CA3159670A1 - Brassica gat event dp-073496-4 and compositions and methods for the identification and/or detection thereof - Google Patents
Brassica gat event dp-073496-4 and compositions and methods for the identification and/or detection thereofInfo
- Publication number
- CA3159670A1 CA3159670A1 CA3159670A CA3159670A CA3159670A1 CA 3159670 A1 CA3159670 A1 CA 3159670A1 CA 3159670 A CA3159670 A CA 3159670A CA 3159670 A CA3159670 A CA 3159670A CA 3159670 A1 CA3159670 A1 CA 3159670A1
- Authority
- CA
- Canada
- Prior art keywords
- herbicide
- plant
- event
- dna
- polynucleotide
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 238000000034 method Methods 0.000 title claims abstract description 135
- 241000219198 Brassica Species 0.000 title claims abstract description 76
- 239000000203 mixture Substances 0.000 title abstract description 72
- 238000001514 detection method Methods 0.000 title abstract description 24
- 235000011331 Brassica Nutrition 0.000 title description 39
- XDDAORKBJWWYJS-UHFFFAOYSA-N glyphosate Chemical compound OC(=O)CNCP(O)(O)=O XDDAORKBJWWYJS-UHFFFAOYSA-N 0.000 claims abstract description 78
- 239000005562 Glyphosate Substances 0.000 claims abstract description 70
- 229940097068 glyphosate Drugs 0.000 claims abstract description 67
- 241000196324 Embryophyta Species 0.000 claims description 383
- 108091033319 polynucleotide Proteins 0.000 claims description 126
- 102000040430 polynucleotide Human genes 0.000 claims description 126
- 239000002157 polynucleotide Substances 0.000 claims description 126
- 230000009466 transformation Effects 0.000 claims description 23
- 230000002068 genetic effect Effects 0.000 claims description 17
- 108010039239 glyphosate N-acetyltransferase Proteins 0.000 claims description 5
- 206010021929 Infertility male Diseases 0.000 claims description 3
- 208000007466 Male Infertility Diseases 0.000 claims description 3
- 230000008635 plant growth Effects 0.000 claims description 2
- 108700019146 Transgenes Proteins 0.000 abstract description 34
- 230000009261 transgenic effect Effects 0.000 abstract description 21
- 238000003780 insertion Methods 0.000 abstract description 14
- 230000037431 insertion Effects 0.000 abstract description 14
- 238000009395 breeding Methods 0.000 abstract description 12
- FWMNVWWHGCHHJJ-SKKKGAJSSA-N 4-amino-1-[(2r)-6-amino-2-[[(2r)-2-[[(2r)-2-[[(2r)-2-amino-3-phenylpropanoyl]amino]-3-phenylpropanoyl]amino]-4-methylpentanoyl]amino]hexanoyl]piperidine-4-carboxylic acid Chemical compound C([C@H](C(=O)N[C@H](CC(C)C)C(=O)N[C@H](CCCCN)C(=O)N1CCC(N)(CC1)C(O)=O)NC(=O)[C@H](N)CC=1C=CC=CC=1)C1=CC=CC=C1 FWMNVWWHGCHHJJ-SKKKGAJSSA-N 0.000 abstract description 11
- 230000001488 breeding effect Effects 0.000 abstract description 8
- 238000012512 characterization method Methods 0.000 abstract description 8
- 230000002759 chromosomal effect Effects 0.000 abstract description 5
- 239000004009 herbicide Substances 0.000 description 313
- 230000002363 herbicidal effect Effects 0.000 description 215
- 108020004414 DNA Proteins 0.000 description 120
- 239000013615 primer Substances 0.000 description 112
- 108090000623 proteins and genes Proteins 0.000 description 81
- -1 but not limited to Substances 0.000 description 75
- 239000000523 sample Substances 0.000 description 69
- 238000003752 polymerase chain reaction Methods 0.000 description 66
- 239000012634 fragment Substances 0.000 description 57
- 230000000694 effects Effects 0.000 description 41
- 210000004027 cell Anatomy 0.000 description 39
- 230000014509 gene expression Effects 0.000 description 39
- 238000009396 hybridization Methods 0.000 description 38
- 239000003112 inhibitor Substances 0.000 description 35
- 239000002773 nucleotide Substances 0.000 description 32
- 125000003729 nucleotide group Chemical group 0.000 description 32
- 238000011282 treatment Methods 0.000 description 32
- 239000000126 substance Substances 0.000 description 31
- 239000000047 product Substances 0.000 description 29
- 239000004480 active ingredient Substances 0.000 description 25
- 238000003199 nucleic acid amplification method Methods 0.000 description 25
- 108091028043 Nucleic acid sequence Proteins 0.000 description 24
- 229940100389 Sulfonylurea Drugs 0.000 description 24
- 244000038559 crop plants Species 0.000 description 24
- 230000009471 action Effects 0.000 description 23
- 230000003321 amplification Effects 0.000 description 23
- 238000006243 chemical reaction Methods 0.000 description 23
- 230000001276 controlling effect Effects 0.000 description 23
- 240000000385 Brassica napus var. napus Species 0.000 description 21
- 230000006378 damage Effects 0.000 description 20
- 108091093088 Amplicon Proteins 0.000 description 19
- 235000014698 Brassica juncea var multisecta Nutrition 0.000 description 19
- 235000006008 Brassica napus var napus Nutrition 0.000 description 19
- 235000006618 Brassica rapa subsp oleifera Nutrition 0.000 description 19
- 235000004977 Brassica sinapistrum Nutrition 0.000 description 19
- 230000001976 improved effect Effects 0.000 description 19
- 239000000463 material Substances 0.000 description 19
- 102000004169 proteins and genes Human genes 0.000 description 19
- YROXIXLRRCOBKF-UHFFFAOYSA-N sulfonylurea Chemical class OC(=N)N=S(=O)=O YROXIXLRRCOBKF-UHFFFAOYSA-N 0.000 description 19
- 230000002195 synergetic effect Effects 0.000 description 18
- 230000000295 complement effect Effects 0.000 description 16
- 238000011161 development Methods 0.000 description 16
- 230000018109 developmental process Effects 0.000 description 16
- JJWLVOIRVHMVIS-UHFFFAOYSA-N isopropylamine Chemical compound CC(C)N JJWLVOIRVHMVIS-UHFFFAOYSA-N 0.000 description 15
- 239000013612 plasmid Substances 0.000 description 15
- 229920001184 polypeptide Polymers 0.000 description 15
- 108090000765 processed proteins & peptides Proteins 0.000 description 15
- 102000004196 processed proteins & peptides Human genes 0.000 description 15
- 210000001519 tissue Anatomy 0.000 description 15
- CAAMSDWKXXPUJR-UHFFFAOYSA-N 3,5-dihydro-4H-imidazol-4-one Chemical compound O=C1CNC=N1 CAAMSDWKXXPUJR-UHFFFAOYSA-N 0.000 description 14
- 239000002253 acid Substances 0.000 description 14
- 238000009472 formulation Methods 0.000 description 14
- 230000001105 regulatory effect Effects 0.000 description 14
- 241000894007 species Species 0.000 description 14
- 229910001369 Brass Inorganic materials 0.000 description 13
- 239000012472 biological sample Substances 0.000 description 13
- 239000010951 brass Substances 0.000 description 13
- 230000005764 inhibitory process Effects 0.000 description 12
- 238000007726 management method Methods 0.000 description 12
- 150000007523 nucleic acids Chemical class 0.000 description 12
- 150000003839 salts Chemical class 0.000 description 12
- 238000012408 PCR amplification Methods 0.000 description 11
- 208000027418 Wounds and injury Diseases 0.000 description 11
- 150000001875 compounds Chemical class 0.000 description 11
- 230000012010 growth Effects 0.000 description 11
- 208000014674 injury Diseases 0.000 description 11
- 230000035772 mutation Effects 0.000 description 11
- 239000002689 soil Substances 0.000 description 11
- 238000004422 calculation algorithm Methods 0.000 description 10
- 239000003795 chemical substances by application Substances 0.000 description 10
- 239000003550 marker Substances 0.000 description 10
- 102000004190 Enzymes Human genes 0.000 description 9
- 108090000790 Enzymes Proteins 0.000 description 9
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 description 9
- 239000008187 granular material Substances 0.000 description 9
- 239000011591 potassium Substances 0.000 description 9
- 229910052700 potassium Inorganic materials 0.000 description 9
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 8
- 239000000654 additive Substances 0.000 description 8
- 239000002671 adjuvant Substances 0.000 description 8
- 238000010367 cloning Methods 0.000 description 8
- 238000005516 engineering process Methods 0.000 description 8
- 238000002866 fluorescence resonance energy transfer Methods 0.000 description 8
- 230000001965 increasing effect Effects 0.000 description 8
- 102000039446 nucleic acids Human genes 0.000 description 8
- 108020004707 nucleic acids Proteins 0.000 description 8
- 239000002987 primer (paints) Substances 0.000 description 8
- 230000001568 sexual effect Effects 0.000 description 8
- 239000000243 solution Substances 0.000 description 8
- 238000001228 spectrum Methods 0.000 description 8
- 238000006467 substitution reaction Methods 0.000 description 8
- 238000013519 translation Methods 0.000 description 8
- 108010000700 Acetolactate synthase Proteins 0.000 description 7
- 108091026890 Coding region Proteins 0.000 description 7
- 102000053602 DNA Human genes 0.000 description 7
- 239000005504 Dicamba Substances 0.000 description 7
- 241000209082 Lolium Species 0.000 description 7
- 108091034117 Oligonucleotide Proteins 0.000 description 7
- 239000005616 Rimsulfuron Substances 0.000 description 7
- 238000007792 addition Methods 0.000 description 7
- 238000004458 analytical method Methods 0.000 description 7
- 150000001558 benzoic acid derivatives Chemical class 0.000 description 7
- 230000015572 biosynthetic process Effects 0.000 description 7
- IWEDIXLBFLAXBO-UHFFFAOYSA-N dicamba Chemical compound COC1=C(Cl)C=CC(Cl)=C1C(O)=O IWEDIXLBFLAXBO-UHFFFAOYSA-N 0.000 description 7
- 230000000967 entomopathogenic effect Effects 0.000 description 7
- 235000013305 food Nutrition 0.000 description 7
- 239000004615 ingredient Substances 0.000 description 7
- 239000002917 insecticide Substances 0.000 description 7
- 230000008569 process Effects 0.000 description 7
- MEFOUWRMVYJCQC-UHFFFAOYSA-N rimsulfuron Chemical compound CCS(=O)(=O)C1=CC=CN=C1S(=O)(=O)NC(=O)NC1=NC(OC)=CC(OC)=N1 MEFOUWRMVYJCQC-UHFFFAOYSA-N 0.000 description 7
- 125000000565 sulfonamide group Chemical group 0.000 description 7
- 241000193830 Bacillus <bacterium> Species 0.000 description 6
- ROSDSFDQCJNGOL-UHFFFAOYSA-N Dimethylamine Chemical compound CNC ROSDSFDQCJNGOL-UHFFFAOYSA-N 0.000 description 6
- ZHNUHDYFZUAESO-UHFFFAOYSA-N Formamide Chemical compound NC=O ZHNUHDYFZUAESO-UHFFFAOYSA-N 0.000 description 6
- 239000005578 Mesotrione Substances 0.000 description 6
- 239000005623 Thifensulfuron-methyl Substances 0.000 description 6
- 241000607479 Yersinia pestis Species 0.000 description 6
- 239000011543 agarose gel Substances 0.000 description 6
- 230000008901 benefit Effects 0.000 description 6
- 238000012217 deletion Methods 0.000 description 6
- 230000037430 deletion Effects 0.000 description 6
- 230000007613 environmental effect Effects 0.000 description 6
- NYPJDWWKZLNGGM-UHFFFAOYSA-N fenvalerate Aalpha Natural products C=1C=C(Cl)C=CC=1C(C(C)C)C(=O)OC(C#N)C(C=1)=CC=CC=1OC1=CC=CC=C1 NYPJDWWKZLNGGM-UHFFFAOYSA-N 0.000 description 6
- 239000000417 fungicide Substances 0.000 description 6
- 239000008202 granule composition Substances 0.000 description 6
- 238000010348 incorporation Methods 0.000 description 6
- 239000011159 matrix material Substances 0.000 description 6
- KPUREKXXPHOJQT-UHFFFAOYSA-N mesotrione Chemical compound [O-][N+](=O)C1=CC(S(=O)(=O)C)=CC=C1C(=O)C1C(=O)CCCC1=O KPUREKXXPHOJQT-UHFFFAOYSA-N 0.000 description 6
- 159000000000 sodium salts Chemical class 0.000 description 6
- UCSJYZPVAKXKNQ-HZYVHMACSA-N streptomycin Chemical compound CN[C@H]1[C@H](O)[C@@H](O)[C@H](CO)O[C@H]1O[C@@H]1[C@](C=O)(O)[C@H](C)O[C@H]1O[C@@H]1[C@@H](NC(N)=N)[C@H](O)[C@@H](NC(N)=N)[C@H](O)[C@H]1O UCSJYZPVAKXKNQ-HZYVHMACSA-N 0.000 description 6
- 238000012360 testing method Methods 0.000 description 6
- AHTPATJNIAFOLR-UHFFFAOYSA-N thifensulfuron-methyl Chemical group S1C=CC(S(=O)(=O)NC(=O)NC=2N=C(OC)N=C(C)N=2)=C1C(=O)OC AHTPATJNIAFOLR-UHFFFAOYSA-N 0.000 description 6
- CLQMBPJKHLGMQK-UHFFFAOYSA-N 2-(4-isopropyl-4-methyl-5-oxo-4,5-dihydro-1H-imidazol-2-yl)nicotinic acid Chemical compound N1C(=O)C(C(C)C)(C)N=C1C1=NC=CC=C1C(O)=O CLQMBPJKHLGMQK-UHFFFAOYSA-N 0.000 description 5
- CABMTIJINOIHOD-UHFFFAOYSA-N 2-[4-methyl-5-oxo-4-(propan-2-yl)-4,5-dihydro-1H-imidazol-2-yl]quinoline-3-carboxylic acid Chemical compound N1C(=O)C(C(C)C)(C)N=C1C1=NC2=CC=CC=C2C=C1C(O)=O CABMTIJINOIHOD-UHFFFAOYSA-N 0.000 description 5
- IAJOBQBIJHVGMQ-UHFFFAOYSA-N 2-amino-4-[hydroxy(methyl)phosphoryl]butanoic acid Chemical compound CP(O)(=O)CCC(N)C(O)=O IAJOBQBIJHVGMQ-UHFFFAOYSA-N 0.000 description 5
- 229930192334 Auxin Natural products 0.000 description 5
- 244000242024 Conyza bonariensis Species 0.000 description 5
- 230000004544 DNA amplification Effects 0.000 description 5
- 241000233866 Fungi Species 0.000 description 5
- 239000005981 Imazaquin Substances 0.000 description 5
