# T ITLE



# Administrative Volume for an Application for a Seed Increase Registration of the Plant-Incorporated

Protectant, Bacillus thuringiensis Cry1F and Cry34/35Ab1 Proteins and the Genetic Material

(PHP27118 T-DNA) Necessary for their Production in 4114 Maize

(OECD Unique Identifier: DP-ØØ4114-3)



# D ATA 

# R EQUIREMENT



# 40 CFR Part 174



# A UTHOR



# Jamie Staley

Pioneer Hi-Bred International, Inc.

7100 NW 62 nd 

# Avenue

Johnston, IA 50131

USA



# S TUDY 

# C OMPLETION 

# D ATE



# April 15, 2011



# S TUDY 

# N UMBER



# PHI-2011-080



# EPA R EGISTRATION 

# N UMBER



# 29964-x

Volume 1 Pioneer Hi-Bred International, Inc.

Study Number: PHI-2011-080 Page 1 of 100

# T ABLE OF 

# C ONTENTS 

# P AGE



# Statement of Data C onfidentiality Claim ....................................................................................... .... 2

Transmittal Do cument .......................................................................................................... ............ 4

Section I. Adminis trative Materials .......................................................................................... ........ 7

Section II. Summary of the Application ....................................................................................... ..73

Section III. Product Label ...................................................................................................... ........77

Section IV. Satisfaction of Data Requirem ents ............................................................................. 84

Section V. Product Characterization

.......................................................................................... ...89

Section VI. Toxicity Data .................................................................................................... ...........92

Section VII. Residue Da ta ...................................................................................................... .......94

Section VIII. Efficacy Data .................................................................................................. ...........95

Confidential Attachm ent ....................................................................................................... ..........96

Tables

Table 1. Currently Registered Maize PIPs Expressing Cry1F and/or Cry34/35Ab1 Proteins ...... 75

Table 2. Product Analysis Data Requirem ents ............................................................................. 84

Table 3. Residue Da ta Require ments ........................................................................................... 84

Table 4. Toxicology Data Requirement s ....................................................................................... 85

Table 5. Non-target Organism Data Requirements .......................................................................87

Table 6. Environmental Ex pression Data Requirem ents .............................................................. 88

Table 7. Mean Expression Levels of the Cry1F, Cry34Ab1 and Cry35Ab1 Proteins

in 4114 Ma ize ................................................................................................................. ................90

Table 8. Summary of Product Characterization Data Submitted for 4114 Maize ......................... 91

Table 9. Environmental Safety Data Submitted for 4114 Maize ................................................... 93 Pioneer Hi-Bred International, Inc.

Study Number: PHI-2011-080 Page 3 of 100

# T RANSMITTAL 

# D OCUMENT



# S UBMITTER



# Pioneer Hi-Bred International, Inc.

7100 NW 62 nd 

# Ave.

Johnston, IA 50131



# R EGULATORY 

# A CTION IN 

# S UPPORT OF 

# W HICH 

# T HIS 

# D OCUMENT IS 

# S UBMITTED



# Application for a New Product Registration with a Plant-Incorporated Protectant

Pursuant to FIFRA §3(c)(5):

Administrative Volume for an Application for a Seed Increase Registration of the Plant-Incorporated

Protectant, Bacillus thuringiensis Cry1F and Cry34/35Ab1 Proteins and the Genetic Material

(PHP27118 T-DNA) Necessary for their Production in 4114 Maize

(OECD Unique Identifier: DP-ØØ4114-3)



# EPA R EGISTRATION 

# N UMBER



# 29964-x



# T RANSMITTAL 

# D ATE



# April 15, 2011



# S TUDY 

# N UMBER



# PHI-2011-080 Pioneer Hi-Bred International, Inc.

Study Number: PHI-2011-080 Page 4 of 100

# L IST OF 

# A CCOMPANYING 

# V OLUMES



# VOL. FIFRA

10(G) VOLUME/STUDY TITLE MRID

1 A Administrative Volume for an Application for a Seed Increase

Registration of the Plant-Incorporated Protectant, Bacillus thuringiensis

Cry1F and Cry34/35Ab1 Proteins and the Genetic Material (PHP27118

T-DNA) Necessary for their Production in 4114 Maize

Staley, J. (2011).

Study Number: PHI-2011-080 April 15, 2011

Pages: 1-100

C Confidential Attachment

2 B Characterization of DP-ØØ4114-3 maize: Insertion integrity, stability,

copy number, and backbone analysis

Brink, K., and J. Weaver. (2011).

Study Number: PHI-2009-015 April 1, 2011

Pages: 1-48 (3 copies)

3 B Equivalency of expressed proteins in 4114 maize to previously

registered events

Cressman, B. et al . (2011).

Study Number: PHI-2011-076 April 15, 2011

Pages: 1-23 (3 copies)

4 B Field and greenhouse insect efficacy for 4114 maize

Nowatzki, T. et al . (2011).

Study Number: PHI-2011-077 April 8, 2011

Pages: 1-31 (3 copies)

5 B Environmental Risk Assessment for the cultivation of maize containing

event DP-ØØ4114-3 (4114 maize)

Califf, K. et al . (2011).

Study Number: PHI-2011-078 April 18, 2011

Pages: 1-39 (3 copies)

6 B Trait Durability Plan supporting a Seed Increase Registration for 4114

maize

Binning, R. et al. (2011).

Study Number: PHI-2011-079 April 14, 2011

Pages: 1-9 (3 copies)

7 B Western blot analysis of maize lines containing event DP-ØØ4114-3

and combined trait product DAS-Ø15Ø7-1xDAS-59122-7

Sult, T and P. Utley. (2011).

Study Number: PHI-2011-067 April 15, 2011

Pages: 1-15 (3 copies) Pioneer Hi-Bred International, Inc.

Study Number: PHI-2011-080 Page 5 of 100

# S ECTION 

# I. A DMINISTRATIVE 

# M ATERIALS



# A.1. Letter of Authorization (Data Access Letter) from Dow AgroSciences LLC Pioneer Hi-Bred International, Inc.

Study Number: PHI-2011-080 Page 7 of 100

# Pioneer Hi-Bred International, Inc.

Study Number: PHI-2011-080 Page 8 of 100

# A.2. EPA Form 8570-1: Application for Pesticide Registration Pioneer Hi-Bred International, Inc.

Study Number: PHI-2011-080 Page 9 of 100

# A.3. EPA Form 8570-4: Confidential Statement of Formula

See CBI Cross Reference Number 1 Pioneer Hi-Bred International, Inc.

