Alexander B. Grannis

Commiss ioner

New York State Department of Environmental ConservationDivision of Solid & Hazardous MaterialsBureau of Pesticides ManagementPesticide Product Registration Section625 Broadway, Albany, New York 12233-7257Phone: 518-402-8768 FAX: 518-402-9024Website: http://www.dec.ny.gov/chemical/298.htmlE-Mail: ppr@gw.dec.state.ny.us

September 4, 2008

CERTIFIED MAIL (Co. No. 524)RETURN RECEIPT REQUESTED

Ms. Christine HullMonsanto Company800 North Lindbergh BlvdSt. Louis, MO 63167

Dear Ms. Hull:

Re: Registration of MON 89034 (EPA Reg. No. 524-575) and MON 89034 x 88017 (EPAReg. No. 524-576) Containing The New Active Ingredients Bacillus thuringiensisCryA.105 Protein and the Genetic Material Necessary for its Production (ActiveIngredient Code 006514) and Bacillus thuringiensis Cry2Ab2 Protein and theGenetic Material Necessary for its Production (Active Ingredient Code 006515)

The New York State Department of Environmental Conservation (Department) hascompleted review of your application and data package received on June 26, 2008 in support ofthe registration of MON 89034 and MON 89034 x MON 88017. Both MON 89034 and MON89034 x MON 88017 are defined as Plant Incorporated Protectants (PIPs), meaning that apesticidal gene has been introduced into the plant’s genetic material.

MON 89034 (EPA Reg. No. 524-575) contains 0.002-0.0056% Bacillus thuringiensisCryA.105 protein and the genetic material necessary for its production and 0.0015-0.0055%Bacillus thuringiensis Cry2Ab2 protein and the genetic material necessary for its production. MON 89034 is intended for use only in the production of corn with the goal of providingprotection from Euoropean corn borers, southwestern corn borers, southern cornstalk borers, cornearworms, fall armyworms, corn stalk borers, and sugarcane borers.

MON 89034 x MON 88017 (EPA Reg. No. 524-576) is produced through traditionalbreeding of MON 89034 with MON 88017. MON 89034 x MON 88017 contains 0.001-0.0024% Bacillus thuringiensis CryA.105 protein and the genetic material necessary for itsproduction, 0.0030-0.0057% Bacillus thuringiensis Cry2Ab2 protein and the genetic materialnecessary for its production, and 0.0037-0.0070% Bacillus thuringiensis Cry3Bb1 protein andthe genetic material necessary for its production. Bacillus thuringiensis Cry3Bb1 protein and thegenetic material necessary for its production has previously undergone a new active ingredientapplication in New York State that was completed on August 2, 2006 with the registration ofMON 88017 (EPA 524-551). MON 89034 x MON 88017 is solely for corn production and isintended for protection from feeding damage from corn rootworm in addition to Euoropean cornborers, southwestern corn borers, southern cornstalk borers, corn earworms, fall armyworms,

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corn stalk borers, and sugarcane borers while providing inherent tolerance to glyphosate-containing herbicides.

The subject application and data package were deemed complete for purposes of technicalreview on August 1, 2008. Pursuant to the review time frame specified in EnvironmentalConservation Law (ECL) §33-0704.2, a registration decision date of December 29, 2008 wasestablished. Due to the substantially similar nature of Bacillus thuringiensis CryA.105 proteinand Bacillus thuringiensis Cry2Ab2 protein to Bacillus thuringiensis subspecies previouslyregistered by the Department, it was determined that neither a human health technical review noran environmental fate technical review would be performed for this application. Hence, only anecological effects technical review was undertaken. Shown below is the technical reviewproduced by the Department’s Bureau of Habitat.

Ecological Effects Technical Review:

Ecological Risk Assessment of MON 89034 and MON 89034 x MON 88017 TransgenicCorn with the Active Ingredient Bacillus thuringiensis (Bt) Cry1A.105 and Bacillus thuringiensisCry2Ab2 proteins:

1. Chemical Description

Cry proteins are endotoxins stored in the form of protein crystals in certain bacteria justbefore sporulation. The bacteria Bacillus thuringiensis has been used as a source of such proteincrystals that are toxic to some varieties of insects such as mosquitoes, blackflies, and agriculturalpests; typically insects with alkaline digestive processes. Once the crystals have been ingested bythe larvae, they are solubilized and cleaved by a gut protease to produce the active toxin: a delta-endotoxin. The latter bore a hole in the insect’s midgut epithelium thus causing chaotic iontransfer and bringing the insect’s pH to a level in which bacteria can flourish and causesepticemia. Currently, almost one hundred different Bt endotoxin crystal proteins are known.One Bt can carry more than one type of endotoxin and thus be effective against a number ofdifferent insects (Protein Spotlight, 2005). As discussed in previous reviews (see references),Cry proteins have been genetically engineered into corn so that the corn plant itself will producetoxins for specific insect pests that feed on it (i.e., Plant-Incorporated Protectants).