- XVOKUMIPKHGGTN-UHFFFAOYSA-N Imazethapyr Chemical compound OC(=O)C1=CC(CC)=CN=C1C1=NC(C)(C(C)C)C(=O)N1 XVOKUMIPKHGGTN-UHFFFAOYSA-N 0.000 description 5
- 108090000848 Ubiquitin Proteins 0.000 description 5
- 102000044159 Ubiquitin Human genes 0.000 description 5
- 240000008042 Zea mays Species 0.000 description 5
- 235000002017 Zea mays subsp mays Nutrition 0.000 description 5
- 150000007513 acids Chemical class 0.000 description 5
- 230000000996 additive effect Effects 0.000 description 5
- 239000003905 agrochemical Substances 0.000 description 5
- 239000002363 auxin Substances 0.000 description 5
- 230000008859 change Effects 0.000 description 5
- 230000010354 integration Effects 0.000 description 5
- 239000012528 membrane Substances 0.000 description 5
- 108020004999 messenger RNA Proteins 0.000 description 5
- 230000004048 modification Effects 0.000 description 5
- 238000012986 modification Methods 0.000 description 5
- 238000012545 processing Methods 0.000 description 5
- 238000003786 synthesis reaction Methods 0.000 description 5
- YMXOXAPKZDWXLY-QWRGUYRKSA-N tribenuron methyl Chemical group COC(=O)[C@H]1CCCC[C@@H]1S(=O)(=O)NC(=O)N(C)C1=NC(C)=NC(OC)=N1 YMXOXAPKZDWXLY-QWRGUYRKSA-N 0.000 description 5
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 4
- 239000005631 2,4-Dichlorophenoxyacetic acid Substances 0.000 description 4
- 240000006122 Chenopodium album Species 0.000 description 4
- 241000537219 Deltabaculovirus Species 0.000 description 4
- 238000002965 ELISA Methods 0.000 description 4
- 239000005950 Oxamyl Substances 0.000 description 4
- 108020001991 Protoporphyrinogen Oxidase Proteins 0.000 description 4
- 102000005135 Protoporphyrinogen oxidase Human genes 0.000 description 4
- 239000005614 Quizalofop-P-ethyl Substances 0.000 description 4
- DBMJMQXJHONAFJ-UHFFFAOYSA-M Sodium laurylsulphate Chemical compound [Na+].CCCCCCCCCCCCOS([O-])(=O)=O DBMJMQXJHONAFJ-UHFFFAOYSA-M 0.000 description 4
- 241000700605 Viruses Species 0.000 description 4
- 125000003275 alpha amino acid group Chemical group 0.000 description 4
- 125000000539 amino acid group Chemical group 0.000 description 4
- NSWAMPCUPHPTTC-UHFFFAOYSA-N chlorimuron-ethyl Chemical group CCOC(=O)C1=CC=CC=C1S(=O)(=O)NC(=O)NC1=NC(Cl)=CC(OC)=N1 NSWAMPCUPHPTTC-UHFFFAOYSA-N 0.000 description 4
- MZZBPDKVEFVLFF-UHFFFAOYSA-N cyanazine Chemical compound CCNC1=NC(Cl)=NC(NC(C)(C)C#N)=N1 MZZBPDKVEFVLFF-UHFFFAOYSA-N 0.000 description 4
- XQUXKZZNEFRCAW-UHFFFAOYSA-N fenpropathrin Chemical compound CC1(C)C(C)(C)C1C(=O)OC(C#N)C1=CC=CC(OC=2C=CC=CC=2)=C1 XQUXKZZNEFRCAW-UHFFFAOYSA-N 0.000 description 4
- WPOOICLZIIBUBM-UHFFFAOYSA-H iron;iron(3+);methyl-dioxido-oxo-$l^{5}-arsane Chemical compound [Fe].[Fe+3].[Fe+3].C[As]([O-])([O-])=O.C[As]([O-])([O-])=O.C[As]([O-])([O-])=O WPOOICLZIIBUBM-UHFFFAOYSA-H 0.000 description 4
- 230000008018 melting Effects 0.000 description 4
- 238000002844 melting Methods 0.000 description 4
- 238000002156 mixing Methods 0.000 description 4
- YTYGAJLZOJPJGH-UHFFFAOYSA-N noviflumuron Chemical compound FC1=C(Cl)C(OC(F)(F)C(C(F)(F)F)F)=C(Cl)C=C1NC(=O)NC(=O)C1=C(F)C=CC=C1F YTYGAJLZOJPJGH-UHFFFAOYSA-N 0.000 description 4
- KZAUOCCYDRDERY-UHFFFAOYSA-N oxamyl Chemical compound CNC(=O)ON=C(SC)C(=O)N(C)C KZAUOCCYDRDERY-UHFFFAOYSA-N 0.000 description 4
- 239000000575 pesticide Substances 0.000 description 4
- 230000008488 polyadenylation Effects 0.000 description 4
- 239000013641 positive control Substances 0.000 description 4
- OSUHJPCHFDQAIT-GFCCVEGCSA-N quizalofop-P-ethyl Chemical group C1=CC(O[C@H](C)C(=O)OCC)=CC=C1OC1=CN=C(C=C(Cl)C=C2)C2=N1 OSUHJPCHFDQAIT-GFCCVEGCSA-N 0.000 description 4
- 230000010076 replication Effects 0.000 description 4
- 230000004044 response Effects 0.000 description 4
- 239000011780 sodium chloride Substances 0.000 description 4
- 235000019333 sodium laurylsulphate Nutrition 0.000 description 4
- XLNZEKHULJKQBA-UHFFFAOYSA-N terbufos Chemical compound CCOP(=S)(OCC)SCSC(C)(C)C XLNZEKHULJKQBA-UHFFFAOYSA-N 0.000 description 4
- 230000002103 transcriptional effect Effects 0.000 description 4
- 239000013598 vector Substances 0.000 description 4
- CKPCAYZTYMHQEX-NBVRZTHBSA-N (e)-1-(2,4-dichlorophenyl)-n-methoxy-2-pyridin-3-ylethanimine Chemical compound C=1C=C(Cl)C=C(Cl)C=1C(=N/OC)/CC1=CC=CN=C1 CKPCAYZTYMHQEX-NBVRZTHBSA-N 0.000 description 3
- MNHVNIJQQRJYDH-UHFFFAOYSA-N 2-[2-(1-chlorocyclopropyl)-3-(2-chlorophenyl)-2-hydroxypropyl]-1,2-dihydro-1,2,4-triazole-3-thione Chemical compound N1=CNC(=S)N1CC(C1(Cl)CC1)(O)CC1=CC=CC=C1Cl MNHVNIJQQRJYDH-UHFFFAOYSA-N 0.000 description 3
- 102100028626 4-hydroxyphenylpyruvate dioxygenase Human genes 0.000 description 3
- 108010068327 4-hydroxyphenylpyruvate dioxygenase Proteins 0.000 description 3
- 239000005660 Abamectin Substances 0.000 description 3
- 108700028369 Alleles Proteins 0.000 description 3
- 244000036975 Ambrosia artemisiifolia Species 0.000 description 3
- 108700003918 Bacillus Thuringiensis insecticidal crystal Proteins 0.000 description 3
- 241000194108 Bacillus licheniformis Species 0.000 description 3
- 241000193388 Bacillus thuringiensis Species 0.000 description 3
- 241000894006 Bacteria Species 0.000 description 3
- 239000005488 Bispyribac Substances 0.000 description 3
- 235000009344 Chenopodium album Nutrition 0.000 description 3
- 239000005944 Chlorpyrifos Substances 0.000 description 3
- 241000207892 Convolvulus Species 0.000 description 3
- 235000004385 Conyza canadensis Nutrition 0.000 description 3
- 235000005853 Cyperus esculentus Nutrition 0.000 description 3
- 239000003155 DNA primer Substances 0.000 description 3
- 102000016928 DNA-directed DNA polymerase Human genes 0.000 description 3
- 108010014303 DNA-directed DNA polymerase Proteins 0.000 description 3
- 239000005507 Diflufenican Substances 0.000 description 3
- 244000025670 Eleusine indica Species 0.000 description 3
- 235000014716 Eleusine indica Nutrition 0.000 description 3
- 239000005533 Fluometuron Substances 0.000 description 3
- 239000005785 Fluquinconazole Substances 0.000 description 3
- 235000014820 Galium aparine Nutrition 0.000 description 3
- 239000005561 Glufosinate Substances 0.000 description 3
- 241000441237 Lolium lowei Species 0.000 description 3
- 239000005584 Metsulfuron-methyl Substances 0.000 description 3
- 238000002105 Southern blotting Methods 0.000 description 3
- 229920002472 Starch Polymers 0.000 description 3
- ATJFFYVFTNAWJD-UHFFFAOYSA-N Tin Chemical compound [Sn] ATJFFYVFTNAWJD-UHFFFAOYSA-N 0.000 description 3
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 description 3
- 230000000895 acaricidal effect Effects 0.000 description 3
- 239000000642 acaricide Substances 0.000 description 3
- 108020002494 acetyltransferase Proteins 0.000 description 3
- 150000001413 amino acids Chemical class 0.000 description 3
- 230000000844 anti-bacterial effect Effects 0.000 description 3
- 238000003556 assay Methods 0.000 description 3
- MXWJVTOOROXGIU-UHFFFAOYSA-N atrazine Chemical compound CCNC1=NC(Cl)=NC(NC(C)C)=N1 MXWJVTOOROXGIU-UHFFFAOYSA-N 0.000 description 3
- 229940097012 bacillus thuringiensis Drugs 0.000 description 3
- 239000003899 bactericide agent Substances 0.000 description 3
- ZOMSMJKLGFBRBS-UHFFFAOYSA-N bentazone Chemical compound C1=CC=C2NS(=O)(=O)N(C(C)C)C(=O)C2=C1 ZOMSMJKLGFBRBS-UHFFFAOYSA-N 0.000 description 3
- GINJFDRNADDBIN-FXQIFTODSA-N bilanafos Chemical compound OC(=O)[C@H](C)NC(=O)[C@H](C)NC(=O)[C@@H](N)CCP(C)(O)=O GINJFDRNADDBIN-FXQIFTODSA-N 0.000 description 3
- 230000004071 biological effect Effects 0.000 description 3
- 239000012620 biological material Substances 0.000 description 3
- RYVIXQCRCQLFCM-UHFFFAOYSA-N bispyribac Chemical compound COC1=CC(OC)=NC(OC=2C(=C(OC=3N=C(OC)C=C(OC)N=3)C=CC=2)C(O)=O)=N1 RYVIXQCRCQLFCM-UHFFFAOYSA-N 0.000 description 3
- 238000004061 bleaching Methods 0.000 description 3
- 235000013877 carbamide Nutrition 0.000 description 3
- 239000003153 chemical reaction reagent Substances 0.000 description 3
- SBPBAQFWLVIOKP-UHFFFAOYSA-N chlorpyrifos Chemical compound CCOP(=S)(OCC)OC1=NC(Cl)=C(Cl)C=C1Cl SBPBAQFWLVIOKP-UHFFFAOYSA-N 0.000 description 3
- 239000011248 coating agent Substances 0.000 description 3
- 238000000576 coating method Methods 0.000 description 3
- 239000012141 concentrate Substances 0.000 description 3
- 235000005822 corn Nutrition 0.000 description 3
- 230000003247 decreasing effect Effects 0.000 description 3
- 238000009795 derivation Methods 0.000 description 3
- WYEHFWKAOXOVJD-UHFFFAOYSA-N diflufenican Chemical compound FC1=CC(F)=CC=C1NC(=O)C1=CC=CN=C1OC1=CC=CC(C(F)(F)F)=C1 WYEHFWKAOXOVJD-UHFFFAOYSA-N 0.000 description 3
- USIUVYZYUHIAEV-UHFFFAOYSA-N diphenyl ether Chemical class C=1C=CC=CC=1OC1=CC=CC=C1 USIUVYZYUHIAEV-UHFFFAOYSA-N 0.000 description 3
- 239000003623 enhancer Substances 0.000 description 3
- 238000000605 extraction Methods 0.000 description 3
- RZILCCPWPBTYDO-UHFFFAOYSA-N fluometuron Chemical compound CN(C)C(=O)NC1=CC=CC(C(F)(F)F)=C1 RZILCCPWPBTYDO-UHFFFAOYSA-N 0.000 description 3
- IJJVMEJXYNJXOJ-UHFFFAOYSA-N fluquinconazole Chemical compound C=1C=C(Cl)C=C(Cl)C=1N1C(=O)C2=CC(F)=CC=C2N=C1N1C=NC=N1 IJJVMEJXYNJXOJ-UHFFFAOYSA-N 0.000 description 3
- RUCAXVJJQQJZGU-UHFFFAOYSA-M hydron;2-(phosphonatomethylamino)acetate;trimethylsulfanium Chemical compound C[S+](C)C.OP(O)(=O)CNCC([O-])=O RUCAXVJJQQJZGU-UHFFFAOYSA-M 0.000 description 3
- SEOVTRFCIGRIMH-UHFFFAOYSA-N indole-3-acetic acid Chemical compound C1=CC=C2C(CC(=O)O)=CNC2=C1 SEOVTRFCIGRIMH-UHFFFAOYSA-N 0.000 description 3
- 230000002045 lasting effect Effects 0.000 description 3
- 125000005647 linker group Chemical group 0.000 description 3
- 238000004519 manufacturing process Methods 0.000 description 3
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 description 3
- RSMUVYRMZCOLBH-UHFFFAOYSA-N metsulfuron methyl Chemical group COC(=O)C1=CC=CC=C1S(=O)(=O)NC(=O)NC1=NC(C)=NC(OC)=N1 RSMUVYRMZCOLBH-UHFFFAOYSA-N 0.000 description 3
- 244000005700 microbiome Species 0.000 description 3
- 238000010369 molecular cloning Methods 0.000 description 3
- 238000012544 monitoring process Methods 0.000 description 3
- 230000001069 nematicidal effect Effects 0.000 description 3
- 239000005645 nematicide Substances 0.000 description 3
- 239000003921 oil Substances 0.000 description 3
- 235000019198 oils Nutrition 0.000 description 3
- FIKAKWIAUPDISJ-UHFFFAOYSA-L paraquat dichloride Chemical compound [Cl-].[Cl-].C1=C[N+](C)=CC=C1C1=CC=[N+](C)C=C1 FIKAKWIAUPDISJ-UHFFFAOYSA-L 0.000 description 3
- 239000002245 particle Substances 0.000 description 3
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 3
- TVLSRXXIMLFWEO-UHFFFAOYSA-N prochloraz Chemical compound C1=CN=CN1C(=O)N(CCC)CCOC1=C(Cl)C=C(Cl)C=C1Cl TVLSRXXIMLFWEO-UHFFFAOYSA-N 0.000 description 3
- 238000010791 quenching Methods 0.000 description 3
- 230000000171 quenching effect Effects 0.000 description 3
- 230000002829 reductive effect Effects 0.000 description 3
- 239000002336 ribonucleotide Substances 0.000 description 3
- GOLXNESZZPUPJE-UHFFFAOYSA-N spiromesifen Chemical compound CC1=CC(C)=CC(C)=C1C(C(O1)=O)=C(OC(=O)CC(C)(C)C)C11CCCC1 GOLXNESZZPUPJE-UHFFFAOYSA-N 0.000 description 3
- 239000007921 spray Substances 0.000 description 3
- 235000019698 starch Nutrition 0.000 description 3
- 239000008107 starch Substances 0.000 description 3
- 239000007858 starting material Substances 0.000 description 3
- 229960005322 streptomycin Drugs 0.000 description 3
- 229940124530 sulfonamide Drugs 0.000 description 3
- 150000003456 sulfonamides Chemical class 0.000 description 3
- 238000012546 transfer Methods 0.000 description 3
- XOPFESVZMSQIKC-UHFFFAOYSA-N triasulfuron Chemical compound COC1=NC(C)=NC(NC(=O)NS(=O)(=O)C=2C(=CC=CC=2)OCCCl)=N1 XOPFESVZMSQIKC-UHFFFAOYSA-N 0.000 description 3
- YWBFPKPWMSWWEA-UHFFFAOYSA-O triazolopyrimidine Chemical compound BrC1=CC=CC(C=2N=C3N=CN[N+]3=C(NCC=3C=CN=CC=3)C=2)=C1 YWBFPKPWMSWWEA-UHFFFAOYSA-O 0.000 description 3
- 238000011144 upstream manufacturing Methods 0.000 description 3
- 235000015112 vegetable and seed oil Nutrition 0.000 description 3
- VIXCLRUCUMWJFF-KGLIPLIRSA-N (1R,5S)-benzobicyclon Chemical compound CS(=O)(=O)c1ccc(C(=O)C2=C(Sc3ccccc3)[C@H]3CC[C@H](C3)C2=O)c(Cl)c1 VIXCLRUCUMWJFF-KGLIPLIRSA-N 0.000 description 2
- XERJKGMBORTKEO-VZUCSPMQSA-N (1e)-2-(ethylcarbamoylamino)-n-methoxy-2-oxoethanimidoyl cyanide Chemical compound CCNC(=O)NC(=O)C(\C#N)=N\OC XERJKGMBORTKEO-VZUCSPMQSA-N 0.000 description 2
- NHOWDZOIZKMVAI-UHFFFAOYSA-N (2-chlorophenyl)(4-chlorophenyl)pyrimidin-5-ylmethanol Chemical compound C=1N=CN=CC=1C(C=1C(=CC=CC=1)Cl)(O)C1=CC=C(Cl)C=C1 NHOWDZOIZKMVAI-UHFFFAOYSA-N 0.000 description 2