Study Number: PHI-2011-080 Page 11 of 100

# A.4. EPA Form 8570-34: Certification with Respect to Citation of Data Pioneer Hi-Bred International, Inc.

Study Number: PHI-2011-080 Page 12 of 100

# A.5. EPA Form 8570-35: Data Matrix for 4114 Maize Pioneer Hi-Bred International, Inc.

Study Number: PHI-2011-080 Page 14 of 100Form Approved OMB No. 2070-0060UNITED STATES ENVIRONMENTAL PROTECTION AGENCY1200 Pennsylvania Avenue, N.W.Washington, D.C. 20460Paperwork Reduction Act Notice:The public reporting burden for this collection of information is estimated to average 0.25 hours per response for registrationactivities and 0.25 hours perresponse for reregistration and special review activities, including time for reading the instructions and completing the necessary forms. Send comments regarding the burden estimate or anyother aspect of this collection of information, including suggestions for reducing the burden to: Director, OPPE Information Management Division (2137), U.S. Environmental Protection Agency,1200 Pennsylvania Avenue, N.W., Washington, DC 20460. Do not send the form to this address.DATA MATRIXDate:April 15, 2011EPA Reg. No. / File Symbol:29964-xPage1of29Applicant's/Registrant's Name & Address:Pioneer Hi-Bred International, Inc., 7100 NW 62ndAvenue, Johnston, IA 50131Product:4114 MaizeIngredientBacillus thuringiensisCry1F and Cry34/35Ab1 insecticidal proteins and the genetic material (PHP27118 T-DNA) necessary for their production in cornGuideline Reference Number Guideline Study Name MRID Number Submitter Status NoteAcute oral toxicity in mice: Cry1FBacillusthuringiensissubsp.aizawaidelta endotoxin 44691101 68467 PEREffectiveness data forBacillus thuringiensisvar.aizawaiCry1F insect control protein as expressed inmaize 44691102 68467 PERBackground document on resistance management 44691103 68467 PERProduct characterization data forBacillusthuringiensisvar.aizawai: Cry1F insect controlprotein as expressed in maize 44714801 68467 PERCharacterization of gene inserts –Bacillusthuringiensis aizawai: Cry1F control protein asexpressed in maize 44714802 68467 PEREquivalency of microbial and maize expressedCry1F protein; characterization of test substancesfor biochemical and toxicological studies. In vitrodigestibility of microbial and maize expressed Cry1Fprotein under simulated gastric conditions 44714803 68467 PERSignatureName and TitleJamie StaleyU.S. Registration ManagerDateApril 15, 2011EPA Form 8570-35 (9-97) Electronic and Paper versions available. Submit only Paper version.Agency Internal Use CopyPioneer Hi-Bred International, Inc.Study Number: PHI-2011-080Page 15 of 100Form Approved OMB No. 2070-0060UNITED STATES ENVIRONMENTAL PROTECTION AGENCY1200 Pennsylvania Avenue, N.W.Washington, D.C. 20460Paperwork Reduction Act Notice:The public reporting burden for this collection of information is estimated to average 0.25 hours per response for registrationactivities and 0.25 hours perresponse for reregistration and special review activities, including time for reading the instructions and completing the necessary forms. Send comments regarding the burden estimate or anyother aspect of this collection of information, including suggestions for reducing the burden to: Director, OPPE Information Management Division (2137), U.S. Environmental Protection Agency,1200 Pennsylvania Avenue, N.W., Washington, DC 20460. Do not send the form to this address.DATA MATRIXDate:April 15, 2011EPA Reg. No. / File Symbol:29964-xPage2of29Applicant's/Registrant's Name & Address:Pioneer Hi-Bred International, Inc., 7100 NW 62ndAvenue, Johnston, IA 50131Product:4114 MaizeIngredientBacillus thuringiensisCry1F and Cry34/35Ab1 insecticidal proteins and the genetic material (PHP27118 T-DNA) necessary for their production in cornGuideline Reference Number Guideline Study Name MRID Number Submitter Status NoteQuantitative ELISA analysis of Cry1F expressionand PAT expression levels in maize MPS inbred andhybrid lines 1360, 1365, 1366 and 1369 (interimreport) 44714804 68467 PERComparison of the amino acid sequence similarity ofCry1F and PAT proteins to KnPER allergen protein 44971701 68467 PERMicrobialB.t.Cry1F (truncated) delta-endotoxin:Maize-insect-pest susceptibility study 45020101 68467 PERCharacterization of inserted genes in Cry1F maizeline 1507 45020102 68467 PERCharacterization of expressed Cry1F protein inmaize tissues (pollen, grain, grain-containing feedand purified maize-expressed Cry1F protein) andmicrobial expressed Cry1F delta-endotoxin bybiological and biochemical procedures 45020103 68467 PERQuantitative ELISA analysis of Cry1F and PATexpression levels in and compositional analysis ofmaize inbred and hybrid line 1362 and 1507 45020104 68467 PERSignatureName and TitleJamie StaleyU.S. Registration ManagerDateApril 15, 2011EPA Form 8570-35 (9-97) Electronic and Paper versions available. Submit only Paper version.Agency Internal Use CopyPioneer Hi-Bred International, Inc.Study Number: PHI-2011-080Page 16 of 100Form Approved OMB No. 2070-0060UNITED STATES ENVIRONMENTAL PROTECTION AGENCY1200 Pennsylvania Avenue, N.W.Washington, D.C. 20460Paperwork Reduction Act Notice:The public reporting burden for this collection of information is estimated to average 0.25 hours per response for registrationactivities and 0.25 hours perresponse for reregistration and special review activities, including time for reading the instructions and completing the necessary forms. Send comments regarding the burden estimate or anyother aspect of this collection of information, including suggestions for reducing the burden to: Director, OPPE Information Management Division (2137), U.S. Environmental Protection Agency,1200 Pennsylvania Avenue, N.W., Washington, DC 20460. Do not send the form to this address.DATA MATRIXDate:April 15, 2011EPA Reg. No. / File Symbol:29964-xPage3of29Applicant's/Registrant's Name & Address:Pioneer Hi-Bred International, Inc., 7100 NW 62ndAvenue, Johnston, IA 50131Product:4114 MaizeIngredientBacillus thuringiensisCry1F and Cry34/35Ab1 insecticidal proteins and the genetic material (PHP27118 T-DNA) necessary for their production in cornGuideline Reference Number Guideline Study Name MRID Number Submitter Status NoteEnvironmental fate of Cry1F protein incorporatedinto soil 45020105 68467 PERCry1FBacillus thuringiensisvar.aizawaideltaendotoxin: an acute toxicity study with the earthwormin an artificial soil substrate 45020106 68467 PERChronic exposure ofFolsomia candidato bacteriallyexpressed Cry1F protein 45020107 68467 PEROECD 202B.t.Cry1F delta endotoxin: A 48-hour static-renewalacute toxicity test with the Cladoceran (Daphniamagna) using bacterially expressedB.t.Cry1F deltaendotoxin, and pollen from maize expressingB.t.Cry1F delta endotoxin 45020108 68467 PER885.4340Cry1FBacillus thuringiensisvar.aizawaideltaendotoxin: A dietary study with green lacewinglarvae 45020109 68467 PER885.4340Cry1FBacillus thuringiensisvar.aizawaideltaendotoxin: A dietary study with the ladybird beetle 45020110 68467 PERSignatureName and TitleJamie StaleyU.S. Registration ManagerDateApril 15, 2011EPA Form 8570-35 (9-97) Electronic and Paper versions available. Submit only Paper version.Agency Internal Use CopyPioneer Hi-Bred International, Inc.Study Number: PHI-2011-080Page 17 of 100Form Approved OMB No. 2070-0060UNITED STATES ENVIRONMENTAL PROTECTION AGENCY1200 Pennsylvania Avenue, N.W.Washington, D.C. 20460Paperwork Reduction Act Notice:The public reporting burden for this collection of information is estimated to average 0.25 hours per response for registrationactivities and 0.25 hours perresponse for reregistration and special review activities, including time for reading the instructions and completing the necessary forms. Send comments regarding the burden estimate or anyother aspect of this collection of information, including suggestions for reducing the burden to: Director, OPPE Information Management Division (2137), U.S. Environmental Protection Agency,1200 Pennsylvania Avenue, N.W., Washington, DC 20460. Do not send the form to this address.DATA MATRIXDate:April 15, 2011EPA Reg. No. / File Symbol:29964-xPage4of29Applicant's/Registrant's Name & Address:Pioneer Hi-Bred International, Inc., 7100 NW 62ndAvenue, Johnston, IA 50131Product:4114 MaizeIngredientBacillus thuringiensisCry1F and Cry34/35Ab1 insecticidal proteins and the genetic (PHP27118 T-DNA) material necessary for their production in cornGuideline Reference Number Guideline Study Name MRID Number Submitter Status Note885.4340Cry1FBacillus thuringiensisvar.aizawaideltaendotoxin: A dietary toxicity study with parasitichymenoptera 45020111 68467 PER71-2Transgenic corn expressingBacillus thuringiensisvar.aizawai(B.t.) Cry1F delta endotoxin: A dietarytoxicity study with Northern bobwhite 45020112 68467 PERField survey of beneficial arthropods associated withBacillus thuringiensisCry1F maize 45020113 68467 PEREfficacy of Cry1F events TC1360 and TC1507 45020114 68467 PERCry1F binding studies 45020115 68467 PERResistance management plan for transgenic maizeexpressing the Cry1F insecticidal protein fromBacillus thuringiensisvar.aizawai45020116 68467 PERSignatureName and TitleJamie StaleyU.S. Registration ManagerDateApril 15, 2011EPA Form 8570-35 (9-97) Electronic and Paper versions available. Submit only Paper version.Agency Internal Use CopyPioneer Hi-Bred International, Inc.Study Number: PHI-2011-080Page 18 of 100Form Approved OMB No. 2070-0060UNITED STATES ENVIRONMENTAL PROTECTION AGENCY1200 Pennsylvania Avenue, N.W.Washington, D.C. 20460Paperwork Reduction Act Notice:The public reporting burden for this collection of information is estimated to average 0.25 hours per response for registrationactivities and 0.25 hours perresponse for reregistration and special review activities, including time for reading the instructions and completing the necessary forms. Send comments regarding the burden estimate or anyother aspect of this collection of information, including suggestions for reducing the burden to: Director, OPPE Information Management Division (2137), U.S. Environmental Protection Agency,1200 Pennsylvania Avenue, N.W., Washington, DC 20460. Do not send the form to this address.DATA MATRIXDate:April 15, 2011EPA Reg. No. / File Symbol:29964-xPage5of29Applicant's/Registrant's Name & Address:Pioneer Hi-Bred International, Inc., 7100 NW 62ndAvenue, Johnston, IA 50131Product:4114 MaizeIngredientBacillus thuringiensisCry1F and Cry34/35Ab1 insecticidal proteins and the genetic material (PHP27118 T-DNA) necessary for their production in cornGuideline Reference Number Guideline Study Name MRID Number Submitter Status NoteSupplement to MRID 44714801: Supplemental data- Product characterization data forBacillusthuringiensisvar.aizawai: Cry1F control protein asexpressed in maize 45020117 68467 PERSupplement to MRID 44691101: Supplemental data- Acute oral toxicity in mice:Bacillus thuringiensisvar.aizawaiCry1F delta endotoxin 45020118 68467 PERPhosphinothricin acetyltransferase (PAT) protein: Invitro digestibility study 45041501 68467 PERNon-target exposure and risk assessment forenvironmental dispersal of Cry1F maize pollen 45041502 68467 PEREvaluation of the dietary effect(s) on honeybeedevelopment using bacterially expressedB.t.Cry1Fdelta endotoxin and pollen from maize expressingB.t.Cry1F delta endotoxin 45041503 68467 PERWaiver request: Fish toxicity test with transgenicmaize (corn) containingBacillus thuringiensisvar.aizawai(B.t.) Cry1F delta endotoxin 45044201 68467 PERSignatureName and TitleJamie StaleyU.S. Registration ManagerDateApril 15, 2011EPA Form 8570-35 (9-97) Electronic and Paper versions available. Submit only Paper version.Agency Internal Use CopyPioneer Hi-Bred International, Inc.Study Number: PHI-2011-080Page 19 of 100Form Approved OMB No. 2070-0060UNITED STATES ENVIRONMENTAL PROTECTION AGENCY1200 Pennsylvania Avenue, N.W.Washington, D.C. 20460Paperwork Reduction Act Notice:The public reporting burden for this collection of information is estimated to average 0.25 hours per response for registrationactivities and 0.25 hours perresponse for reregistration and special review activities, including time for reading the instructions and completing the necessary forms. Send comments regarding the burden estimate or anyother aspect of this collection of information, including suggestions for reducing the burden to: Director, OPPE Information Management Division (2137), U.S. Environmental Protection Agency,1200 Pennsylvania Avenue, N.W., Washington, DC 20460. Do not send the form to this address.DATA MATRIXDate:April 15, 2011EPA Reg. No. / File Symbol:29964-xPage6of29Applicant's/Registrant's Name & Address:Pioneer Hi-Bred International, Inc., 7100 NW 62ndAvenue, Johnston, IA 50131Product:4114 MaizeIngredientBacillus thuringiensisCry1F and Cry34/35Ab1 insecticidal proteins and the genetic material (PHP27118 T-DNA) necessary for their production in cornGuideline Reference Number Guideline Study Name MRID Number Submitter Status NoteHigh dose demonstration of Cry1F events TC1360and TC1507: European corn borer 45131101 68467 PERToxicity of the Cry1F protein to neonate larvae of themonarch buttefly (Danaus plexippus(Linneaus)) 45131102 68467 PERPublic interest document for Cry1F-protected corn 45131103 29964 OWNThermolability of Cry1F (truncated) delta endotoxin 45274801 68467 PERCompositional analysis of maize MPS hybrid line1507 45274802 68467 PERCry1F lateral flow test kit procedure for analyzingCry1F corn grain 45279301 68467 PERMethod validation report for the determination ofCry1F delta endotoxin protein in grain by EnzymeLinked Immunosorbent Assay 45279302 68467 PERSignatureName and TitleJamie StaleyU.S. Registration ManagerDateApril 15, 2011EPA Form 8570-35 (9-97) Electronic and Paper versions available. Submit only Paper version.Agency Internal Use CopyPioneer Hi-Bred International, Inc.Study Number: PHI-2011-080Page 20 of 100Form Approved OMB No. 2070-0060UNITED STATES ENVIRONMENTAL PROTECTION AGENCY1200 Pennsylvania Avenue, N.W.Washington, D.C. 20460Paperwork Reduction Act Notice:The public reporting burden for this collection of information is estimated to average 0.25 hours per response for registrationactivities and 0.25 hours perresponse for reregistration and special review activities, including time for reading the instructions and completing the necessary forms. Send comments regarding the burden estimate or anyother aspect of this collection of information, including suggestions for reducing the burden to: Director, OPPE Information Management Division (2137), U.S. Environmental Protection Agency,1200 Pennsylvania Avenue, N.W., Washington, DC 20460. Do not send the form to this address.DATA MATRIXDate:April 15, 2011EPA Reg. No. / File Symbol:29964-xPage7of29Applicant's/Registrant's Name & Address:Pioneer Hi-Bred International, Inc., 7100 NW 62ndAvenue, Johnston, IA 50131Product:4114 MaizeIngredientBacillus thuringiensisCry1F and Cry34/35Ab1 insecticidal proteins and the genetic material (PHP27118 T-DNA) necessary for their production in cornGuideline Reference Number Guideline Study Name MRID Number Submitter Status NotePublic interest document for Cry1F protected corn 45298101 29964 OWNSupplement to MRID 45131102: Supplemental data- High dose demonstration of Cry1F events TC1360and TC1507: European corn borer 45307701 68467 PERWaiver request: Fish toxicity test to assess thepotential effects of maize containingBacillusthuringiensisvar.aizawai(Bt) Cry1F insecticidalprotein (ICP) in native fish 45307702 68467 PERSupplement to MRID 45020109: Cry1FBacillusthuringiensisvar.aizawaidelta endotoxin: A dietarytoxicity study with green lacewing larvae 45307801 68467 PERSupplement to MRID 45020110: Cry1FBacillusthuringiensisvar.aizawaidelta endotoxin: A dietarytoxicity study with green ladybird beetle 45307802 68467 PERSupplement to MRID 45020111: Cry1FBacillusthuringiensisvar.aizawaidelta endotoxin: A dietarytoxicity study with parasitic hymenoptera 45307803 68467 PERSignatureName and TitleJamie StaleyU.S. Registration ManagerDateApril 15, 2011EPA Form 8570-35 (9-97) Electronic and Paper versions available. Submit only Paper version.Agency Internal Use CopyPioneer Hi-Bred International, Inc.Study Number: PHI-2011-080Page 21 of 100Form Approved OMB No. 2070-0060UNITED STATES ENVIRONMENTAL PROTECTION AGENCY1200 Pennsylvania Avenue, N.W.Washington, D.C. 20460Paperwork Reduction Act Notice:The public reporting burden for this collection of information is estimated to average 0.25 hours per response for registrationactivities and 0.25 hours perresponse for reregistration and special review activities, including time for reading the instructions and completing the necessary forms. Send comments regarding the burden estimate or anyother aspect of this collection of information, including suggestions for reducing the burden to: Director, OPPE Information Management Division (2137), U.S. Environmental Protection Agency,1200 Pennsylvania Avenue, N.W., Washington, DC 20460. Do not send the form to this address.DATA MATRIXDate:April 15, 2011EPA Reg. No. / File Symbol:29964-xPage8of29Applicant's/Registrant's Name & Address:Pioneer Hi-Bred International, Inc., 7100 NW 62ndAvenue, Johnston, IA 50131Product:4114 MaizeIngredientBacillus thuringiensisCry1F and Cry34/35Ab1 insecticidal proteins and the genetic material (PHP27118 T-DNA) necessary for their production in cornGuideline Reference Number Guideline Study Name MRID Number Submitter Status NoteSupplement to MRID 45020106: Cry1FBacillusthuringiensisvar.aizawaidelta endotoxin: An acutetoxicity study with the earthworm in an artificial soilsubstrate 45307804 68467 PERSupplement to MRID 45041503: Evaluation of thedietary effect(s) on honeybee development usingbacterially expressedB.t.Cry1F delta endotoxin andpollen from maize expressingB.t.Cry1F deltaendotoxin 45307805 68467 PERSupplement to MRID 45298101: Supplemental data- Public interest document for Cry1F protected corn 45309401 29964 OWNExposure and risk assessment of Herculex I Bt fieldcorn pollen to Karner blue butterfly 45512901 68467 PERNutritional equivalency ofB.t.Cry1F maize - poultryfeeding study 45622001 68467 PERField survey of beneficial arthropods associated withBacillus thuringiensisCry1F maize 45648001 68467 PERSignatureName and TitleJamie StaleyU.S. Registration ManagerDateApril 15, 2011EPA Form 8570-35 (9-97) Electronic and Paper versions available. Submit only Paper version.Agency Internal Use CopyPioneer Hi-Bred International, Inc.Study Number: PHI-2011-080Page 22 of 100Form Approved OMB No. 2070-0060UNITED STATES ENVIRONMENTAL PROTECTION AGENCY1200 Pennsylvania Avenue, N.W.Washington, D.C. 20460Paperwork Reduction Act Notice:The public reporting burden for this collection of information is estimated to average 0.25 hours per response for registrationactivities and 0.25 hours perresponse for reregistration and special review activities, including time for reading the instructions and completing the necessary forms. Send comments regarding the burden estimate or anyother aspect of this collection of information, including suggestions for reducing the burden to: Director, OPPE Information Management Division (2137), U.S. Environmental Protection Agency,1200 Pennsylvania Avenue, N.W., Washington, DC 20460. Do not send the form to this address.DATA MATRIXDate:April 15, 2011EPA Reg. No. / File Symbol:29964-xPage9of29Applicant's/Registrant's Name & Address:Pioneer Hi-Bred International, Inc., 7100 NW 62ndAvenue, Johnston, IA 50131Product:4114 MaizeIngredientBacillus thuringiensisCry1F and Cry34/35Ab1 insecticidal proteins and the genetic material (PHP27118 T-DNA) necessary for their production in cornGuideline Reference Number Guideline Study Name MRID Number Submitter Status NoteField surveys of non-target invertebrate populationsinB.t.corn 45652001 68467 PERDevelopment and characterization of EnzymeLinked Immunosorbent Assay (ELISA) for detectionof Cry1F protein 45685601 68467 PERIndependent laboratory validation of method GRM02.13, determination of Cry1F delta endotoxinprotein in corn grain by an of Enzyme-LinkedImmunosorbent Assay 45685602 68467 PERSupplement to MRID 45131102: Toxicity of theCry1F protein to neonate larvae of the monarchbutterfly (Danaus plexippus(Linneaus)) 45759701 68467 PERStewardship of Herculex I Insect Protection withrespect to the secondary lepidopteran pest, westernbean cutworm (Richia albicostaSmith) 45896601 29964 OWNLack of cross reactivity between Cry1F protein inHerculex I maize and the dust mite Der p7 proteinwith human sera positive for Der p7-IgE 46444001 68467 PERSignatureName and TitleJamie StaleyU.S. Registration ManagerDateApril 15, 2011EPA Form 8570-35 (9-97) Electronic and Paper versions available. Submit only Paper version.Agency Internal Use CopyPioneer Hi-Bred International, Inc.Study Number: PHI-2011-080Page 23 of 100Form Approved OMB No. 2070-0060UNITED STATES ENVIRONMENTAL PROTECTION AGENCY1200 Pennsylvania Avenue, N.W.Washington, D.C. 20460Paperwork Reduction Act Notice:The public reporting burden for this collection of information is estimated to average 0.25 hours per response for registrationactivities and 0.25 hours perresponse for reregistration and special review activities, including time for reading the instructions and completing the necessary forms. Send comments regarding the burden estimate or anyother aspect of this collection of information, including suggestions for reducing the burden to: Director, OPPE Information Management Division (2137), U.S. Environmental Protection Agency,1200 Pennsylvania Avenue, N.W., Washington, DC 20460. Do not send the form to this address.DATA MATRIXDate:April 15, 2011EPA Reg. No. / File Symbol:29964-xPage10of29Applicant's/Registrant's Name & Address:Pioneer Hi-Bred International, Inc., 7100 NW 62ndAvenue, Johnston, IA 50131Product:4114 MaizeIngredientBacillus thuringiensisCry1F and Cry34/35Ab1 insecticidal proteins and the genetic material (PHP27118 T-DNA) necessary for their production in cornGuideline Reference Number Guideline Study Name MRID Number Submitter Status NoteMonitoring the susceptibility of European corn borerto Cry1Ab and Cry1F Bt proteins: Results from the2004 collections and diapausing larvae collected in2003 46583101 80778 PERStewardship of event TC1507 maize with respect tothe secondary lepidopteran pests lesser corn stalkborer (Elasmopalpus lignosellusZeller), southerncorn stalk borer (Diatraea crambidiodesGrote), andsugarcane borer (Biatraea saccharalisFabricius) 46600201 68467 PERSlide presentation summarizing European corn borerand Cry1F resistance monitoring update 46695801 68467 PERInsect resistance management complianceassurance program report for corn borer-protectedBt corn 46747801 80778 PERField surveys of non-target invertebrate populationsin Bt corn: Supplement to MRID No. 45652001 46784601 80778 PERMonitoring the susceptibility of corn lepidopteranpests to Cry1Ab and Cry1F proteins: 2005monitoring results 46874901 80778 PERSignatureName and TitleJamie StaleyU.S. Registration ManagerDateApril 15, 2011EPA Form 8570-35 (9-97) Electronic and Paper versions available. Submit only Paper version.Agency Internal Use CopyPioneer Hi-Bred International, Inc.Study Number: PHI-2011-080Page 24 of 100Form Approved OMB No. 2070-0060UNITED STATES ENVIRONMENTAL PROTECTION AGENCY1200 Pennsylvania Avenue, N.W.Washington, D.C. 20460Paperwork Reduction Act Notice:The public reporting burden for this collection of information is estimated to average 0.25 hours per response for registrationactivities and 0.25 hours perresponse for reregistration and special review activities, including time for reading the instructions and completing the necessary forms. Send comments regarding the burden estimate or anyother aspect of this collection of information, including suggestions for reducing the burden to: Director, OPPE Information Management Division (2137), U.S. Environmental Protection Agency,1200 Pennsylvania Avenue, N.W., Washington, DC 20460. Do not send the form to this address.DATA MATRIXDate:April 15, 2011EPA Reg. No. / File Symbol:29964-xPage11of29Applicant's/Registrant's Name & Address:Pioneer Hi-Bred International, Inc., 7100 NW 62ndAvenue, Johnston, IA 50131Product:4114 MaizeIngredientBacillus thuringiensisCry1F and Cry34/35Ab1 insecticidal proteins and the genetic material (PHP27118 T-DNA) necessary for their production in cornGuideline Reference Number Guideline Study Name MRID Number Submitter Status NoteResearch results on 2004 European corn borercollections from Hamilton County, Iowa: Cry1F 47011201 68467 PERInsect resistance management complianceassurance program report for corn borer-protectedBt corn, corn rootworm-protected Bt corn, and cornborer/corn rootworm protected stacked Bt corn 47044401 80778 PERMonitoring the susceptibility of corn lepidopteranpests to Cry1Ab and Cry1F proteins: 2006monitoring results 47118401 80778 PERSoil accumulation of Cry1F after three years ofcropping with Herculex 1 corn 47120701 68467 PERTC1507 maize and fall armyworm in Puerto Rico 47176001 68467 PERProposed revisions to IRM-related registrationrequirements for Cry1 plant-incorporated protectantsin field corn 47407001 80778 PERSignatureName and TitleJamie StaleyU.S. Registration ManagerDateApril 15, 2011EPA Form 8570-35 (9-97) Electronic and Paper versions available. Submit only Paper version.Agency Internal Use CopyPioneer Hi-Bred International, Inc.Study Number: PHI-2011-080Page 25 of 100Form Approved OMB No. 2070-0060UNITED STATES ENVIRONMENTAL PROTECTION AGENCY1200 Pennsylvania Avenue, N.W.Washington, D.C. 20460Paperwork Reduction Act Notice:The public reporting burden for this collection of information is estimated to average 0.25 hours per response for registrationactivities and 0.25 hours perresponse for reregistration and special review activities, including time for reading the instructions and completing the necessary forms. Send comments regarding the burden estimate or anyother aspect of this collection of information, including suggestions for reducing the burden to: Director, OPPE Information Management Division (2137), U.S. Environmental Protection Agency,1200 Pennsylvania Avenue, N.W., Washington, DC 20460. Do not send the form to this address.DATA MATRIXDate:April 15, 2011EPA Reg. No. / File Symbol:29964-xPage12of29Applicant's/Registrant's Name & Address:Pioneer Hi-Bred International, Inc., 7100 NW 62ndAvenue, Johnston, IA 50131Product:4114 MaizeIngredientBacillus thuringiensisCry1F and Cry34/35Ab1 insecticidal proteins and the genetic material (PHP27118 T-DNA) necessary for their production in cornGuideline Reference Number Guideline Study Name MRID Number Submitter Status NoteMonitoring the susceptibility of corn lepidopteranpests to Cry1Ab and Cry1F proteins: 2007monitoring results 47413901 80778 PERProposed revisions to IRM-related registrationrequirements for Cry1 plant-incorporated protectantsin field corn 47543901 80778 PERMonitoring the susceptibility of corn lepidopteranpests to Cry1Ab and Cry1F proteins: 2008monitoring results 47841801 80778 PERMonitoring the susceptibility of corn lepidopteranpests to Cry1Ab and Cry1F proteins: 2009monitoring results 47971001 80778 PERProduct characterization data forBacillusthuringiensisPS149B1 13.6 kDa and 43.8 kDainsecticidal crystal proteins expressed in transgenicmaize plants 45242201 68467 PEREquivalency of microbial and maize-expressedPS149B1 proteins 45242203 68467 PERSignatureName and TitleJamie StaleyU.S. Registration