Past transgenic corn products have been primarily aimed at protecting corn from variouscoleopteran and lepidopteran pests that attack corn roots, that is, corn rootworms. MON 89034 isdesigned to be effective against insects that attack the leaves and stalk of corn plants, such asEuropean corn borer, corn earworm, fall armyworm, and the southwestern corn borer. MON89034 contains two transgenic Bti proteins, Cry1A.105 and Cry2Ab2. While the Cry2Ab2 incorn has been shown in several analyses to be practically identical to the Cry2Ab2 produced inbacteria, Cry1A.105 is itself a product of genetic engineering. It is a “chimeric” or “fusion”protein. It does not occur naturally. Rather, it is constructed from Domain I, Domain II, and theC terminus of the Cry1Ac protein and Domain III of the Cry1Fa protein. The Cry1A.105chimeric protein is more effective against the target pests than the proteins from which it wasmade.

MON 88017 is a transgenic corn (already reviewed and registered for use in New YorkState) that contains the Cry3Bb1 protein, which is effective against several species of cornrootworms. When MON 89034 and MON 88017 are cross bred, the resulting product is a cornthat is protected against both rootworms and the leaf and stalk pests described above.

The leaf and stalk pests of corn do not inflict the same level of damage to corn yield ascorn rootworms, however, they can inflict significant damage. Some stalk pests are also very

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difficult to control with conventional pesticides, because they are only present on the outside ofthe stalk for a short period of time. One the insect has bored into the stalk, it is protected frominsecticides that are applied to the surface of the plant.

Leaf and stalk pests can inflict damage to corn, particularly the edible grain, that makesthe corn much more susceptible to fungal infections. Corn impacted with fungal infections canaccumulate significant levels of mycotoxins and can be dangerous to consume. In field trials,MON 89034 was found to contain much lower levels of mycotoxins than non-Bt corn. Thelevels of the mycotoxin aflatoxin were much lower in MON 89034 corn than non-Bt corn, andthere was less ear-rot damage to MON 89034 corn from the fungus Fusarium, there was not asignificant decrease in the concentration of the mycotoxin fumonisin.

2. Toxicity

MON 89034 and the MON 89034 x MON 88017 cross were tested extensively withnumerous non-target invertebrates, as well as birds and mammals. Test species included ladybeetles, earthworms, parasitic wasps, collembola, honeybee adults and larvae. Generally, notreatment effects were noted and no observed effects concentrations (NOECs) were greater thanthe highest concentration tested. The minute pirate bug (Orius insidiosus) is a beneficial insectthat preys on spider mites and thrips. Survival was reduced to 56% for pirate bugs exposed to240 µg Cry1A.105 /g diet. However, percent survival was not statistically significantly differentfrom controls for pirate bugs exposed to the next highest test concentration of 120 µgCry1A.105/g diet. With Cry2Ab2 proteins, only concentrations as high as 100 µg/g diet weretested with minute pirate bugs, and no adverse impacts were noted. The lack of non-targettoxicity observed for the Bt proteins in corns was consistent with results observed anddocumented in past reviews.

50A 48 hour Daphnia EC test was conducted by exposing Daphnia magna to pollen fromMON 89034 corn. However, this test was not accepted by the U.S. EPA. EPA reviewers notedthat 48 hours was too short a period to detect toxicity from Bti endotoxins. A 7 to 14 day test isrequired as a condition of registration, and is due to the U.S. EPA by 4/12/2009.

3. Exposure

Tissue levels of the transgenic proteins in MON 89034 and MON 89034 x MON 88017corn are listed in Tables 1 and 2, respectively:

Table 1. Tissue residue concentrations (and ranges) of transgenic proteins in MON 89034 corn

Tissue Type

Cry1A.105

µg/g dry weight ± Standard Dev.

Cry2Ab2

µg/g dry weight ± Standard Dev.

Leaf 72 ± 14 to 520 ± 130 130 ± 34 to 180 ± 59

Root 11 ± 1.4 to 79 ± 17 21 ± 5.9 to 58 ± 18

Whole plant 100 ± 26 to 380 ± 90 39 ± 16 to 130 ± 51

Pollen 12 ± 1.7 0.64 ± 0.091

Silk 26 ± 3.9 71 ± 35

Forage 42 ± 9.4 38 ± 14

Grain 5.9 ± 0.77 1.3 ± 0.36

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Table 2. Tissue residue concentrations of transgenic proteins in MON 89034 x MN 88017 corn

Tissue Type

Cry1A.105µg/g dry weight ±

Standard Dev.

Cry2Ab2µg/g dry weight± Standard Dev.

Cry3Bb1µg/g dry weight ±

Standard Dev.

CP4EPSPS*µg/g dry weight± Standard Dev.