- IPPAUTOBDWNELX-UHFFFAOYSA-N (2-ethoxy-2-oxoethyl) 5-[2-chloro-4-(trifluoromethyl)phenoxy]-2-nitrobenzoate Chemical group C1=C([N+]([O-])=O)C(C(=O)OCC(=O)OCC)=CC(OC=2C(=CC(=CC=2)C(F)(F)F)Cl)=C1 IPPAUTOBDWNELX-UHFFFAOYSA-N 0.000 description 2
- ZMYFCFLJBGAQRS-IRXDYDNUSA-N (2R,3S)-epoxiconazole Chemical compound C1=CC(F)=CC=C1[C@@]1(CN2N=CN=C2)[C@H](C=2C(=CC=CC=2)Cl)O1 ZMYFCFLJBGAQRS-IRXDYDNUSA-N 0.000 description 2
- RYAUSSKQMZRMAI-ALOPSCKCSA-N (2S,6R)-4-[3-(4-tert-butylphenyl)-2-methylpropyl]-2,6-dimethylmorpholine Chemical compound C=1C=C(C(C)(C)C)C=CC=1CC(C)CN1C[C@H](C)O[C@H](C)C1 RYAUSSKQMZRMAI-ALOPSCKCSA-N 0.000 description 2
- CXNPLSGKWMLZPZ-GIFSMMMISA-N (2r,3r,6s)-3-[[(3s)-3-amino-5-[carbamimidoyl(methyl)amino]pentanoyl]amino]-6-(4-amino-2-oxopyrimidin-1-yl)-3,6-dihydro-2h-pyran-2-carboxylic acid Chemical compound O1[C@@H](C(O)=O)[C@H](NC(=O)C[C@@H](N)CCN(C)C(N)=N)C=C[C@H]1N1C(=O)N=C(N)C=C1 CXNPLSGKWMLZPZ-GIFSMMMISA-N 0.000 description 2
- LDVVMCZRFWMZSG-OLQVQODUSA-N (3ar,7as)-2-(trichloromethylsulfanyl)-3a,4,7,7a-tetrahydroisoindole-1,3-dione Chemical compound C1C=CC[C@H]2C(=O)N(SC(Cl)(Cl)Cl)C(=O)[C@H]21 LDVVMCZRFWMZSG-OLQVQODUSA-N 0.000 description 2
- PPDBOQMNKNNODG-NTEUORMPSA-N (5E)-5-(4-chlorobenzylidene)-2,2-dimethyl-1-(1,2,4-triazol-1-ylmethyl)cyclopentanol Chemical compound C1=NC=NN1CC1(O)C(C)(C)CC\C1=C/C1=CC=C(Cl)C=C1 PPDBOQMNKNNODG-NTEUORMPSA-N 0.000 description 2
- WRIDQFICGBMAFQ-UHFFFAOYSA-N (E)-8-Octadecenoic acid Natural products CCCCCCCCCC=CCCCCCCC(O)=O WRIDQFICGBMAFQ-UHFFFAOYSA-N 0.000 description 2
- ZFHGXWPMULPQSE-SZGBIDFHSA-N (Z)-(1S)-cis-tefluthrin Chemical compound FC1=C(F)C(C)=C(F)C(F)=C1COC(=O)[C@@H]1C(C)(C)[C@@H]1\C=C(/Cl)C(F)(F)F ZFHGXWPMULPQSE-SZGBIDFHSA-N 0.000 description 2
- QNBTYORWCCMPQP-JXAWBTAJSA-N (Z)-dimethomorph Chemical compound C1=C(OC)C(OC)=CC=C1C(\C=1C=CC(Cl)=CC=1)=C/C(=O)N1CCOCC1 QNBTYORWCCMPQP-JXAWBTAJSA-N 0.000 description 2
- OVXMBIVWNJDDSM-UHFFFAOYSA-N (benzhydrylideneamino) 2,6-bis[(4,6-dimethoxypyrimidin-2-yl)oxy]benzoate Chemical compound COC1=CC(OC)=NC(OC=2C(=C(OC=3N=C(OC)C=C(OC)N=3)C=CC=2)C(=O)ON=C(C=2C=CC=CC=2)C=2C=CC=CC=2)=N1 OVXMBIVWNJDDSM-UHFFFAOYSA-N 0.000 description 2
- IAKOZHOLGAGEJT-UHFFFAOYSA-N 1,1,1-trichloro-2,2-bis(p-methoxyphenyl)-Ethane Chemical compound C1=CC(OC)=CC=C1C(C(Cl)(Cl)Cl)C1=CC=C(OC)C=C1 IAKOZHOLGAGEJT-UHFFFAOYSA-N 0.000 description 2
- JWUCHKBSVLQQCO-UHFFFAOYSA-N 1-(2-fluorophenyl)-1-(4-fluorophenyl)-2-(1H-1,2,4-triazol-1-yl)ethanol Chemical compound C=1C=C(F)C=CC=1C(C=1C(=CC=CC=1)F)(O)CN1C=NC=N1 JWUCHKBSVLQQCO-UHFFFAOYSA-N 0.000 description 2
- PXMNMQRDXWABCY-UHFFFAOYSA-N 1-(4-chlorophenyl)-4,4-dimethyl-3-(1H-1,2,4-triazol-1-ylmethyl)pentan-3-ol Chemical compound C1=NC=NN1CC(O)(C(C)(C)C)CCC1=CC=C(Cl)C=C1 PXMNMQRDXWABCY-UHFFFAOYSA-N 0.000 description 2
- LQDARGUHUSPFNL-UHFFFAOYSA-N 1-[2-(2,4-dichlorophenyl)-3-(1,1,2,2-tetrafluoroethoxy)propyl]1,2,4-triazole Chemical compound C=1C=C(Cl)C=C(Cl)C=1C(COC(F)(F)C(F)F)CN1C=NC=N1 LQDARGUHUSPFNL-UHFFFAOYSA-N 0.000 description 2
- MGNFYQILYYYUBS-UHFFFAOYSA-N 1-[3-(4-tert-butylphenyl)-2-methylpropyl]piperidine Chemical compound C=1C=C(C(C)(C)C)C=CC=1CC(C)CN1CCCCC1 MGNFYQILYYYUBS-UHFFFAOYSA-N 0.000 description 2
- YIKWKLYQRFRGPM-UHFFFAOYSA-N 1-dodecylguanidine acetate Chemical compound CC(O)=O.CCCCCCCCCCCCN=C(N)N YIKWKLYQRFRGPM-UHFFFAOYSA-N 0.000 description 2
- NFGXHKASABOEEW-UHFFFAOYSA-N 1-methylethyl 11-methoxy-3,7,11-trimethyl-2,4-dodecadienoate Chemical compound COC(C)(C)CCCC(C)CC=CC(C)=CC(=O)OC(C)C NFGXHKASABOEEW-UHFFFAOYSA-N 0.000 description 2
- AAILEWXSEQLMNI-UHFFFAOYSA-N 1h-pyridazin-6-one Chemical class OC1=CC=CN=N1 AAILEWXSEQLMNI-UHFFFAOYSA-N 0.000 description 2
- YOYAIZYFCNQIRF-UHFFFAOYSA-N 2,6-dichlorobenzonitrile Chemical compound ClC1=CC=CC(Cl)=C1C#N YOYAIZYFCNQIRF-UHFFFAOYSA-N 0.000 description 2
- BDQWWOHKFDSADC-UHFFFAOYSA-N 2-(2,4-dichloro-3-methylphenoxy)-n-phenylpropanamide Chemical compound C=1C=CC=CC=1NC(=O)C(C)OC1=CC=C(Cl)C(C)=C1Cl BDQWWOHKFDSADC-UHFFFAOYSA-N 0.000 description 2
- WNTGYJSOUMFZEP-UHFFFAOYSA-N 2-(4-chloro-2-methylphenoxy)propanoic acid Chemical compound OC(=O)C(C)OC1=CC=C(Cl)C=C1C WNTGYJSOUMFZEP-UHFFFAOYSA-N 0.000 description 2
- UFNOUKDBUJZYDE-UHFFFAOYSA-N 2-(4-chlorophenyl)-3-cyclopropyl-1-(1H-1,2,4-triazol-1-yl)butan-2-ol Chemical compound C1=NC=NN1CC(O)(C=1C=CC(Cl)=CC=1)C(C)C1CC1 UFNOUKDBUJZYDE-UHFFFAOYSA-N 0.000 description 2
- NUPJIGQFXCQJBK-UHFFFAOYSA-N 2-(4-isopropyl-4-methyl-5-oxo-4,5-dihydro-1H-imidazol-2-yl)-5-(methoxymethyl)nicotinic acid Chemical compound OC(=O)C1=CC(COC)=CN=C1C1=NC(C)(C(C)C)C(=O)N1 NUPJIGQFXCQJBK-UHFFFAOYSA-N 0.000 description 2
- WNZQDUSMALZDQF-UHFFFAOYSA-N 2-benzofuran-1(3H)-one Chemical compound C1=CC=C2C(=O)OCC2=C1 WNZQDUSMALZDQF-UHFFFAOYSA-N 0.000 description 2
- WVQBLGZPHOPPFO-UHFFFAOYSA-N 2-chloro-N-(2-ethyl-6-methylphenyl)-N-(1-methoxypropan-2-yl)acetamide Chemical compound CCC1=CC=CC(C)=C1N(C(C)COC)C(=O)CCl WVQBLGZPHOPPFO-UHFFFAOYSA-N 0.000 description 2
- IRCMYGHHKLLGHV-UHFFFAOYSA-N 2-ethoxy-3,3-dimethyl-2,3-dihydro-1-benzofuran-5-yl methanesulfonate Chemical compound C1=C(OS(C)(=O)=O)C=C2C(C)(C)C(OCC)OC2=C1 IRCMYGHHKLLGHV-UHFFFAOYSA-N 0.000 description 2
- ZDOOQPFIGYHZFV-UHFFFAOYSA-N 2-ethyl-4-[(4-phenoxyphenoxy)methyl]-1,3-dioxolane Chemical compound O1C(CC)OCC1COC(C=C1)=CC=C1OC1=CC=CC=C1 ZDOOQPFIGYHZFV-UHFFFAOYSA-N 0.000 description 2
- LAUKAWOMRTYHJK-UHFFFAOYSA-N 2-pyrimidin-2-yloxysulfanylbenzoic acid Chemical compound OC(=O)C1=CC=CC=C1SOC1=NC=CC=N1 LAUKAWOMRTYHJK-UHFFFAOYSA-N 0.000 description 2
- XFTOZRASFIPMOY-UHFFFAOYSA-N 2-pyrimidin-2-ylsulfanylbenzoic acid Chemical class OC(=O)C1=CC=CC=C1SC1=NC=CC=N1 XFTOZRASFIPMOY-UHFFFAOYSA-N 0.000 description 2
- LQJBNNIYVWPHFW-UHFFFAOYSA-N 20:1omega9c fatty acid Natural products CCCCCCCCCCC=CCCCCCCCC(O)=O LQJBNNIYVWPHFW-UHFFFAOYSA-N 0.000 description 2
- UPMXNNIRAGDFEH-UHFFFAOYSA-N 3,5-dibromo-4-hydroxybenzonitrile Chemical compound OC1=C(Br)C=C(C#N)C=C1Br UPMXNNIRAGDFEH-UHFFFAOYSA-N 0.000 description 2
- SOUGWDPPRBKJEX-UHFFFAOYSA-N 3,5-dichloro-N-(1-chloro-3-methyl-2-oxopentan-3-yl)-4-methylbenzamide Chemical compound ClCC(=O)C(C)(CC)NC(=O)C1=CC(Cl)=C(C)C(Cl)=C1 SOUGWDPPRBKJEX-UHFFFAOYSA-N 0.000 description 2
- XMTQQYYKAHVGBJ-UHFFFAOYSA-N 3-(3,4-DICHLOROPHENYL)-1,1-DIMETHYLUREA Chemical compound CN(C)C(=O)NC1=CC=C(Cl)C(Cl)=C1 XMTQQYYKAHVGBJ-UHFFFAOYSA-N 0.000 description 2
- KKADPXVIOXHVKN-UHFFFAOYSA-M 3-(4-hydroxyphenyl)pyruvate Chemical compound OC1=CC=C(CC(=O)C([O-])=O)C=C1 KKADPXVIOXHVKN-UHFFFAOYSA-M 0.000 description 2
- ZXVONLUNISGICL-UHFFFAOYSA-N 4,6-dinitro-o-cresol Chemical compound CC1=CC([N+]([O-])=O)=CC([N+]([O-])=O)=C1O ZXVONLUNISGICL-UHFFFAOYSA-N 0.000 description 2
- RQDJADAKIFFEKQ-UHFFFAOYSA-N 4-(4-chlorophenyl)-2-phenyl-2-(1,2,4-triazol-1-ylmethyl)butanenitrile Chemical compound C1=CC(Cl)=CC=C1CCC(C=1C=CC=CC=1)(C#N)CN1N=CN=C1 RQDJADAKIFFEKQ-UHFFFAOYSA-N 0.000 description 2
- ZOGDSYNXUXQGHF-UHFFFAOYSA-N 5-(3-butanoyl-2,4,6-trimethylphenyl)-2-[(Z)-N-ethoxy-C-ethylcarbonimidoyl]-3-hydroxycyclohex-2-en-1-one Chemical compound C(C)ON=C(CC)/C=1C(CC(CC1O)C1=C(C(=C(C=C1C)C)C(CCC)=O)C)=O ZOGDSYNXUXQGHF-UHFFFAOYSA-N 0.000 description 2
- NYRMIJKDBAQCHC-UHFFFAOYSA-N 5-(methylamino)-2-phenyl-4-[3-(trifluoromethyl)phenyl]furan-3(2H)-one Chemical compound O1C(NC)=C(C=2C=C(C=CC=2)C(F)(F)F)C(=O)C1C1=CC=CC=C1 NYRMIJKDBAQCHC-UHFFFAOYSA-N 0.000 description 2
- CTSLUCNDVMMDHG-UHFFFAOYSA-N 5-bromo-3-(butan-2-yl)-6-methylpyrimidine-2,4(1H,3H)-dione Chemical compound CCC(C)N1C(=O)NC(C)=C(Br)C1=O CTSLUCNDVMMDHG-UHFFFAOYSA-N 0.000 description 2
- XJFIKRXIJXAJGH-UHFFFAOYSA-N 5-chloro-1,3-dihydroimidazo[4,5-b]pyridin-2-one Chemical group ClC1=CC=C2NC(=O)NC2=N1 XJFIKRXIJXAJGH-UHFFFAOYSA-N 0.000 description 2
- PCCSBWNGDMYFCW-UHFFFAOYSA-N 5-methyl-5-(4-phenoxyphenyl)-3-(phenylamino)-1,3-oxazolidine-2,4-dione Chemical compound O=C1C(C)(C=2C=CC(OC=3C=CC=CC=3)=CC=2)OC(=O)N1NC1=CC=CC=C1 PCCSBWNGDMYFCW-UHFFFAOYSA-N 0.000 description 2
- PRZRAMLXTKZUHF-UHFFFAOYSA-N 5-oxo-n-sulfonyl-4h-triazole-1-carboxamide Chemical compound O=S(=O)=NC(=O)N1N=NCC1=O PRZRAMLXTKZUHF-UHFFFAOYSA-N 0.000 description 2
- QSBYPNXLFMSGKH-UHFFFAOYSA-N 9-Heptadecensaeure Natural products CCCCCCCC=CCCCCCCCC(O)=O QSBYPNXLFMSGKH-UHFFFAOYSA-N 0.000 description 2
- 240000006995 Abutilon theophrasti Species 0.000 description 2
- 102000000452 Acetyl-CoA carboxylase Human genes 0.000 description 2
- 108010016219 Acetyl-CoA carboxylase Proteins 0.000 description 2
- 239000002890 Aclonifen Substances 0.000 description 2
- XKJMBINCVNINCA-UHFFFAOYSA-N Alfalone Chemical compound CON(C)C(=O)NC1=CC=C(Cl)C(Cl)=C1 XKJMBINCVNINCA-UHFFFAOYSA-N 0.000 description 2
- 241000219318 Amaranthus Species 0.000 description 2
- 235000003129 Ambrosia artemisiifolia var elatior Nutrition 0.000 description 2
- 235000009051 Ambrosia paniculata var. peruviana Nutrition 0.000 description 2
- 239000003666 Amidosulfuron Substances 0.000 description 2
- CTTHWASMBLQOFR-UHFFFAOYSA-N Amidosulfuron Chemical compound COC1=CC(OC)=NC(NC(=O)NS(=O)(=O)N(C)S(C)(=O)=O)=N1 CTTHWASMBLQOFR-UHFFFAOYSA-N 0.000 description 2
- QGZKDVFQNNGYKY-UHFFFAOYSA-O Ammonium Chemical compound [NH4+] QGZKDVFQNNGYKY-UHFFFAOYSA-O 0.000 description 2
- 241001666377 Apera Species 0.000 description 2
- 239000005878 Azadirachtin Substances 0.000 description 2
- 239000005469 Azimsulfuron Substances 0.000 description 2
- QGQSRQPXXMTJCM-UHFFFAOYSA-N Benfuresate Chemical compound CCS(=O)(=O)OC1=CC=C2OCC(C)(C)C2=C1 QGQSRQPXXMTJCM-UHFFFAOYSA-N 0.000 description 2
- JDWQITFHZOBBFE-UHFFFAOYSA-N Benzofenap Chemical compound C=1C=C(Cl)C(C)=C(Cl)C=1C(=O)C=1C(C)=NN(C)C=1OCC(=O)C1=CC=C(C)C=C1 JDWQITFHZOBBFE-UHFFFAOYSA-N 0.000 description 2
- 239000005874 Bifenthrin Substances 0.000 description 2
- 108010018763 Biotin carboxylase Proteins 0.000 description 2
- 239000005739 Bordeaux mixture Substances 0.000 description 2
- 239000005489 Bromoxynil Substances 0.000 description 2
- 241001148727 Bromus tectorum Species 0.000 description 2
- 239000005885 Buprofezin Substances 0.000 description 2
- SPNQRCTZKIBOAX-UHFFFAOYSA-N Butralin Chemical compound CCC(C)NC1=C([N+]([O-])=O)C=C(C(C)(C)C)C=C1[N+]([O-])=O SPNQRCTZKIBOAX-UHFFFAOYSA-N 0.000 description 2
- JFLRKDZMHNBDQS-UCQUSYKYSA-N CC[C@H]1CCC[C@@H]([C@H](C(=O)C2=C[C@H]3[C@@H]4C[C@@H](C[C@H]4C(=C[C@H]3[C@@H]2CC(=O)O1)C)O[C@H]5[C@@H]([C@@H]([C@H]([C@@H](O5)C)OC)OC)OC)C)O[C@H]6CC[C@@H]([C@H](O6)C)N(C)C.CC[C@H]1CCC[C@@H]([C@H](C(=O)C2=C[C@H]3[C@@H]4C[C@@H](C[C@H]4C=C[C@H]3C2CC(=O)O1)O[C@H]5[C@@H]([C@@H]([C@H]([C@@H](O5)C)OC)OC)OC)C)O[C@H]6CC[C@@H]([C@H](O6)C)N(C)C Chemical compound CC[C@H]1CCC[C@@H]([C@H](C(=O)C2=C[C@H]3[C@@H]4C[C@@H](C[C@H]4C(=C[C@H]3[C@@H]2CC(=O)O1)C)O[C@H]5[C@@H]([C@@H]([C@H]([C@@H](O5)C)OC)OC)OC)C)O[C@H]6CC[C@@H]([C@H](O6)C)N(C)C.CC[C@H]1CCC[C@@H]([C@H](C(=O)C2=C[C@H]3[C@@H]4C[C@@H](C[C@H]4C=C[C@H]3C2CC(=O)O1)O[C@H]5[C@@H]([C@@H]([C@H]([C@@H](O5)C)OC)OC)OC)C)O[C@H]6CC[C@@H]([C@H](O6)C)N(C)C JFLRKDZMHNBDQS-UCQUSYKYSA-N 0.000 description 2
- 239000005745 Captan Substances 0.000 description 2
- 239000005490 Carbetamide Substances 0.000 description 2
- 239000005492 Carfentrazone-ethyl Substances 0.000 description 2
- 244000277285 Cassia obtusifolia Species 0.000 description 2
- 235000006719 Cassia obtusifolia Nutrition 0.000 description 2
- HSSBORCLYSCBJR-UHFFFAOYSA-N Chloramben Chemical compound NC1=CC(Cl)=CC(C(O)=O)=C1Cl HSSBORCLYSCBJR-UHFFFAOYSA-N 0.000 description 2
- RAPBNVDSDCTNRC-UHFFFAOYSA-N Chlorobenzilate Chemical compound C=1C=C(Cl)C=CC=1C(O)(C(=O)OCC)C1=CC=C(Cl)C=C1 RAPBNVDSDCTNRC-UHFFFAOYSA-N 0.000 description 2
- 239000005747 Chlorothalonil Substances 0.000 description 2
- 239000005494 Chlorotoluron Substances 0.000 description 2
- 239000005945 Chlorpyrifos-methyl Substances 0.000 description 2
- 239000005496 Chlorsulfuron Substances 0.000 description 2
- 239000005887 Chromafenozide Substances 0.000 description 2
- 239000005499 Clomazone Substances 0.000 description 2
- 241000233838 Commelina Species 0.000 description 2
- 241000132542 Conyza Species 0.000 description 2
- VYNOULHXXDFBLU-UHFFFAOYSA-N Cumyluron Chemical compound C=1C=CC=CC=1C(C)(C)NC(=O)NCC1=CC=CC=C1Cl VYNOULHXXDFBLU-UHFFFAOYSA-N 0.000 description 2
- 241001135545 Cydia pomonella granulovirus Species 0.000 description 2
- 239000005502 Cyhalofop-butyl Substances 0.000 description 2
- TYIYMOAHACZAMQ-CQSZACIVSA-N Cyhalofop-butyl Chemical group C1=CC(O[C@H](C)C(=O)OCCCC)=CC=C1OC1=CC=C(C#N)C=C1F TYIYMOAHACZAMQ-CQSZACIVSA-N 0.000 description 2
- 239000005756 Cymoxanil Substances 0.000 description 2
- 244000052363 Cynodon dactylon Species 0.000 description 2
- 239000005946 Cypermethrin Substances 0.000 description 2
- 240000001505 Cyperus odoratus Species 0.000 description 2
- 239000005757 Cyproconazole Substances 0.000 description 2