ManagerDateApril 15, 2011EPA Form 8570-35 (9-97) Electronic and Paper versions available. Submit only Paper version.Agency Internal Use CopyPioneer Hi-Bred International, Inc.Study Number: PHI-2011-080Page 26 of 100Form Approved OMB No. 2070-0060UNITED STATES ENVIRONMENTAL PROTECTION AGENCY1200 Pennsylvania Avenue, N.W.Washington, D.C. 20460Paperwork Reduction Act Notice:The public reporting burden for this collection of information is estimated to average 0.25 hours per response for registrationactivities and 0.25 hours perresponse for reregistration and special review activities, including time for reading the instructions and completing the necessary forms. Send comments regarding the burden estimate or anyother aspect of this collection of information, including suggestions for reducing the burden to: Director, OPPE Information Management Division (2137), U.S. Environmental Protection Agency,1200 Pennsylvania Avenue, N.W., Washington, DC 20460. Do not send the form to this address.DATA MATRIXDate:April 15, 2011EPA Reg. No. / File Symbol:29964-xPage13of29Applicant's/Registrant's Name & Address:Pioneer Hi-Bred International, Inc., 7100 NW 62nd Avenue, Johnston, IA 50131Product:4114 MaizeIngredientBacillus thuringiensisCry1F and Cry34/35Ab1 insecticidal proteins and the genetic material (PHP27118 T-DNA) necessary for their production in cornGuideline Reference Number Guideline Study Name MRID Number Submitter Status NoteMicrobial PS149B1 binary delta endotoxin: Maizeinsect-pest susceptibility study 45242204 68467 PERComparison of the amino acid sequence of theBacillus thuringiensisstrain PS149B1 13.6 kDa and43.8 kDa insecticidal crystal proteins to KnPERprotein allergens 45242205 68467 PERCharacterization ofPseudomonasproduced andtransgenic maize expressed phosphinothricinacetyltransferase (PAT) protein 45242206 68467 PERPS149B1 14 kDa protein: Acute oral toxicity study inCD-1 mice 45242207 68467 PERPS149B1 44 kDa protein: Acute oral toxicity study inCD-1 mice 45242208 68467 PERPS149B1 14 kDa and 44 kDa proteins: Acute oraltoxicity study in CD-1 mice 45242209 68467 PERSignatureName and TitleJamie StaleyU.S. Registration ManagerDateApril 15, 2011EPA Form 8570-35 (9-97) Electronic and Paper versions available. Submit only Paper version.Agency Internal Use CopyPioneer Hi-Bred International, Inc.Study Number: PHI-2011-080Page 27 of 100Form Approved OMB No. 2070-0060UNITED STATES ENVIRONMENTAL PROTECTION AGENCY1200 Pennsylvania Avenue, N.W.Washington, D.C. 20460Paperwork Reduction Act Notice:The public reporting burden for this collection of information is estimated to average 0.25 hours per response for registrationactivities and 0.25 hours perresponse for reregistration and special review activities, including time for reading the instructions and completing the necessary forms. Send comments regarding the burden estimate or anyother aspect of this collection of information, including suggestions for reducing the burden to: Director, OPPE Information Management Division (2137), U.S. Environmental Protection Agency,1200 Pennsylvania Avenue, N.W., Washington, DC 20460. Do not send the form to this address.DATA MATRIXDate:April 15, 2011EPA Reg. No. / File Symbol:29964-xPage14of29Applicant's/Registrant's Name & Address:Pioneer Hi-Bred International, Inc., 7100 NW 62nd Avenue, Johnston, IA 50131Product:4114 MaizeIngredientBacillus thuringiensisCry1F and Cry34/35Ab1 insecticidal proteins and the genetic material (PHP27118 T-DNA) necessary for their production in cornGuideline Reference Number Guideline Study Name MRID Number Submitter Status NotePS149B1 binary insecticidal crystal protein: A dietarytoxicity study with ladybird beetle 45242210 68467 PERThe tri-trophic interaction between PS149B1transformed maize, corn leaf aphid and ladybirdbeetle 45242211 68467 PERIn vitro digestibility of PS149B1 proteins 45242212 68467 PERQuantitative ELISA analysis of PS149B1 proteinexpression levels in hybrid and inbred lines of maizeevent TC5639 (interim report) 45242213 68467 PERDegradation of microbial binary PS149B1 deltaendotoxin in a representative soil from the midWestern USA maize growing region 45242214 68467 PERProduct durability plan for transgenic maizeexpressing insecticidal crystal protein fromBacillusthuringiensisstrain PS149B1 during theexperimental use period 45242215 68467 PERSignatureName and TitleJamie StaleyU.S. Registration ManagerDateApril 15, 2011EPA Form 8570-35 (9-97) Electronic and Paper versions available. Submit only Paper version.Agency Internal Use CopyPioneer Hi-Bred International, Inc.Study Number: PHI-2011-080Page 28 of 100Form Approved OMB No. 2070-0060UNITED STATES ENVIRONMENTAL PROTECTION AGENCY1200 Pennsylvania Avenue, N.W.Washington, D.C. 20460Paperwork Reduction Act Notice:The public reporting burden for this collection of information is estimated to average 0.25 hours per response for registrationactivities and 0.25 hours perresponse for reregistration and special review activities, including time for reading the instructions and completing the necessary forms. Send comments regarding the burden estimate or anyother aspect of this collection of information, including suggestions for reducing the burden to: Director, OPPE Information Management Division (2137), U.S. Environmental Protection Agency,1200 Pennsylvania Avenue, N.W., Washington, DC 20460. Do not send the form to this address.DATA MATRIXDate:April 15, 2011EPA Reg. No. / File Symbol:29964-xPage15of29Applicant's/Registrant's Name & Address:Pioneer Hi-Bred International, Inc., 7100 NW 62nd Avenue, Johnston, IA 50131Product:4114 MaizeIngredientBacillus thuringiensisCry1F and Cry34/35Ab1 insecticidal proteins and the genetic material (PHP27118 T-DNA) necessary for their production in cornGuideline Reference Number Guideline Study Name MRID Number Submitter Status NoteField efficacy of PS149B1 maize events against cornrootworms 45242216 68467 PERMicrobial PS149B1 binary insecticidal crystalproteins, pollen expressing PS149B1 binaryinsecticidal crystal proteins, and individual PS149B114 kDa and 44 kDa insecticidal crystal proteins onhoneybee development 45340701 68467 PERThermolability of PS149B1 binary delta endotoxin 45358401 68467 PERPS149B1binary insecticidal crystal protein: Acutetoxicity study to the earthworm in an artificialsubstrate 45360201 68464 PERLateral flow test kit method validation for thedetection of the PS149B1 14 kDa and 44 kDa proteinin maize grain 45383401 68467 PERHeat lability of individual proteins of the PS149B1binary ICP 45584501 68467 PERSignatureName and TitleJamie StaleyU.S. Registration ManagerDateApril 15, 2011EPA Form 8570-35 (9-97) Electronic and Paper versions available. Submit only Paper version.Agency Internal Use CopyPioneer Hi-Bred International, Inc.Study Number: PHI-2011-080Page 29 of 100Form Approved OMB No. 2070-0060UNITED STATES ENVIRONMENTAL PROTECTION AGENCY1200 Pennsylvania Avenue, N.W.Washington, D.C. 20460Paperwork Reduction Act Notice:The public reporting burden for this collection of information is estimated to average 0.25 hours per response for registrationactivities and 0.25 hours perresponse for reregistration and special review activities, including time for reading the instructions and completing the necessary forms. Send comments regarding the burden estimate or anyother aspect of this collection of information, including suggestions for reducing the burden to: Director, OPPE Information Management Division (2137), U.S. Environmental Protection Agency,1200 Pennsylvania Avenue, N.W., Washington, DC 20460. Do not send the form to this address.DATA MATRIXDate:April 15, 2011EPA Reg. No. / File Symbol:29964-xPage16of29Applicant's/Registrant's Name & Address:Pioneer Hi-Bred International, Inc., 7100 NW 62nd Avenue, Johnston, IA 50131Product:4114 MaizeIngredientBacillus thuringiensisCry1F and Cry34/35Ab1 insecticidal proteins and the (PHP27118 T-DNA) genetic material necessary for their production in cornGuideline Reference Number Guideline Study Name MRID Number Submitter Status NoteIn vitro simulated gastric fluid digestibility study ofmicrobially derived Cry34Ab1 protein 45584502 68467 PERCharacterization of Cry34Ab1 and Cry35Ab1 fromrecombinantPseudomonas fluorescensandtransgenic maize 45790401 68467 PERCharacterization of DNA inserted into transgeniccorn events (Cry34Ab1 and Cry35Ab1) 45790402 68467 PERPS149B1 binary insecticidal crystal protein: An 8-day dietary study with rainbow trout,OncorhynchusmykissWallbaum 45790403 68467 PERPS149B1 binary insecticidal crystal protein: Acutetoxicity study with Daphnid,Daphnia magnaStraus 45790404 68467 PERPS149B1 binary insecticidal crystal protein: Dietarytoxicity to parasitic hymenoptera (Nasoniavitripennis)45790405 68467 PERSignatureName and TitleJamie StaleyU.S. Registration ManagerDateApril 15, 2011EPA Form 8570-35 (9-97) Electronic and Paper versions available. Submit only Paper version.Agency Internal Use CopyPioneer Hi-Bred International, Inc.Study Number: PHI-2011-080Page 30 of 100Form Approved OMB No. 2070-0060UNITED STATES ENVIRONMENTAL PROTECTION AGENCY1200 Pennsylvania Avenue, N.W.Washington, D.C. 20460Paperwork Reduction Act Notice:The public reporting burden for this collection of information is estimated to average 0.25 hours per response for registrationactivities and 0.25 hours perresponse for reregistration and special review activities, including time for reading the instructions and completing the necessary forms. Send comments regarding the burden estimate or anyother aspect of this collection of information, including suggestions for reducing the burden to: Director, OPPE Information Management Division (2137), U.S. Environmental Protection Agency,1200 Pennsylvania Avenue, N.W., Washington, DC 20460. Do not send the form to this address.DATA MATRIXDate:April 15, 2011EPA Reg. No. / File Symbol:29964-xPage17of29Applicant's/Registrant's Name & Address:Pioneer Hi-Bred International, Inc., 7100 NW 62nd Avenue, Johnston, IA 50131Product:4114 MaizeIngredientBacillus thuringiensisCry1F and Cry34/35Ab1 insecticidal proteins and the genetic material (PHP27118 T-DNA) necessary for their production in cornGuideline Reference Number Guideline Study Name MRID Number Submitter Status NoteAssessment of the chronic toxicity of diet containingBacillus thuringiensisPS149B1 insecticidal crystalprotein to collembolan (Folsomia candida) 45790406 68467 PERPS149B1 binary insecticidal crystal protein: Dietarytoxicity to green lacewing (Chrysoperla carnea) 45790407 68467 PERSDS-PAGE sensitivity analysis for Cry35Ab1 insupport of the simulated gastric fluid digestibilitystudy MRID 45242212 45790408 68467 PERTrait durability and experimental use of transgenicmaize expressing the insecticidal crystalline proteinsCry34Ab1and Cry35Ab1 45790509 68467 PERField efficacy of Cry34Ab1/Cry35Ab1 maize eventsagainst corn rootworms 45790410 68467 PERProduct characterization data forBacillusthuringiensisCry34Ab1 and Cry35Ab1 proteinsexpressed in transgenic maize plants (PHP17658) 45790501 68467 PERSignatureName and TitleJamie StaleyU.S. Registration ManagerDateApril 15, 2011EPA Form 8570-35 (9-97) Electronic and Paper versions available. Submit only Paper version.Agency Internal Use CopyPioneer Hi-Bred International, Inc.Study Number: PHI-2011-080Page 31 of 100Form Approved OMB No. 2070-0060UNITED STATES ENVIRONMENTAL PROTECTION AGENCY1200 Pennsylvania Avenue, N.W.Washington, D.C. 20460Paperwork Reduction Act Notice:The public reporting burden for this collection of information is estimated to average 0.25 hours per response for registrationactivities and 0.25 hours perresponse for reregistration and special review activities, including time for reading the instructions and completing the necessary forms. Send comments regarding the burden estimate or anyother aspect of this collection of information, including suggestions for reducing the burden to: Director, OPPE Information Management Division (2137), U.S. Environmental Protection Agency,1200 Pennsylvania Avenue, N.W., Washington, DC 20460. Do not send the form to this address.DATA MATRIXDate:April 15, 2011EPA Reg. No. / File Symbol:29964-xPage18of29Applicant's/Registrant's Name & Address:Pioneer Hi-Bred International, Inc., 7100 NW 62nd Avenue, Johnston, IA 50131Product:4114 MaizeIngredientBacillus thuringiensisCry1F and Cry34/35Ab1 insecticidal proteins and the genetic material (PHP27118 T-DNA) necessary for their production in cornGuideline Reference Number Guideline Study Name MRID Number Submitter Status NoteProduct characterization data forBacillusthuringiensisCry34Ab1 and Cry35Ab1 proteinsexpressed in transgenic maize plants (PHP17662) 45790601 68467 PERSummary heat lability studies with Cry34Ab1/Cry35Ab1 45808601 68467 PERQuantitative ELISA analysis of Cry34Ab1 andCry35Ab1 proteins expressed in maize plantstransformed with the vector PHP17658 45833101 68467 PERQuantitative ELISA analysis of Cry34Ab1 andCry35Ab1 proteins expressed in maize plantstransformed with the vector PHP17662 45833102 68467 PERSlide presentation summarizing Cry34Ab1/Cry35Ab1 heat inactivation studies 45860201 68467 PERProbe MOA studies to assess potential for proteinsynthesis inhibition byBacillus thuringiensisPS149B1 Cry34Ab1/Cry35Ab1 proteins in rabbitreticulocyte assay: Re-examination of lab notebookdata 45942801 68467 PERSignatureName and TitleJamie StaleyU.S. Registration ManagerDateApril 15, 2011EPA Form 8570-35 (9-97) Electronic and Paper versions available. Submit only Paper version.Agency Internal Use CopyPioneer Hi-Bred International, Inc.Study Number: PHI-2011-080Page 32 of 100Form Approved OMB No. 2070-0060UNITED STATES ENVIRONMENTAL PROTECTION AGENCY1200 Pennsylvania Avenue, N.W.Washington, D.C. 20460Paperwork Reduction Act Notice:The public reporting burden for this collection of information is estimated to average 0.25 hours per response for registrationactivities and 0.25 hours perresponse for reregistration and special review activities, including time for reading the instructions and completing the necessary forms. Send comments regarding the burden estimate or anyother aspect of this collection of information, including suggestions for reducing the burden to: Director, OPPE Information Management Division (2137), U.S. Environmental Protection Agency,1200 Pennsylvania Avenue, N.W., Washington, DC 20460. Do not send the form to this address.DATA MATRIXDate:April 15, 2011EPA Reg. No. / File Symbol:29964-xPage19of29Applicant's/Registrant's Name & Address:Pioneer Hi-Bred International, Inc., 7100 NW 62nd Avenue, Johnston, IA 50131Product:4114 MaizeIngredientBacillus thuringiensisCry1F and Cry34/35Ab1 insecticidal proteins and the genetic material (PHP27118 T-DNA) necessary for their production in cornGuideline Reference Number Guideline Study Name MRID Number Submitter Status NoteProduct characterization data forBacillusthuringiensisCry34Ab1 and Cry35Ab1 proteinsexpressed in transgenic maize plants (PHP17662) 46030001 68467 PERPublic interest document for Cry34/35Ab1 cornrootworm-protected corn 46122701 29964 OWNIndependent laboratory validation Pioneer Hi-BredInternational, Inc. ELISA method for thequantification of Cry34Ab1 protein from transgenicplants 46123901 68467 PERIndependent laboratory validation of DowAgroSciences method GRM 03.13, “ Determinationof Cry35Ab1 insecticidal crystal protein in maizetissue by Enzyme-Linked Immunosorbent Assay” 46123902 68467 PERCry34/35 protein distribution and familiarity 46123903 68467 PERAgronomic characteristics, quantitative ELISA andnutrient composition analysis of hybrid maize linescontaining cry34Ab1, cry35Ab1 and pat genes: Chilelocations 46123904 68467 PERSignatureName and TitleJamie StaleyU.S. Registration ManagerDateApril 15, 2011EPA Form 8570-35 (9-97) Electronic and Paper versions available. Submit only Paper version.Agency Internal Use CopyPioneer Hi-Bred International, Inc.Study Number: PHI-2011-080Page 33 of 100Form Approved OMB No. 2070-0060UNITED STATES ENVIRONMENTAL PROTECTION AGENCY1200 Pennsylvania Avenue, N.W.Washington, D.C. 20460Paperwork Reduction Act Notice:The public reporting burden for this collection of information is estimated to average 0.25 hours per response for registrationactivities and 0.25 hours perresponse for reregistration and special review activities, including time for reading the instructions and completing the necessary forms. Send comments regarding the burden estimate or anyother aspect of this collection of information, including suggestions for reducing the burden to: Director, OPPE Information Management Division (2137), U.S. Environmental Protection Agency,1200 Pennsylvania Avenue, N.W., Washington, DC 20460. Do not send the form to this address.DATA MATRIXDate:April 15, 2011EPA Reg. No. / File Symbol:29964-xPage20of29Applicant's/Registrant's Name & Address:Pioneer Hi-Bred International, Inc., 7100 NW 62nd Avenue, Johnston, IA 50131Product:4114 MaizeIngredientBacillus thuringiensisCry1F and Cry34/35Ab1 insecticidal proteins and the genetic material (PHP27118 T-DNA) necessary for their production in cornGuideline Reference Number Guideline Study Name MRID Number Submitter Status NoteBiological equivalency of Cry34/35Ab1 insecticidalcrystal protein in transgenic plants derived fromtransgenicPseudomonas flourscens46123905 68467 PERCharacterization of Cry34Ab and Cry35Ab1 proteinsderived from transgenic maize event E4497.59.1.22(DAS-59122-7) 46123906 68467 PERCharacterization of phosphinothricinacetyltransferase (PAT) derived from transgenicmaize event E4497.59.1.22 46123907 68467 PERCharacterization of DNA inserted into transgeniccorn events DAS-45216-6 and DAS-59122-7 46123908 68467 PERDetailed characterization of DNA into transgeniccorn events DAS-45216-6 and DAS-59122-7 46123909 68467 PEREvaluation of microbe derived Cry34Ab1 andCry35Ab1 proteins for protein synthesis inhibitionactivity 46123910 68467 PERSignatureName and TitleJamie StaleyU.S. Registration ManagerDateApril 15, 2011EPA Form 8570-35 (9-97) Electronic and Paper versions available. Submit only Paper version.Agency Internal Use CopyPioneer Hi-Bred International, Inc.Study Number: PHI-2011-080Page 34 of 100Form Approved OMB No. 2070-0060UNITED STATES ENVIRONMENTAL PROTECTION AGENCY1200 Pennsylvania Avenue, N.W.Washington, D.C. 20460Paperwork Reduction Act Notice:The public reporting burden for this collection of information is estimated to average 0.25 hours per response for registrationactivities and 0.25 hours perresponse for reregistration and special review activities, including time for reading the instructions and completing the necessary forms. Send comments regarding the burden estimate or anyother aspect of this collection of information, including suggestions for reducing the burden to: Director, OPPE Information Management Division (2137), U.S. Environmental Protection Agency,1200 Pennsylvania Avenue, N.W., Washington, DC 20460. Do not send the form to this address.DATA MATRIXDate:April 15, 2011EPA Reg. No. / File Symbol:29964-xPage21of29Applicant's/Registrant's Name & Address:Pioneer Hi-Bred International, Inc., 7100 NW 62nd Avenue, Johnston, IA 50131Product:4114 MaizeIngredientBacillus thuringiensisCry1F and Cry34/35Ab1 insecticidal proteins and the genetic material (PHP27118 T-DNA) necessary for their production in cornGuideline Reference Number Guideline Study Name MRID Number Submitter Status NoteNutritional equivalency study of maize containingCry34Ab1 and Cry35Ab1: Poultry feeding study 46123911 68467 PERThe effect of Cry34Ab1/Cry35Ab1 proteins on thedevelopment and mortality of the ladybird beetleColeomegilla maculataDeGeer 46123912 68467 PERNon-target invertebrate ecological risk assessmentfor field corn expressing Cry34Ab1 and Cry35Ab1insecticidal crystal proteins in event DAS-59122-7 46123913 68467 PEREvaluation of the impact corn rootworm controlstrategies on non-target arthropods 46123914 68467 PERInvestigations into dose of Cry34Ab1/Cry35Ab1rootworm-resistant maize event DAS-59122-7against western and northern corn rootworms insupport of trait durability plans 46123915 68467 PEREffect on western corn rootworm adults feeding onCry34/35Ab1 corn rootworm-protected corn tissueand implication for product durability 46123916 68467 PERSignatureName and TitleJamie StaleyU.S. Registration ManagerDateApril 15, 2011EPA Form 8570-35 (9-97) Electronic and Paper versions available. Submit only Paper version.Agency Internal Use CopyPioneer Hi-Bred International, Inc.Study Number: PHI-2011-080Page 35 of 100Form Approved OMB No. 2070-0060UNITED STATES ENVIRONMENTAL PROTECTION AGENCY1200 Pennsylvania Avenue, N.W.Washington, D.C. 20460Paperwork Reduction Act Notice:The public reporting burden for this collection of information is estimated to average 0.25 hours per response for registrationactivities and 0.25 hours perresponse for reregistration and special review activities, including time for reading the instructions and completing the necessary forms. Send comments regarding the burden estimate or anyother aspect of this collection of information, including suggestions for reducing the burden to: Director, OPPE Information Management Division (2137), U.S. Environmental Protection Agency,1200 Pennsylvania Avenue, N.W., Washington, DC 20460. Do not send the form to this address.DATA MATRIXDate:April 15, 2011EPA Reg. No. / File Symbol:29964-xPage22of29Applicant's/Registrant's Name & Address:Pioneer Hi-Bred International, Inc., 7100 NW 62nd Avenue, Johnston, IA 50131Product:4114 MaizeIngredientBacillus thuringiensisCry1F and Cry34/35Ab1 insecticidal proteins and the genetic material (PHP27118 T-DNA) necessary for their production in cornGuideline Reference Number Guideline Study Name MRID Number Submitter Status NoteEvaluation of endangered/threatened insect speciesrelative to the use of Cry34/35Ab1 corn rootwormresistant maize hybrids 46123917 68467 PERTrait durability plan for Cry34/35 corn rootwormprotected corn event DAS-59122-7 followingcommercialization 46123918 68467 PERSimulations of corn rootworm adaptation to Cry34/35corn rootworm protected corn in support of traitdurability plans for event DAS-59122-7 46123919 68467 PERDigestion of allergenic and non-allergenic proteins insimulated gastric fluid 46123920 68467 PERDigestion efficiency of allergens and non-allergens insimulated gastric fluid:Bacillus thuringiensisCry34/35Ab1 construct PHP17662 46388601 68467 PERLack of Cry34Ab1/Cry35Ab1 co-association insolution 46556801 68467 PERSignatureName and TitleJamie StaleyU.S. Registration ManagerDateApril 15, 2011EPA Form 8570-35 (9-97) Electronic and Paper versions available. Submit only Paper version.Agency Internal Use CopyPioneer Hi-Bred International, Inc.Study Number: PHI-2011-080Page 36 of 100Form Approved OMB No. 2070-0060UNITED STATES ENVIRONMENTAL PROTECTION AGENCY1200 Pennsylvania Avenue, N.W.Washington, D.C. 20460Paperwork Reduction Act Notice:The public reporting burden for this collection of information is estimated to average 0.25 hours per response for registrationactivities and 0.25 hours perresponse for reregistration and special review activities, including time for reading the instructions and completing the necessary forms. Send comments regarding the burden estimate or anyother aspect of this collection of information, including suggestions for reducing the burden to: Director, OPPE Information Management Division (2137), U.S. Environmental Protection Agency,1200 Pennsylvania Avenue, N.W., Washington, DC 20460. Do not send the form to this address.DATA MATRIXDate:April 15, 2011EPA Reg. No. / File Symbol:29964-xPage23of29Applicant's/Registrant's Name & Address:Pioneer Hi-Bred International, Inc., 7100 NW 62nd Avenue, Johnston, IA 50131Product:4114 MaizeIngredientBacillus thuringiensisCry1F and Cry34/35Ab1 insecticidal proteins and the genetic material (PHP27118 T-DNA) necessary for their production in cornGuideline Reference Number Guideline Study Name MRID Number Submitter Status NoteSummary report of a carabid beetle laboratorytoxicity study using Cry34Ab1 and Cry35Ab1including copies of references 46714101 68467 PERDetailed resistance monitoring plan for Cry34/35Ab1corn event DAS-59122-7 47334201 68467 PEREvaluation of potential dietary effects ofCry34/35Ab1 protein on insidious flower bugs,Oriusinsidiosus(Hemiptera: Anthocoridae) 47436701 68467 PERMonitoring corn rootworm susceptibility toCry34/35Ab1 event DAS-59122-7: 2007 insectcollections 47522501 68467 PERMonitoring corn rootworm susceptibility toCry34/35Ab1 event DAS-59122-7: 2008 insectcollections 47900801 68467 PERSoil accumulation of Cry34Ab1 and Cry35Ab1proteins after three years of cropping with DAS59122-7 corn 47959501 68467 PERSignatureName and TitleJamie StaleyU.S. Registration ManagerDateApril 15, 2011EPA Form 8570-35 (9-97) Electronic and Paper versions available. Submit only Paper version.Agency Internal Use CopyPioneer Hi-Bred International, Inc.Study Number: PHI-2011-080Page 37 of 100Form Approved OMB No. 2070-0060UNITED STATES ENVIRONMENTAL PROTECTION AGENCY1200 Pennsylvania Avenue, N.W.Washington, D.C. 20460Paperwork Reduction Act Notice:The public reporting burden for this collection of information is estimated to average 0.25 hours per response for registrationactivities and 0.25 hours perresponse for reregistration and special review activities, including time for reading the instructions and completing the necessary forms. Send comments regarding the burden estimate or anyother aspect of this collection of information, including suggestions for reducing the burden to: Director, OPPE Information Management Division (2137), U.S. Environmental Protection Agency,1200 Pennsylvania Avenue, N.W., Washington, DC 20460. Do not send the form to this address.DATA MATRIXDate:April 15, 2011EPA Reg. No. / File Symbol:29964-xPage24of29Applicant's/Registrant's Name & Address:Pioneer Hi-Bred International, Inc., 7100 NW 62nd Avenue, Johnston, IA 50131Product:4114 MaizeIngredientBacillus thuringiensisCry1F and Cry34/35Ab1 insecticidal proteins and the (PHP27118 T-DNA) genetic material necessary for their production in cornGuideline Reference Number Guideline Study Name MRID Number Submitter Status NoteThree-year field monitoring of Cry34/35Ab1 andCry1FxCry34/35Ab1 maize hybrids for non-targetarthropod effects 47870301 68467 PERInvestigating performance of HerculexTM RW andHerculexTM XTRA under commercial use 48045501 68467 PERMonitoring corn rootworm susceptibility toCry34/35Ab1 event DAS-59122-7: 2009 growingseason 48279701 68467 PERSummary information in support of the Section 3registration of 1507x59122 corn 46297101 68467 PERCharacterization of a stacked hybrid of transgeniccorn lines DAS-01507-1 and DAS-59122-7 bycomparison to the individual lines 46297102 68467 PERAgronomic characteristics, quantitative ELISA andnutrient composition analysis of hybrid corn linescontaining cry1F, cry34Ab1, cry35Ab1 and patgenes: US and Canada locations 46297103 68467 PERSignatureName and TitleJamie StaleyU.S. Registration ManagerDateApril 15, 2011EPA Form 8570-35 (9-97) Electronic and Paper versions available. Submit only Paper version.Agency Internal Use CopyPioneer Hi-Bred International, Inc.Study Number: PHI-2011-080Page 38 of 100Form Approved OMB No. 2070-0060UNITED STATES ENVIRONMENTAL PROTECTION AGENCY1200 Pennsylvania Avenue, N.W.Washington, D.C. 20460Paperwork Reduction Act Notice:The public reporting burden for this collection of information is estimated to average 0.25 hours per response for