.

Schedule Planting dates: February - June Harvest dates: July - November

Activities and Agronomic Practices Plots will be harvested by hand or mechanically. If by hand, the bolls will be placed in cloth or paper bags of such construction to avoid loss of seed outside of the bags. If by machine, seed cotton will be harvested, transported and processe d under conditions appropriate for the handling of regulated material. This includes separate, redundant labeled packaging of all regulated material leaving the location.

Use of GPS coordinates, stakes, markers or othe r methods will be used to identify the area where the transgenic plants are grown, and such an area will be subsequently monitored for volunteers for an appropriate period of time. Volunteer plan ts will be terminated by hand weeding, disking, herbicide spraying or other method.

Containment EUP test plants will be isolated in accordance with USDA-APHIS Performance Standards for regulated cotton trials. Isolation methods will incl ude one or more of the following: (1) EUP test plants will be located at least 660 feet from other pa rties’ sexually receptiv e cotton; (2) a 40 footwide perimeter of non-transgenic cotton will surround the transgenic plants planted at a density within 20% (greater or less than) the trial density to act as pollen sink for insect pollinators (the perimeter cotton would be disposed of by harvest ing, disking and monitoring); (3) temporal isolation, where the flowering period for the EUP plants will not coincide with the presence of other parties’ sexually receptive cotton within 660 feet of the EUP test plants. Open flowering EUP test plants may be located within 660 feet of sexually receptive co tton provided such other cotton is used only for experimental purposes and/or destroyed.

Following the trial completion, all remaining plant debris will be destroyed by incorporation in the soil. All equipment used during crop destruction pr actices will be inspected and cleaned before leaving the field. Seed cotton not destined fo r further experimentation will be destroyed by incineration or deep burial.

If harvested material is to be ginned, seed cotton will securely be transported to a gin. Processing of harvested seed cotton will consist of either hand ginning or research/commercial scale ginning. Hand ginning will be on small, tabl e-top gins. Machine ginning will occur at the ginning facility on-site on a limited number of re search/commercial scale gi ns. All packaging and waste will be destroyed by devitalization. No seed cotton or gi nning by-products will be used for food or feed. Ginned seed will be stored under containment practices for regulated materials.

Agronomic Data Collection If available, the following data may be collected from the plots, usin g a 1-9 scale, where applicable:

 Strain uniformity: 1=uniform, 9=highly variable

 Leaf pubescence: 1=highly pubesce nt, 5=semi-smooth, 9=glabrousBayer CropScience Cry1Ab Cotton EUP Amendment Application 264-E UP-140, Page 33 of 36

 Disease reaction (verticillium wilt, bacterial blight, bronze wilt, etc. IF applicable): 1=no symptoms, 5=some symptoms apparent, 9=severe

 Stalk lodging: 1=upright, 9=severely lodged

 % open bolls as a visual average when un infested recurrent parent is 40-60% open

 Yield in lb lint per acre

 % lint

 # seed per boll

 Boll size

 Seed index

 Fiber properties: length, length uniform ity, strength, micronaire, elongation

 Plant mapping: plant map 10 plants per plot, each of 3 reps at maturity shortly before defoliation. Data will include plant height, number of nodes, and boll position. Boll damage ratings may be a part of the mapping data. Information collected will reflect overall plant architecture and maturity.

Target Insect Evaluation Data Collection Data will be taken on insect damag e for 6-8 weeks to measure resist ance to the infested insect. Insect infestation and damage data are collected by examining the terminal and one square or boll on 10 plants per row on each of the two center rows in each plot. Data collected may include:

 Number of live larvae in squares

 Number of live larvae in bolls

 Number of squares damaged by larvae

 Number of bolls damaged by larvae

 Number of damaged white flowers

 Number of live larvae in white flowers

Quality Control Sample Collection Leaf samples are to be taken from two individual plants per plot for QC purposes.Bayer CropScience Cry1Ab Cotton EUP Amendment Application 264-E UP-140, Page 34 of 36

Experimental Program: PRODUCTION OF SAMPLE MATERIAL FOR USE IN REGULATORY STUDIES

Objectives The purpose of these trials is to produce sample materials for regulatory trials, as part of the requirement of various regulatory agencies for t he submission of registration packages in different countries.

Cooperators and participants The Participants and Cooperators who will supervise the experimental work conducted under this EUP are listed in the Confidential Attachment A.

{CBI Cross Reference Number 1}

Locations The total maximum acreage by state planted with seed containing Cry1Ab protein for production of regulatory samples is listed below.

Regulatory Study Sample Material Generation Locations and Acreage



### State Non-PIP Acres Cry1Ab Cotton Acres Total Acreage by State Max. lb of Cry1Ab Cotton Seed

AL
- - - N/A 2


### AR 10.5 4.75 15.25 57 AZ 75 7 82 84 CA 28 3 31 36 FL 3.5 0.25 3.75 3 GA 8 2 10 24 LA 15 2.5 17.5 30 MS 105 8.5 113.5 102 NC 16 4 20 48 PR 8 2 10 24 SC
- - - - TN 8 2 10 24 TX 158.5 16.25 174.75 195
TOTAL 435.5 52.25 487.75 627

1 Calculated as: Cry1Ab acres x 12 lb of seed/acre
2 Not Applicable

Genotypes and vectors Transgenic cotton plants expressing the cry1ab gene (event T304-40 and T303-3) Experimental lines with different backgrounds Non transgenic plants Coker 315, Coker 312 or equivalents Transgenic commercial control Non transgenic commercial controlBayer CropScience Cry1Ab Cotton EUP Amendment Application 264-E UP-140, Page 35 of 36

Trial Design For seed increased to plant future regulatory studi es, a block consisting of transgenic seed will be planted with rows of non transgenic border rows of cotton planted on all four sides of the perimeter of the seed block. The spacing bet ween each row is approximately 38 inches.