- 239000005891 Cyromazine Substances 0.000 description 2
- ZAKOWWREFLAJOT-CEFNRUSXSA-N D-alpha-tocopherylacetate Chemical compound CC(=O)OC1=C(C)C(C)=C2O[C@@](CCC[C@H](C)CCC[C@H](C)CCCC(C)C)(C)CCC2=C1C ZAKOWWREFLAJOT-CEFNRUSXSA-N 0.000 description 2
- 108020001019 DNA Primers Proteins 0.000 description 2
- NNYRZQHKCHEXSD-UHFFFAOYSA-N Daimuron Chemical compound C1=CC(C)=CC=C1NC(=O)NC(C)(C)C1=CC=CC=C1 NNYRZQHKCHEXSD-UHFFFAOYSA-N 0.000 description 2
- 239000005644 Dazomet Substances 0.000 description 2
- 239000005892 Deltamethrin Substances 0.000 description 2
- 239000005503 Desmedipham Substances 0.000 description 2
- HCRWJJJUKUVORR-UHFFFAOYSA-N Desmetryn Chemical compound CNC1=NC(NC(C)C)=NC(SC)=N1 HCRWJJJUKUVORR-UHFFFAOYSA-N 0.000 description 2
- QNXAVFXEJCPCJO-UHFFFAOYSA-N Diclosulam Chemical compound N=1N2C(OCC)=NC(F)=CC2=NC=1S(=O)(=O)NC1=C(Cl)C=CC=C1Cl QNXAVFXEJCPCJO-UHFFFAOYSA-N 0.000 description 2
- LWLJUMBEZJHXHV-UHFFFAOYSA-N Dienochlor Chemical compound ClC1=C(Cl)C(Cl)=C(Cl)C1(Cl)C1(Cl)C(Cl)=C(Cl)C(Cl)=C1Cl LWLJUMBEZJHXHV-UHFFFAOYSA-N 0.000 description 2
- 239000005760 Difenoconazole Substances 0.000 description 2
- LBGPXIPGGRQBJW-UHFFFAOYSA-N Difenzoquat Chemical compound C[N+]=1N(C)C(C=2C=CC=CC=2)=CC=1C1=CC=CC=C1 LBGPXIPGGRQBJW-UHFFFAOYSA-N 0.000 description 2
- DHWRNDJOGMTCPB-UHFFFAOYSA-N Dimefuron Chemical compound ClC1=CC(NC(=O)N(C)C)=CC=C1N1C(=O)OC(C(C)(C)C)=N1 DHWRNDJOGMTCPB-UHFFFAOYSA-N 0.000 description 2
- 239000005508 Dimethachlor Substances 0.000 description 2
- 239000005947 Dimethoate Substances 0.000 description 2
- 239000005761 Dimethomorph Substances 0.000 description 2
- 239000005762 Dimoxystrobin Substances 0.000 description 2
- OFDYMSKSGFSLLM-UHFFFAOYSA-N Dinitramine Chemical compound CCN(CC)C1=C([N+]([O-])=O)C=C(C(F)(F)F)C(N)=C1[N+]([O-])=O OFDYMSKSGFSLLM-UHFFFAOYSA-N 0.000 description 2
- IIPZYDQGBIWLBU-UHFFFAOYSA-N Dinoterb Chemical compound CC(C)(C)C1=CC([N+]([O-])=O)=CC([N+]([O-])=O)=C1O IIPZYDQGBIWLBU-UHFFFAOYSA-N 0.000 description 2
- YUBJPYNSGLJZPQ-UHFFFAOYSA-N Dithiopyr Chemical compound CSC(=O)C1=C(C(F)F)N=C(C(F)(F)F)C(C(=O)SC)=C1CC(C)C YUBJPYNSGLJZPQ-UHFFFAOYSA-N 0.000 description 2
- 239000005510 Diuron Substances 0.000 description 2
- 239000005766 Dodine Substances 0.000 description 2
- GUVLYNGULCJVDO-UHFFFAOYSA-N EPTC Chemical compound CCCN(CCC)C(=O)SCC GUVLYNGULCJVDO-UHFFFAOYSA-N 0.000 description 2
- 239000005894 Emamectin Substances 0.000 description 2
- 239000005767 Epoxiconazole Substances 0.000 description 2
- 239000005895 Esfenvalerate Substances 0.000 description 2
- BXEHUCNTIZGSOJ-UHFFFAOYSA-N Esprocarb Chemical compound CC(C)C(C)N(CC)C(=O)SCC1=CC=CC=C1 BXEHUCNTIZGSOJ-UHFFFAOYSA-N 0.000 description 2
- FNELVJVBIYMIMC-UHFFFAOYSA-N Ethiprole Chemical compound N1=C(C#N)C(S(=O)CC)=C(N)N1C1=C(Cl)C=C(C(F)(F)F)C=C1Cl FNELVJVBIYMIMC-UHFFFAOYSA-N 0.000 description 2
- 239000005512 Ethofumesate Substances 0.000 description 2
- 239000005897 Etoxazole Substances 0.000 description 2
- 239000005772 Famoxadone Substances 0.000 description 2
- 239000005774 Fenamidone Substances 0.000 description 2
- 239000005958 Fenamiphos (aka phenamiphos) Substances 0.000 description 2
- 239000005656 Fenazaquin Substances 0.000 description 2
- 239000005775 Fenbuconazole Substances 0.000 description 2
- 239000005898 Fenoxycarb Substances 0.000 description 2
- 239000005777 Fenpropidin Substances 0.000 description 2
- 239000005778 Fenpropimorph Substances 0.000 description 2
- 239000005657 Fenpyroximate Substances 0.000 description 2
- LLQPHQFNMLZJMP-UHFFFAOYSA-N Fentrazamide Chemical compound N1=NN(C=2C(=CC=CC=2)Cl)C(=O)N1C(=O)N(CC)C1CCCCC1 LLQPHQFNMLZJMP-UHFFFAOYSA-N 0.000 description 2
- 239000005514 Flazasulfuron Substances 0.000 description 2
- HWATZEJQIXKWQS-UHFFFAOYSA-N Flazasulfuron Chemical compound COC1=CC(OC)=NC(NC(=O)NS(=O)(=O)C=2C(=CC=CN=2)C(F)(F)F)=N1 HWATZEJQIXKWQS-UHFFFAOYSA-N 0.000 description 2
- 239000005900 Flonicamid Substances 0.000 description 2
- 239000005529 Florasulam Substances 0.000 description 2
- QZXATCCPQKOEIH-UHFFFAOYSA-N Florasulam Chemical compound N=1N2C(OC)=NC=C(F)C2=NC=1S(=O)(=O)NC1=C(F)C=CC=C1F QZXATCCPQKOEIH-UHFFFAOYSA-N 0.000 description 2
- 239000005780 Fluazinam Substances 0.000 description 2
- 239000005781 Fludioxonil Substances 0.000 description 2
- 239000005531 Flufenacet Substances 0.000 description 2
- RXCPQSJAVKGONC-UHFFFAOYSA-N Flumetsulam Chemical compound N1=C2N=C(C)C=CN2N=C1S(=O)(=O)NC1=C(F)C=CC=C1F RXCPQSJAVKGONC-UHFFFAOYSA-N 0.000 description 2
- IRECWLYBCAZIJM-UHFFFAOYSA-N Flumiclorac pentyl Chemical group C1=C(Cl)C(OCC(=O)OCCCCC)=CC(N2C(C3=C(CCCC3)C2=O)=O)=C1F IRECWLYBCAZIJM-UHFFFAOYSA-N 0.000 description 2
- 239000005784 Fluoxastrobin Substances 0.000 description 2
- AOQMRUTZEYVDIL-UHFFFAOYSA-N Flupoxam Chemical compound C=1C=C(Cl)C(COCC(F)(F)C(F)(F)F)=CC=1N1N=C(C(=O)N)N=C1C1=CC=CC=C1 AOQMRUTZEYVDIL-UHFFFAOYSA-N 0.000 description 2
- 239000005534 Flupyrsulfuron-methyl Substances 0.000 description 2
- YWBVHLJPRPCRSD-UHFFFAOYSA-N Fluridone Chemical compound O=C1C(C=2C=C(C=CC=2)C(F)(F)F)=CN(C)C=C1C1=CC=CC=C1 YWBVHLJPRPCRSD-UHFFFAOYSA-N 0.000 description 2
- 239000005559 Flurtamone Substances 0.000 description 2
- 239000005786 Flutolanil Substances 0.000 description 2
- 239000005787 Flutriafol Substances 0.000 description 2
- 239000005789 Folpet Substances 0.000 description 2
- 239000005560 Foramsulfuron Substances 0.000 description 2
- 101150075567 Glyat gene Proteins 0.000 description 2
- NYHBQMYGNKIUIF-UUOKFMHZSA-N Guanosine Chemical compound C1=NC=2C(=O)NC(N)=NC=2N1[C@@H]1O[C@H](CO)[C@@H](O)[C@H]1O NYHBQMYGNKIUIF-UUOKFMHZSA-N 0.000 description 2
- 239000005564 Halosulfuron methyl Substances 0.000 description 2
- FMGZEUWROYGLAY-UHFFFAOYSA-N Halosulfuron-methyl Chemical group ClC1=NN(C)C(S(=O)(=O)NC(=O)NC=2N=C(OC)C=C(OC)N=2)=C1C(=O)OC FMGZEUWROYGLAY-UHFFFAOYSA-N 0.000 description 2
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 description 2
- 239000005566 Imazamox Substances 0.000 description 2
- 239000005567 Imazosulfuron Substances 0.000 description 2
- NAGRVUXEKKZNHT-UHFFFAOYSA-N Imazosulfuron Chemical compound COC1=CC(OC)=NC(NC(=O)NS(=O)(=O)C=2N3C=CC=CC3=NC=2Cl)=N1 NAGRVUXEKKZNHT-UHFFFAOYSA-N 0.000 description 2
- 239000005906 Imidacloprid Substances 0.000 description 2
- 239000005907 Indoxacarb Substances 0.000 description 2
- 239000005796 Ipconazole Substances 0.000 description 2
- 240000001549 Ipomoea eriocarpa Species 0.000 description 2
- 235000005146 Ipomoea eriocarpa Nutrition 0.000 description 2
- JLLJHQLUZAKJFH-UHFFFAOYSA-N Isouron Chemical compound CN(C)C(=O)NC=1C=C(C(C)(C)C)ON=1 JLLJHQLUZAKJFH-UHFFFAOYSA-N 0.000 description 2
- 239000005570 Isoxaben Substances 0.000 description 2
- 239000005571 Isoxaflutole Substances 0.000 description 2
- 241000110847 Kochia Species 0.000 description 2
- 239000005800 Kresoxim-methyl Substances 0.000 description 2
- 239000005573 Linuron Substances 0.000 description 2
- 240000004296 Lolium perenne Species 0.000 description 2
- 239000005912 Lufenuron Substances 0.000 description 2
- 208000030984 MIRAGE syndrome Diseases 0.000 description 2
- 239000005949 Malathion Substances 0.000 description 2
- 239000005802 Mancozeb Substances 0.000 description 2
- 239000005807 Metalaxyl Substances 0.000 description 2
- 239000005956 Metaldehyde Substances 0.000 description 2
- 239000002169 Metam Substances 0.000 description 2
- 239000005579 Metamitron Substances 0.000 description 2
- 239000005580 Metazachlor Substances 0.000 description 2
- 239000005868 Metconazole Substances 0.000 description 2
- RRVIAQKBTUQODI-UHFFFAOYSA-N Methabenzthiazuron Chemical compound C1=CC=C2SC(N(C)C(=O)NC)=NC2=C1 RRVIAQKBTUQODI-UHFFFAOYSA-N 0.000 description 2
- 239000005916 Methomyl Substances 0.000 description 2
- 239000005917 Methoxyfenozide Substances 0.000 description 2
- 239000005581 Metobromuron Substances 0.000 description 2
- WLFDQEVORAMCIM-UHFFFAOYSA-N Metobromuron Chemical compound CON(C)C(=O)NC1=CC=C(Br)C=C1 WLFDQEVORAMCIM-UHFFFAOYSA-N 0.000 description 2
- VGHPMIFEKOFHHQ-UHFFFAOYSA-N Metosulam Chemical compound N1=C2N=C(OC)C=C(OC)N2N=C1S(=O)(=O)NC1=C(Cl)C=CC(C)=C1Cl VGHPMIFEKOFHHQ-UHFFFAOYSA-N 0.000 description 2
- 239000005582 Metosulam Substances 0.000 description 2
- 239000005810 Metrafenone Substances 0.000 description 2
- 235000003805 Musa ABB Group Nutrition 0.000 description 2
- 240000005561 Musa balbisiana Species 0.000 description 2
- WXZVAROIGSFCFJ-UHFFFAOYSA-N N,N-diethyl-2-(naphthalen-1-yloxy)propanamide Chemical compound C1=CC=C2C(OC(C)C(=O)N(CC)CC)=CC=CC2=C1 WXZVAROIGSFCFJ-UHFFFAOYSA-N 0.000 description 2
- XFOXDUJCOHBXRC-UHFFFAOYSA-N N-Ethyl-N-methyl-4-(trifluoromethyl)-2-(3,4-dimethoxyphenyl)benzamide Chemical compound CCN(C)C(=O)C1=CC=C(C(F)(F)F)C=C1C1=CC=C(OC)C(OC)=C1 XFOXDUJCOHBXRC-UHFFFAOYSA-N 0.000 description 2
- GRSMWKLPSNHDHA-UHFFFAOYSA-N Naphthalic anhydride Chemical compound C1=CC(C(=O)OC2=O)=C3C2=CC=CC3=C1 GRSMWKLPSNHDHA-UHFFFAOYSA-N 0.000 description 2
- 239000005585 Napropamide Substances 0.000 description 2
- 239000005586 Nicosulfuron Substances 0.000 description 2
- 239000004677 Nylon Substances 0.000 description 2
- 239000005642 Oleic acid Substances 0.000 description 2
- ZQPPMHVWECSIRJ-UHFFFAOYSA-N Oleic acid Natural products CCCCCCCCC=CCCCCCCCC(O)=O ZQPPMHVWECSIRJ-UHFFFAOYSA-N 0.000 description 2
- 108020005187 Oligonucleotide Probes Proteins 0.000 description 2
- 108700026244 Open Reading Frames Proteins 0.000 description 2
- 239000005588 Oxadiazon Substances 0.000 description 2
- CHNUNORXWHYHNE-UHFFFAOYSA-N Oxadiazon Chemical compound C1=C(Cl)C(OC(C)C)=CC(N2C(OC(=N2)C(C)(C)C)=O)=C1Cl CHNUNORXWHYHNE-UHFFFAOYSA-N 0.000 description 2
- 239000005589 Oxasulfuron Substances 0.000 description 2
- 239000005590 Oxyfluorfen Substances 0.000 description 2
- OQMBBFQZGJFLBU-UHFFFAOYSA-N Oxyfluorfen Chemical compound C1=C([N+]([O-])=O)C(OCC)=CC(OC=2C(=CC(=CC=2)C(F)(F)F)Cl)=C1 OQMBBFQZGJFLBU-UHFFFAOYSA-N 0.000 description 2
- 239000012807 PCR reagent Substances 0.000 description 2
- SGEJQUSYQTVSIU-UHFFFAOYSA-N Pebulate Chemical compound CCCCN(CC)C(=O)SCCC SGEJQUSYQTVSIU-UHFFFAOYSA-N 0.000 description 2
- 239000005814 Pencycuron Substances 0.000 description 2
- 239000005591 Pendimethalin Substances 0.000 description 2
- 239000005592 Penoxsulam Substances 0.000 description 2
- SYJGKVOENHZYMQ-UHFFFAOYSA-N Penoxsulam Chemical compound N1=C2C(OC)=CN=C(OC)N2N=C1NS(=O)(=O)C1=C(OCC(F)F)C=CC=C1C(F)(F)F SYJGKVOENHZYMQ-UHFFFAOYSA-N 0.000 description 2
- WGVWLKXZBUVUAM-UHFFFAOYSA-N Pentanochlor Chemical compound CCCC(C)C(=O)NC1=CC=C(C)C(Cl)=C1 WGVWLKXZBUVUAM-UHFFFAOYSA-N 0.000 description 2
- 239000005921 Phosmet Substances 0.000 description 2
- 108010060806 Photosystem II Protein Complex Proteins 0.000 description 2
- 239000005595 Picloram Substances 0.000 description 2
- 239000005923 Pirimicarb Substances 0.000 description 2
- 241001127637 Plantago Species 0.000 description 2
- 235000015266 Plantago major Nutrition 0.000 description 2
- YLPGTOIOYRQOHV-UHFFFAOYSA-N Pretilachlor Chemical compound CCCOCCN(C(=O)CCl)C1=C(CC)C=CC=C1CC YLPGTOIOYRQOHV-UHFFFAOYSA-N 0.000 description 2
- 239000005821 Propamocarb Substances 0.000 description 2
- 239000005600 Propaquizafop Substances 0.000 description 2
- 239000005822 Propiconazole Substances 0.000 description 2
- 239000005824 Proquinazid Substances 0.000 description 2
- 239000005825 Prothioconazole Substances 0.000 description 2
- 239000005605 Pyraflufen-ethyl Substances 0.000 description 2
- BGNQYGRXEXDAIQ-UHFFFAOYSA-N Pyrazosulfuron-ethyl Chemical group C1=NN(C)C(S(=O)(=O)NC(=O)NC=2N=C(OC)C=C(OC)N=2)=C1C(=O)OCC BGNQYGRXEXDAIQ-UHFFFAOYSA-N 0.000 description 2
- 239000005828 Pyrimethanil Substances 0.000 description 2
- 239000005927 Pyriproxyfen Substances 0.000 description 2
- 239000005607 Pyroxsulam Substances 0.000 description 2
- 239000005831 Quinoxyfen Substances 0.000 description 2
- 108091028664 Ribonucleotide Proteins 0.000 description 2
- CSPPKDPQLUUTND-NBVRZTHBSA-N Sethoxydim Chemical compound CCO\N=C(/CCC)C1=C(O)CC(CC(C)SCC)CC1=O CSPPKDPQLUUTND-NBVRZTHBSA-N 0.000 description 2
- JXVIIQLNUPXOII-UHFFFAOYSA-N Siduron Chemical compound CC1CCCCC1NC(=O)NC1=CC=CC=C1 JXVIIQLNUPXOII-UHFFFAOYSA-N 0.000 description 2
- 240000002439 Sorghum halepense Species 0.000 description 2
- 239000005930 Spinosad Substances 0.000 description 2
- 239000005837 Spiroxamine Substances 0.000 description 2
- 240000006694 Stellaria media Species 0.000 description 2
- 239000005618 Sulcotrione Substances 0.000 description 2
- NINIDFKCEFEMDL-UHFFFAOYSA-N Sulfur Chemical compound [S] NINIDFKCEFEMDL-UHFFFAOYSA-N 0.000 description 2
- 239000005839 Tebuconazole Substances 0.000 description 2