registrationactivities and 0.25 hours perresponse for reregistration and special review activities, including time for reading the instructions and completing the necessary forms. Send comments regarding the burden estimate or anyother aspect of this collection of information, including suggestions for reducing the burden to: Director, OPPE Information Management Division (2137), U.S. Environmental Protection Agency,1200 Pennsylvania Avenue, N.W., Washington, DC 20460. Do not send the form to this address.DATA MATRIXDate:April 15, 2011EPA Reg. No. / File Symbol:29964-xPage25of29Applicant's/Registrant's Name & Address:Pioneer Hi-Bred International, Inc., 7100 NW 62ndAvenue, Johnston, IA 50131Product:4114 MaizeIngredientBacillus thuringiensisCry1F and Cry34/35Ab1 insecticidal proteins and the genetic material (PHP27118 T-DNA) necessary for their production in cornGuideline Reference Number Guideline Study Name MRID Number Submitter Status NoteAmended report: Agronomic characteristics,quantitative ELISA and nutrient composition analysisof hybrid corn lines containing cry1F, cry34Ab1,cry35Ab1 and pat genes: US and Canada locations 46343801 68467 PERNutritional equivalency study of stacked hybrid oftransgenic corn line 1507 (event DAS-01507-1) and59122 (event DAS-59122-7): Poultry feeding study 46343803 68467 PERComparison of insect efficacy and expression inCry34/35 corn events DAS-59122-7 and DAS59132-8 expressed as a single and a stack withDAS-01507-1 46343804 68467 PERField efficacy of Herculex XTRA (DAS-01507-1xDAS-59122-7) insect resistant corn 46343805 68467 PERInvestigation of potential interaction between Cry1Fand the binary Cry34Ab1/Cry35Ab1 proteins 46343806 68467 PERThe effect of corn pollen expressing the Cry1F andCry34Ab1/Cry35Ab1 proteins on the developmentand mortality of the ladybird beetleColeomegillamaculataDeGeer 46343807 68467 PERSignatureName and TitleJamie StaleyU.S. Registration ManagerDateApril 15, 2011EPA Form 8570-35 (9-97) Electronic and Paper versions available. Submit only Paper version.Agency Internal Use CopyPioneer Hi-Bred International, Inc.Study Number: PHI-2011-080Page 39 of 100Form Approved OMB No. 2070-0060UNITED STATES ENVIRONMENTAL PROTECTION AGENCY1200 Pennsylvania Avenue, N.W.Washington, D.C. 20460Paperwork Reduction Act Notice:The public reporting burden for this collection of information is estimated to average 0.25 hours per response for registrationactivities and 0.25 hours perresponse for reregistration and special review activities, including time for reading the instructions and completing the necessary forms. Send comments regarding the burden estimate or anyother aspect of this collection of information, including suggestions for reducing the burden to: Director, OPPE Information Management Division (2137), U.S. Environmental Protection Agency,1200 Pennsylvania Avenue, N.W., Washington, DC 20460. Do not send the form to this address.DATA MATRIXDate:April 15, 2011EPA Reg. No. / File Symbol:29964-xPage26of29Applicant's/Registrant's Name & Address:Pioneer Hi-Bred International, Inc., 7100 NW 62ndAvenue, Johnston, IA 50131Product:4114 MaizeIngredientBacillus thuringiensisCry1F and Cry34/35Ab1 insecticidal proteins and the genetic material (PHP27118 T-DNA) necessary for their production in cornGuideline Reference Number Guideline Study Name MRID Number Submitter Status NoteLaboratory bioassay assessment of corn pollenprotein (Cry34/35 and Cry1F, stacked transgenes),and their effect on the larvae of monarch butterfly 46343808 68467 PEREquimolar ratio of Cry1F (tr) + Cry35Ab1 +Cry34Ab1: Acute oral toxicity study in CD-1 mice 46384301 68467 PERComparison of the amino acid sequence identitybetween the Cry1F protein and known proteinallergens 47786401 29964 OWNComparison of the amino acid sequence identitybetween the Cry34/35 proteins and known proteinallergens 47786402 29964 OWNComparison of the amino acid sequence identitybetween the PAT protein and known proteinallergens 47786403 29964 OWNEvaluation of the amino acid sequence similarity ofthe Cry1F protein to the NCBI protein sequencedatasets 47786405 29964 OWNSignatureName and TitleJamie StaleyU.S. Registration ManagerDateApril 15, 2011EPA Form 8570-35 (9-97) Electronic and Paper versions available. Submit only Paper version.Agency Internal Use CopyPioneer Hi-Bred International, Inc.Study Number: PHI-2011-080Page 40 of 100Form Approved OMB No. 2070-0060UNITED STATES ENVIRONMENTAL PROTECTION AGENCY1200 Pennsylvania Avenue, N.W.Washington, D.C. 20460Paperwork Reduction Act Notice:The public reporting burden for this collection of information is estimated to average 0.25 hours per response for registrationactivities and 0.25 hours perresponse for reregistration and special review activities, including time for reading the instructions and completing the necessary forms. Send comments regarding the burden estimate or anyother aspect of this collection of information, including suggestions for reducing the burden to: Director, OPPE Information Management Division (2137), U.S. Environmental Protection Agency,1200 Pennsylvania Avenue, N.W., Washington, DC 20460. Do not send the form to this address.DATA MATRIXDate:April 15, 2011EPA Reg. No. / File Symbol:29964-xPage27of29Applicant's/Registrant's Name & Address:Pioneer Hi-Bred International, Inc., 7100 NW 62ndAvenue, Johnston, IA 50131Product:4114 MaizeIngredientBacillus thuringiensisCry1F and Cry34/35Ab1 insecticidal proteins and the genetic material (PHP27118 T-DNA) necessary for their production in cornGuideline Reference Number Guideline Study Name MRID Number Submitter Status NoteEvaluation of the amino acid sequence similarity ofthe PAT protein to the NCBI protein sequencedatasets 47786407 29964 OWNEvaluation of the amino acid sequence similarity ofthe Cry34/35Ab1 proteins to the NCBI proteinsequence datasets 47786408 29964 OWNMonitoring corn rootworm susceptibility to eventDAS-59122-7 using the sub-lethal seedling assay:2009 growing season 48309902 29964 OWNMonitoring the susceptibility of corn lepidopteranpests to Cry1Ab and Cry1F proteins: 2009monitoring results 47971001 80778 PERMonitoring corn rootworm susceptibility toCry34/35Ab1 event DAS-59122-7: 2009 growingseason 48279701 68467 PERCharacterization of DP-ØØ4114-3 maize: Insertionintegrity, stability, copy number and backboneanalysis Submitted 29964 OWNSignatureName and TitleJamie StaleyU.S. Registration ManagerDateApril 15, 2011EPA Form 8570-35 (9-97) Electronic and Paper versions available. Submit only Paper version.Agency Internal Use CopyPioneer Hi-Bred International, Inc.Study Number: PHI-2011-080Page 41 of 100Form Approved OMB No. 2070-0060UNITED STATES ENVIRONMENTAL PROTECTION AGENCY1200 Pennsylvania Avenue, N.W.Washington, D.C. 20460Paperwork Reduction Act Notice:The public reporting burden for this collection of information is estimated to average 0.25 hours per response for registrationactivities and 0.25 hours perresponse for reregistration and special review activities, including time for reading the instructions and completing the necessary forms. Send comments regarding the burden estimate or anyother aspect of this collection of information, including suggestions for reducing the burden to: Director, OPPE Information Management Division (2137), U.S. Environmental Protection Agency,1200 Pennsylvania Avenue, N.W., Washington, DC 20460. Do not send the form to this address.DATA MATRIXDate:April 15, 2011EPA Reg. No. / File Symbol:29964-xPage28of29Applicant's/Registrant's Name & Address:Pioneer Hi-Bred International, Inc., 7100 NW 62ndAvenue, Johnston, IA 50131Product:4114 MaizeIngredientBacillus thuringiensisCry1F and Cry34/35Ab1 insecticidal proteins and the genetic material (PHP27118 T-DNA) necessary for their production in cornGuideline Reference Number Guideline Study Name MRID Number Submitter Status NoteEquivalency of expressed proteins in 4114 maize topreviously registered events Submitted 29964 OWNField and greenhouse insect efficacy for 4114 maize Submitted 29964 OWNEnvironmental Risk Assessment for the cultivation ofmaize containing event DP-ØØ4114-3 (4114 maize) Submitted 29964 OWNTrait Durability Plan supporting a Seed IncreaseRegistration for 4114 maize Submitted 29964 OWNWestern blot analysis of maize lines containingevent DP-ØØ4114-3 and combined trait productDAS-Ø15Ø7-1xDAS-59122-7 Submitted 29964 OWNExpressed Trait Protein Concentration of a MaizeLine Containing Events DP-ØØ4114-3,DAS-Ø15Ø7-1, DAS-59122-7, and Combined TraitProduct DAS-Ø1507-1xDAS-59122-7: US andCanada Test Sites Submitted 29964 OWNSignatureName and TitleJamie StaleyU.S. Registration ManagerDateApril 15, 2011EPA Form 8570-35 (9-97) Electronic and Paper versions available. Submit only Paper version.Agency Internal Use CopyPioneer Hi-Bred International, Inc.Study Number: PHI-2011-080Page 42 of 100Form Approved OMB No. 2070-0060UNITED STATES ENVIRONMENTAL PROTECTION AGENCY1200 Pennsylvania Avenue, N.W.Washington, D.C. 20460Paperwork Reduction Act Notice:The public reporting burden for this collection of information is estimated to average 0.25 hours per response for registrationactivities and 0.25 hours perresponse for reregistration and special review activities, including time for reading the instructions and completing the necessary forms. Send comments regarding the burden estimate or anyother aspect of this collection of information, including suggestions for reducing the burden to: Director, OPPE Information Management Division (2137), U.S. Environmental Protection Agency,1200 Pennsylvania Avenue, N.W., Washington, DC 20460. Do not send the form to this address.DATA MATRIXDate:April 15, 2011EPA Reg. No. / File Symbol:29964-xPage29of29Applicant's/Registrant's Name & Address:Pioneer Hi-Bred International, Inc., 7100 NW 62ndAvenue, Johnston, IA 50131Product:4114 MaizeIngredientBacillus thuringiensisCry1F and Cry34/35Ab1 insecticidal proteins and the genetic material (PHP27118 T-DNA) necessary for their production in cornGuideline Reference Number Guideline Study Name MRID Number Submitter Status NoteLiterature references supporting a Seed IncreaseRegistration for 4114 maize Submitted 29964 OWNSignatureName and TitleJamie StaleyU.S. Registration ManagerDateApril 15, 2011EPA Form 8570-35 (9-97) Electronic and Paper versions available. Submit only Paper version.Agency Internal Use CopyPioneer Hi-Bred International, Inc.Study Number: PHI-2011-080Page 43 of 100Form Approved OMB No. 2070-0060UNITED STATES ENVIRONMENTAL PROTECTION AGENCY1200 Pennsylvania Avenue, N.W.Washington, D.C. 20460Paperwork Reduction Act Notice:The public reporting burden for this collection of information is estimated to average 0.25 hours per response for registrationactivities and 0.25 hours perresponse for reregistration and special review activities, including time for reading the instructions and completing the necessary forms. Send comments regarding the burden estimate or anyother aspect of this collection of information, including suggestions for reducing the burden to: Director, OPPE Information Management Division (2137), U.S. Environmental Protection Agency,1200 Pennsylvania Avenue, N.W., Washington, DC 20460. Do not send the form to this address.DATA MATRIXDate:April 15, 2011EPA Reg. No. / File Symbol:29964-xPage1of29Applicant's/Registrant's Name & Address:Pioneer Hi-Bred International, Inc., 7100 NW 62nd Avenue, Johnston, IA 50131Product:4114 MaizeIngredientBacillus thuringiensisCry1F and Cry34/35Ab1 insecticidal proteins and the genetic material (PHP27118 T-DNA) necessary for their production in cornGuideline Reference Number Guideline Study Name MRID Number Submitter Status Note68467 PER68467 PER68467 PER68467 PER68467 PER68467 PERSignatureName and TitleJamie StaleyU.S. Registration ManagerDateApril 15, 2011EPA Form 8570-35 (9-97) Electronic and Paper versions available. Submit only Paper version.Public File CopyPioneer Hi-Bred International, Inc.Study Number: PHI-2011-080Page 44 of 100Form Approved OMB No. 2070-0060UNITED STATES ENVIRONMENTAL PROTECTION AGENCY1200 Pennsylvania Avenue, N.W.Washington, D.C. 20460Paperwork Reduction Act Notice:The public reporting burden for this collection of information is estimated to average 0.25 hours per response for registrationactivities and 0.25 hours perresponse for reregistration and special review activities, including time for reading the instructions and completing the necessary forms. Send comments regarding the burden estimate or anyother aspect of this collection of information, including suggestions for reducing the burden to: Director, OPPE Information Management Division (2137), U.S. Environmental Protection Agency,1200 Pennsylvania Avenue, N.W., Washington, DC 20460. Do not send the form to this address.DATA MATRIXDate:April 15, 2011EPA Reg. No. / File Symbol:29964-xPage2of29Applicant's/Registrant's Name & Address:Pioneer Hi-Bred International, Inc., 7100 NW 62nd Avenue, Johnston, IA 50131Product:4114 MaizeIngredientBacillus thuringiensisCry1F and Cry34/35Ab1 insecticidal proteins and the genetic material (PHP27118 T-DNA) necessary for their production in cornGuideline Reference Number Guideline Study Name MRID Number Submitter Status Note68467 PER68467 PER68467 PER68467 PER68467 PER68467 PERSignatureName and TitleJamie StaleyU.S. Registration ManagerDateApril 15, 2011EPA Form 8570-35 (9-97) Electronic and Paper versions available. Submit only Paper versionPublic File CopyPioneer Hi-Bred International, Inc.Study Number: PHI-2011-080Page 45 of 100Form Approved OMB No. 2070-0060UNITED STATES ENVIRONMENTAL PROTECTION AGENCY1200 Pennsylvania Avenue, N.W.Washington, D.C. 20460Paperwork Reduction Act Notice:The public reporting burden for this collection of information is estimated to average 0.25 hours per response for registrationactivities and 0.25 hours perresponse for reregistration and special review activities, including time for reading the instructions and completing the necessary forms. Send comments regarding the burden estimate or anyother aspect of this collection of information, including suggestions for reducing the burden to: Director, OPPE Information Management Division (2137), U.S. Environmental Protection Agency,1200 Pennsylvania Avenue, N.W., Washington, DC 20460. Do not send the form to this address.DATA MATRIXDate:April 15, 2011EPA Reg. No. / File Symbol:29964-xPage3of29Applicant's/Registrant's Name & Address:Pioneer Hi-Bred International, Inc., 7100 NW 62nd Avenue, Johnston, IA 50131Product:4114 MaizeIngredientBacillus thuringiensisCry1F and Cry34/35Ab1 insecticidal proteins and the genetic material (PHP27118 T-DNA) necessary for their production in cornGuideline Reference Number Guideline Study Name MRID Number Submitter Status Note68467 PER68467 PER68467 PER68467 PER68467 PER68467 PERSignatureName and TitleJamie StaleyU.S. Registration ManagerDateApril 15, 2011EPA Form 8570-35 (9-97) Electronic and Paper versions available. Submit only Paper versionPublic File CopyPioneer Hi-Bred International, Inc.Study Number: PHI-2011-080Page 46 of 100Form Approved OMB No. 2070-0060UNITED STATES ENVIRONMENTAL PROTECTION AGENCY1200 Pennsylvania Avenue, N.W.Washington, D.C. 20460Paperwork Reduction Act Notice:The public reporting burden for this collection of information is estimated to average 0.25 hours per response for registrationactivities and 0.25 hours perresponse for reregistration and special review activities, including time for reading the instructions and completing the necessary forms. Send comments regarding the burden estimate or anyother aspect of this collection of information, including suggestions for reducing the burden to: Director, OPPE Information Management Division (2137), U.S. Environmental Protection Agency,1200 Pennsylvania Avenue, N.W., Washington, DC 20460. Do not send the form to this address.DATA MATRIXDate:April 15, 2011EPA Reg. No. / File Symbol:29964-xPage4of29Applicant's/Registrant's Name & Address:Pioneer Hi-Bred International, Inc., 7100 NW 62nd Avenue, Johnston, IA 50131Product:4114 MaizeIngredientBacillus thuringiensisCry1F and Cry34/35Ab1 insecticidal proteins and the genetic material (PHP27118 T-DNA) necessary for their production in cornGuideline Reference Number Guideline Study Name MRID Number Submitter Status Note68467 PER68467 PER68467 PER68467 PER68467 PER68467 PERSignatureName and TitleJamie StaleyU.S. Registration ManagerDateApril 15, 2011EPA Form 8570-35 (9-97) Electronic and Paper versions available. Submit only Paper versionPublic File CopyPioneer Hi-Bred International, Inc.Study Number: PHI-2011-080Page 47 of 100Form Approved OMB No. 2070-0060UNITED STATES ENVIRONMENTAL PROTECTION AGENCY1200 Pennsylvania Avenue, N.W.Washington, D.C. 20460Paperwork Reduction Act Notice:The public reporting burden for this collection of information is estimated to average 0.25 hours per response for registrationactivities and 0.25 hours perresponse for reregistration and special review activities, including time for reading the instructions and completing the necessary forms. Send comments regarding the burden estimate or anyother aspect of this collection of information, including suggestions for reducing the burden to: Director, OPPE Information Management Division (2137), U.S. Environmental Protection Agency,1200 Pennsylvania Avenue, N.W., Washington, DC 20460. Do not send the form to this address.DATA MATRIXDate:April 15, 2011EPA Reg. No. / File Symbol:29964-xPage5of29Applicant's/Registrant's Name & Address:Pioneer Hi-Bred International, Inc., 7100 NW 62nd Avenue, Johnston, IA 50131Product:4114 MaizeIngredientBacillus thuringiensisCry1F and Cry34/35Ab1 insecticidal proteins and the genetic material (PHP27118 T-DNA) necessary for their production in cornGuideline Reference Number Guideline Study Name MRID Number Submitter Status Note68467 PER68467 PER68467 PER68467 PER68467 PER68467 PERSignatureName and TitleJamie StaleyU.S. Registration ManagerDateApril 15, 2011EPA Form 8570-35 (9-97) Electronic and Paper versions available. Submit only Paper versionPublic File CopyPioneer Hi-Bred International, Inc.Study Number: PHI-2011-080Page 48 of 100Form Approved OMB No. 2070-0060UNITED STATES ENVIRONMENTAL PROTECTION AGENCY1200 Pennsylvania Avenue, N.W.Washington, D.C. 20460Paperwork Reduction Act Notice:The public reporting burden for this collection of information is estimated to average 0.25 hours per response for registrationactivities and 0.25 hours perresponse for reregistration and special review activities, including time for reading the instructions and completing the necessary forms. Send comments regarding the burden estimate or anyother aspect of this collection of information, including suggestions for reducing the burden to: Director, OPPE Information Management Division (2137), U.S. Environmental Protection Agency,1200 Pennsylvania Avenue, N.W., Washington, DC 20460. Do not send the form to this address.DATA MATRIXDate:April 15, 2011EPA Reg. No. / File Symbol:29964-xPage6of29Applicant's/Registrant's Name & Address:Pioneer Hi-Bred International, Inc., 7100 NW 62nd Avenue, Johnston, IA 50131Product:4114 MaizeIngredientBacillus thuringiensisCry1F and Cry34/35Ab1 insecticidal proteins and the genetic material (PHP27118 T-DNA) necessary for their production in cornGuideline Reference Number Guideline Study Name MRID Number Submitter Status Note68467 PER68467 PER29964 OWN68467 PER68467 PER68467 PER68467 PERSignatureName and TitleJamie StaleyU.S. Registration ManagerDateApril 15, 2011EPA Form 8570-35 (9-97) Electronic and Paper versions available. Submit only Paper versionPublic File CopyPioneer Hi-Bred International, Inc.Study Number: PHI-2011-080Page 49 of 100Form Approved OMB No. 2070-0060UNITED STATES ENVIRONMENTAL PROTECTION AGENCY1200 Pennsylvania Avenue, N.W.Washington, D.C. 20460Paperwork Reduction Act Notice:The public reporting burden for this collection of information is estimated to average 0.25 hours per response for registrationactivities and 0.25 hours perresponse for reregistration and special review activities, including time for reading the instructions and completing the necessary forms. Send comments regarding the burden estimate or anyother aspect of this collection of information, including suggestions for reducing the burden to: Director, OPPE Information Management Division (2137), U.S. Environmental Protection Agency,1200 Pennsylvania Avenue, N.W., Washington, DC 20460. Do not send the form to this address.DATA MATRIXDate:April 15, 2011EPA Reg. No. / File Symbol:29964-xPage7of29Applicant's/Registrant's Name & Address:Pioneer Hi-Bred International, Inc., 7100 NW 62nd Avenue, Johnston, IA 50131Product:4114 MaizeIngredientBacillus thuringiensisCry1F and Cry34/35Ab1 insecticidal proteins and the genetic material (PHP27118 T-DNA) necessary for their production in cornGuideline Reference Number Guideline Study Name MRID Number Submitter Status Note29964 OWN68467 PER68467 PER68467 PER68467 PER68467 PERSignatureName and TitleJamie StaleyU.S. Registration ManagerDateApril 15, 2011EPA Form 8570-35 (9-97) Electronic and Paper versions available. Submit only Paper versionPublic File CopyPioneer Hi-Bred International, Inc.Study Number: PHI-2011-080Page 50 of 100Form Approved OMB No. 2070-0060UNITED STATES ENVIRONMENTAL PROTECTION AGENCY1200 Pennsylvania Avenue, N.W.Washington, D.C. 20460Paperwork Reduction Act Notice:The public reporting burden for this collection of information is estimated to average 0.25 hours per response for registrationactivities and 0.25 hours perresponse for reregistration and special review activities, including time for reading the instructions and completing the necessary forms. Send comments regarding the burden estimate or anyother aspect of this collection of information, including suggestions for reducing the burden to: Director, OPPE Information Management Division (2137), U.S. Environmental Protection Agency,1200 Pennsylvania Avenue, N.W., Washington, DC 20460. Do not send the form to this address.DATA MATRIXDate:April 15, 2011EPA Reg. No. / File Symbol:29964-xPage8of29Applicant's/Registrant's Name & Address:Pioneer Hi-Bred International, Inc., 7100 NW 62nd Avenue, Johnston, IA 50131Product:4114 MaizeIngredientBacillus thuringiensisCry1F and Cry34/35Ab1 insecticidal proteins and the genetic material (PHP27118 T-DNA) necessary for their production in cornGuideline Reference Number Guideline Study Name MRID Number Submitter Status Note68467 PER68467 PER29964 OWN68467 PER68467 PER68467 PERSignatureName and TitleJamie StaleyU.S. Registration ManagerDateApril 15, 2011EPA Form 8570-35 (9-97) Electronic and Paper versions available. Submit only Paper versionPublic File CopyPioneer Hi-Bred International, Inc.Study Number: PHI-2011-080Page 51 of 100Form Approved OMB No. 2070-0060UNITED STATES ENVIRONMENTAL PROTECTION AGENCY1200 Pennsylvania Avenue, N.W.Washington, D.C. 20460Paperwork Reduction Act Notice:The public reporting burden for this collection of information is estimated to average 0.25 hours per response for registrationactivities and 0.25 hours perresponse for reregistration and special review activities, including time for reading the instructions and completing the necessary forms. Send comments regarding the burden estimate or anyother aspect of this collection of information, including suggestions for reducing the burden to: Director, OPPE Information Management Division (2137), U.S. Environmental Protection Agency,1200 Pennsylvania Avenue, N.W., Washington, DC 20460. Do not send the form to this address.DATA MATRIXDate:April 15, 2011EPA Reg. No. / File Symbol:29964-xPage9of29Applicant's/Registrant's Name & Address:Pioneer Hi-Bred International, Inc., 7100 NW 62nd Avenue, Johnston, IA 50131Product:4114 MaizeIngredientBacillus thuringiensisCry1F and Cry34/35Ab1 insecticidal proteins and the genetic material (PHP27118 T-DNA) necessary for their production in cornGuideline Reference Number Guideline Study Name MRID Number Submitter Status Note68467 PER68467 PER68467 PER68467 PER29964 OWN68467 PERSignatureName and TitleJamie StaleyU.S. Registration ManagerDateApril 15, 2011EPA Form 8570-35 (9-97) Electronic and Paper versions available. Submit only Paper versionPublic File CopyPioneer Hi-Bred International, Inc.Study Number: PHI-2011-080Page 52 of 100Form Approved OMB No. 2070-0060UNITED STATES ENVIRONMENTAL PROTECTION AGENCY1200 Pennsylvania Avenue, N.W.Washington, D.C. 20460Paperwork Reduction Act Notice:The public reporting burden for this collection of information is estimated to average 0.25 hours per response for registrationactivities and 0.25 hours perresponse for reregistration and special review activities, including time for reading the instructions and completing the necessary forms. Send comments regarding the burden estimate or anyother aspect of this collection of information, including suggestions for reducing the burden to: Director, OPPE Information Management Division (2137), U.S. Environmental Protection Agency,1200 Pennsylvania Avenue, N.W., Washington, DC 20460. Do not send the form to this address.DATA MATRIXDate:April 15, 2011EPA Reg. No. / File Symbol:29964-xPage10of29Applicant's/Registrant's Name & Address:Pioneer Hi-Bred International, Inc., 7100 NW 62nd Avenue, Johnston, IA 50131Product:4114 MaizeIngredientBacillus thuringiensisCry1F and Cry34/35Ab1 insecticidal proteins and the genetic material (PHP27118 T-DNA) necessary for their production in cornGuideline Reference Number Guideline Study Name MRID Number Submitter Status Note80778 PER68467 PER68467 PER80778 PER80778 PER80778 PERSignatureName and TitleJamie StaleyU.S. Registration ManagerDateApril 15, 2011EPA Form 8570-35 (9-97) Electronic and Paper versions available. Submit only Paper versionPublic File CopyPioneer Hi-Bred International, Inc.Study Number: PHI-2011-080Page 53 of 100Form Approved OMB No. 2070-0060UNITED STATES ENVIRONMENTAL PROTECTION AGENCY1200 Pennsylvania Avenue, N.W.Washington, D.C. 20460Paperwork Reduction Act Notice:The public reporting burden for this collection of information is estimated to average 0.25 hours per response for registrationactivities and 0.25 hours perresponse for reregistration and special review activities, including time for reading the instructions and completing the necessary forms. Send comments regarding the burden estimate or anyother aspect of this collection of information, including suggestions for reducing the burden to: Director, OPPE Information Management Division (2137), U.S. Environmental Protection Agency,1200 Pennsylvania Avenue, N.W., Washington, DC 20460. Do not send the form to this address.DATA MATRIXDate:April 15, 2011EPA Reg. No. / File Symbol:29964-xPage11of29Applicant's/Registrant's Name & Address:Pioneer Hi-Bred International, Inc., 7100 NW 62nd Avenue, Johnston, IA 50131Product:4114 MaizeIngredientBacillus thuringiensisCry1F and Cry34/35Ab1 insecticidal proteins and the genetic material (PHP27118 T-DNA) necessary for their production in cornGuideline Reference Number Guideline Study Name MRID Number Submitter Status Note68467 PER80778 PER80778 PER68467 PER68467 PER80778 PERSignatureName and TitleJamie StaleyU.S. Registration ManagerDateApril 15, 2011EPA Form 8570-35 (9-97) Electronic and Paper versions available. Submit only Paper versionPublic File CopyPioneer Hi-Bred International, Inc.Study Number: PHI-2011-080Page 54 of 100Form Approved OMB No. 2070-0060UNITED STATES ENVIRONMENTAL PROTECTION AGENCY1200 Pennsylvania Avenue, N.W.Washington, D.C. 20460Paperwork Reduction Act Notice:The