Leaf 430 ± 71 170 ± 69 220 ± 66 200 ± 32

Root 83 ± 18 53 ± 27 200 ± 83 75 ± 38

Whole plant 140 ± 28 54 ± 15 160 ± 29 150 ± 26

Pollen 16 ± 1.7 0.62 ± 0.13 15 ± 3.4 320 ± 89

Forage-root 13 ± 3.5 24 ± 9.3 69 ± 21 30 ± 11

Forage 48 ± 13 44 ± 7.4 50 ± 9.1 55 ± 8.9

Grain 5.6 ± 1.3 1.3 ± 0.26 4.1 ± 2.3 3.4 ± 0.68

* Gene for glyphosate resistance

In soil, the Cry1A.105 and Cry2Ab2 proteins degraded rapidly. The half-life of theCry1A.105 protein has been found to range from 2 - 7 days in three different soils, and the half-life of Cry2Ab2 ranged from 0.5 - 3 days. However, (Saxena, et al. 2004) reported thattransgenic proteins can be exuded from corn roots, and can be bound up by surface-activeparticles such as clay particles and humic substances. In the bound state, they degrade muchmore slowly and retain their toxicity.

The environmental fate of Bti Cry proteins has been discussed in previous reviews. Alsoevaluated and discussed in previous reviews is the potential for transgenic traits to be transferredto native plants. The potential for this type of lateral transfer is considered to be very low, andthere are no receptive wild plants likely to cross breed with corn in New York.

4. Risk Assessment

The information provided with the MON 89034 and the MON 89034 x MON 88017registration applications is consistent with all of the other Bti Corn products reviewed. It is non-toxic to birds, mammals, fish, and most insects besides the targeted varieties. Database searchesshow that it has no amino acid sequences in common with known toxins or allergens. Itdisassociates very rapidly when exposed to simulated gastric juices, demonstrating that it israpidly and completely digested in a mammalian stomach

The potential for toxicity of transgenic Bti proteins in corn to non-target organisms hasbeen evaluated and reviewed, and considered to be slight. Some susceptible, non-targetorganisms can be exposed to corn pollen, but corn pollen breaks down very quickly, and isheavy, so it is not blown far from corn fields by the wind. Thus, any non-target impacts arelimited to the fringes of corn fields. The concentration of transgenic Bti proteins in pollen isalso much lower than in most other corn tissues (see Tables 1 and 2, above).

Studies submitted by the applicant showed that in 2005, the planting of transgenic cornreduced the usage of conventional corn insecticides by 4.85 million pounds nationwide. Thereduction in the use of conventional chemical insecticides might be perceived as a significantbenefit of the use of transgenic corn.

Bureau of Habitat staff have no objection to the registration of MON 89034 or MON89034 x MON 88017 in New York State.

Ms. Christine Hull 5.

5. Literature Cited

Protein Spotlight, 2005. The cry proteins. September 4, 2005. Protein Spotlight (ISSN 1424-4721), a monthly review of the Swiss Institute of Bioinformatics.(http://www.expasy.org/spotlight/snapshot/2005/09/the_cry_protein.shtml)

Saxena, D., C.N. Stewart, I. Altosaar, Q. Shu, and G. Stotzky, 2005. Larvicidal Cry proteinsfrom Bacillus thuringiensis are released in root exudates of transgenic B. Thuringiensis corn,potato, and rice but not of B. Thuringiensis canola, cotton, and tobacco. Plant Physiology andBiochemistry 42(2004):383-387. Doi:10.1016/j.plaphy.2004.03.004

REGISTRATION SUMMARY

The Department has registered MON 89034 (EPA Reg. No. 524-575) and MON89034 x MON 88017 (EPA Reg. No. 524-576) for use in New York State.

Enclosed for your record is a copy of the stamped “Accepted For Registration” label andthe New York State Certificate of Pesticide Registration for MON 89034 and MON 89034 xMON 88017. Please note that a proposal by Monsanto Company or any other registrant toregister a product that contains Bacillus thuringiensis CryA.105 protein and the genetic materialnecessary for its production and/or Bacillus thuringiensis Cry2Ab2 protein and the geneticmaterial necessary for its production, and whose labeled uses are likely to increase the potentialfor significant impact to humans, non-target organisms, or the environment, would constitute amajor change in labeled use pattern. Such an application must be accompanied by a newapplication fee and meet the requirements listed in Appendix 1.B. of “New York State PesticideProduct Registration Procedures: (September 2005). Such information, as well as forms, can beaccessed at our website as listed in our letterhead.

Please be aware that any unregistered product may not be sold, offered for sale,distributed, or used in New York State.

If you have any questions regarding this matter, please contact our Pesticide ProductRegistration Section, at (518) 402-8768.

Sincerely,

Maureen P. Serafini

Maureen P. SerafiniDirectorBureau of Pesticides Management

Enclosures

ecc: Bob Mungari, NYS Department of Agriculture and MarketsWill Smith, Cornell University, PMEP