- 239000005937 Tebufenozide Substances 0.000 description 2
- 239000005658 Tebufenpyrad Substances 0.000 description 2
- HBPDKDSFLXWOAE-UHFFFAOYSA-N Tebuthiuron Chemical compound CNC(=O)N(C)C1=NN=C(C(C)(C)C)S1 HBPDKDSFLXWOAE-UHFFFAOYSA-N 0.000 description 2
- 239000005938 Teflubenzuron Substances 0.000 description 2
- 239000005939 Tefluthrin Substances 0.000 description 2
- NBQCNZYJJMBDKY-UHFFFAOYSA-N Terbacil Chemical compound CC=1NC(=O)N(C(C)(C)C)C(=O)C=1Cl NBQCNZYJJMBDKY-UHFFFAOYSA-N 0.000 description 2
- 239000005621 Terbuthylazine Substances 0.000 description 2
- 239000005840 Tetraconazole Substances 0.000 description 2
- 239000005941 Thiamethoxam Substances 0.000 description 2
- YIJZJEYQBAAWRJ-UHFFFAOYSA-N Thiazopyr Chemical compound N1=C(C(F)F)C(C(=O)OC)=C(CC(C)C)C(C=2SCCN=2)=C1C(F)(F)F YIJZJEYQBAAWRJ-UHFFFAOYSA-N 0.000 description 2
- QHTQREMOGMZHJV-UHFFFAOYSA-N Thiobencarb Chemical compound CCN(CC)C(=O)SCC1=CC=C(Cl)C=C1 QHTQREMOGMZHJV-UHFFFAOYSA-N 0.000 description 2
- 239000005842 Thiophanate-methyl Substances 0.000 description 2
- PHSUVQBHRAWOQD-UHFFFAOYSA-N Tiocarbazil Chemical compound CCC(C)N(C(C)CC)C(=O)SCC1=CC=CC=C1 PHSUVQBHRAWOQD-UHFFFAOYSA-N 0.000 description 2
- 108700029229 Transcriptional Regulatory Elements Proteins 0.000 description 2
- 239000005848 Tribasic copper sulfate Substances 0.000 description 2
- 239000005857 Trifloxystrobin Substances 0.000 description 2
- 239000005942 Triflumuron Substances 0.000 description 2
- 239000005859 Triticonazole Substances 0.000 description 2
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 description 2
- 241000972221 Urochloa decumbens Species 0.000 description 2
- 229930195482 Validamycin Natural products 0.000 description 2
- 244000067505 Xanthium strumarium Species 0.000 description 2
- 235000016383 Zea mays subsp huehuetenangensis Nutrition 0.000 description 2
- AMRQXHFXNZFDCH-SECBINFHSA-N [(2r)-1-(ethylamino)-1-oxopropan-2-yl] n-phenylcarbamate Chemical compound CCNC(=O)[C@@H](C)OC(=O)NC1=CC=CC=C1 AMRQXHFXNZFDCH-SECBINFHSA-N 0.000 description 2
- YASYVMFAVPKPKE-UHFFFAOYSA-N acephate Chemical compound COP(=O)(SC)NC(C)=O YASYVMFAVPKPKE-UHFFFAOYSA-N 0.000 description 2
- CGIHPACLZJDCBQ-UHFFFAOYSA-N acibenzolar Chemical compound SC(=O)C1=CC=CC2=C1SN=N2 CGIHPACLZJDCBQ-UHFFFAOYSA-N 0.000 description 2
- DDBMQDADIHOWIC-UHFFFAOYSA-N aclonifen Chemical compound C1=C([N+]([O-])=O)C(N)=C(Cl)C(OC=2C=CC=CC=2)=C1 DDBMQDADIHOWIC-UHFFFAOYSA-N 0.000 description 2
- QGLZXHRNAYXIBU-WEVVVXLNSA-N aldicarb Chemical compound CNC(=O)O\N=C\C(C)(C)SC QGLZXHRNAYXIBU-WEVVVXLNSA-N 0.000 description 2
- FPIPGXGPPPQFEQ-OVSJKPMPSA-N all-trans-retinol Chemical compound OC\C=C(/C)\C=C\C=C(/C)\C=C\C1=C(C)CCCC1(C)C FPIPGXGPPPQFEQ-OVSJKPMPSA-N 0.000 description 2
- RQVYBGPQFYCBGX-UHFFFAOYSA-N ametryn Chemical compound CCNC1=NC(NC(C)C)=NC(SC)=N1 RQVYBGPQFYCBGX-UHFFFAOYSA-N 0.000 description 2
- ORFPWVRKFLOQHK-UHFFFAOYSA-N amicarbazone Chemical compound CC(C)C1=NN(C(=O)NC(C)(C)C)C(=O)N1N ORFPWVRKFLOQHK-UHFFFAOYSA-N 0.000 description 2
- 238000000137 annealing Methods 0.000 description 2
- 235000003484 annual ragweed Nutrition 0.000 description 2
- 238000013459 approach Methods 0.000 description 2
- VGPYEHKOIGNJKV-UHFFFAOYSA-N asulam Chemical compound COC(=O)NS(=O)(=O)C1=CC=C(N)C=C1 VGPYEHKOIGNJKV-UHFFFAOYSA-N 0.000 description 2
- 239000005667 attractant Substances 0.000 description 2
- RRZXIRBKKLTSOM-XPNPUAGNSA-N avermectin B1a Chemical compound C1=C[C@H](C)[C@@H]([C@@H](C)CC)O[C@]11O[C@H](C\C=C(C)\[C@@H](O[C@@H]2O[C@@H](C)[C@H](O[C@@H]3O[C@@H](C)[C@H](O)[C@@H](OC)C3)[C@@H](OC)C2)[C@@H](C)\C=C\C=C/2[C@]3([C@H](C(=O)O4)C=C(C)[C@@H](O)[C@H]3OC\2)O)C[C@H]4C1 RRZXIRBKKLTSOM-XPNPUAGNSA-N 0.000 description 2
- VEHPJKVTJQSSKL-UHFFFAOYSA-N azadirachtin Natural products O1C2(C)C(C3(C=COC3O3)O)CC3C21C1(C)C(O)C(OCC2(OC(C)=O)C(CC3OC(=O)C(C)=CC)OC(C)=O)C2C32COC(C(=O)OC)(O)C12 VEHPJKVTJQSSKL-UHFFFAOYSA-N 0.000 description 2
- FTNJWQUOZFUQQJ-IRYYUVNJSA-N azadirachtin A Natural products C([C@@H]([C@]1(C=CO[C@H]1O1)O)[C@]2(C)O3)[C@H]1[C@]23[C@]1(C)[C@H](O)[C@H](OC[C@@]2([C@@H](C[C@@H]3OC(=O)C(\C)=C/C)OC(C)=O)C(=O)OC)[C@@H]2[C@]32CO[C@@](C(=O)OC)(O)[C@@H]12 FTNJWQUOZFUQQJ-IRYYUVNJSA-N 0.000 description 2
- FTNJWQUOZFUQQJ-NDAWSKJSSA-N azadirachtin A Chemical compound C([C@@H]([C@]1(C=CO[C@H]1O1)O)[C@]2(C)O3)[C@H]1[C@]23[C@]1(C)[C@H](O)[C@H](OC[C@@]2([C@@H](C[C@@H]3OC(=O)C(\C)=C\C)OC(C)=O)C(=O)OC)[C@@H]2[C@]32CO[C@@](C(=O)OC)(O)[C@@H]12 FTNJWQUOZFUQQJ-NDAWSKJSSA-N 0.000 description 2
- OTSAMNSACVKIOJ-UHFFFAOYSA-N azane;carbamoyl(ethoxy)phosphinic acid Chemical compound [NH4+].CCOP([O-])(=O)C(N)=O OTSAMNSACVKIOJ-UHFFFAOYSA-N 0.000 description 2
- MAHPNPYYQAIOJN-UHFFFAOYSA-N azimsulfuron Chemical compound COC1=CC(OC)=NC(NC(=O)NS(=O)(=O)C=2N(N=CC=2C2=NN(C)N=N2)C)=N1 MAHPNPYYQAIOJN-UHFFFAOYSA-N 0.000 description 2
- CJJOSEISRRTUQB-UHFFFAOYSA-N azinphos-methyl Chemical group C1=CC=C2C(=O)N(CSP(=S)(OC)OC)N=NC2=C1 CJJOSEISRRTUQB-UHFFFAOYSA-N 0.000 description 2
- WQRCEBAZAUAUQC-UHFFFAOYSA-N benazolin-ethyl Chemical group C1=CC=C2SC(=O)N(CC(=O)OCC)C2=C1Cl WQRCEBAZAUAUQC-UHFFFAOYSA-N 0.000 description 2
- RIOXQFHNBCKOKP-UHFFFAOYSA-N benomyl Chemical compound C1=CC=C2N(C(=O)NCCCC)C(NC(=O)OC)=NC2=C1 RIOXQFHNBCKOKP-UHFFFAOYSA-N 0.000 description 2
- MITFXPHMIHQXPI-UHFFFAOYSA-N benzoxaprofen Natural products N=1C2=CC(C(C(O)=O)C)=CC=C2OC=1C1=CC=C(Cl)C=C1 MITFXPHMIHQXPI-UHFFFAOYSA-N 0.000 description 2
- OMFRMAHOUUJSGP-IRHGGOMRSA-N bifenthrin Chemical compound C1=CC=C(C=2C=CC=CC=2)C(C)=C1COC(=O)[C@@H]1[C@H](\C=C(/Cl)C(F)(F)F)C1(C)C OMFRMAHOUUJSGP-IRHGGOMRSA-N 0.000 description 2
- 239000003124 biologic agent Substances 0.000 description 2
- OIPMQULDKWSNGX-UHFFFAOYSA-N bis[[ethoxy(oxo)phosphaniumyl]oxy]alumanyloxy-ethoxy-oxophosphanium Chemical compound [Al+3].CCO[P+]([O-])=O.CCO[P+]([O-])=O.CCO[P+]([O-])=O OIPMQULDKWSNGX-UHFFFAOYSA-N 0.000 description 2
- CXNPLSGKWMLZPZ-UHFFFAOYSA-N blasticidin-S Natural products O1C(C(O)=O)C(NC(=O)CC(N)CCN(C)C(N)=N)C=CC1N1C(=O)N=C(N)C=C1 CXNPLSGKWMLZPZ-UHFFFAOYSA-N 0.000 description 2
- WYEMLYFITZORAB-UHFFFAOYSA-N boscalid Chemical compound C1=CC(Cl)=CC=C1C1=CC=CC=C1NC(=O)C1=CC=CN=C1Cl WYEMLYFITZORAB-UHFFFAOYSA-N 0.000 description 2
- WZDDLAZXUYIVMU-UHFFFAOYSA-N bromobutide Chemical compound CC(C)(C)C(Br)C(=O)NC(C)(C)C1=CC=CC=C1 WZDDLAZXUYIVMU-UHFFFAOYSA-N 0.000 description 2
- PRLVTUNWOQKEAI-VKAVYKQESA-N buprofezin Chemical compound O=C1N(C(C)C)\C(=N\C(C)(C)C)SCN1C1=CC=CC=C1 PRLVTUNWOQKEAI-VKAVYKQESA-N 0.000 description 2
- HKPHPIREJKHECO-UHFFFAOYSA-N butachlor Chemical compound CCCCOCN(C(=O)CCl)C1=C(CC)C=CC=C1CC HKPHPIREJKHECO-UHFFFAOYSA-N 0.000 description 2
- HFEJHAAIJZXXRE-UHFFFAOYSA-N cafenstrole Chemical compound CCN(CC)C(=O)N1C=NC(S(=O)(=O)C=2C(=CC(C)=CC=2C)C)=N1 HFEJHAAIJZXXRE-UHFFFAOYSA-N 0.000 description 2
- 229940117949 captan Drugs 0.000 description 2
- 150000004657 carbamic acid derivatives Chemical class 0.000 description 2
- TWFZGCMQGLPBSX-UHFFFAOYSA-N carbendazim Chemical compound C1=CC=C2NC(NC(=O)OC)=NC2=C1 TWFZGCMQGLPBSX-UHFFFAOYSA-N 0.000 description 2
- DUEPRVBVGDRKAG-UHFFFAOYSA-N carbofuran Chemical compound CNC(=O)OC1=CC=CC2=C1OC(C)(C)C2 DUEPRVBVGDRKAG-UHFFFAOYSA-N 0.000 description 2
- 235000021466 carotenoid Nutrition 0.000 description 2
- 150000001747 carotenoids Chemical class 0.000 description 2
- RXDMAYSSBPYBFW-UHFFFAOYSA-N carpropamid Chemical compound C=1C=C(Cl)C=CC=1C(C)NC(=O)C1(CC)C(C)C1(Cl)Cl RXDMAYSSBPYBFW-UHFFFAOYSA-N 0.000 description 2
- WYKYKTKDBLFHCY-UHFFFAOYSA-N chloridazon Chemical compound O=C1C(Cl)=C(N)C=NN1C1=CC=CC=C1 WYKYKTKDBLFHCY-UHFFFAOYSA-N 0.000 description 2
- PFIADAMVCJPXSF-UHFFFAOYSA-N chloroneb Chemical compound COC1=CC(Cl)=C(OC)C=C1Cl PFIADAMVCJPXSF-UHFFFAOYSA-N 0.000 description 2
- CRQQGFGUEAVUIL-UHFFFAOYSA-N chlorothalonil Chemical compound ClC1=C(Cl)C(C#N)=C(Cl)C(C#N)=C1Cl CRQQGFGUEAVUIL-UHFFFAOYSA-N 0.000 description 2
- JXCGFZXSOMJFOA-UHFFFAOYSA-N chlorotoluron Chemical compound CN(C)C(=O)NC1=CC=C(C)C(Cl)=C1 JXCGFZXSOMJFOA-UHFFFAOYSA-N 0.000 description 2
- VJYIFXVZLXQVHO-UHFFFAOYSA-N chlorsulfuron Chemical compound COC1=NC(C)=NC(NC(=O)NS(=O)(=O)C=2C(=CC=CC=2)Cl)=N1 VJYIFXVZLXQVHO-UHFFFAOYSA-N 0.000 description 2
- HPNSNYBUADCFDR-UHFFFAOYSA-N chromafenozide Chemical compound CC1=CC(C)=CC(C(=O)N(NC(=O)C=2C(=C3CCCOC3=CC=2)C)C(C)(C)C)=C1 HPNSNYBUADCFDR-UHFFFAOYSA-N 0.000 description 2
- KIEDNEWSYUYDSN-UHFFFAOYSA-N clomazone Chemical compound O=C1C(C)(C)CON1CC1=CC=CC=C1Cl KIEDNEWSYUYDSN-UHFFFAOYSA-N 0.000 description 2
- BIKACRYIQSLICJ-UHFFFAOYSA-N cloransulam-methyl Chemical group N=1N2C(OCC)=NC(F)=CC2=NC=1S(=O)(=O)NC1=C(Cl)C=CC=C1C(=O)OC BIKACRYIQSLICJ-UHFFFAOYSA-N 0.000 description 2
- 235000003488 common ragweed Nutrition 0.000 description 2
- 239000002299 complementary DNA Substances 0.000 description 2
- 238000007796 conventional method Methods 0.000 description 2
- 150000001879 copper Chemical class 0.000 description 2
- 238000003967 crop rotation Methods 0.000 description 2
- 125000004122 cyclic group Chemical group 0.000 description 2
- 229960001591 cyfluthrin Drugs 0.000 description 2
- QQODLKZGRKWIFG-QSFXBCCZSA-N cyfluthrin Chemical compound CC1(C)[C@@H](C=C(Cl)Cl)[C@H]1C(=O)O[C@@H](C#N)C1=CC=C(F)C(OC=2C=CC=CC=2)=C1 QQODLKZGRKWIFG-QSFXBCCZSA-N 0.000 description 2
- WCMMILVIRZAPLE-UHFFFAOYSA-M cyhexatin Chemical compound C1CCCCC1[Sn](C1CCCCC1)(O)C1CCCCC1 WCMMILVIRZAPLE-UHFFFAOYSA-M 0.000 description 2
- 229960005424 cypermethrin Drugs 0.000 description 2
- KAATUXNTWXVJKI-UHFFFAOYSA-N cypermethrin Chemical compound CC1(C)C(C=C(Cl)Cl)C1C(=O)OC(C#N)C1=CC=CC(OC=2C=CC=CC=2)=C1 KAATUXNTWXVJKI-UHFFFAOYSA-N 0.000 description 2
- 229950000775 cyromazine Drugs 0.000 description 2
- LVQDKIWDGQRHTE-UHFFFAOYSA-N cyromazine Chemical compound NC1=NC(N)=NC(NC2CC2)=N1 LVQDKIWDGQRHTE-UHFFFAOYSA-N 0.000 description 2
- QAYICIQNSGETAS-UHFFFAOYSA-N dazomet Chemical compound CN1CSC(=S)N(C)C1 QAYICIQNSGETAS-UHFFFAOYSA-N 0.000 description 2
- 229960002483 decamethrin Drugs 0.000 description 2
- 230000007423 decrease Effects 0.000 description 2
- 230000007547 defect Effects 0.000 description 2
- OWZREIFADZCYQD-NSHGMRRFSA-N deltamethrin Chemical compound CC1(C)[C@@H](C=C(Br)Br)[C@H]1C(=O)O[C@H](C#N)C1=CC=CC(OC=2C=CC=CC=2)=C1 OWZREIFADZCYQD-NSHGMRRFSA-N 0.000 description 2
- 238000004925 denaturation Methods 0.000 description 2
- 230000036425 denaturation Effects 0.000 description 2
- BIXZHMJUSMUDOQ-UHFFFAOYSA-N dichloran Chemical compound NC1=C(Cl)C=C([N+]([O-])=O)C=C1Cl BIXZHMJUSMUDOQ-UHFFFAOYSA-N 0.000 description 2
- YEJGPFZQLRMXOI-PKEIRNPWSA-N diclocymet Chemical compound N#CC(C(C)(C)C)C(=O)N[C@H](C)C1=CC=C(Cl)C=C1Cl YEJGPFZQLRMXOI-PKEIRNPWSA-N 0.000 description 2
- UWQMKVBQKFHLCE-UHFFFAOYSA-N diclomezine Chemical compound C1=C(Cl)C(C)=C(Cl)C=C1C1=NNC(=O)C=C1 UWQMKVBQKFHLCE-UHFFFAOYSA-N 0.000 description 2
- 229940004812 dicloran Drugs 0.000 description 2
- JXSJBGJIGXNWCI-UHFFFAOYSA-N diethyl 2-[(dimethoxyphosphorothioyl)thio]succinate Chemical compound CCOC(=O)CC(SP(=S)(OC)OC)C(=O)OCC JXSJBGJIGXNWCI-UHFFFAOYSA-N 0.000 description 2
- BQYJATMQXGBDHF-UHFFFAOYSA-N difenoconazole Chemical compound O1C(C)COC1(C=1C(=CC(OC=2C=CC(Cl)=CC=2)=CC=1)Cl)CN1N=CN=C1 BQYJATMQXGBDHF-UHFFFAOYSA-N 0.000 description 2
- 230000029087 digestion Effects 0.000 description 2
- 239000003085 diluting agent Substances 0.000 description 2
- BWUPSGJXXPATLU-UHFFFAOYSA-N dimepiperate Chemical compound C=1C=CC=CC=1C(C)(C)SC(=O)N1CCCCC1 BWUPSGJXXPATLU-UHFFFAOYSA-N 0.000 description 2
- SCCDDNKJYDZXMM-UHFFFAOYSA-N dimethachlor Chemical compound COCCN(C(=O)CCl)C1=C(C)C=CC=C1C SCCDDNKJYDZXMM-UHFFFAOYSA-N 0.000 description 2
- MCWXGJITAZMZEV-UHFFFAOYSA-N dimethoate Chemical compound CNC(=O)CSP(=S)(OC)OC MCWXGJITAZMZEV-UHFFFAOYSA-N 0.000 description 2
- WXUZAHCNPWONDH-DYTRJAOYSA-N dimoxystrobin Chemical compound CNC(=O)C(=N\OC)\C1=CC=CC=C1COC1=CC(C)=CC=C1C WXUZAHCNPWONDH-DYTRJAOYSA-N 0.000 description 2
- FBOUIAKEJMZPQG-BLXFFLACSA-N diniconazole-M Chemical compound C1=NC=NN1/C([C@H](O)C(C)(C)C)=C/C1=CC=C(Cl)C=C1Cl FBOUIAKEJMZPQG-BLXFFLACSA-N 0.000 description 2
- ZUOUZKKEUPVFJK-UHFFFAOYSA-N diphenyl Chemical compound C1=CC=CC=C1C1=CC=CC=C1 ZUOUZKKEUPVFJK-UHFFFAOYSA-N 0.000 description 2
- 238000002845 discoloration Methods 0.000 description 2
- 238000009826 distribution Methods 0.000 description 2
- GCKZANITAMOIAR-XWVCPFKXSA-N dsstox_cid_14566 Chemical compound [O-]C(=O)C1=CC=CC=C1.C1=C[C@H](C)[C@@H]([C@@H](C)CC)O[C@]11O[C@H](C\C=C(C)\[C@@H](O[C@@H]2O[C@@H](C)[C@H](O[C@@H]3O[C@@H](C)[C@H]([NH2+]C)[C@@H](OC)C3)[C@@H](OC)C2)[C@@H](C)\C=C\C=C/2[C@]3([C@H](C(=O)O4)C=C(C)[C@@H](O)[C@H]3OC\2)O)C[C@H]4C1 GCKZANITAMOIAR-XWVCPFKXSA-N 0.000 description 2
- 239000000975 dye Substances 0.000 description 2