public reporting burden for this collection of information is estimated to average 0.25 hours per response for registrationactivities and 0.25 hours perresponse for reregistration and special review activities, including time for reading the instructions and completing the necessary forms. Send comments regarding the burden estimate or anyother aspect of this collection of information, including suggestions for reducing the burden to: Director, OPPE Information Management Division (2137), U.S. Environmental Protection Agency,1200 Pennsylvania Avenue, N.W., Washington, DC 20460. Do not send the form to this address.DATA MATRIXDate:April 15, 2011EPA Reg. No. / File Symbol:29964-xPage12of29Applicant's/Registrant's Name & Address:Pioneer Hi-Bred International, Inc., 7100 NW 62nd Avenue, Johnston, IA 50131Product:4114 MaizeIngredientBacillus thuringiensisCry1F and Cry34/35Ab1 insecticidal proteins and the genetic material (PHP27118 T-DNA) necessary for their production in cornGuideline Reference Number Guideline Study Name MRID Number Submitter Status Note80778 PER80778 PER80778 PER80778 PER68467 PER68467 PERSignatureName and TitleJamie StaleyU.S. Registration ManagerDateApril 15, 2011EPA Form 8570-35 (9-97) Electronic and Paper versions available. Submit only Paper versionPublic File CopyPioneer Hi-Bred International, Inc.Study Number: PHI-2011-080Page 55 of 100Form Approved OMB No. 2070-0060UNITED STATES ENVIRONMENTAL PROTECTION AGENCY1200 Pennsylvania Avenue, N.W.Washington, D.C. 20460Paperwork Reduction Act Notice:The public reporting burden for this collection of information is estimated to average 0.25 hours per response for registrationactivities and 0.25 hours perresponse for reregistration and special review activities, including time for reading the instructions and completing the necessary forms. Send comments regarding the burden estimate or anyother aspect of this collection of information, including suggestions for reducing the burden to: Director, OPPE Information Management Division (2137), U.S. Environmental Protection Agency,1200 Pennsylvania Avenue, N.W., Washington, DC 20460. Do not send the form to this address.DATA MATRIXDate:April 15, 2011EPA Reg. No. / File Symbol:29964-xPage13of29Applicant's/Registrant's Name & Address:Pioneer Hi-Bred International, Inc., 7100 NW 62nd Avenue, Johnston, IA 50131Product:4114 MaizeIngredientBacillus thuringiensisCry1F and Cry34/35Ab1 insecticidal proteins and the genetic material (PHP27118 T-DNA) necessary for their production in cornGuideline Reference Number Guideline Study Name MRID Number Submitter Status Note68467 PER68467 PER68467 PER68467 PER68467 PER68467 PERSignatureName and TitleJamie StaleyU.S. Registration ManagerDateApril 15, 2011EPA Form 8570-35 (9-97) Electronic and Paper versions available. Submit only Paper versionPublic File CopyPioneer Hi-Bred International, Inc.Study Number: PHI-2011-080Page 56 of 100Form Approved OMB No. 2070-0060UNITED STATES ENVIRONMENTAL PROTECTION AGENCY1200 Pennsylvania Avenue, N.W.Washington, D.C. 20460Paperwork Reduction Act Notice:The public reporting burden for this collection of information is estimated to average 0.25 hours per response for registrationactivities and 0.25 hours perresponse for reregistration and special review activities, including time for reading the instructions and completing the necessary forms. Send comments regarding the burden estimate or anyother aspect of this collection of information, including suggestions for reducing the burden to: Director, OPPE Information Management Division (2137), U.S. Environmental Protection Agency,1200 Pennsylvania Avenue, N.W., Washington, DC 20460. Do not send the form to this address.DATA MATRIXDate:April 15, 2011EPA Reg. No. / File Symbol:29964-xPage14of29Applicant's/Registrant's Name & Address:Pioneer Hi-Bred International, Inc., 7100 NW 62nd Avenue, Johnston, IA 50131Product:4114 MaizeIngredientBacillus thuringiensisCry1F and Cry34/35Ab1 insecticidal proteins and the genetic material (PHP27118 T-DNA) necessary for their production in cornGuideline Reference Number Guideline Study Name MRID Number Submitter Status Note68467 PER68467 PER68467 PER68467 PER68467 PER68467 PERSignatureName and TitleJamie StaleyU.S. Registration ManagerDateApril 15, 2011EPA Form 8570-35 (9-97) Electronic and Paper versions available. Submit only Paper versionPublic File CopyPioneer Hi-Bred International, Inc.Study Number: PHI-2011-080Page 57 of 100Form Approved OMB No. 2070-0060UNITED STATES ENVIRONMENTAL PROTECTION AGENCY1200 Pennsylvania Avenue, N.W.Washington, D.C. 20460Paperwork Reduction Act Notice:The public reporting burden for this collection of information is estimated to average 0.25 hours per response for registrationactivities and 0.25 hours perresponse for reregistration and special review activities, including time for reading the instructions and completing the necessary forms. Send comments regarding the burden estimate or anyother aspect of this collection of information, including suggestions for reducing the burden to: Director, OPPE Information Management Division (2137), U.S. Environmental Protection Agency,1200 Pennsylvania Avenue, N.W., Washington, DC 20460. Do not send the form to this address.DATA MATRIXDate:April 15, 2011EPA Reg. No. / File Symbol:29964-xPage15of29Applicant's/Registrant's Name & Address:Pioneer Hi-Bred International, Inc., 7100 NW 62nd Avenue, Johnston, IA 50131Product:4114 MaizeIngredientBacillus thuringiensisCry1F and Cry34/35Ab1 insecticidal proteins and the genetic material (PHP27118 T-DNA) necessary for their production in cornGuideline Reference Number Guideline Study Name MRID Number Submitter Status Note68467 PER68467 PER68467 PER68467 PER68467 PER68467 PERSignatureName and TitleJamie StaleyU.S. Registration ManagerDateApril 15, 2011EPA Form 8570-35 (9-97) Electronic and Paper versions available. Submit only Paper versionPublic File CopyPioneer Hi-Bred International, Inc.Study Number: PHI-2011-080Page 58 of 100Form Approved OMB No. 2070-0060UNITED STATES ENVIRONMENTAL PROTECTION AGENCY1200 Pennsylvania Avenue, N.W.Washington, D.C. 20460Paperwork Reduction Act Notice:The public reporting burden for this collection of information is estimated to average 0.25 hours per response for registrationactivities and 0.25 hours perresponse for reregistration and special review activities, including time for reading the instructions and completing the necessary forms. Send comments regarding the burden estimate or anyother aspect of this collection of information, including suggestions for reducing the burden to: Director, OPPE Information Management Division (2137), U.S. Environmental Protection Agency,1200 Pennsylvania Avenue, N.W., Washington, DC 20460. Do not send the form to this address.DATA MATRIXDate:April 15, 2011EPA Reg. No. / File Symbol:29964-xPage16of29Applicant's/Registrant's Name & Address:Pioneer Hi-Bred International, Inc., 7100 NW 62nd Avenue, Johnston, IA 50131Product:4114 MaizeIngredientBacillus thuringiensisCry1F and Cry34/35Ab1 insecticidal proteins and the genetic material (PHP27118 T-DNA) necessary for their production in cornGuideline Reference Number Guideline Study Name MRID Number Submitter Status Note68467 PER68467 PER68467 PER68467 PER68467 PER68467 PERSignatureName and TitleJamie StaleyU.S. Registration ManagerDateApril 15, 2011EPA Form 8570-35 (9-97) Electronic and Paper versions available. Submit only Paper versionPublic File CopyPioneer Hi-Bred International, Inc.Study Number: PHI-2011-080Page 59 of 100Form Approved OMB No. 2070-0060UNITED STATES ENVIRONMENTAL PROTECTION AGENCY1200 Pennsylvania Avenue, N.W.Washington, D.C. 20460Paperwork Reduction Act Notice:The public reporting burden for this collection of information is estimated to average 0.25 hours per response for registrationactivities and 0.25 hours perresponse for reregistration and special review activities, including time for reading the instructions and completing the necessary forms. Send comments regarding the burden estimate or anyother aspect of this collection of information, including suggestions for reducing the burden to: Director, OPPE Information Management Division (2137), U.S. Environmental Protection Agency,1200 Pennsylvania Avenue, N.W., Washington, DC 20460. Do not send the form to this address.DATA MATRIXDate:April 15, 2011EPA Reg. No. / File Symbol:29964-xPage17of29Applicant's/Registrant's Name & Address:Pioneer Hi-Bred International, Inc., 7100 NW 62nd Avenue, Johnston, IA 50131Product:4114 MaizeIngredientBacillus thuringiensisCry1F and Cry34/35Ab1 insecticidal proteins and the genetic material (PHP27118 T-DNA) necessary for their production in cornGuideline Reference Number Guideline Study Name MRID Number Submitter Status Note68467 PER68467 PER68467 PER68467 PER68467 PER68467 PERSignatureName and TitleJamie StaleyU.S. Registration ManagerDateApril 15, 2011EPA Form 8570-35 (9-97) Electronic and Paper versions available. Submit only Paper versionPublic File CopyPioneer Hi-Bred International, Inc.Study Number: PHI-2011-080Page 60 of 100Form Approved OMB No. 2070-0060UNITED STATES ENVIRONMENTAL PROTECTION AGENCY1200 Pennsylvania Avenue, N.W.Washington, D.C. 20460Paperwork Reduction Act Notice:The public reporting burden for this collection of information is estimated to average 0.25 hours per response for registrationactivities and 0.25 hours perresponse for reregistration and special review activities, including time for reading the instructions and completing the necessary forms. Send comments regarding the burden estimate or anyother aspect of this collection of information, including suggestions for reducing the burden to: Director, OPPE Information Management Division (2137), U.S. Environmental Protection Agency,1200 Pennsylvania Avenue, N.W., Washington, DC 20460. Do not send the form to this address.DATA MATRIXDate:April 15, 2011EPA Reg. No. / File Symbol:29964-xPage18of29Applicant's/Registrant's Name & Address:Pioneer Hi-Bred International, Inc., 7100 NW 62nd Avenue, Johnston, IA 50131Product:4114 MaizeIngredientBacillus thuringiensisCry1F and Cry34/35Ab1 insecticidal proteins and the genetic material (PHP27118 T-DNA) necessary for their production in cornGuideline Reference Number Guideline Study Name MRID Number Submitter Status Note68467 PER68467 PER68467 PER68467 PER68467 PER68467 PERSignatureName and TitleJamie StaleyU.S. Registration ManagerDateApril 15, 2011EPA Form 8570-35 (9-97) Electronic and Paper versions available. Submit only Paper versionPublic File CopyPioneer Hi-Bred International, Inc.Study Number: PHI-2011-080Page 61 of 100Form Approved OMB No. 2070-0060UNITED STATES ENVIRONMENTAL PROTECTION AGENCY1200 Pennsylvania Avenue, N.W.Washington, D.C. 20460Paperwork Reduction Act Notice:The public reporting burden for this collection of information is estimated to average 0.25 hours per response for registrationactivities and 0.25 hours perresponse for reregistration and special review activities, including time for reading the instructions and completing the necessary forms. Send comments regarding the burden estimate or anyother aspect of this collection of information, including suggestions for reducing the burden to: Director, OPPE Information Management Division (2137), U.S. Environmental Protection Agency,1200 Pennsylvania Avenue, N.W., Washington, DC 20460. Do not send the form to this address.DATA MATRIXDate:April 15, 2011EPA Reg. No. / File Symbol:29964-xPage19of29Applicant's/Registrant's Name & Address:Pioneer Hi-Bred International, Inc., 7100 NW 62nd Avenue, Johnston, IA 50131Product:4114 MaizeIngredientBacillus thuringiensisCry1F and Cry34/35Ab1 insecticidal proteins and the genetic material (PHP27118 T-DNA) necessary for their production in cornGuideline Reference Number Guideline Study Name MRID Number Submitter Status Note68467 PER29964 OWN68467 PER68467 PER68467 PER68467 PERSignatureName and TitleJamie StaleyU.S. Registration ManagerDateApril 15, 2011EPA Form 8570-35 (9-97) Electronic and Paper versions available. Submit only Paper versionPublic File CopyPioneer Hi-Bred International, Inc.Study Number: PHI-2011-080Page 62 of 100Form Approved OMB No. 2070-0060UNITED STATES ENVIRONMENTAL PROTECTION AGENCY1200 Pennsylvania Avenue, N.W.Washington, D.C. 20460Paperwork Reduction Act Notice:The public reporting burden for this collection of information is estimated to average 0.25 hours per response for registrationactivities and 0.25 hours perresponse for reregistration and special review activities, including time for reading the instructions and completing the necessary forms. Send comments regarding the burden estimate or anyother aspect of this collection of information, including suggestions for reducing the burden to: Director, OPPE Information Management Division (2137), U.S. Environmental Protection Agency,1200 Pennsylvania Avenue, N.W., Washington, DC 20460. Do not send the form to this address.DATA MATRIXDate:April 15, 2011EPA Reg. No. / File Symbol:29964-xPage20of29Applicant's/Registrant's Name & Address:Pioneer Hi-Bred International, Inc., 7100 NW 62nd Avenue, Johnston, IA 50131Product:4114 MaizeIngredientBacillus thuringiensisCry1F and Cry34/35Ab1 insecticidal proteins and the genetic material (PHP27118 T-DNA) necessary for their production in cornGuideline Reference Number Guideline Study Name MRID Number Submitter Status Note68467 PER68467 PER68467 PER68467 PER68467 PER68467 PERSignatureName and TitleJamie StaleyU.S. Registration ManagerDateApril 15, 2011EPA Form 8570-35 (9-97) Electronic and Paper versions available. Submit only Paper versionPublic File CopyPioneer Hi-Bred International, Inc.Study Number: PHI-2011-080Page 63 of 100Form Approved OMB No. 2070-0060UNITED STATES ENVIRONMENTAL PROTECTION AGENCY1200 Pennsylvania Avenue, N.W.Washington, D.C. 20460Paperwork Reduction Act Notice:The public reporting burden for this collection of information is estimated to average 0.25 hours per response for registrationactivities and 0.25 hours perresponse for reregistration and special review activities, including time for reading the instructions and completing the necessary forms. Send comments regarding the burden estimate or anyother aspect of this collection of information, including suggestions for reducing the burden to: Director, OPPE Information Management Division (2137), U.S. Environmental Protection Agency,1200 Pennsylvania Avenue, N.W., Washington, DC 20460. Do not send the form to this address.DATA MATRIXDate:April 15, 2011EPA Reg. No. / File Symbol:29964-xPage21of29Applicant's/Registrant's Name & Address:Pioneer Hi-Bred International, Inc., 7100 NW 62nd Avenue, Johnston, IA 50131Product:4114 MaizeIngredientBacillus thuringiensisCry1F and Cry34/35Ab1 insecticidal proteins and the genetic material (PHP27118 T-DNA) necessary for their production in cornGuideline Reference Number Guideline Study Name MRID Number Submitter Status Note68467 PER68467 PER68467 PER68467 PER68467 PER68467 PERSignatureName and TitleJamie StaleyU.S. Registration ManagerDateApril 15, 2011EPA Form 8570-35 (9-97) Electronic and Paper versions available. Submit only Paper versionPublic File CopyPioneer Hi-Bred International, Inc.Study Number: PHI-2011-080Page 64 of 100Form Approved OMB No. 2070-0060UNITED STATES ENVIRONMENTAL PROTECTION AGENCY1200 Pennsylvania Avenue, N.W.Washington, D.C. 20460Paperwork Reduction Act Notice:The public reporting burden for this collection of information is estimated to average 0.25 hours per response for registrationactivities and 0.25 hours perresponse for reregistration and special review activities, including time for reading the instructions and completing the necessary forms. Send comments regarding the burden estimate or anyother aspect of this collection of information, including suggestions for reducing the burden to: Director, OPPE Information Management Division (2137), U.S. Environmental Protection Agency,1200 Pennsylvania Avenue, N.W., Washington, DC 20460. Do not send the form to this address.DATA MATRIXDate:April 15, 2011EPA Reg. No. / File Symbol:29964-xPage22of29Applicant's/Registrant's Name & Address:Pioneer Hi-Bred International, Inc., 7100 NW 62nd Avenue, Johnston, IA 50131Product:4114 MaizeIngredientBacillus thuringiensisCry1F and Cry34/35Ab1 insecticidal proteins and the genetic material (PHP27118 T-DNA) necessary for their production in cornGuideline Reference Number Guideline Study Name MRID Number Submitter Status Note68467 PER68467 PER68467 PER68467 PER68467 PER68467 PERSignatureName and TitleJamie StaleyU.S. Registration ManagerDateApril 15, 2011EPA Form 8570-35 (9-97) Electronic and Paper versions available. Submit only Paper versionPublic File CopyPioneer Hi-Bred International, Inc.Study Number: PHI-2011-080Page 65 of 100Form Approved OMB No. 2070-0060UNITED STATES ENVIRONMENTAL PROTECTION AGENCY1200 Pennsylvania Avenue, N.W.Washington, D.C. 20460Paperwork Reduction Act Notice:The public reporting burden for this collection of information is estimated to average 0.25 hours per response for registrationactivities and 0.25 hours perresponse for reregistration and special review activities, including time for reading the instructions and completing the necessary forms. Send comments regarding the burden estimate or anyother aspect of this collection of information, including suggestions for reducing the burden to: Director, OPPE Information Management Division (2137), U.S. Environmental Protection Agency,1200 Pennsylvania Avenue, N.W., Washington, DC 20460. Do not send the form to this address.DATA MATRIXDate:April 15, 2011EPA Reg. No. / File Symbol:29964-xPage23of29Applicant's/Registrant's Name & Address:Pioneer Hi-Bred International, Inc., 7100 NW 62nd Avenue, Johnston, IA 50131Product:4114 MaizeIngredientBacillus thuringiensisCry1F and Cry34/35Ab1 insecticidal proteins and the genetic material (PHP27118 T-DNA) necessary for their production in cornGuideline Reference Number Guideline Study Name MRID Number Submitter Status Note68467 PER68467 PER68467 PER68467 PER68467 PER68467 PERSignatureName and TitleJamie StaleyU.S. Registration ManagerDateApril 15, 2011EPA Form 8570-35 (9-97) Electronic and Paper versions available. Submit only Paper versionPublic File CopyPioneer Hi-Bred International, Inc.Study Number: PHI-2011-080Page 66 of 100Form Approved OMB No. 2070-0060UNITED STATES ENVIRONMENTAL PROTECTION AGENCY1200 Pennsylvania Avenue, N.W.Washington, D.C. 20460Paperwork Reduction Act Notice:The public reporting burden for this collection of information is estimated to average 0.25 hours per response for registrationactivities and 0.25 hours perresponse for reregistration and special review activities, including time for reading the instructions and completing the necessary forms. Send comments regarding the burden estimate or anyother aspect of this collection of information, including suggestions for reducing the burden to: Director, OPPE Information Management Division (2137), U.S. Environmental Protection Agency,1200 Pennsylvania Avenue, N.W., Washington, DC 20460. Do not send the form to this address.DATA MATRIXDate:April 15, 2011EPA Reg. No. / File Symbol:29964-xPage24of29Applicant's/Registrant's Name & Address:Pioneer Hi-Bred International, Inc., 7100 NW 62nd Avenue, Johnston, IA 50131Product:4114 MaizeIngredientBacillus thuringiensisCry1F and Cry34/35Ab1 insecticidal proteins and the genetic material (PHP27118 T-DNA) necessary for their production in cornGuideline Reference Number Guideline Study Name MRID Number Submitter Status Note68467 PER68467 PER68467 PER68467 PER68467 PER68467 PERSignatureName and TitleJamie StaleyU.S. Registration ManagerDateApril 15, 2011EPA Form 8570-35 (9-97) Electronic and Paper versions available. Submit only Paper versionPublic File CopyPioneer Hi-Bred International, Inc.Study Number: PHI-2011-080Page 67 of 100Form Approved OMB No. 2070-0060UNITED STATES ENVIRONMENTAL PROTECTION AGENCY1200 Pennsylvania Avenue, N.W.Washington, D.C. 20460Paperwork Reduction Act Notice:The public reporting burden for this collection of information is estimated to average 0.25 hours per response for registrationactivities and 0.25 hours perresponse for reregistration and special review activities, including time for reading the instructions and completing the necessary forms. Send comments regarding the burden estimate or anyother aspect of this collection of information, including suggestions for reducing the burden to: Director, OPPE Information Management Division (2137), U.S. Environmental Protection Agency,1200 Pennsylvania Avenue, N.W., Washington, DC 20460. Do not send the form to this address.DATA MATRIXDate:April 15, 2011EPA Reg. No. / File Symbol:29964-xPage25of29Applicant's/Registrant's Name & Address:Pioneer Hi-Bred International, Inc., 7100 NW 62nd Avenue, Johnston, IA 50131Product:4114 MaizeIngredientBacillus thuringiensisCry1F and Cry34/35Ab1 insecticidal proteins and the genetic material (PHP27118 T-DNA) necessary for their production in cornGuideline Reference Number Guideline Study Name MRID Number Submitter Status Note68467 PER68467 PER68467 PER68467 PER68467 PER68467 PERSignatureName and TitleJamie StaleyU.S. Registration ManagerDateApril 15, 2011EPA Form 8570-35 (9-97) Electronic and Paper versions available. Submit only Paper versionPublic File CopyPioneer Hi-Bred International, Inc.Study Number: PHI-2011-080Page 68 of 100Form Approved OMB No. 2070-0060UNITED STATES ENVIRONMENTAL PROTECTION AGENCY1200 Pennsylvania Avenue, N.W.Washington, D.C. 20460Paperwork Reduction Act Notice:The public reporting burden for this collection of information is estimated to average 0.25 hours per response for registrationactivities and 0.25 hours perresponse for reregistration and special review activities, including time for reading the instructions and completing the necessary forms. Send comments regarding the burden estimate or anyother aspect of this collection of information, including suggestions for reducing the burden to: Director, OPPE Information Management Division (2137), U.S. Environmental Protection Agency,1200 Pennsylvania Avenue, N.W., Washington, DC 20460. Do not send the form to this address.DATA MATRIXDate:April 15, 2011EPA Reg. No. / File Symbol:29964-xPage26of29Applicant's/Registrant's Name & Address:Pioneer Hi-Bred International, Inc., 7100 NW 62nd Avenue, Johnston, IA 50131Product:4114 MaizeIngredientBacillus thuringiensisCry1F and Cry34/35Ab1 insecticidal proteins and the genetic material (PHP27118 T-DNA) necessary for their production in cornGuideline Reference Number Guideline Study Name MRID Number Submitter Status Note68467 PER68467 PER29964 OWN29964 OWN29964 OWN29964 OWNSignatureName and TitleJamie StaleyU.S. Registration ManagerDateApril 15, 2011EPA Form 8570-35 (9-97) Electronic and Paper versions available. Submit only Paper versionPublic File CopyPioneer Hi-Bred International, Inc.Study Number: PHI-2011-080Page 69 of 100Form Approved OMB No. 2070-0060UNITED STATES ENVIRONMENTAL PROTECTION AGENCY1200 Pennsylvania Avenue, N.W.Washington, D.C. 20460Paperwork Reduction Act Notice:The public reporting burden for this collection of information is estimated to average 0.25 hours per response for registrationactivities and 0.25 hours perresponse for reregistration and special review activities, including time for reading the instructions and completing the necessary forms. Send comments regarding the burden estimate or anyother aspect of this collection of information, including suggestions for reducing the burden to: Director, OPPE Information Management Division (2137), U.S. Environmental Protection Agency,1200 Pennsylvania Avenue, N.W., Washington, DC 20460. Do not send the form to this address.DATA MATRIXDate:April 15, 2011EPA Reg. No. / File Symbol:29964-xPage27of29Applicant's/Registrant's Name & Address:Pioneer Hi-Bred International, Inc., 7100 NW 62nd Avenue, Johnston, IA 50131Product:4114 MaizeIngredientBacillus thuringiensisCry1F and Cry34/35Ab1 insecticidal proteins and the genetic material (PHP27118 T-DNA) necessary for their production in cornGuideline Reference Number Guideline Study Name MRID Number Submitter Status Note29964 OWN29964 OWN29964 OWN80778 PER68467 PER29964 OWNSignatureName and TitleJamie StaleyU.S. Registration ManagerDateApril 15, 2011EPA Form 8570-35 (9-97) Electronic and Paper versions available. Submit only Paper versionPublic File CopyPioneer Hi-Bred International, Inc.Study Number: PHI-2011-080Page 70 of 100Form Approved OMB No. 2070-0060UNITED STATES ENVIRONMENTAL PROTECTION AGENCY1200 Pennsylvania Avenue, N.W.Washington, D.C. 20460Paperwork Reduction Act Notice:The public reporting burden for this collection of information is estimated to average 0.25 hours per response for registrationactivities and 0.25 hours perresponse for reregistration and special review activities, including time for reading the instructions and completing the necessary forms. Send comments regarding the burden estimate or anyother aspect of this collection of information, including suggestions for reducing the burden to: Director, OPPE Information Management Division (2137), U.S. Environmental Protection Agency,1200 Pennsylvania Avenue, N.W., Washington, DC 20460. Do not send the form to this address.DATA MATRIXDate:April 15, 2011EPA Reg. No. / File Symbol:29964-xPage28of29Applicant's/Registrant's Name & Address:Pioneer Hi-Bred International, Inc., 7100 NW 62nd Avenue, Johnston, IA 50131Product:4114 MaizeIngredientBacillus thuringiensisCry1F and Cry34/35Ab1 insecticidal proteins and the genetic material (PHP27118 T-DNA) necessary for their production in cornGuideline Reference Number Guideline Study Name MRID Number Submitter Status Note29964 OWN29964 OWN29964 OWN29964 OWN29964 OWN29964 OWNSignatureName and TitleJamie StaleyU.S. Registration ManagerDateApril 15, 2011EPA Form 8570-35 (9-97) Electronic and Paper versions available. Submit only Paper versionPublic File CopyPioneer Hi-Bred International, Inc.Study Number: PHI-2011-080Page 71 of 100Form Approved OMB No. 2070-0060UNITED STATES ENVIRONMENTAL PROTECTION AGENCY1200 Pennsylvania Avenue, N.W.Washington, D.C. 20460Paperwork Reduction Act Notice:The public reporting burden for this collection of information is estimated to average 0.25 hours per response for registrationactivities and 0.25 hours perresponse for reregistration and special review activities, including time for reading the instructions and completing the necessary forms. Send comments regarding the burden estimate or anyother aspect of this collection of information, including suggestions for reducing the burden to: Director, OPPE Information Management Division (2137), U.S. Environmental Protection Agency,1200 Pennsylvania Avenue, N.W., Washington, DC 20460. Do not send the form to this address.DATA MATRIXDate:April 15, 2011EPA Reg. No. / File Symbol:29964-xPage29of29Applicant's/Registrant's Name & Address:Pioneer Hi-Bred International, Inc., 7100 NW 62nd Avenue, Johnston, IA 50131Product:4114 MaizeIngredientBacillus thuringiensisCry1F and Cry34/35Ab1 insecticidal proteins and the genetic material (PHP27118 T-DNA) necessary for their production in cornGuideline Reference Number Guideline Study Name MRID Number Submitter Status Note29964 OWNSignatureName and TitleJamie StaleyU.S. Registration ManagerDateApril 15, 2011EPA Form 8570-35 (9-97) Electronic and Paper versions available. Submit only Paper versionPublic File CopyPioneer Hi-Bred International, Inc.Study Number: PHI-2011-080Page 72 of 100