Agronomic Treatments Typical agronomic inputs for conventionally grown cotton for the area, including, but not limited to:

 Conventional herbicide treatments, both pre- and post-planting

 Granular insecticide and/or fungicide application at planting

 Fertilizer applications

 Necessary in-season insecticide applications

 Growth regulator application (this should be done sparingly if at all)

 Additional hand weeding as necessary

 Chemical defoliation without boll-opening desiccants as this will mask maturity

Border rows The EUP test plants or the trial will be surrounded by one or more border rows of cotton.

Schedule Planting dates: February - June Harvest dates: July - November PR – Planting dates: October –November Harvest dates: April-May

Activities and Agronomic Practices After planting, transgenic and control seeds that were not planted will be collected. The transgenic seeds will be returned to the attent ion of the Study Director. Any non-transgenic seeds that are not planted will be disposed of at th e discretion of the Coop erator. The disposition of the seeds w ill be documented.

Plots will be harvested by hand or mechanically. The harvested seed cotton will be ginned at or near the field location to obtain required fuzzy seed samples. Lint obtained from the ginning will be disposed of. Seed samples will be packed, placed in frozen storage and ultimately shipped, frozen, to the Study Director and/or to a de signated analytical laboratory for compositional analyses.

After the field phase of the study is complete, the remaining transgenic and non-transgenic cotton plants will be destroyed by plowi ng the crop into the ground.

Containment EUP test plants will be isolat ed in accordance with USDA-APH IS Performance Standards for regulated cotton trials. Isolation methods will incl ude one or more of the following: (1) EUP test plants will be located at least 660 feet from other pa rties’ sexually receptiv e cotton; (2) a 40 footwide perimeter of non-tra nsgenic cotton will surroun d the transgenic plants planted at a density within 20% (greater or less than) the trial densit y to act as pollen sink for insect pollinators (the perimeter cotton would be disposed of by har vesting, disking and monitoring); (3) temporal isolation, where the flowering period for the EUP plants will not coincide with the presence of other parties’ sexually receptive cotton within 66 0 feet of the EUP test plants. Open flowering EUP test plants may be located within 660 feet of sexually receptive cotton provided such other cotton is used only for experimen tal purposes and/or destroyed.

Following the trial completion, all remaining plant debris will be destroyed by incorporation in the soil. All equipment used during crop destruction pr actices will be inspected and cleaned beforeBayer CropScience Cry1Ab Cotton EUP Amendment Application 264-EUP-140, Page 36 of 36

leaving the field. Seed cotton not destined fo r further experimentation will be destroyed by incineration or deep burial.

After completion of sampling, all perimeter cotton, if perimeters are planted, will be disposed of by mowing, followed by disking into the soil. The tr ansgenic plots and any perimeter cotton fields will be monitored the following season for volunteers. Volunteer plants will be terminated by hand weeding, disking, herbicide spraying or other method. If harvested material is to be ginned, seed cotton will securely be transported to a gin. All packaging and waste will be destroyed by devital ization. Ginned seed will be stored under containment practices for regulated materials.

Data Requirement Depending upon the study, one or two treatments of Cry1Ab cotton seed will be produced. If two treatments are produced, one treatment will be from plants that were treated with glufosinate ammonium and one treatment will be from plants that were not treated with glufosinate ammonium. In addition to the Cry1Ab Cotton material, a non-genetically modified line of the same genetic backgro und will also be produced to provide ma terial for comparative treatments.

Cotton seed will be harvest ed from each plot, including edge rows, at normal maturity. Material from the control non-transgenic plots will be har vested before harvesting t he transgenic plots, if they were established in the same field. The harvested seed cotton will be ginned to obtain the required seed samples to prepare meal for feeding studies. Ginning will be done at or near the field site. If the seed cotton is transported to the gin, the samples will be contained in an enclosed or covered vehicle or module. Ginning may be performed on a commercial gin, under the supervision of the Cooperator.

The ginned seed samples will be stored for tr ansport at ambient temperature in suitable containers such as large bulk bags. Sub-sample s will be sent to BCS. The bulk samples will be shipped to a processing facility for processing into meal. Transgenic cotton lint may be retained for BCS after ginning for immediate storage by BCS and possible future commercial use after registration. Alternatively, it will be disposed of. Any “waste” transgenic lint (e.g., spillage or clean-out waste) will be disposed of.

After samples are collected, remaining transgenic crop (left in the field a fter harvesting) will be destroyed.

Future Plans Transgenic seed harvested from trial may be used for regulatory trials at a later date.