- AWZOLILCOUMRDG-UHFFFAOYSA-N edifenphos Chemical compound C=1C=CC=CC=1SP(=O)(OCC)SC1=CC=CC=C1 AWZOLILCOUMRDG-UHFFFAOYSA-N 0.000 description 2
- 230000000408 embryogenic effect Effects 0.000 description 2
- 210000002257 embryonic structure Anatomy 0.000 description 2
- RDYMFSUJUZBWLH-SVWSLYAFSA-N endosulfan Chemical compound C([C@@H]12)OS(=O)OC[C@@H]1[C@]1(Cl)C(Cl)=C(Cl)[C@@]2(Cl)C1(Cl)Cl RDYMFSUJUZBWLH-SVWSLYAFSA-N 0.000 description 2
- 230000002708 enhancing effect Effects 0.000 description 2
- NYPJDWWKZLNGGM-RPWUZVMVSA-N esfenvalerate Chemical compound C=1C([C@@H](C#N)OC(=O)[C@@H](C(C)C)C=2C=CC(Cl)=CC=2)=CC=CC=1OC1=CC=CC=C1 NYPJDWWKZLNGGM-RPWUZVMVSA-N 0.000 description 2
- 150000002148 esters Chemical class 0.000 description 2
- PQKBPHSEKWERTG-LLVKDONJSA-N ethyl (2r)-2-[4-[(6-chloro-1,3-benzoxazol-2-yl)oxy]phenoxy]propanoate Chemical group C1=CC(O[C@H](C)C(=O)OCC)=CC=C1OC1=NC2=CC=C(Cl)C=C2O1 PQKBPHSEKWERTG-LLVKDONJSA-N 0.000 description 2
- MLKCGVHIFJBRCD-UHFFFAOYSA-N ethyl 2-chloro-3-{2-chloro-5-[4-(difluoromethyl)-3-methyl-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl]-4-fluorophenyl}propanoate Chemical group C1=C(Cl)C(CC(Cl)C(=O)OCC)=CC(N2C(N(C(F)F)C(C)=N2)=O)=C1F MLKCGVHIFJBRCD-UHFFFAOYSA-N 0.000 description 2
- IXSZQYVWNJNRAL-UHFFFAOYSA-N etoxazole Chemical compound CCOC1=CC(C(C)(C)C)=CC=C1C1N=C(C=2C(=CC=CC=2F)F)OC1 IXSZQYVWNJNRAL-UHFFFAOYSA-N 0.000 description 2
- 239000013604 expression vector Substances 0.000 description 2
- 239000011536 extraction buffer Substances 0.000 description 2
- LMVPQMGRYSRMIW-KRWDZBQOSA-N fenamidone Chemical compound O=C([C@@](C)(N=C1SC)C=2C=CC=CC=2)N1NC1=CC=CC=C1 LMVPQMGRYSRMIW-KRWDZBQOSA-N 0.000 description 2
- ZCJPOPBZHLUFHF-UHFFFAOYSA-N fenamiphos Chemical compound CCOP(=O)(NC(C)C)OC1=CC=C(SC)C(C)=C1 ZCJPOPBZHLUFHF-UHFFFAOYSA-N 0.000 description 2
- DMYHGDXADUDKCQ-UHFFFAOYSA-N fenazaquin Chemical compound C1=CC(C(C)(C)C)=CC=C1CCOC1=NC=NC2=CC=CC=C12 DMYHGDXADUDKCQ-UHFFFAOYSA-N 0.000 description 2
- HJUFTIJOISQSKQ-UHFFFAOYSA-N fenoxycarb Chemical compound C1=CC(OCCNC(=O)OCC)=CC=C1OC1=CC=CC=C1 HJUFTIJOISQSKQ-UHFFFAOYSA-N 0.000 description 2
- FKLFBQCQQYDUAM-UHFFFAOYSA-N fenpiclonil Chemical compound ClC1=CC=CC(C=2C(=CNC=2)C#N)=C1Cl FKLFBQCQQYDUAM-UHFFFAOYSA-N 0.000 description 2
- YYJNOYZRYGDPNH-MFKUBSTISA-N fenpyroximate Chemical compound C=1C=C(C(=O)OC(C)(C)C)C=CC=1CO/N=C/C=1C(C)=NN(C)C=1OC1=CC=CC=C1 YYJNOYZRYGDPNH-MFKUBSTISA-N 0.000 description 2
- BFWMWWXRWVJXSE-UHFFFAOYSA-M fentin hydroxide Chemical compound C=1C=CC=CC=1[Sn](C=1C=CC=CC=1)(O)C1=CC=CC=C1 BFWMWWXRWVJXSE-UHFFFAOYSA-M 0.000 description 2
- XXOYNJXVWVNOOJ-UHFFFAOYSA-N fenuron Chemical compound CN(C)C(=O)NC1=CC=CC=C1 XXOYNJXVWVNOOJ-UHFFFAOYSA-N 0.000 description 2
- 239000003337 fertilizer Substances 0.000 description 2
- RLQJEEJISHYWON-UHFFFAOYSA-N flonicamid Chemical compound FC(F)(F)C1=CC=NC=C1C(=O)NCC#N RLQJEEJISHYWON-UHFFFAOYSA-N 0.000 description 2
- VAIZTNZGPYBOGF-CYBMUJFWSA-N fluazifop-P-butyl Chemical group C1=CC(O[C@H](C)C(=O)OCCCC)=CC=C1OC1=CC=C(C(F)(F)F)C=N1 VAIZTNZGPYBOGF-CYBMUJFWSA-N 0.000 description 2
- UZCGKGPEKUCDTF-UHFFFAOYSA-N fluazinam Chemical compound [O-][N+](=O)C1=CC(C(F)(F)F)=C(Cl)C([N+]([O-])=O)=C1NC1=NC=C(C(F)(F)F)C=C1Cl UZCGKGPEKUCDTF-UHFFFAOYSA-N 0.000 description 2
- GINFBXXYGUODAT-UHFFFAOYSA-N flucarbazone Chemical compound O=C1N(C)C(OC)=NN1C(=O)NS(=O)(=O)C1=CC=CC=C1OC(F)(F)F GINFBXXYGUODAT-UHFFFAOYSA-N 0.000 description 2
- GBIHOLCMZGAKNG-CGAIIQECSA-N flucythrinate Chemical compound O=C([C@@H](C(C)C)C=1C=CC(OC(F)F)=CC=1)OC(C#N)C(C=1)=CC=CC=1OC1=CC=CC=C1 GBIHOLCMZGAKNG-CGAIIQECSA-N 0.000 description 2
- MUJOIMFVNIBMKC-UHFFFAOYSA-N fludioxonil Chemical compound C=12OC(F)(F)OC2=CC=CC=1C1=CNC=C1C#N MUJOIMFVNIBMKC-UHFFFAOYSA-N 0.000 description 2
- IANUJLZYFUDJIH-UHFFFAOYSA-N flufenacet Chemical compound C=1C=C(F)C=CC=1N(C(C)C)C(=O)COC1=NN=C(C(F)(F)F)S1 IANUJLZYFUDJIH-UHFFFAOYSA-N 0.000 description 2
- RYLHNOVXKPXDIP-UHFFFAOYSA-N flufenoxuron Chemical compound C=1C=C(NC(=O)NC(=O)C=2C(=CC=CC=2F)F)C(F)=CC=1OC1=CC=C(C(F)(F)F)C=C1Cl RYLHNOVXKPXDIP-UHFFFAOYSA-N 0.000 description 2
- UFEODZBUAFNAEU-NLRVBDNBSA-N fluoxastrobin Chemical compound C=1C=CC=C(OC=2C(=C(OC=3C(=CC=CC=3)Cl)N=CN=2)F)C=1C(=N/OC)\C1=NOCCO1 UFEODZBUAFNAEU-NLRVBDNBSA-N 0.000 description 2
- FQKUGOMFVDPBIZ-UHFFFAOYSA-N flusilazole Chemical compound C=1C=C(F)C=CC=1[Si](C=1C=CC(F)=CC=1)(C)CN1C=NC=N1 FQKUGOMFVDPBIZ-UHFFFAOYSA-N 0.000 description 2
- ZCNQYNHDVRPZIH-UHFFFAOYSA-N fluthiacet-methyl Chemical group C1=C(Cl)C(SCC(=O)OC)=CC(N=C2N3CCCCN3C(=O)S2)=C1F ZCNQYNHDVRPZIH-UHFFFAOYSA-N 0.000 description 2
- PTCGDEVVHUXTMP-UHFFFAOYSA-N flutolanil Chemical compound CC(C)OC1=CC=CC(NC(=O)C=2C(=CC=CC=2)C(F)(F)F)=C1 PTCGDEVVHUXTMP-UHFFFAOYSA-N 0.000 description 2
- HKIOYBQGHSTUDB-UHFFFAOYSA-N folpet Chemical compound C1=CC=C2C(=O)N(SC(Cl)(Cl)Cl)C(=O)C2=C1 HKIOYBQGHSTUDB-UHFFFAOYSA-N 0.000 description 2
- BGZZWXTVIYUUEY-UHFFFAOYSA-N fomesafen Chemical compound C1=C([N+]([O-])=O)C(C(=O)NS(=O)(=O)C)=CC(OC=2C(=CC(=CC=2)C(F)(F)F)Cl)=C1 BGZZWXTVIYUUEY-UHFFFAOYSA-N 0.000 description 2
- KVGLBTYUCJYMND-UHFFFAOYSA-N fonofos Chemical compound CCOP(=S)(CC)SC1=CC=CC=C1 KVGLBTYUCJYMND-UHFFFAOYSA-N 0.000 description 2
- PXDNXJSDGQBLKS-UHFFFAOYSA-N foramsulfuron Chemical compound COC1=CC(OC)=NC(NC(=O)NS(=O)(=O)C=2C(=CC=C(NC=O)C=2)C(=O)N(C)C)=N1 PXDNXJSDGQBLKS-UHFFFAOYSA-N 0.000 description 2
- 239000012737 fresh medium Substances 0.000 description 2
- 239000000446 fuel Substances 0.000 description 2
- 239000000499 gel Substances 0.000 description 2
- 230000004077 genetic alteration Effects 0.000 description 2
- 231100000118 genetic alteration Toxicity 0.000 description 2
- ZDXPYRJPNDTMRX-UHFFFAOYSA-N glutamine Natural products OC(=O)C(N)CCC(N)=O ZDXPYRJPNDTMRX-UHFFFAOYSA-N 0.000 description 2
- 102000007434 glycine N-acyltransferase Human genes 0.000 description 2
- 108020005567 glycine N-acyltransferase Proteins 0.000 description 2
- ZEKANFGSDXODPD-UHFFFAOYSA-N glyphosate-isopropylammonium Chemical compound CC(C)N.OC(=O)CNCP(O)(O)=O ZEKANFGSDXODPD-UHFFFAOYSA-N 0.000 description 2
- 208000037824 growth disorder Diseases 0.000 description 2
- 239000001963 growth medium Substances 0.000 description 2
- CNKHSLKYRMDDNQ-UHFFFAOYSA-N halofenozide Chemical compound C=1C=CC=CC=1C(=O)N(C(C)(C)C)NC(=O)C1=CC=C(Cl)C=C1 CNKHSLKYRMDDNQ-UHFFFAOYSA-N 0.000 description 2
- RGNPBRKPHBKNKX-UHFFFAOYSA-N hexaflumuron Chemical compound C1=C(Cl)C(OC(F)(F)C(F)F)=C(Cl)C=C1NC(=O)NC(=O)C1=C(F)C=CC=C1F RGNPBRKPHBKNKX-UHFFFAOYSA-N 0.000 description 2
- IGMNYECMUMZDDF-UHFFFAOYSA-N homogentisic acid Chemical compound OC(=O)CC1=CC(O)=CC=C1O IGMNYECMUMZDDF-UHFFFAOYSA-N 0.000 description 2
- 229940056881 imidacloprid Drugs 0.000 description 2
- YWTYJOPNNQFBPC-UHFFFAOYSA-N imidacloprid Chemical compound [O-][N+](=O)\N=C1/NCCN1CC1=CC=C(Cl)N=C1 YWTYJOPNNQFBPC-UHFFFAOYSA-N 0.000 description 2
- VBCVPMMZEGZULK-NRFANRHFSA-N indoxacarb Chemical compound C([C@@]1(OC2)C(=O)OC)C3=CC(Cl)=CC=C3C1=NN2C(=O)N(C(=O)OC)C1=CC=C(OC(F)(F)F)C=C1 VBCVPMMZEGZULK-NRFANRHFSA-N 0.000 description 2
- NRXQIUSYPAHGNM-UHFFFAOYSA-N ioxynil Chemical compound OC1=C(I)C=C(C#N)C=C1I NRXQIUSYPAHGNM-UHFFFAOYSA-N 0.000 description 2
- QTYCMDBMOLSEAM-UHFFFAOYSA-N ipconazole Chemical compound C1=NC=NN1CC1(O)C(C(C)C)CCC1CC1=CC=C(Cl)C=C1 QTYCMDBMOLSEAM-UHFFFAOYSA-N 0.000 description 2
- FCOAHACKGGIURQ-UHFFFAOYSA-N iprobenfos Chemical compound CC(C)OP(=O)(OC(C)C)SCC1=CC=CC=C1 FCOAHACKGGIURQ-UHFFFAOYSA-N 0.000 description 2
- NWUWYYSKZYIQAE-WMCAAGNKSA-N iprovalicarb Chemical compound CC(C)OC(=O)N[C@@H](C(C)C)C(=O)NC(C)C1=CC=C(C)C=C1 NWUWYYSKZYIQAE-WMCAAGNKSA-N 0.000 description 2
- HOQADATXFBOEGG-UHFFFAOYSA-N isofenphos Chemical compound CCOP(=S)(NC(C)C)OC1=CC=CC=C1C(=O)OC(C)C HOQADATXFBOEGG-UHFFFAOYSA-N 0.000 description 2
- QXJSBBXBKPUZAA-UHFFFAOYSA-N isooleic acid Natural products CCCCCCCC=CCCCCCCCCC(O)=O QXJSBBXBKPUZAA-UHFFFAOYSA-N 0.000 description 2
- JJWLVOIRVHMVIS-UHFFFAOYSA-O isopropylaminium Chemical compound CC(C)[NH3+] JJWLVOIRVHMVIS-UHFFFAOYSA-O 0.000 description 2
- UFHLMYOGRXOCSL-UHFFFAOYSA-N isoprothiolane Chemical compound CC(C)OC(=O)C(C(=O)OC(C)C)=C1SCCS1 UFHLMYOGRXOCSL-UHFFFAOYSA-N 0.000 description 2
- PUIYMUZLKQOUOZ-UHFFFAOYSA-N isoproturon Chemical compound CC(C)C1=CC=C(NC(=O)N(C)C)C=C1 PUIYMUZLKQOUOZ-UHFFFAOYSA-N 0.000 description 2
- PMHURSZHKKJGBM-UHFFFAOYSA-N isoxaben Chemical compound O1N=C(C(C)(CC)CC)C=C1NC(=O)C1=C(OC)C=CC=C1OC PMHURSZHKKJGBM-UHFFFAOYSA-N 0.000 description 2
- OYIKARCXOQLFHF-UHFFFAOYSA-N isoxaflutole Chemical compound CS(=O)(=O)C1=CC(C(F)(F)F)=CC=C1C(=O)C1=C(C2CC2)ON=C1 OYIKARCXOQLFHF-UHFFFAOYSA-N 0.000 description 2
- 229940088649 isoxaflutole Drugs 0.000 description 2
- ZOTBXTZVPHCKPN-HTXNQAPBSA-N kresoxim-methyl Chemical compound CO\N=C(\C(=O)OC)C1=CC=CC=C1COC1=CC=CC=C1C ZOTBXTZVPHCKPN-HTXNQAPBSA-N 0.000 description 2
- CONWAEURSVPLRM-UHFFFAOYSA-N lactofen Chemical compound C1=C([N+]([O-])=O)C(C(=O)OC(C)C(=O)OCC)=CC(OC=2C(=CC(=CC=2)C(F)(F)F)Cl)=C1 CONWAEURSVPLRM-UHFFFAOYSA-N 0.000 description 2
- 230000000670 limiting effect Effects 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 229960000521 lufenuron Drugs 0.000 description 2
- PWPJGUXAGUPAHP-UHFFFAOYSA-N lufenuron Chemical compound C1=C(Cl)C(OC(F)(F)C(C(F)(F)F)F)=CC(Cl)=C1NC(=O)NC(=O)C1=C(F)C=CC=C1F PWPJGUXAGUPAHP-UHFFFAOYSA-N 0.000 description 2
- 235000009973 maize Nutrition 0.000 description 2
- 229960000453 malathion Drugs 0.000 description 2
- YKSNLCVSTHTHJA-UHFFFAOYSA-L maneb Chemical compound [Mn+2].[S-]C(=S)NCCNC([S-])=S YKSNLCVSTHTHJA-UHFFFAOYSA-L 0.000 description 2
- 229920000940 maneb Polymers 0.000 description 2
- 230000007246 mechanism Effects 0.000 description 2
- 230000010534 mechanism of action Effects 0.000 description 2
- XIGAUIHYSDTJHW-UHFFFAOYSA-N mefenacet Chemical compound N=1C2=CC=CC=C2SC=1OCC(=O)N(C)C1=CC=CC=C1 XIGAUIHYSDTJHW-UHFFFAOYSA-N 0.000 description 2
- BCTQJXQXJVLSIG-UHFFFAOYSA-N mepronil Chemical compound CC(C)OC1=CC=CC(NC(=O)C=2C(=CC=CC=2)C)=C1 BCTQJXQXJVLSIG-UHFFFAOYSA-N 0.000 description 2
- ZQEIXNIJLIKNTD-GFCCVEGCSA-N metalaxyl-M Chemical compound COCC(=O)N([C@H](C)C(=O)OC)C1=C(C)C=CC=C1C ZQEIXNIJLIKNTD-GFCCVEGCSA-N 0.000 description 2
- GKKDCARASOJPNG-UHFFFAOYSA-N metaldehyde Chemical compound CC1OC(C)OC(C)OC(C)O1 GKKDCARASOJPNG-UHFFFAOYSA-N 0.000 description 2
- VHCNQEUWZYOAEV-UHFFFAOYSA-N metamitron Chemical compound O=C1N(N)C(C)=NN=C1C1=CC=CC=C1 VHCNQEUWZYOAEV-UHFFFAOYSA-N 0.000 description 2
- STEPQTYSZVCJPV-UHFFFAOYSA-N metazachlor Chemical compound CC1=CC=CC(C)=C1N(C(=O)CCl)CN1N=CC=C1 STEPQTYSZVCJPV-UHFFFAOYSA-N 0.000 description 2
- XWPZUHJBOLQNMN-UHFFFAOYSA-N metconazole Chemical compound C1=NC=NN1CC1(O)C(C)(C)CCC1CC1=CC=C(Cl)C=C1 XWPZUHJBOLQNMN-UHFFFAOYSA-N 0.000 description 2
- NNKVPIKMPCQWCG-UHFFFAOYSA-N methamidophos Chemical compound COP(N)(=O)SC NNKVPIKMPCQWCG-UHFFFAOYSA-N 0.000 description 2
- MYWUZJCMWCOHBA-VIFPVBQESA-N methamphetamine Chemical compound CN[C@@H](C)CC1=CC=CC=C1 MYWUZJCMWCOHBA-VIFPVBQESA-N 0.000 description 2
- MEBQXILRKZHVCX-UHFFFAOYSA-N methidathion Chemical compound COC1=NN(CSP(=S)(OC)OC)C(=O)S1 MEBQXILRKZHVCX-UHFFFAOYSA-N 0.000 description 2
- UHXUZOCRWCRNSJ-QPJJXVBHSA-N methomyl Chemical compound CNC(=O)O\N=C(/C)SC UHXUZOCRWCRNSJ-QPJJXVBHSA-N 0.000 description 2
- 229950003442 methoprene Drugs 0.000 description 2
- 229930002897 methoprene Natural products 0.000 description 2
- QCAWEPFNJXQPAN-UHFFFAOYSA-N methoxyfenozide Chemical compound COC1=CC=CC(C(=O)NN(C(=O)C=2C=C(C)C=C(C)C=2)C(C)(C)C)=C1C QCAWEPFNJXQPAN-UHFFFAOYSA-N 0.000 description 2
- RBNIGDFIUWJJEV-LLVKDONJSA-N methyl (2r)-2-(n-benzoyl-3-chloro-4-fluoroanilino)propanoate Chemical group C=1C=C(F)C(Cl)=CC=1N([C@H](C)C(=O)OC)C(=O)C1=CC=CC=C1 RBNIGDFIUWJJEV-LLVKDONJSA-N 0.000 description 2
- NIFKBBMCXCMCAO-UHFFFAOYSA-N methyl 2-[(4,6-dimethoxypyrimidin-2-yl)carbamoylsulfamoyl]-4-(methanesulfonamidomethyl)benzoate Chemical group COC(=O)C1=CC=C(CNS(C)(=O)=O)C=C1S(=O)(=O)NC(=O)NC1=NC(OC)=CC(OC)=N1 NIFKBBMCXCMCAO-UHFFFAOYSA-N 0.000 description 2
- DTVOKYWXACGVGO-UHFFFAOYSA-N methyl 2-[(4,6-dimethoxypyrimidin-2-yl)carbamoylsulfamoyl]-6-(trifluoromethyl)pyridine-3-carboxylate Chemical group COC(=O)C1=CC=C(C(F)(F)F)N=C1S(=O)(=O)NC(=O)NC1=NC(OC)=CC(OC)=N1 DTVOKYWXACGVGO-UHFFFAOYSA-N 0.000 description 2
- JTHMVYBOQLDDIY-UHFFFAOYSA-N methyl 2-[(4-methyl-5-oxo-3-propoxy-1,2,4-triazole-1-carbonyl)sulfamoyl]benzoate Chemical compound O=C1N(C)C(OCCC)=NN1C(=O)NS(=O)(=O)C1=CC=CC=C1C(=O)OC JTHMVYBOQLDDIY-UHFFFAOYSA-N 0.000 description 2
- KBHDSWIXRODKSZ-UHFFFAOYSA-N methyl 5-chloro-2-(trifluoromethylsulfonylamino)benzoate Chemical compound COC(=O)C1=CC(Cl)=CC=C1NS(=O)(=O)C(F)(F)F KBHDSWIXRODKSZ-UHFFFAOYSA-N 0.000 description 2