# S ECTION 

# II. S UMMARY OF APPLICATION



# Pioneer Hi-Bred International, Inc. (Pioneer) is requesting a Seed Increase Registration for a plantincorporated protectant (PIP) containing Pioneer’s new active ingredient Bacillus thuringiensis Cry1F and

Cry34/35Ab1 insecticidal proteins and the genetic material necessary for their production (via elements of

vector PHP27118) in event 4114 maize (Organization for Economic Cooperation and Development

[OECD] Unique Identifier DP-ØØ4114-3). 4114 maize will not be sold or planted commercially for the

purposes of grain production and controlling corn insect pests. 4114 maize will be used only for breeding

purposes, agronomic testing, increasing inbred seed, and producing hybrid seed on a limited amount of

acreage per county and per year. 4114 maize will be used to create inbred lines that can be crossed with

other inbred lines to create hybrid maize products with multiple modes of action for control of pest insects.

The new PIP, 4114 maize, produces its own insecticidal proteins within the corn plant. The insecticidal

proteins were derived from Bacillus thuringiensis ( Bt ), a naturally occurring soil bacterium. The Cry1F

and Cry34/35Ab1 insecticidal proteins expressed in 4114 maize control certain lepidopteran and corn

rootworm pests of corn, respectively.

4114 maize is a new genetic event that is a “molecular stack” (DNA inserted at one locus in maize

genome) of the previously registered “breeding stack” (DAS-Ø15Ø7-1 x DAS-59122-7; EPA registration

29964-5). The Cry1F, Cry34/35Ab1 and PAT proteins expressed in 4114 maize are identical to those

expressed in 1507 (EPA Registration 29964-3), 59122 (EPA Registration 29964-4) and/or 1507x59122

maize. Cry1F, Cry34/35Ab1 and PAT have tolerance exemptions at EPA and are components of

commercial Bt maize products currently being grown on a large number of U.S. corn acres.

A. Use Profile

Active Ingredient Name: Bacillus thuringiensis Cry1F and Cry34/35Ab1 insecticidal proteins and the

genetic material necessary for their production (via elements of vector

PHP27118) in Event 4114 maize (OECD Unique Identifier DP-ØØ4114-3)

Trade and Other Name(s): 4114 Maize

OPP Chemical Codes: 006481 (Cry1F corn) and 006490 (Cry34/35Ab1 corn)

Basic Manufacturer: Pioneer Hi-Bred International, Inc.

7300 NW 62 nd 

# Ave.

Johnston, IA 50131

Type of Pesticide: Plant-Incorporated Protectant

Uses: Seed Increase in Field Corn

Target Pests for 4114 maize: Black cutworm ( Agrotis ipsilon) , corn earworm ( Helicoverpa zea)

, European corn borer ( Ostrinia nubilalis) , fall armyworm ( Spodoptera frugiperda ), Southwestern corn

borer ( Diatraea grandiosella), western bean cutworm ( Striacosta albicosta ), lesser corn stalk borer

( Elasmopalpus lignosellus ), southern corn stalk borer ( Diatraea crambidoides ), sugarcane borer

( Diatraea saccharalis ), western corn rootworm ( Diabrotica virgifera virgifera ), northern corn rootworm

( Diabrotica barberi ), and Mexican corn rootworm ( Diabrotica virgifera zeae ). Pioneer Hi-Bred International, Inc.

Study Number: PHI-2011-080 Page 73 of 100

# B. Regulatory History

4114 maize is a new genetic event containing previously approved active ingredients in the same crop

(field corn). No Experimental Use Permit (EUP) application for event 4114 maize was submitted. 4114

maize has been grown in small-scale field tests under the 10-acre EUP exemption, as described in 40

CFR §172.3(c)(1).

4114 maize is also regulated by the United States Department of Agriculture-Animal and Plant Health and

Inspection Service, Biotechnology Regulatory Services (USDA-APHIS BRS). A petition for deregulation

of 4114 maize is being submitted to USDA-APHIS BRS in the third quarter of 2011. A safety and

nutritional assessment of 4114 maize will also be submitted to the Food and Drug Administration’s Center

for Food Safety and Applied Nutrition (FDA CFSAN) in a similar time period.

4114 maize was developed by Agrobacterium -mediated transformation of corn using the DNA plasmid

vector PHP27118. The T-DNA of PHP27118 contains the cry 1F, cry 34Ab1, cry 35Ab1 and pat genes.

4114 maize produces two Bacillus thuringiensis ( Bt ) proteins, Cry1F and Cry34/35Ab1. The Cry1F

protein confers resistance to certain lepidopteran pests, including the European corn borer ( Ostrinia

nubilalis ). The Cry34Ab1 and Cry35Ab1 proteins together comprise an active insecticidal crystal protein

that confers resistance to certain coleopteran pests, including western corn rootworm ( Diabrotica virgifera

virgifera) . The PAT protein confers tolerance to the herbicidal active ingredient glufosinate-ammonium.

4114 maize is a single transformation event that is intended to replace the previously registered breeding

stack DAS-Ø15Ø7-1 (1507 maize) x DAS-59122-7 (59122 maize); EPA registration number 29964-5.

1507 and 59122 maize are also individually registered (EPA registrations 29964-3 and 29964-4,

respectively).

The Cry1F and Cry34/35Ab1 proteins in 4114 maize are identical to the Cry1F and Cry34/35Ab1 proteins

in 1507 and 59122 maize, respectively. The Cry1F and Cry34/35Ab1 proteins are components of many

registered maize PIP products (Table 1). The Cry1F protein in 1507 and 4114 maize is from the

requirement of a tolerance in maize (40CFR §174.520). The Cry34/35Ab1 protein from 59122 and 4114

maize is also exempt from the requirement of a tolerance in maize (40CFR §174.506). In addition, the

PAT protein in 1507, 59122 and 4114 maize is exempt from the requirement of a tolerance in all plants

(40CFR §174.522). A summary of the regulatory history is as follows:

On April 11, 1997 (62 FR No. 70), in response to a petition from Dekalb Genetics Corporation, EPA

established a permanent exemption from the requirement of a tolerance for phosphinothricin

acetyltransferase (PAT) in all plant raw agricultural commodities.

On June 6, 2001 (66 FR No. 109), in response to a petition from Mycogen Seeds c/o Dow AgroSciences

LLC, EPA established a permanent exemption from the requirement of a tolerance for Cry1F in the food

and feed commodities of field corn, sweet corn and pop corn.

On July 7, 2003 (68 FR No. 129), in response to a petition from Mycogen Seeds c/o Dow AgroSciences

LLC, EPA established a time-limited exemption from the requirement of a tolerance for Cry34/35Ab1 in

the food and feed commodities of field corn, sweet corn and pop corn.

On September 21, 2005 (70 FR No. 182), in response to a petition from Mycogen Seeds c/o Dow

AgroSciences LLC, EPA established a permanent exemption from the requirement of a tolerance for

Cry34/35Ab1 in the food and feed commodities of field corn, sweet corn and pop corn.

On April 25, 2007 (72 FR 20435), the tolerance exemptions for the PAT, Cry1F, and Cry34/35Ab1

proteins were redesignated from 40 CFR §180 to 40 CFR §174. Pioneer Hi-Bred International, Inc.

Study Number: PHI-2011-080 Page 74 of 100

# Table 1. Currently Registered Maize PIPs Expressing Cry1F and/or Cry34/35Ab1 Proteins

EPA Reg.

Number Registration Name Company Active

Ingredient(s) Initial Date of

Registration

29964-3

TC1507

(Herculex I) Pioneer Hi-Bred Int’l, Inc.

Cry1F May 18, 2001

68467-2 Mycogen Seeds (c/o Dow

Agrosciences LLC)

29964-4

DAS-59122-7

(Herculex Rootworm) Pioneer Hi-Bred Int’l, Inc.

Cry34/35Ab1 August 31, 2005

68467-5 Mycogen Seeds (c/o Dow

Agrosciences LLC)

68467-6 TC1507 x DAS-59122-7

(Herculex XTRA) Mycogen Seeds (c/o Dow

Agrosciences LLC) Cry1F

Cry34/35Ab1 October 27, 2005

29964-5 Pioneer Hi-Bred Int’l, Inc.