- ZQEIXNIJLIKNTD-UHFFFAOYSA-N methyl N-(2,6-dimethylphenyl)-N-(methoxyacetyl)alaninate Chemical compound COCC(=O)N(C(C)C(=O)OC)C1=C(C)C=CC=C1C ZQEIXNIJLIKNTD-UHFFFAOYSA-N 0.000 description 2
- CIEXPHRYOLIQQD-UHFFFAOYSA-N methyl N-(2,6-dimethylphenyl)-N-2-furoylalaninate Chemical compound CC=1C=CC=C(C)C=1N(C(C)C(=O)OC)C(=O)C1=CC=CO1 CIEXPHRYOLIQQD-UHFFFAOYSA-N 0.000 description 2
- QYPPRTNMGCREIM-UHFFFAOYSA-N methylarsonic acid Chemical compound C[As](O)(O)=O QYPPRTNMGCREIM-UHFFFAOYSA-N 0.000 description 2
- DSRNRYQBBJQVCW-UHFFFAOYSA-N metoxuron Chemical compound COC1=CC=C(NC(=O)N(C)C)C=C1Cl DSRNRYQBBJQVCW-UHFFFAOYSA-N 0.000 description 2
- AMSPWOYQQAWRRM-UHFFFAOYSA-N metrafenone Chemical compound COC1=CC=C(Br)C(C)=C1C(=O)C1=C(C)C=C(OC)C(OC)=C1OC AMSPWOYQQAWRRM-UHFFFAOYSA-N 0.000 description 2
- 229960001952 metrifonate Drugs 0.000 description 2
- DEDOPGXGGQYYMW-UHFFFAOYSA-N molinate Chemical compound CCSC(=O)N1CCCCCC1 DEDOPGXGGQYYMW-UHFFFAOYSA-N 0.000 description 2
- KRTSDMXIXPKRQR-AATRIKPKSA-N monocrotophos Chemical compound CNC(=O)\C=C(/C)OP(=O)(OC)OC KRTSDMXIXPKRQR-AATRIKPKSA-N 0.000 description 2
- 230000000877 morphologic effect Effects 0.000 description 2
- GLBLPMUBLHYFCW-UHFFFAOYSA-N n-(5,7-dimethoxy-[1,2,4]triazolo[1,5-a]pyrimidin-2-yl)-2-methoxy-4-(trifluoromethyl)pyridine-3-sulfonamide Chemical compound N1=C2N=C(OC)C=C(OC)N2N=C1NS(=O)(=O)C1=C(OC)N=CC=C1C(F)(F)F GLBLPMUBLHYFCW-UHFFFAOYSA-N 0.000 description 2
- JXTHEWSKYLZVJC-UHFFFAOYSA-N naptalam Chemical compound OC(=O)C1=CC=CC=C1C(=O)NC1=CC=CC2=CC=CC=C12 JXTHEWSKYLZVJC-UHFFFAOYSA-N 0.000 description 2
- RTCOGUMHFFWOJV-UHFFFAOYSA-N nicosulfuron Chemical compound COC1=CC(OC)=NC(NC(=O)NS(=O)(=O)C=2C(=CC=CN=2)C(=O)N(C)C)=N1 RTCOGUMHFFWOJV-UHFFFAOYSA-N 0.000 description 2
- 150000002825 nitriles Chemical class 0.000 description 2
- FBUKVWPVBMHYJY-UHFFFAOYSA-N nonanoic acid Chemical compound CCCCCCCCC(O)=O FBUKVWPVBMHYJY-UHFFFAOYSA-N 0.000 description 2
- NJPPVKZQTLUDBO-UHFFFAOYSA-N novaluron Chemical compound C1=C(Cl)C(OC(F)(F)C(OC(F)(F)F)F)=CC=C1NC(=O)NC(=O)C1=C(F)C=CC=C1F NJPPVKZQTLUDBO-UHFFFAOYSA-N 0.000 description 2
- 229920001778 nylon Polymers 0.000 description 2
- ZQPPMHVWECSIRJ-KTKRTIGZSA-N oleic acid Chemical compound CCCCCCCC\C=C/CCCCCCCC(O)=O ZQPPMHVWECSIRJ-KTKRTIGZSA-N 0.000 description 2
- 239000002751 oligonucleotide probe Substances 0.000 description 2
- LLLFASISUZUJEQ-UHFFFAOYSA-N orbencarb Chemical compound CCN(CC)C(=O)SCC1=CC=CC=C1Cl LLLFASISUZUJEQ-UHFFFAOYSA-N 0.000 description 2
- 239000005416 organic matter Substances 0.000 description 2
- JHIPUJPTQJYEQK-ZLHHXESBSA-N orysastrobin Chemical compound CNC(=O)C(=N\OC)\C1=CC=CC=C1CO\N=C(/C)\C(=N\OC)\C(\C)=N\OC JHIPUJPTQJYEQK-ZLHHXESBSA-N 0.000 description 2
- 238000009401 outcrossing Methods 0.000 description 2
- UWVQIROCRJWDKL-UHFFFAOYSA-N oxadixyl Chemical compound CC=1C=CC=C(C)C=1N(C(=O)COC)N1CCOC1=O UWVQIROCRJWDKL-UHFFFAOYSA-N 0.000 description 2
- IOXAXYHXMLCCJJ-UHFFFAOYSA-N oxetan-3-yl 2-[(4,6-dimethylpyrimidin-2-yl)carbamoylsulfamoyl]benzoate Chemical compound CC1=CC(C)=NC(NC(=O)NS(=O)(=O)C=2C(=CC=CC=2)C(=O)OC2COC2)=N1 IOXAXYHXMLCCJJ-UHFFFAOYSA-N 0.000 description 2
- 239000005022 packaging material Substances 0.000 description 2
- LCCNCVORNKJIRZ-UHFFFAOYSA-N parathion Chemical compound CCOP(=S)(OCC)OC1=CC=C([N+]([O-])=O)C=C1 LCCNCVORNKJIRZ-UHFFFAOYSA-N 0.000 description 2
- RLBIQVVOMOPOHC-UHFFFAOYSA-N parathion-methyl Chemical group COP(=S)(OC)OC1=CC=C([N+]([O-])=O)C=C1 RLBIQVVOMOPOHC-UHFFFAOYSA-N 0.000 description 2
- OGYFATSSENRIKG-UHFFFAOYSA-N pencycuron Chemical compound C1=CC(Cl)=CC=C1CN(C(=O)NC=1C=CC=CC=1)C1CCCC1 OGYFATSSENRIKG-UHFFFAOYSA-N 0.000 description 2
- CHIFOSRWCNZCFN-UHFFFAOYSA-N pendimethalin Chemical compound CCC(CC)NC1=C([N+]([O-])=O)C=C(C)C(C)=C1[N+]([O-])=O CHIFOSRWCNZCFN-UHFFFAOYSA-N 0.000 description 2
- JZPKLLLUDLHCEL-UHFFFAOYSA-N pentoxazone Chemical compound O=C1C(=C(C)C)OC(=O)N1C1=CC(OC2CCCC2)=C(Cl)C=C1F JZPKLLLUDLHCEL-UHFFFAOYSA-N 0.000 description 2
- 229960000490 permethrin Drugs 0.000 description 2
- RLLPVAHGXHCWKJ-UHFFFAOYSA-N permethrin Chemical compound CC1(C)C(C=C(Cl)Cl)C1C(=O)OCC1=CC=CC(OC=2C=CC=CC=2)=C1 RLLPVAHGXHCWKJ-UHFFFAOYSA-N 0.000 description 2
- 125000001997 phenyl group Chemical group [H]C1=C([H])C([H])=C(*)C([H])=C1[H] 0.000 description 2
- BULVZWIRKLYCBC-UHFFFAOYSA-N phorate Chemical compound CCOP(=S)(OCC)SCSCC BULVZWIRKLYCBC-UHFFFAOYSA-N 0.000 description 2
- IOUNQDKNJZEDEP-UHFFFAOYSA-N phosalone Chemical compound C1=C(Cl)C=C2OC(=O)N(CSP(=S)(OCC)OCC)C2=C1 IOUNQDKNJZEDEP-UHFFFAOYSA-N 0.000 description 2
- LMNZTLDVJIUSHT-UHFFFAOYSA-N phosmet Chemical compound C1=CC=C2C(=O)N(CSP(=S)(OC)OC)C(=O)C2=C1 LMNZTLDVJIUSHT-UHFFFAOYSA-N 0.000 description 2
- RGCLLPNLLBQHPF-HJWRWDBZSA-N phosphamidon Chemical compound CCN(CC)C(=O)C(\Cl)=C(/C)OP(=O)(OC)OC RGCLLPNLLBQHPF-HJWRWDBZSA-N 0.000 description 2
- 108010082527 phosphinothricin N-acetyltransferase Proteins 0.000 description 2
- 108010001545 phytoene dehydrogenase Proteins 0.000 description 2
- NQQVFXUMIDALNH-UHFFFAOYSA-N picloram Chemical compound NC1=C(Cl)C(Cl)=NC(C(O)=O)=C1Cl NQQVFXUMIDALNH-UHFFFAOYSA-N 0.000 description 2
- YFGYUFNIOHWBOB-UHFFFAOYSA-N pirimicarb Chemical compound CN(C)C(=O)OC1=NC(N(C)C)=NC(C)=C1C YFGYUFNIOHWBOB-UHFFFAOYSA-N 0.000 description 2
- 239000005648 plant growth regulator Substances 0.000 description 2
- 230000010287 polarization Effects 0.000 description 2
- 230000010152 pollination Effects 0.000 description 2
- 239000005014 poly(hydroxyalkanoate) Substances 0.000 description 2
- 229920000903 polyhydroxyalkanoate Polymers 0.000 description 2
- 229920000642 polymer Polymers 0.000 description 2
- LIOPHZNMBKHGAV-UHFFFAOYSA-M potassium;2-(phosphonomethylamino)acetate Chemical compound [K+].OC(=O)CNCP(O)([O-])=O LIOPHZNMBKHGAV-UHFFFAOYSA-M 0.000 description 2
- WHHIPMZEDGBUCC-UHFFFAOYSA-N probenazole Chemical compound C1=CC=C2C(OCC=C)=NS(=O)(=O)C2=C1 WHHIPMZEDGBUCC-UHFFFAOYSA-N 0.000 description 2
- QYMMJNLHFKGANY-UHFFFAOYSA-N profenofos Chemical compound CCCSP(=O)(OCC)OC1=CC=C(Br)C=C1Cl QYMMJNLHFKGANY-UHFFFAOYSA-N 0.000 description 2
- 230000035755 proliferation Effects 0.000 description 2
- ISEUFVQQFVOBCY-UHFFFAOYSA-N prometon Chemical compound COC1=NC(NC(C)C)=NC(NC(C)C)=N1 ISEUFVQQFVOBCY-UHFFFAOYSA-N 0.000 description 2
- AAEVYOVXGOFMJO-UHFFFAOYSA-N prometryn Chemical compound CSC1=NC(NC(C)C)=NC(NC(C)C)=N1 AAEVYOVXGOFMJO-UHFFFAOYSA-N 0.000 description 2
- MFOUDYKPLGXPGO-UHFFFAOYSA-N propachlor Chemical compound ClCC(=O)N(C(C)C)C1=CC=CC=C1 MFOUDYKPLGXPGO-UHFFFAOYSA-N 0.000 description 2
- WZZLDXDUQPOXNW-UHFFFAOYSA-N propamocarb Chemical compound CCCOC(=O)NCCCN(C)C WZZLDXDUQPOXNW-UHFFFAOYSA-N 0.000 description 2
- LFULEKSKNZEWOE-UHFFFAOYSA-N propanil Chemical compound CCC(=O)NC1=CC=C(Cl)C(Cl)=C1 LFULEKSKNZEWOE-UHFFFAOYSA-N 0.000 description 2
- FROBCXTULYFHEJ-OAHLLOKOSA-N propaquizafop Chemical compound C1=CC(O[C@H](C)C(=O)OCCON=C(C)C)=CC=C1OC1=CN=C(C=C(Cl)C=C2)C2=N1 FROBCXTULYFHEJ-OAHLLOKOSA-N 0.000 description 2
- ZYHMJXZULPZUED-UHFFFAOYSA-N propargite Chemical compound C1=CC(C(C)(C)C)=CC=C1OC1C(OS(=O)OCC#C)CCCC1 ZYHMJXZULPZUED-UHFFFAOYSA-N 0.000 description 2
- VXPLXMJHHKHSOA-UHFFFAOYSA-N propham Chemical compound CC(C)OC(=O)NC1=CC=CC=C1 VXPLXMJHHKHSOA-UHFFFAOYSA-N 0.000 description 2
- STJLVHWMYQXCPB-UHFFFAOYSA-N propiconazole Chemical compound O1C(CCC)COC1(C=1C(=CC(Cl)=CC=1)Cl)CN1N=CN=C1 STJLVHWMYQXCPB-UHFFFAOYSA-N 0.000 description 2
- PHNUZKMIPFFYSO-UHFFFAOYSA-N propyzamide Chemical compound C#CC(C)(C)NC(=O)C1=CC(Cl)=CC(Cl)=C1 PHNUZKMIPFFYSO-UHFFFAOYSA-N 0.000 description 2
- FLVBXVXXXMLMOX-UHFFFAOYSA-N proquinazid Chemical compound C1=C(I)C=C2C(=O)N(CCC)C(OCCC)=NC2=C1 FLVBXVXXXMLMOX-UHFFFAOYSA-N 0.000 description 2
- APTZNLHMIGJTEW-UHFFFAOYSA-N pyraflufen-ethyl Chemical group C1=C(Cl)C(OCC(=O)OCC)=CC(C=2C(=C(OC(F)F)N(C)N=2)Cl)=C1F APTZNLHMIGJTEW-UHFFFAOYSA-N 0.000 description 2
- ASRAWSBMDXVNLX-UHFFFAOYSA-N pyrazolynate Chemical compound C=1C=C(Cl)C=C(Cl)C=1C(=O)C=1C(C)=NN(C)C=1OS(=O)(=O)C1=CC=C(C)C=C1 ASRAWSBMDXVNLX-UHFFFAOYSA-N 0.000 description 2
- FKERUJTUOYLBKB-UHFFFAOYSA-N pyrazoxyfen Chemical compound C=1C=C(Cl)C=C(Cl)C=1C(=O)C=1C(C)=NN(C)C=1OCC(=O)C1=CC=CC=C1 FKERUJTUOYLBKB-UHFFFAOYSA-N 0.000 description 2
- VTRWMTJQBQJKQH-UHFFFAOYSA-N pyributicarb Chemical compound COC1=CC=CC(N(C)C(=S)OC=2C=C(C=CC=2)C(C)(C)C)=N1 VTRWMTJQBQJKQH-UHFFFAOYSA-N 0.000 description 2
- ZLIBICFPKPWGIZ-UHFFFAOYSA-N pyrimethanil Chemical compound CC1=CC(C)=NC(NC=2C=CC=CC=2)=N1 ZLIBICFPKPWGIZ-UHFFFAOYSA-N 0.000 description 2
- NHDHVHZZCFYRSB-UHFFFAOYSA-N pyriproxyfen Chemical compound C=1C=CC=NC=1OC(C)COC(C=C1)=CC=C1OC1=CC=CC=C1 NHDHVHZZCFYRSB-UHFFFAOYSA-N 0.000 description 2
- QJBZDBLBQWFTPZ-UHFFFAOYSA-N pyrrolnitrin Chemical compound [O-][N+](=O)C1=C(Cl)C=CC=C1C1=CNC=C1Cl QJBZDBLBQWFTPZ-UHFFFAOYSA-N 0.000 description 2
- FFSSWMQPCJRCRV-UHFFFAOYSA-N quinclorac Chemical compound ClC1=CN=C2C(C(=O)O)=C(Cl)C=CC2=C1 FFSSWMQPCJRCRV-UHFFFAOYSA-N 0.000 description 2
- FBQQHUGEACOBDN-UHFFFAOYSA-N quinomethionate Chemical compound N1=C2SC(=O)SC2=NC2=CC(C)=CC=C21 FBQQHUGEACOBDN-UHFFFAOYSA-N 0.000 description 2
- WRPIRSINYZBGPK-UHFFFAOYSA-N quinoxyfen Chemical compound C1=CC(F)=CC=C1OC1=CC=NC2=CC(Cl)=CC(Cl)=C12 WRPIRSINYZBGPK-UHFFFAOYSA-N 0.000 description 2
- 230000006798 recombination Effects 0.000 description 2
- 238000005215 recombination Methods 0.000 description 2
- 238000011084 recovery Methods 0.000 description 2
- 230000022532 regulation of transcription, DNA-dependent Effects 0.000 description 2
- 238000009877 rendering Methods 0.000 description 2
- 239000005871 repellent Substances 0.000 description 2
- 230000002940 repellent Effects 0.000 description 2
- 108091008146 restriction endonucleases Proteins 0.000 description 2
- 125000002652 ribonucleotide group Chemical group 0.000 description 2
- 229940080817 rotenone Drugs 0.000 description 2
- JUVIOZPCNVVQFO-UHFFFAOYSA-N rotenone Natural products O1C2=C3CC(C(C)=C)OC3=CC=C2C(=O)C2C1COC1=C2C=C(OC)C(OC)=C1 JUVIOZPCNVVQFO-UHFFFAOYSA-N 0.000 description 2
- ODCWYMIRDDJXKW-UHFFFAOYSA-N simazine Chemical compound CCNC1=NC(Cl)=NC(NCC)=N1 ODCWYMIRDDJXKW-UHFFFAOYSA-N 0.000 description 2
- 239000011734 sodium Substances 0.000 description 2
- 229910052708 sodium Inorganic materials 0.000 description 2
- 239000001509 sodium citrate Substances 0.000 description 2
- 229910001415 sodium ion Inorganic materials 0.000 description 2
- 239000007787 solid Substances 0.000 description 2
- 238000011895 specific detection Methods 0.000 description 2
- 229940014213 spinosad Drugs 0.000 description 2
- PUYXTUJWRLOUCW-UHFFFAOYSA-N spiroxamine Chemical compound O1C(CN(CC)CCC)COC11CCC(C(C)(C)C)CC1 PUYXTUJWRLOUCW-UHFFFAOYSA-N 0.000 description 2
- 125000001424 substituent group Chemical group 0.000 description 2
- 125000003107 substituted aryl group Chemical group 0.000 description 2
- PQTBTIFWAXVEPB-UHFFFAOYSA-N sulcotrione Chemical compound ClC1=CC(S(=O)(=O)C)=CC=C1C(=O)C1C(=O)CCCC1=O PQTBTIFWAXVEPB-UHFFFAOYSA-N 0.000 description 2
- ZDXMLEQEMNLCQG-UHFFFAOYSA-N sulfometuron methyl Chemical group COC(=O)C1=CC=CC=C1S(=O)(=O)NC(=O)NC1=NC(C)=CC(C)=N1 ZDXMLEQEMNLCQG-UHFFFAOYSA-N 0.000 description 2
- 239000011593 sulfur Substances 0.000 description 2
- 229910052717 sulfur Inorganic materials 0.000 description 2
- 239000004094 surface-active agent Substances 0.000 description 2
- 230000008685 targeting Effects 0.000 description 2
- 239000005936 tau-Fluvalinate Substances 0.000 description 2
- INISTDXBRIBGOC-XMMISQBUSA-N tau-fluvalinate Chemical compound N([C@H](C(C)C)C(=O)OC(C#N)C=1C=C(OC=2C=CC=CC=2)C=CC=1)C1=CC=C(C(F)(F)F)C=C1Cl INISTDXBRIBGOC-XMMISQBUSA-N 0.000 description 2
- QYPNKSZPJQQLRK-UHFFFAOYSA-N tebufenozide Chemical compound C1=CC(CC)=CC=C1C(=O)NN(C(C)(C)C)C(=O)C1=CC(C)=CC(C)=C1 QYPNKSZPJQQLRK-UHFFFAOYSA-N 0.000 description 2
- ZZYSLNWGKKDOML-UHFFFAOYSA-N tebufenpyrad Chemical compound CCC1=NN(C)C(C(=O)NCC=2C=CC(=CC=2)C(C)(C)C)=C1Cl ZZYSLNWGKKDOML-UHFFFAOYSA-N 0.000 description 2
- RJKCKKDSSSRYCB-UHFFFAOYSA-N tebutam Chemical compound CC(C)(C)C(=O)N(C(C)C)CC1=CC=CC=C1 RJKCKKDSSSRYCB-UHFFFAOYSA-N 0.000 description 2
- CJDWRQLODFKPEL-UHFFFAOYSA-N teflubenzuron Chemical compound FC1=CC=CC(F)=C1C(=O)NC(=O)NC1=CC(Cl)=C(F)C(Cl)=C1F CJDWRQLODFKPEL-UHFFFAOYSA-N 0.000 description 2
- BCQMBFHBDZVHKU-UHFFFAOYSA-N terbumeton Chemical compound CCNC1=NC(NC(C)(C)C)=NC(OC)=N1 BCQMBFHBDZVHKU-UHFFFAOYSA-N 0.000 description 2
- IROINLKCQGIITA-UHFFFAOYSA-N terbutryn Chemical compound CCNC1=NC(NC(C)(C)C)=NC(SC)=N1 IROINLKCQGIITA-UHFFFAOYSA-N 0.000 description 2
- FZXISNSWEXTPMF-UHFFFAOYSA-N terbutylazine Chemical compound CCNC1=NC(Cl)=NC(NC(C)(C)C)=N1 FZXISNSWEXTPMF-UHFFFAOYSA-N 0.000 description 2