68467-4 DAS-06275-8 Mycogen Seeds (c/o Dow

Agrosciences LLC) Cry1F May 27, 2005

29964-6 Optimum AcreMax 1 Pioneer Hi-Bred Int’l, Inc. Cry1F

Cry34/35Ab1 April 30, 2010

29964-7 TC1507 x MON810

(Optimum Intrasect) Pioneer Hi-Bred Int’l, Inc. Cry1Ab

Cry1F February 24, 2010

29964-8 TC1507 x DAS-59122-7 x

MON810 Pioneer Hi-Bred Int’l, Inc. Cry1Ab

Cry1F

Cry34/35Ab1 February 24, 2010

29964-9 DAS-59122-7 x MON810 Pioneer Hi-Bred Int’l, Inc. Cry1Ab

Cry34/35Ab1 February 24, 2010

29964-10 Optimum AcreMax RW Pioneer Hi-Bred Int’l, Inc. Cry34/35Ab1 April 30, 2010

524-581 MON 89034 x TC1507 x

MON 88017 x DAS-59122-7

(SmartStax) Monsanto Company Cry1A.105

Cry2Ab2

Cry1F

Cry3Bb1

Cry34/35Ab1 July 20, 2009

68467-7 Mycogen Seeds (c/o Dow

Agrosciences LLC)

524-584 MON 89034 x DAS-59122-7* Monsanto Company Cry1A.105

Cry2Ab2

Cry34/35Ab1 December 14,

2009

524-586 MON 88017 x DAS-59122-7* Monsanto Company Cry3Bb1

Cry34/35Ab1 December 14,

2009

524-588 MON 89034 x TC1507 x

DAS-59122-7* Monsanto Company Cry1A.105

Cry2Ab2

Cry1F

Cry34/35Ab1 December 14,

2009

524-589 MON 89034 x MON 88017 x

DAS-59122-7* Monsanto Company Cry1A.105

Cry2Ab2

Cry3Bb1

Cry34/35Ab1 December 14,

2009

524-590 TC1507 x MON 88017 x

DAS-59122-7* Monsanto Company Cry1F

Cry3Bb1

Cry34/35Ab1 December 14,

2009



# *Note: Registration is for breeding purposes, agronomic testing, increasing inbred seed stocks, and

producing hybrid seed only. Pioneer Hi-Bred International, Inc.

Study Number: PHI-2011-080 Page 75 of 100

# C. Benefits of 4114 Maize

The corn seed industry is transitioning from Bt products with a single mode of action to products with

multiple modes of action, in order to extend the durability of important Bt traits that many growers rely on

to manage pests. In addition, growers are requiring an increasingly sophisticated suite of traits to meet

their changing farming needs, which include transgenic herbicide resistance as well as native traits such

as disease tolerance, drought tolerance, and higher yields.

Furthermore, as new transgenic traits are developed to provide improved nutrition or agronomic

performance, growers will continue to demand multiple Bt traits for protection against pest damage in

addition to the new traits. It is imperative for growers to continue to increase their overall farm

productivity to meet the food security needs of a growing global population. 4114 maize, as one

component of future Bt breeding stack products with multiple modes of action, will provide benefits to

growers in the form of multi-trait products with enhanced durability, higher yields and more diverse

germplasm suited for diverse growing regions.

Corn breeders have suspected for some time that there is a finite number of transgenic events that can

be combined in one maize plant before starting to reduce the plant’s agronomic performance and yield.

Precise quantitative data on the yield impact of multiple transgenic events in a maize product are

impossible to obtain, primarily due to the much larger variation in yield due to the specific genetic

background of the maize hybrid and the exact environment in which it grows. 4114 maize is a “molecular

stack” of Cry1F and Cry34/35Ab1 in one transgenic event. It will replace the previously registered

“breeding stack” of 1507x59122 (Cry1F and Cry34/35Ab1 in separate transgenic events), thereby

reducing the number of events in complex breeding stacks of multiple traits.

Breeding multiple traits in one maize product becomes increasingly complex and expensive with each

breeding locus added. In 4114 maize, the cry 1F and cry 34/35Ab1 genes are at one locus in maize

genome, whereas in 1507x59122, the cry 1F and cry 34/35Ab1 genes are at separate loci. Molecular

stacks such as 4114 maize reduce the number of loci required for insect traits, thus increasing the speed

at which new high-yielding products are available to growers. For example, the effort required to breed

two transgenic loci into multiple elite inbred lines is double the effort required for one transgenic locus. In

addition, each trait (locus) must be in a homozygous state in inbreds. Based on simple Mendelian

segregation of multiple loci that do not segregate together (as is the case with unlinked traits in breeding

stacks), the proportion of plants that are homozygous for all traits of interest becomes smaller with each

trait added, resulting in more seed discard during the breeding process.

4114 maize is expected to be able to be bred into a much wider variety of maize genetic backgrounds

than 1507x59122, thus giving growers more choice in high-yielding products offering in-plant insect

production as well as more products customized to their local growing areas and agronomic needs

D. Nature of Registration Being Requested

An application for a Seed Increase Registration of plant-incorporated protectants pursuant to FIFRA

§3(c)(5) is being sought for Bacillus thuringiensis Cry1F and Cry34/35Ab1 proteins and the genetic

material (PHP27118 T-DNA) necessary for their production in 4114 maize. Pioneer Hi-Bred International, Inc.

Study Number: PHI-2011-080 Page 76 of 100

# S ECTION 

# III. P RODUCT 

# L ABEL



# Three (3) copies of the label for 4114 maize are attached. Pioneer Hi-Bred International, Inc.

Study Number: PHI-2011-080 Page 77 of 100

# 4114 Maize

(OECD Unique Identifier: DP-ØØ4114-3)

Active Ingredients:

Bacillus thuringiensis Cry1F protein and the genetic material (PHP27118 T-DNA)

necessary for its production in corn event DP-ØØ4114- 3 ............................................................ < 0.0015%*

Bacillus thuringiensis Cry34Ab1 protein and the genetic material (PHP27118 T-DNA)

necessary for its production in corn event DP-ØØ4114- 3 ............................................................ < 0.0064%*

Bacillus thuringiensis Cry35Ab1 protein and the genetic material (PHP27118 T-DNA)

necessary for its production in corn event DP-ØØ4114- 3 ............................................................ < 0.0100%*

Inert Ingredients:

Phosphinothricin acetyltransferase (PAT) protein and the genetic material (PHP27118 T-DNA)

necessary for its production in corn events DP-ØØ4114- 3 .......................................................... < 0.0011%*

* Percentage (wt/wt) on a dry wt. basis for whole plant

KEEP OUT OF REACH OF CHILDREN

CAUTION

NET CONTENTS ________________

EPA REGISTRATION NUMBER: 29964-xx

EPA ESTABLISHMENT NUMBER: 029964-IA-001

Pioneer Hi-Bred International, Inc.

7300 NW 62 nd 

# Avenue

Johnston, IA 50131 Pioneer Hi-Bred International, Inc.

Study Number: PHI-2011-080 Page 78 of 100

# DIRECTIONS FOR USE

It is a violation of Federal law to use this product in any manner inconsistent with its labeling.

This plant-incorporated protectant (PIP) may be combined through conventional breeding with other

registered plant-incorporated protectants that are similarly approved for use in combination, through

conventional breeding, with other registered plant-incorporated protectants to produce inbred corn lines

and hybrid corn varieties with combined pesticidal traits.

4114 maize may be used for breeding purposes, agronomic testing, increasing inbred seed and

producing hybrid seed corn up to a total of 20,000 acres per county and up to a combined United States

(U.S.) total of 250,000 acres per plant-incorporated protectant active ingredient per registrant per year.

Commercial plantings of this product for the purposes of grain production and controlling corn insect

pests are prohibited.



# Use Pattern

Crop Pests

Field corn black cutworm

corn earworm

European corn borer

fall armyworm

lesser corn stalk borer

southern corn stalk borer

southwestern corn borer

sugarcane borer

stalk borer

western bean cutworm

western corn rootworm

northern corn rootworm

Mexican corn rootworm Pioneer Hi-Bred International, Inc.

Study Number: PHI-2011-080 Page 79 of 100

# 4114 Maize

(OECD Unique Identifier: DP-ØØ4114-3)

Active Ingredients:

Bacillus thuringiensis Cry1F protein and the genetic material (PHP27118 T-DNA)

necessary for its production in corn event DP-ØØ4114- 3 ............................................................ < 0.0015%*

Bacillus thuringiensis Cry34Ab1 protein and the genetic material (PHP27118 T-DNA)

necessary for its production in corn event DP-ØØ4114- 3 ............................................................ < 0.0064%*

Bacillus thuringiensis Cry35Ab1 protein and the genetic material (PHP27118 T-DNA)

necessary for its production in corn event DP-ØØ4114- 3 ............................................................ < 0.0100%*

Inert Ingredients:

Phosphinothricin acetyltransferase (PAT) protein and the genetic material (PHP27118 T-DNA)

necessary for its production in corn events DP-ØØ4114- 3 .......................................................... < 0.0011%*

* Percentage (wt/wt) on a dry wt. basis for whole plant.

KEEP OUT OF REACH OF CHILDREN

CAUTION

NET CONTENTS ________________

EPA REGISTRATION NUMBER: 29964-xx

EPA ESTABLISHMENT NUMBER: 029964-IA-001

Pioneer Hi-Bred International, Inc.

7300 NW 62 nd 

# Avenue

Johnston, IA 50131 Pioneer Hi-Bred International, Inc.

Study Number: PHI-2011-080 Page 80 of 100

# DIRECTIONS FOR USE

It is a violation of Federal law to use this product in any manner inconsistent with its labeling.

This plant-incorporated protectant (PIP) may be combined through conventional breeding with other

registered plant-incorporated protectants that are similarly approved for use in combination, through

conventional breeding, with other registered plant-incorporated protectants to produce inbred corn lines

and hybrid corn varieties with combined pesticidal traits.

4114 maize may be used for breeding purposes, agronomic testing, increasing inbred seed and

producing hybrid seed corn up to a total of 20,000 acres per county and up to a combined United States

(U.S.) total of 250,000 acres per plant-incorporated protectant active ingredient per registrant per year.

Commercial plantings of this product for the purposes of grain production and controlling corn insect

pests are prohibited.



# Use Pattern

Crop Pests

Field corn black cutworm

corn earworm

European corn borer

fall armyworm

lesser corn stalk borer

southern corn stalk borer

southwestern corn borer

sugarcane borer

stalk borer

western bean cutworm

western corn rootworm

northern corn rootworm

Mexican corn rootworm Pioneer Hi-Bred International, Inc.

Study Number: PHI-2011-080 Page 81 of 100

# 4114 Maize

(OECD Unique Identifier: DP-ØØ4114-3)

Active Ingredients:

Bacillus thuringiensis Cry1F protein and the genetic material (PHP27118 T-DNA)

necessary for its production in corn event DP-ØØ4114- 3 ............................................................ < 0.0015%*

Bacillus thuringiensis Cry34Ab1 protein and the genetic material (PHP27118 T-DNA)

necessary for its production in corn event DP-ØØ4114- 3 ............................................................ < 0.0064%*

Bacillus thuringiensis Cry35Ab1 protein and the genetic material (PHP27118 T-DNA)

necessary for its production in corn event DP-ØØ4114- 3 ............................................................ < 0.0100%*

Inert Ingredients:

Phosphinothricin acetyltransferase (PAT) protein and the genetic material (PHP27118 T-DNA)

necessary for its production in corn events DP-ØØ4114- 3 .......................................................... < 0.0011%*

* Percentage (wt/wt) on a dry wt. basis for whole plant.

KEEP OUT OF REACH OF CHILDREN

CAUTION

NET CONTENTS ________________

EPA REGISTRATION NUMBER: 29964-xx

EPA ESTABLISHMENT NUMBER: 029964-IA-001

Pioneer Hi-Bred International, Inc.

7300 NW 62 nd 

# Avenue

Johnston, IA 50131 Pioneer Hi-Bred International, Inc.

Study Number: PHI-2011-080 Page 82 of 100

# DIRECTIONS FOR USE

It is a violation of Federal law to use this product in any manner inconsistent with its labeling.

This plant-incorporated protectant (PIP) may be combined through conventional breeding with other

registered plant-incorporated protectants that are similarly approved for use in combination, through

conventional breeding, with other registered plant-incorporated protectants to produce inbred corn lines

and hybrid corn varieties with combined pesticidal traits.

4114 maize may be used for breeding purposes, agronomic testing, increasing inbred seed and

producing hybrid seed corn up to a total of 20,000 acres per county and up to a combined United States

(U.S.) total of 250,000 acres per plant-incorporated protectant active ingredient per registrant per year.

Commercial plantings of this product for the purposes of grain production and controlling corn insect

pests are prohibited.



# Use Pattern

Crop Pests

Field corn black cutworm

corn earworm

European corn borer

fall armyworm

lesser corn stalk borer

southern corn stalk borer

southwestern corn borer

sugarcane borer

stalk borer

western bean cutworm

western corn rootworm

northern corn rootworm

Mexican corn rootworm Pioneer Hi-Bred International, Inc.

Study Number: PHI-2011-080 Page 83 of 100

# S ECTION 

# IV. S ATISFACTION OF 

# D ATA 

# R EQUIREMENTS



# Specific data requirements for obtaining registration of a plant-incorporated protectant (PIP) have not

been promulgated under 40 CFR §174 Subpart H. In the absence of PIP-specific data requirements,

EPA has utilized those requirements delineated for microbial pesticides (40 CFR §158.2120) as a partial

guide for determining if an Applicant has provided all of the required data. In addition to the microbial

data requirements there is a core set of studies more pertinent to PIPs that EPA has been requiring of

applicants in recent years. These latter data requirements are not published and became known to the

Applicant through previous consultations with the Agency.

The tables in this section provide a listing of the data requirements applicable to registration of a plantincorporated protectant as known to Pioneer. An explanation of how each data requirement is satisfied in

this application is included in the table. Specific study reports addressing a data requirement are cited

either by the volume number within this submission or by MRID if previously submitted. Not all of the

microbial pesticide data requirements are applicable to a PIP given the inherent differences between the

two types of products. Where appropriate, a brief explanation is provided in the table for those data

requirements that are not applicable. The data requirements are organized by OPPTS Microbial

Pesticide Test Guideline number, using the 885- guideline series. OPPTS guideline numbers are not

assigned (N/A) to some PIP data requirements.

Table 2. Product Analysis Data Requirements

OPPTS # Data Requirement Method of Satisfaction

885.1100 Product identity CSF (PHI-2011-080), Volume 2 (PHI-2009-015),

MRID 447148-03, 450201-03, 457904-01, 461239-03,

461239-05, 461239-06, 452422-06, 461239-07,

885.1200 Manufacturing process Volume 2 (PHI-2009-015)

885.1300 Formation of unintentional

ingredients MRID 477864-01, 447864-05, 477864-02, 477864-08,

477864-03, 477864-07,

885.1400 Analysis of samples Volume 8 (PHI-2010-059)

885.1500 Certification of limits CSF (PHI-2011-080) and Volume 8 (PHI-2010-059)

Table 3. Residue Data Requirements

OPPTS # Data Requirement Method of Satisfaction

885.2100 Chemical identity CSF (PHI-2011-080) and Volume 8 (PHI-2010-059)

885.2200 Nature of residue in plants CSF (PHI-2011-080)

885.2250 Nature of residue in

animals No adverse effects associated with mammalian

exposure to Cry1F, Cry34/35Ab1, or the PAT protein

have been observed; thus they are not of toxicological

concern and do not trigger this data requirement.

885.2300 Analytical methods - plants Volume 8 (PHI-2010-059), MRID 452793-01 and

453834-01 Pioneer Hi-Bred International, Inc.

Study Number: PHI-2011-080 Page 84 of 100

# OPPTS # Data Requirement Method of Satisfaction

885.2350 Analytical methods -

animals Cry1F, Cry34/35Ab1, and the PAT proteins in 4114

maize are exempt from the requirement of tolerances,

thus an analytical enforcement method is not needed.

885.2400 Storage stability Cry1F, Cry34/35Ab1, and the PAT proteins in 4114

maize are exempt from the requirement of tolerances,

thus storage stability data is not needed

885.2500 Magnitude of residues in

plants Volume 8 (PHI-2010-059)

885.2550 Magnitude of residues in

meat, milk, poultry, and

eggs Cry1F, Cry34/35Ab1, and the PAT proteins in 4114

maize are exempt from the requirement of tolerances;

thus residue studies are not required.

885.2600 Magnitude of residues in

potable water, fish, and

irrigated crops The Cry1F, Cry34/35Ab1, and PAT proteins are plantincorporated and will not be used in aquatic sites.

4114 maize seed may be planted near aquatic sites

and there is the potential for pollen drift into bodies of

water. However, there are no residues of toxicological

concern in 4114 maize; thus, aquatic residue studies

are not required.

Table 4. Toxicology Data Requirements

OPPTS # Data Requirement Method of Satisfaction

885.3050 Acute oral

toxicity/pathogenicity MRID 446911-01, 450201-18, 452422-07, 452422-08,

452422-09 and 436843-01

885.3100 Acute dermal

toxicity/pathogenicity The Cry1F, Cry34/35Ab1, and PAT proteins are plantincorporated, contained within individual plant cells.

There is minimal opportunity for human dermal

contact because corn is not a hand harvested crop.

Furthermore, Cry1F, Cry34/35Ab1, and PAT proteins

are of no toxicological concern; thus, dermal toxicity

data are not required. As the proteins are not living

organisms, a pathogenicity determination is not

applicable. Pioneer Hi-Bred International, Inc.

Study Number: PHI-2011-080 Page 85 of 100

# OPPTS # Data Requirement Method of Satisfaction

885.3150 Acute pulmonary

toxicity/pathogenicity The Cry1F, Cry34/35Ab1, and PAT proteins are plantincorporated and are not volatile. There is minimal

opportunity for human inhalation exposure. The

proteins are of no toxicological concern; thus,

inhalation toxicity data are not required. As the

proteins are not a living organism, a pathogenicity

determination is not applicable.

885.3200 Acute injection

toxicity/pathogenicity The Cry1F, Cry34/35Ab1, and PAT proteins are plantincorporated and of no toxicological concern. There is

minimal opportunity for direct entry of the proteins into

the blood stream via injury during handling because

corn is not a hand-harvested crop. As the proteins

are not living organisms, a pathogenicity

determination is not applicable.

885.3400 Hypersensitivity incidents Pioneer is aware of no incidents of hypersensitivity

reactions to Cry1F, Cry34/35Ab1, or PAT protein in

4114 maize.

885.3500 Cell culture Cry1F, Cry34/35Ab1 and PAT are not living

organisms and therefore, have no infectivity potential.

A cell culture study is not required.

885.3550 Acute toxicology, Tier II No adverse effects were observed at the maximum

attainable oral doses for Cry1F, Cry34/35Ab1, and

PAT; LD 50 

# determinations are therefore, not possible.

Criteria needed to trigger this Tier II study were not

met in the Tier I studies.

885.3600 Subchronic

toxicity/pathogenicity No evidence of infectivity, pathogenicity, or toxicity

was observed in Tier I studies with Cry1F,

Cry34/35Ab1, and PAT in 4114 maize. Criteria

needed to trigger this Tier II study were not met in Tier

I studies.

885.3650 Reproductive/fertility effects No evidence of infectivity, pathogenicity, or toxicity

was observed in Tier I studies with Cry1F,

Cry34/35Ab1, and PAT in 4114 maize. Criteria

needed to trigger this Tier II study were not met in Tier

I studies. Pioneer Hi-Bred International, Inc.

Study Number: PHI-2011-080 Page 86 of 100

# Table 5. Non-target Organism Data Requirements

OPPTS # Data Requirement Method of Satisfaction

885.4050 Avian oral, Tier I MRID 450201-12, 456220-01, 461239-11, and

463438-03.

885.4100 Avian inhalation, Tier I The Cry1F, Cry34/35Ab1, and PAT proteins are plantincorporated and are not ‘applied’ in aerosol or dust

formulations as microbial pesticide products would be.

The only airborne route of exposure for Cry1F,

Cry34/35Ab1, and PAT proteins would be via pollen

drift, and corn pollen is not of a size that is respirable.

885.4150 Wild mammal testing, Tier I MRID 446911-01, 450201-18, 452422-07, 452422-08,

452422-09, and 463843-01.

885.4200 Freshwater fish testing, Tier

I MRID 450442-01, 453077-02, and 457904-03.

885.4240 Freshwater aquatic

invertebrate testing, Tier I MRID 450201-08 and 457904-04

885.4280 Estuarine and marine

animal testing, Tier I There will be no ‘direct application’ of Cry1F,

Cry34/35Ab1, and PAT proteins to fresh, estuarine, or

marine waters. Furthermore, Cry1F, Cry34/35Ab1,

and PAT proteins do not display broad spectrums of

activity. This data requirement does not apply.

885.4300 Nontarget plant studies,

Tier I Cry1F, Cry34/35Ab1, and PAT are not living

organisms and therefore, are not pathogenic to plants.

885.4340 Non-target insect testing,

Tier I MRID 450201-07, 450201-09, 450201-10, 450201-11,

451311-02, 453078-01, 453078-02, 453078-03,

455129-01, 457597-01, 452422-10, 457904-05,

457904-06, 457904-07, 461239-12, 467141-01,

474367-01, 463438-08, and 463438-09

885.4380 Honey bee testing, Tier I MRID 450415-03, 453078-05, and 453407-01

885.4600 Avian chronic pathogenicity

and reproduction test, Tier

III No adverse effects were observed in Tier I studies

and Cry1F, Cry34/35Ab1, and PAT proteins are not

persistent in the environment. Criteria needed to

trigger higher tier testing are not met. Pioneer Hi-Bred International, Inc.

Study Number: PHI-2011-080 Page 87 of 100

# OPPTS # Data Requirement Method of Satisfaction

885.4650 Aquatic invertebrate range

testing, Tier III No adverse effects were observed in Tier I studies

and Cry1F, Cry34/35Ab1, and PAT proteins are not

persistent in the environment. Criteria needed to

trigger higher tier testing are not met.

885.4700 Fish life cycle studies, Tier

III No adverse effects were observed in Tier I studies

and Cry1F, Cry34/35Ab1, and PAT proteins are not

persistent in the environment. Criteria needed to

trigger higher tier testing are not met.

885.4750 Aquatic ecosystem test No adverse effects were observed in Tier I studies

and Cry1F, Cry34/35Ab1, and PAT proteins are not

persistent in the environment. Criteria needed to

trigger higher tier testing are not met.

N/A Other non-target organism

tests MRID 450201-06, 450201-13, 450415-02, 453078-04,

456482-01, 456520-01, 467846-01, 452422-11,

453602-01, 461239-13, 461239-14, 461239-17, and

478477-01.

Table 6. Environmental Expression Data Requirements

OPPTS # Data Requirement Method of Satisfaction

885.5200 Expression in a terrestrial

environment MRID 450201-05, 471207-01, 452422-14, and

479595-01.

885.5300 Expression in a freshwater

environment Cry1F, Cry34/35Ab1, and PAT protein in 4114 maize

do not have the ability to invade, survive, or replicate

in a freshwater environment. Criteria needed to

trigger higher tier testing are not met.

885.5400 Expression in a marine or

estuarine environment Cry1F, Cry34/35Ab1, and PAT protein in 4114 maize

do not have the ability to invade, survive, or replicate

in a marine or estuarine environment. Criteria needed

to trigger higher tier testing are not met. Pioneer Hi-Bred International, Inc.

Study Number: PHI-2011-080 Page 88 of 100

# S ECTION 

# V. P RODUCT 

# C HARACTERIZATION



# 4114 maize was created using Agrobacterium -mediated transformation of a select maize line. The

plasmid PHP27118 was used to produce this event. The T-DNA region of plasmid PHP27118

incorporated into 4114 maize contained the cry1F

, pat, cry34Ab1 and cry35Ab1 genes and the regulatory

sequences necessary for their expression. The regulatory elements (as well as the coding sequences)

for the cry1F

, pat

, cry34Ab1 and cry35Ab1 genes in 4114 maize are identical to those in 1507 and/or

59122 maize.

A. Transformation System

Immature embryos of maize ( Zea mays L.) were aseptically removed from the developing caryopsis nine

to eleven days after pollination and inoculated with Agrobacterium tumefaciens strain LBA4404

containing plasmid PHP27118, essentially as described in Zhao et al ., 2001. After three to six days of

embryo and Agrobacterium co-cultivation on solid culture medium with no selection, the embryos were

then transferred to a medium without herbicide selection but containing carbenicillin for selection against

Agrobacterium . After three to five days on this medium, embryos were then transferred to selective

medium that was stimulatory to maize somatic embryogenesis and contained bialaphos for selection of

cells expressing the pat transgene. The medium also contained carbenicillin to select against any

remaining Agrobacterium . After six to eight weeks on the selective medium, healthy, growing calli that

demonstrated resistance to bialaphos were identified. The putative transgenic calli were subsequently

regenerated to produce T0 plantlets.

B. Characterization of the DNA Inserted in the Plant and Inheritance and Stability

Samples were taken from the T0 plantlets for PCR analysis to verify the presence and copy number of

the PHP27118 T-DNA by screening for the cry 1F, cry 34Ab1, cry 35Ab1, and/or pat genes. In addition to

this analysis, the T0 plantlets were analyzed for the presence of certain Agrobacterium binary vector

backbone sequences by PCR. Plants that were determined to be single copy for the inserted genes and

negative for vector backbone sequences were selected for further greenhouse propagation. These

selected T0 plants were additionally screened to confirm trait efficacy. The T0 plants meeting all PCR

and trait efficacy criteria, including maize event 4114, were advanced and crossed to inbred lines to

produce seed for further testing.

Subsequent characterization of the DNA isolated from maize line 4114 using restriction enzyme digests

and southern blots indicated that the T-DNA from plasmid PHP27118 was integrated as a single

insertion. In addition, DNA analysis indicated stability and inheritance of the inserted DNA within and

across several generations. The Southern blot analysis also confirmed that 4114 maize does not contain

plasmid vector backbone DNA (Volume 2; PHI-2009-015).

C. Analytical Detection Methods

Validated methods for extraction and direct ELISA analysis of Cry1F, Cry34Ab1 and Cry35Ab1 in corn

grain have been previously submitted and found acceptable by the Agency (MRIDs 45279302, 46123901

and 46123902, respectively).

D. Bridging Protein Equivalency/Identity

The Cry1F, Cry34Ab1, Cry35Ab1 and PAT proteins expressed in 4114 maize are identical to the proteins

expressed in the previously registered 1507, 59122 and/or 1507x59122 maize products (Volume 3: PHI2010-076). This equivalency was established as follows:

The same gene expression cassettes (promoters, protein coding sequences and terminators)

were used in 4114 maize as in previously registered 1507 and 59122 maize, so all genetic

elements are identical. Pioneer Hi-Bred International, Inc.

Study Number: PHI-2011-080 Page 89 of 100

# Western blot analysis demonstrated the Cry1F, Cry34Ab1, Cry35Ab1 and PAT proteins in 4114

maize migrate with the equivalent molecular weight as and have similar relative immunoreactivity

to the proteins expressed in previously registered 1507x59122 maize, indicating equivalency.

The inserted DNA in 4114 maize was sequenced and the translated amino acid sequences of the

encoded Cry1F, Cry34Ab1, Cry35Ab1 and PAT proteins were determined. The translated amino

acid sequences were compared to the amino acid sequence of those proteins in previously

registered 1507, 59122 and 1507x59122 maize and found to be identical.

Bioequivalency of the expressed Cry1F and Cry34/35Ab1 proteins was established via side-byside efficacy testing of 4114 maize with 1507x59122 maize.

The expressed proteins in 4114 maize are identical to proteins expressed in previously registered

products. Therefore, previously submitted safety data and full tolerance exemptions for Cry1F,

Cry34Ab1, Cry35Ab1 and PAT proteins are also applicable for 4114 maize.

E. Bridging Protein Expression

In order to measure and compare the concentrations of Cry1F, Cry34/35Ab1 and PAT proteins in 4114

maize to previously registered products, tissue samples were collected from field-grown plants at five

locations in the United States and Canada (Volume 8; PHI-2010-059).

Expression levels of Cry1F, Cry34Ab1, Cry35Ab1, and PAT protein were measured in various tissues

(including leaf, root, whole plant, pollen, stalk, forage and grain) from 4114 maize, 1507 maize, 59122

maize, 1507x59122 maize, and near isoline control maize using specific quantitative ELISA methods.

For each protein, the concentrations in each 4114 maize tissue were statistically compared to the

concentrations in 1507, 59122, and 1507x59122 maize in order to determine the statistical significance of

any differences.

The expression levels of Cry1F protein in 4114 maize were comparable to or lower than levels in 1507

maize and 1507x59122 maize in all tissues measured except for pollen and late-season leaf tissues.

Concentrations of Cry1F in pollen and late-season leaf tissues were significantly higher in 4114 maize.

The expression levels of Cry34Ab1 protein in 4114 maize were comparable to or lower than levels in

59122 maize and 1507x59122 maize in all tissues measured except for late-season leaf tissue. The

concentration of Cry34Ab1 in late-season leaf tissue was significantly higher in 4114 maize.

The expression levels of Cry35Ab1 protein in 4114 maize were comparable to or lower than levels in

59122 maize and 1507x59122 maize in all tissues measured.

The expression levels of PAT protein in 4114 maize were comparable to or higher than levels in 1507

maize and were comparable to or lower than levels in 59122 maize and 1507x59122 maize.

Table 7. Mean Expression Levels of the Cry1F, Cry34Ab1 and Cry35Ab1 Proteins in 4114 Maize a



# Tissue (ng/mg tissue dry weight) b



# Cry1F Cry34Ab1 Cry35Ab1

Leaf 2.0-34 19-110 27-90

Root 3.8-5.5 17-23 6.4-13

Whole Plant c 

# 4.1-12 23-52 21-76

Pollen 35 9.2 0.34

Stalk 6.0 25 9.1

Grain 3.3 24 1.1

a 

# Summarized from PHI-2010-059 (Volume 8)

b 

# Ranges reflect the range of means at different growth stages.

c 

# Whole plant ranges include forage Pioneer Hi-Bred International, Inc.

Study Number: PHI-2011-080 Page 90 of 100

# Table 8. Summary of Product Characterization Data Submitted for 4114 Maize

Study Title Summary Study No.

Characterization of

DP-ØØ4114-3

Maize Insertion

Integrity, Stability,

Copy Number and

Backbone Analysis Southern blot analyses indicate that 4114 maize has the

introduced DNA inserted in the corn genome at a single locus

and contains one copy each of the cry1F

, cry34Ab1

, cry35Ab1 and pat expression cassettes. All expression

elements are shown to be present in the insert, and there are

no other elements detectable other than those associated

with the respective cassettes. No backbone sequences from

plasmid PHP27118 were detected in the 4114 maize

genome. PHI-2009-015

(Volume 2)

Equivalency of

Expressed Proteins

in 4114 Maize to

Previously

Registered Events The Cry1F, Cry34Ab1, Cry35Ab1 and PAT proteins

expressed in 4114 maize are identical to the proteins

expressed in the previously registered 1507, 59122 and/or

1507x59122 maize products. This equivalency was

confirmed by use of the same genetic elements, Western blot

analysis, translated amino acid sequence of the DNA insert,

and insect efficacy. PHI-2010-076

(Volume 3)

Equivalency of

Expressed Proteins

in 4114 Maize to

Previously

Registered Events Western blot analysis of 4114 and 1507x59122 leaf tissue

demonstrated equivalency between the Cry1F, Cry34Ab1,

Cry35Ab1 and PAT proteins. The translated amino acid

sequence of the 4114 maize DNA insert showed 100%

identity of the Cry1F, Cry34Ab1, Cry35Ab1 and PAT proteins

in 4114 maize with the previously published amino acid

sequences of those proteins. PHI-2011-067

(Volume 7)

Expressed Trait

Protein

Concentration of a

Maize Line

Containing Events

DP-ØØ4114-3,

DAS-Ø15Ø7-1,

DAS-59122-7, and

Combined Trait

Product DASØ1507-1xDAS59122-7: US and

Canada Test Sites The expression levels of Cry1F protein in 4114 maize were

comparable to or lower than levels in 1507 maize and

1507x59122 maize in all tissues measured except for pollen

and late-season leaf tissues. Concentrations of Cry1F in

pollen and late-season leaf tissues were significantly higher in

4114 maize. The expression levels of Cry34Ab1 protein in

4114 maize were comparable to or lower than levels in 59122

maize and 1507x59122 maize in all tissues measured except

for late-season leaf tissue. The concentration of Cry34Ab1 in

late-season leaf tissue was significantly higher in 4114 maize.

The expression levels of Cry35Ab1 protein in 4114 maize

were comparable to or lower than levels in 59122 maize and

1507x59122 maize in all tissues measured. PHI-2010-059

(Volume 8)

References

Zhao, Z., Gu, W., Cai, T., Tagliani, L., Hondred, D., Bond, D., Schroeder, S., Rudert, M. and Pierce, D.

2001. High throughput genetic transformation mediated by Agrobacterium tumefaciens in maize.

Molecular Biology 8: 323-333. Pioneer Hi-Bred International, Inc.

Study Number: PHI-2011-080 Page 91 of 100

# S ECTION 

# VI. T OXICITY 

# D ATA



# A. Human Health Assessment of 4114 Maize

Quantitative expression analysis of 4114 maize grain demonstrates that Cry1F, Cry34/35Ab1 and PAT

protein concentrations are comparable to concentrations in grain of 1507, 59122 and/or 1507x59122

maize (Volume 8; PHI-2010-059). Western blots of plant extracts and DNA sequence data of the 4114

maize insert demonstrate that the Cry1F and Cry34/35Ab1 proteins expressed in 4114 maize are

identical to those produced in 1507, 5912 and/or 1507x59122. 4114 maize is a replacement event for

1507x59122 maize; overall acreage of Cry1F and Cry34/35Ab1 is not expected to increase because of

the availability of 4114 maize as a product component. Therefore, the exposure to Cry1F and

Cry34/35Ab1 in human and animal diets due to the incorporation of products containing 4114 maize is

not expected to increase over current levels. Since the proteins and exposure are identical to previous

events, the tolerance exemptions and human health assessments conducted for 1507 and 59122 maize

are applicable to 4114 maize. The tolerance exemptions for Cry1F; Cry34/35Ab1 and PAT proteins are

as follows:

Cry1F protein is exempt from the requirement of a tolerance in corn: 40CFR §174.520.

Cry34Ab1 and Cry35Ab1 proteins are exempt for the requirement of a tolerance in corn: 40CFR

§174.506.

Phosphinothricin acetyltransferase (PAT) is exempt from the requirement of a tolerance in all

plants: 40CFR §174.522.

There is a reasonable certainty that no harm will result from aggregate exposure to the U.S. population,

including infants and children, to the Cry1F and Cry34/35Ab1 proteins and the genetic material necessary

for their production (as expressed in 4114 maize). This includes all anticipated dietary exposures. No

toxicity to mammals has been observed, nor any indication of allergenicity potential for Cry1F or

Cry34/35Ab1.

B. Endocrine Disruptors

The pesticidal active ingredients Cry1F and Cry34/35Ab1 are proteins, derived from sources that are not

known to exert an influence on the endocrine system.

As required under FFDCA section 408(p), EPA has developed the Endocrine Disruptor Screening

Program (EDSP) to determine whether certain substances (including pesticide active and other

ingredients) may have an effect in humans or wildlife similar to an effect produced by a “naturally

occurring estrogen, or other such endocrine effects as the Administrator may designate.” Between

October 2009 and February 2010, EPA issued test orders/data call-ins for the first group of 67 chemicals,

which contains 58 pesticide active ingredients and 9 inert ingredients. This list of chemicals was selected

based on the potential for human exposure through pathways such as food and water, residential activity,

and certain post-application agricultural scenarios.

The Cry1F and Cry34/35Ab1 insect control proteins (as expressed in 4114 maize) are not among the

group of 58 pesticide active ingredients on the initial list to be screened under the EDSP.

C. Potential for Cry1F and Cry34/35Ab1 Interactions

Herman and Storer (2004) tested for an interaction between Cry1F and Cry34/35Ab1 when they were

combined in artificial diet bioassays using sensitive insect species. No synergism or antagonism between

Cry1F and Cry34/35Ab1 was seen in the tests. Therefore, the absence of antagonistic or synergistic

effects in sensitive organisms makes it extremely unlikely that the Cry1F x Cry34/35Ab1 proteins

produced in Herculex® XTRA (EPA Reg. No: 29964-5) will have any adverse effect on non-target

organisms different from what was seen in individual Cry protein testing (BPPD 2010). These data and

conclusions are equally applicable to 4114 maize. Pioneer Hi-Bred International, Inc.

Study Number: PHI-2011-080 Page 92 of 100

# D. Environmental Assessment of 4114 Maize (Volume 5)

Pioneer has requested a seed increase registration for 4114 maize containing Bacillus thuringiensis

Cry1F and Cry34/35Ab1 proteins and the genetic material necessary for their production in all corn lines

and varieties. The pat selectable marker gene was used in the transformation process, and the PAT

protein confers tolerance to the herbicidal active ingredient glufosinate-ammonium.

The Cry proteins expressed in 4114 maize are intended, when combined in future products with other Bt

traits to provide multiple modes of action, to control the lepidopteran pests such as European corn borer

(ECB, Ostrinia nubilalis ), corn earworm (CEW, Helicoverpa zea ), fall armyworm (FAW, Spodoptera

frugiperda ), and black cutworm (BCW, Agrotis ipsilon ), and coleopteran pests such as western corn

rootworm ( Diabrotica virgifera virgifera ) and northern corn rootworm ( Diabrotica barberi ). These are all

pests of corn in the United States. These pests feed on various parts of the corn plant, including the base

of seedlings and on the stalk, leaf, and ear tissue of corn plants. They can destroy the entire plant,

weaken the stalk, and/or damage the ear. In areas where one or more of these pests is prevalent,

significant financial losses are realized from decreased corn yields and increased expenditures on

chemical pest control agents, including organophosphate, carbamate and pyrethroid insecticides.

Pioneer has conducted an environmental risk assessment of Cry1F and Cry34/35Ab1 when expressed in

4114 maize (Volume 5; PHI-2011-078). General topics covered in this assessment include:

specificity of Cry1F and Cry34/35Ab1 to target insect pests;

fate of Cry1F and Cry34/35Ab1 in the soil environment;

exposure assessment to soil dwelling organisms, predators, and pollen feeders;

hazard assessment of 4114 maize to soil ecology and function, soil-dwelling organisms, nontarget lepidoptera, non-target coleoptera, and other non-target organisms;

impact on wildlife; and

gene flow to related wild plants and its potential effects.

Because the concentration of Cry1F in 4114 maize pollen is ~1.5 times that of Cry1F in 1507 or

1507x59122 (Volume 8; PHI-2010-059), special consideration was given in the environmental risk

assessment to the impact of 4114 maize on non-target and beneficial organisms.

Table 9. Environmental Safety Data Submitted for 4114 Maize

Study Title Summary Study No.

Environmental Risk

Assessment for the

Cultivation of Maize

Containing Event DPØØ4114-3 (4114

maize) The risk of 4114 maize cultivation resulting in adverse

effects on non-target organisms and wildlife is expected to

be low, based on a comprehensive risk assessment which

includes exposure and hazard assessments. PHI-2011-078

(Volume 5)

References

Herman, R and N. Storer. 2004. Investigation of potential interaction between Cry1F and the binary

Cry34Ab1/Cry35Ab1 proteins. MRID 463438-06

BPPD 2010. Biopesticides Registration Action Document: Bacillus thuringiensis Cry34Ab1 and

Cry35Ab1 Proteins and the Genetic Material Necessary for Their Production (PHP17662 T-DNA) in Event

DAS-59122-7 Corn (OECD Unique Identifier: DAS-59122-7). September 2010. Pioneer Hi-Bred International, Inc.

Study Number: PHI-2011-080 Page 93 of 100

# S ECTION 

# VII. R ESIDUE 

# D ATA



# No Codex maximum residue levels exist for the plant-pesticides Bacillus thuringiensis Cry1F or

Cry34/35Ab1 proteins and the genetic material necessary for their production in corn. Furthermore, EPA

has established an exemption from the requirement of a tolerance for Cry1F, Cry34/35Ab1 and PAT

proteins. Therefore, not residue data is required for 4114 maize. Pioneer Hi-Bred International, Inc.

Study Number: PHI-2011-080 Page 94 of 100

# S ECTION 

# VIII. E FFICACY 

# D ATA



# Insect efficacy data of 4114 maize against key target pests are provided in Volume 4 (PHI-2011-077).

The results demonstrate that efficacy of 4114 maize is comparable with 1507x59122 maize against ECB,

SWCB, FAW, CRW, and BCW. 4114 maize is expected to have activity against the same pest spectrum

as 1507x59122 maize. Pioneer Hi-Bred International, Inc.

Study Number: PHI-2011-080 Page 95 of 100

# CONFIDENTIAL ATTACHMENT

{CBI Cross Reference Number 1}

EPA Form 8570-4

Confidential Statement of Formula

The Certified Limits presented in the attached CSFs represent the highest and lowest levels of

individual proteins measured during the course of studies on events or event combinations for

each plant tissue type. No entries are being made for the Nominal Concentration (column 13) of

each protein because their levels vary significantly by tissue type and by growth stages of the

plants. There is not an expected amount of each ingredient in a typical plant sample.

Pioneer is proposing a set of non-standard Certified Limits as allowed under 40 CFR §

158.175(c). These proteins are plant-incorporated protectants or marker proteins produced by

the plant, not in a traditional closely monitored and controlled manufacturing facility. Multiple

factors beyond the control of the Applicant could influence the levels of these proteins

synthesized by individual plants, including but not limited to environmental conditions, plant

growth stage, and the presence of biotic or abiotic stressors. For these reasons, the Applicant is

not providing the Certification required by 40 CFR § 158.175(e). Pioneer Hi-Bred International, Inc.

Study Number: PHI-2011-080 Page 96 of 100