- UBCKGWBNUIFUST-YHYXMXQVSA-N tetrachlorvinphos Chemical compound COP(=O)(OC)O\C(=C/Cl)C1=CC(Cl)=C(Cl)C=C1Cl UBCKGWBNUIFUST-YHYXMXQVSA-N 0.000 description 2
- LITQZINTSYBKIU-UHFFFAOYSA-F tetracopper;hexahydroxide;sulfate Chemical compound [OH-].[OH-].[OH-].[OH-].[OH-].[OH-].[Cu+2].[Cu+2].[Cu+2].[Cu+2].[O-]S([O-])(=O)=O LITQZINTSYBKIU-UHFFFAOYSA-F 0.000 description 2
- NWWZPOKUUAIXIW-FLIBITNWSA-N thiamethoxam Chemical compound [O-][N+](=O)\N=C/1N(C)COCN\1CC1=CN=C(Cl)S1 NWWZPOKUUAIXIW-FLIBITNWSA-N 0.000 description 2
- WOSNCVAPUOFXEH-UHFFFAOYSA-N thifluzamide Chemical compound S1C(C)=NC(C(F)(F)F)=C1C(=O)NC1=C(Br)C=C(OC(F)(F)F)C=C1Br WOSNCVAPUOFXEH-UHFFFAOYSA-N 0.000 description 2
- BAKXBZPQTXCKRR-UHFFFAOYSA-N thiodicarb Chemical compound CSC(C)=NOC(=O)NSNC(=O)ON=C(C)SC BAKXBZPQTXCKRR-UHFFFAOYSA-N 0.000 description 2
- QGHREAKMXXNCOA-UHFFFAOYSA-N thiophanate-methyl Chemical compound COC(=O)NC(=S)NC1=CC=CC=C1NC(=S)NC(=O)OC QGHREAKMXXNCOA-UHFFFAOYSA-N 0.000 description 2
- QSOHVSNIQHGFJU-UHFFFAOYSA-L thiosultap disodium Chemical compound [Na+].[Na+].[O-]S(=O)(=O)SCC(N(C)C)CSS([O-])(=O)=O QSOHVSNIQHGFJU-UHFFFAOYSA-L 0.000 description 2
- 238000003971 tillage Methods 0.000 description 2
- 231100000331 toxic Toxicity 0.000 description 2
- 230000002588 toxic effect Effects 0.000 description 2
- YWSCPYYRJXKUDB-KAKFPZCNSA-N tralomethrin Chemical compound CC1(C)[C@@H](C(Br)C(Br)(Br)Br)[C@H]1C(=O)O[C@H](C#N)C1=CC=CC(OC=2C=CC=CC=2)=C1 YWSCPYYRJXKUDB-KAKFPZCNSA-N 0.000 description 2
- 238000013518 transcription Methods 0.000 description 2
- 230000035897 transcription Effects 0.000 description 2
- 230000001131 transforming effect Effects 0.000 description 2
- NFACJZMKEDPNKN-UHFFFAOYSA-N trichlorfon Chemical compound COP(=O)(OC)C(O)C(Cl)(Cl)Cl NFACJZMKEDPNKN-UHFFFAOYSA-N 0.000 description 2
- DQJCHOQLCLEDLL-UHFFFAOYSA-N tricyclazole Chemical compound CC1=CC=CC2=C1N1C=NN=C1S2 DQJCHOQLCLEDLL-UHFFFAOYSA-N 0.000 description 2
- ONCZDRURRATYFI-TVJDWZFNSA-N trifloxystrobin Chemical compound CO\N=C(\C(=O)OC)C1=CC=CC=C1CO\N=C(/C)C1=CC=CC(C(F)(F)F)=C1 ONCZDRURRATYFI-TVJDWZFNSA-N 0.000 description 2
- XAIPTRIXGHTTNT-UHFFFAOYSA-N triflumuron Chemical compound C1=CC(OC(F)(F)F)=CC=C1NC(=O)NC(=O)C1=CC=CC=C1Cl XAIPTRIXGHTTNT-UHFFFAOYSA-N 0.000 description 2
- ZSDSQXJSNMTJDA-UHFFFAOYSA-N trifluralin Chemical compound CCCN(CCC)C1=C([N+]([O-])=O)C=C(C(F)(F)F)C=C1[N+]([O-])=O ZSDSQXJSNMTJDA-UHFFFAOYSA-N 0.000 description 2
- IMEVJVISCHQJRM-UHFFFAOYSA-N triflusulfuron-methyl Chemical group COC(=O)C1=CC=CC(C)=C1S(=O)(=O)NC(=O)NC1=NC(OCC(F)(F)F)=NC(N(C)C)=N1 IMEVJVISCHQJRM-UHFFFAOYSA-N 0.000 description 2
- 241000701447 unidentified baculovirus Species 0.000 description 2
- 150000003672 ureas Chemical class 0.000 description 2
- JARYYMUOCXVXNK-IMTORBKUSA-N validamycin Chemical compound N([C@H]1C[C@@H]([C@H]([C@H](O)[C@H]1O)O[C@H]1[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O1)O)CO)[C@H]1C=C(CO)[C@H](O)[C@H](O)[C@H]1O JARYYMUOCXVXNK-IMTORBKUSA-N 0.000 description 2
- 239000008158 vegetable oil Substances 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- YNWVFADWVLCOPU-MDWZMJQESA-N (1E)-1-(4-chlorophenyl)-4,4-dimethyl-2-(1H-1,2,4-triazol-1-yl)pent-1-en-3-ol Chemical compound C1=NC=NN1/C(C(O)C(C)(C)C)=C/C1=CC=C(Cl)C=C1 YNWVFADWVLCOPU-MDWZMJQESA-N 0.000 description 1
- ZXQYGBMAQZUVMI-RDDWSQKMSA-N (1S)-cis-(alphaR)-cyhalothrin Chemical compound CC1(C)[C@H](\C=C(/Cl)C(F)(F)F)[C@@H]1C(=O)O[C@@H](C#N)C1=CC=CC(OC=2C=CC=CC=2)=C1 ZXQYGBMAQZUVMI-RDDWSQKMSA-N 0.000 description 1
- GXEKYRXVRROBEV-FBXFSONDSA-N (1r,2s,3r,4s)-7-oxabicyclo[2.2.1]heptane-2,3-dicarboxylic acid Chemical compound C1C[C@@H]2[C@@H](C(O)=O)[C@@H](C(=O)O)[C@H]1O2 GXEKYRXVRROBEV-FBXFSONDSA-N 0.000 description 1
- AGMMRUPNXPWLGF-AATRIKPKSA-N (2,3,5,6-tetrafluoro-4-methylphenyl)methyl 2,2-dimethyl-3-[(e)-prop-1-enyl]cyclopropane-1-carboxylate Chemical compound CC1(C)C(/C=C/C)C1C(=O)OCC1=C(F)C(F)=C(C)C(F)=C1F AGMMRUPNXPWLGF-AATRIKPKSA-N 0.000 description 1
- DQKWXTIYGWPGOO-UHFFFAOYSA-N (2,6-dibromo-4-cyanophenyl) octanoate Chemical compound CCCCCCCC(=O)OC1=C(Br)C=C(C#N)C=C1Br DQKWXTIYGWPGOO-UHFFFAOYSA-N 0.000 description 1
- UDPGUMQDCGORJQ-UHFFFAOYSA-N (2-chloroethyl)phosphonic acid Chemical compound OP(O)(=O)CCCl UDPGUMQDCGORJQ-UHFFFAOYSA-N 0.000 description 1
- SAPGTCDSBGMXCD-UHFFFAOYSA-N (2-chlorophenyl)-(4-fluorophenyl)-pyrimidin-5-ylmethanol Chemical compound C=1N=CN=CC=1C(C=1C(=CC=CC=1)Cl)(O)C1=CC=C(F)C=C1 SAPGTCDSBGMXCD-UHFFFAOYSA-N 0.000 description 1
- LZTIMERBDGGAJD-SNAWJCMRSA-N (2e)-2-(nitromethylidene)-1,3-thiazinane Chemical compound [O-][N+](=O)\C=C1/NCCCS1 LZTIMERBDGGAJD-SNAWJCMRSA-N 0.000 description 1
- NYHLMHAKWBUZDY-QMMMGPOBSA-N (2s)-2-[2-chloro-5-[2-chloro-4-(trifluoromethyl)phenoxy]benzoyl]oxypropanoic acid Chemical compound C1=C(Cl)C(C(=O)O[C@@H](C)C(O)=O)=CC(OC=2C(=CC(=CC=2)C(F)(F)F)Cl)=C1 NYHLMHAKWBUZDY-QMMMGPOBSA-N 0.000 description 1
- LNGRZPZKVUBWQV-UHFFFAOYSA-N (4-chloro-2-methylsulfonylphenyl)-(5-cyclopropyl-1,2-oxazol-4-yl)methanone Chemical compound CS(=O)(=O)C1=CC(Cl)=CC=C1C(=O)C1=C(C2CC2)ON=C1 LNGRZPZKVUBWQV-UHFFFAOYSA-N 0.000 description 1
- XUNYDVLIZWUPAW-UHFFFAOYSA-N (4-chlorophenyl) n-(4-methylphenyl)sulfonylcarbamate Chemical compound C1=CC(C)=CC=C1S(=O)(=O)NC(=O)OC1=CC=C(Cl)C=C1 XUNYDVLIZWUPAW-UHFFFAOYSA-N 0.000 description 1
- WCXDHFDTOYPNIE-RIYZIHGNSA-N (E)-acetamiprid Chemical compound N#C/N=C(\C)N(C)CC1=CC=C(Cl)N=C1 WCXDHFDTOYPNIE-RIYZIHGNSA-N 0.000 description 1
- PGOOBECODWQEAB-UHFFFAOYSA-N (E)-clothianidin Chemical compound [O-][N+](=O)\N=C(/NC)NCC1=CN=C(Cl)S1 PGOOBECODWQEAB-UHFFFAOYSA-N 0.000 description 1
- CFRPSFYHXJZSBI-DHZHZOJOSA-N (E)-nitenpyram Chemical compound [O-][N+](=O)/C=C(\NC)N(CC)CC1=CC=C(Cl)N=C1 CFRPSFYHXJZSBI-DHZHZOJOSA-N 0.000 description 1
- MPIPASJGOJYODL-SFHVURJKSA-N (R)-isoconazole Chemical compound ClC1=CC(Cl)=CC=C1[C@@H](OCC=1C(=CC=CC=1Cl)Cl)CN1C=NC=C1 MPIPASJGOJYODL-SFHVURJKSA-N 0.000 description 1
- WNTGYJSOUMFZEP-SSDOTTSWSA-N (R)-mecoprop Chemical compound OC(=O)[C@@H](C)OC1=CC=C(Cl)C=C1C WNTGYJSOUMFZEP-SSDOTTSWSA-N 0.000 description 1
- ADDQHLREJDZPMT-AWEZNQCLSA-N (S)-metamifop Chemical compound O=C([C@@H](OC=1C=CC(OC=2OC3=CC(Cl)=CC=C3N=2)=CC=1)C)N(C)C1=CC=CC=C1F ADDQHLREJDZPMT-AWEZNQCLSA-N 0.000 description 1
- WVQBLGZPHOPPFO-LBPRGKRZSA-N (S)-metolachlor Chemical compound CCC1=CC=CC(C)=C1N([C@@H](C)COC)C(=O)CCl WVQBLGZPHOPPFO-LBPRGKRZSA-N 0.000 description 1
- XGWIJUOSCAQSSV-XHDPSFHLSA-N (S,S)-hexythiazox Chemical compound S([C@H]([C@@H]1C)C=2C=CC(Cl)=CC=2)C(=O)N1C(=O)NC1CCCCC1 XGWIJUOSCAQSSV-XHDPSFHLSA-N 0.000 description 1
- RMOGWMIKYWRTKW-UONOGXRCSA-N (S,S)-paclobutrazol Chemical compound C([C@@H]([C@@H](O)C(C)(C)C)N1N=CN=C1)C1=CC=C(Cl)C=C1 RMOGWMIKYWRTKW-UONOGXRCSA-N 0.000 description 1
- DARPYRSDRJYGIF-PTNGSMBKSA-N (Z)-3-ethoxy-2-naphthalen-2-ylsulfonylprop-2-enenitrile Chemical compound C1=CC=CC2=CC(S(=O)(=O)C(\C#N)=C/OCC)=CC=C21 DARPYRSDRJYGIF-PTNGSMBKSA-N 0.000 description 1
- HOKKPVIRMVDYPB-UVTDQMKNSA-N (Z)-thiacloprid Chemical compound C1=NC(Cl)=CC=C1CN1C(=N/C#N)/SCC1 HOKKPVIRMVDYPB-UVTDQMKNSA-N 0.000 description 1
- USGUVNUTPWXWBA-JRIXXDKMSA-N (e,2s)-2-amino-4-(2-aminoethoxy)but-3-enoic acid Chemical compound NCCO\C=C\[C@H](N)C(O)=O USGUVNUTPWXWBA-JRIXXDKMSA-N 0.000 description 1
- JYEUMXHLPRZUAT-UHFFFAOYSA-N 1,2,3-triazine Chemical group C1=CN=NN=C1 JYEUMXHLPRZUAT-UHFFFAOYSA-N 0.000 description 1
- DHYXNIKICPUXJI-UHFFFAOYSA-N 1-(2,4-dichlorophenyl)-n-(2,4-difluorophenyl)-5-oxo-n-propan-2-yl-1,2,4-triazole-4-carboxamide Chemical compound C=1C=C(F)C=C(F)C=1N(C(C)C)C(=O)N(C1=O)C=NN1C1=CC=C(Cl)C=C1Cl DHYXNIKICPUXJI-UHFFFAOYSA-N 0.000 description 1
- DAGDLSRRQJATCV-UHFFFAOYSA-N 1-(2-bromoethoxy)-2-propan-2-ylbenzene Chemical compound CC(C)C1=CC=CC=C1OCCBr DAGDLSRRQJATCV-UHFFFAOYSA-N 0.000 description 1
- WURBVZBTWMNKQT-UHFFFAOYSA-N 1-(4-chlorophenoxy)-3,3-dimethyl-1-(1,2,4-triazol-1-yl)butan-2-one Chemical compound C1=NC=NN1C(C(=O)C(C)(C)C)OC1=CC=C(Cl)C=C1 WURBVZBTWMNKQT-UHFFFAOYSA-N 0.000 description 1
- VGPIBGGRCVEHQZ-UHFFFAOYSA-N 1-(biphenyl-4-yloxy)-3,3-dimethyl-1-(1,2,4-triazol-1-yl)butan-2-ol Chemical compound C1=NC=NN1C(C(O)C(C)(C)C)OC(C=C1)=CC=C1C1=CC=CC=C1 VGPIBGGRCVEHQZ-UHFFFAOYSA-N 0.000 description 1
- WKBPZYKAUNRMKP-UHFFFAOYSA-N 1-[2-(2,4-dichlorophenyl)pentyl]1,2,4-triazole Chemical compound C=1C=C(Cl)C=C(Cl)C=1C(CCC)CN1C=NC=N1 WKBPZYKAUNRMKP-UHFFFAOYSA-N 0.000 description 1
- BXKKQFGRMSOANI-UHFFFAOYSA-N 1-methoxy-3-[4-[(2-methoxy-2,4,4-trimethyl-3h-chromen-7-yl)oxy]phenyl]-1-methylurea Chemical compound C1=CC(NC(=O)N(C)OC)=CC=C1OC1=CC=C2C(C)(C)CC(C)(OC)OC2=C1 BXKKQFGRMSOANI-UHFFFAOYSA-N 0.000 description 1
- PFFIDZXUXFLSSR-UHFFFAOYSA-N 1-methyl-N-[2-(4-methylpentan-2-yl)-3-thienyl]-3-(trifluoromethyl)pyrazole-4-carboxamide Chemical compound S1C=CC(NC(=O)C=2C(=NN(C)C=2)C(F)(F)F)=C1C(C)CC(C)C PFFIDZXUXFLSSR-UHFFFAOYSA-N 0.000 description 1
- IBXNCJKFFQIKKY-UHFFFAOYSA-N 1-pentyne Chemical compound CCCC#C IBXNCJKFFQIKKY-UHFFFAOYSA-N 0.000 description 1
- LEZWWPYKPKIXLL-UHFFFAOYSA-N 1-{2-(4-chlorobenzyloxy)-2-(2,4-dichlorophenyl)ethyl}imidazole Chemical compound C1=CC(Cl)=CC=C1COC(C=1C(=CC(Cl)=CC=1)Cl)CN1C=NC=C1 LEZWWPYKPKIXLL-UHFFFAOYSA-N 0.000 description 1
- MCNOFYBITGAAGM-UHFFFAOYSA-N 2,2-dichloro-1-[5-(furan-2-yl)-2,2-dimethyl-1,3-oxazolidin-3-yl]ethanone Chemical compound C1N(C(=O)C(Cl)Cl)C(C)(C)OC1C1=CC=CO1 MCNOFYBITGAAGM-UHFFFAOYSA-N 0.000 description 1
- XZIDTOHMJBOSOX-UHFFFAOYSA-N 2,3,6-TBA Chemical compound OC(=O)C1=C(Cl)C=CC(Cl)=C1Cl XZIDTOHMJBOSOX-UHFFFAOYSA-N 0.000 description 1
- CGNBQYFXGQHUQP-UHFFFAOYSA-N 2,3-dinitroaniline Chemical class NC1=CC=CC([N+]([O-])=O)=C1[N+]([O-])=O CGNBQYFXGQHUQP-UHFFFAOYSA-N 0.000 description 1
- MHKBMNACOMRIAW-UHFFFAOYSA-N 2,3-dinitrophenol Chemical class OC1=CC=CC([N+]([O-])=O)=C1[N+]([O-])=O MHKBMNACOMRIAW-UHFFFAOYSA-N 0.000 description 1
- KGKGSIUWJCAFPX-UHFFFAOYSA-N 2,6-dichlorothiobenzamide Chemical compound NC(=S)C1=C(Cl)C=CC=C1Cl KGKGSIUWJCAFPX-UHFFFAOYSA-N 0.000 description 1
- YTOPFCCWCSOHFV-UHFFFAOYSA-N 2,6-dimethyl-4-tridecylmorpholine Chemical compound CCCCCCCCCCCCCN1CC(C)OC(C)C1 YTOPFCCWCSOHFV-UHFFFAOYSA-N 0.000 description 1
- MZHCENGPTKEIGP-UHFFFAOYSA-N 2-(2,4-dichlorophenoxy)propanoic acid Chemical compound OC(=O)C(C)OC1=CC=C(Cl)C=C1Cl MZHCENGPTKEIGP-UHFFFAOYSA-N 0.000 description 1
- STMIIPIFODONDC-UHFFFAOYSA-N 2-(2,4-dichlorophenyl)-1-(1H-1,2,4-triazol-1-yl)hexan-2-ol Chemical compound C=1C=C(Cl)C=C(Cl)C=1C(O)(CCCC)CN1C=NC=N1 STMIIPIFODONDC-UHFFFAOYSA-N 0.000 description 1
- ROKVVMOXSZIDEG-UHFFFAOYSA-N 2-(3,5,6-trichloropyridin-2-yl)oxyacetate;triethylazanium Chemical compound CCN(CC)CC.OC(=O)COC1=NC(Cl)=C(Cl)C=C1Cl ROKVVMOXSZIDEG-UHFFFAOYSA-N 0.000 description 1
- PKAUICCNAWQPAU-UHFFFAOYSA-N 2-(4-chloro-2-methylphenoxy)acetic acid;n-methylmethanamine Chemical compound CNC.CC1=CC(Cl)=CC=C1OCC(O)=O PKAUICCNAWQPAU-UHFFFAOYSA-N 0.000 description 1
- HZJKXKUJVSEEFU-UHFFFAOYSA-N 2-(4-chlorophenyl)-2-(1H-1,2,4-triazol-1-ylmethyl)hexanenitrile Chemical compound C=1C=C(Cl)C=CC=1C(CCCC)(C#N)CN1C=NC=N1 HZJKXKUJVSEEFU-UHFFFAOYSA-N 0.000 description 1
- KWLVWJPJKJMCSH-UHFFFAOYSA-N 2-(4-chlorophenyl)-N-{2-[3-methoxy-4-(prop-2-yn-1-yloxy)phenyl]ethyl}-2-(prop-2-yn-1-yloxy)acetamide Chemical compound C1=C(OCC#C)C(OC)=CC(CCNC(=O)C(OCC#C)C=2C=CC(Cl)=CC=2)=C1 KWLVWJPJKJMCSH-UHFFFAOYSA-N 0.000 description 1
- YABFPHSQTSFWQB-UHFFFAOYSA-N 2-(4-fluorophenyl)-1-(1,2,4-triazol-1-yl)-3-(trimethylsilyl)propan-2-ol Chemical compound C=1C=C(F)C=CC=1C(O)(C[Si](C)(C)C)CN1C=NC=N1 YABFPHSQTSFWQB-UHFFFAOYSA-N 0.000 description 1
- OWZPCEFYPSAJFR-UHFFFAOYSA-N 2-(butan-2-yl)-4,6-dinitrophenol Chemical compound CCC(C)C1=CC([N+]([O-])=O)=CC([N+]([O-])=O)=C1O OWZPCEFYPSAJFR-UHFFFAOYSA-N 0.000 description 1
- KRQUFUKTQHISJB-YYADALCUSA-N 2-[(E)-N-[2-(4-chlorophenoxy)propoxy]-C-propylcarbonimidoyl]-3-hydroxy-5-(thian-3-yl)cyclohex-2-en-1-one Chemical compound CCC\C(=N/OCC(C)OC1=CC=C(Cl)C=C1)C1=C(O)CC(CC1=O)C1CCCSC1 KRQUFUKTQHISJB-YYADALCUSA-N 0.000 description 1
- IRJQWZWMQCVOLA-ZBKNUEDVSA-N 2-[(z)-n-[(3,5-difluorophenyl)carbamoylamino]-c-methylcarbonimidoyl]pyridine-3-carboxylic acid Chemical compound N=1C=CC=C(C(O)=O)C=1C(/C)=N\NC(=O)NC1=CC(F)=CC(F)=C1 IRJQWZWMQCVOLA-ZBKNUEDVSA-N 0.000 description 1
- BOTNFCTYKJBUMU-UHFFFAOYSA-N 2-[4-(2-methylpropyl)piperazin-4-ium-1-yl]-2-oxoacetate Chemical compound CC(C)C[NH+]1CCN(C(=O)C([O-])=O)CC1 BOTNFCTYKJBUMU-UHFFFAOYSA-N 0.000 description 1
- IQWMUTFHGXREBR-UHFFFAOYSA-N 2-[5-(5-tert-butyl-2-oxo-1,3,4-oxadiazol-3-yl)-2,4-dichlorophenoxy]acetonitrile Chemical group O=C1OC(C(C)(C)C)=NN1C1=CC(OCC#N)=C(Cl)C=C1Cl IQWMUTFHGXREBR-UHFFFAOYSA-N 0.000 description 1
- IOYNQIMAUDJVEI-ZFNPBRLTSA-N 2-[N-[(E)-3-chloroprop-2-enoxy]-C-ethylcarbonimidoyl]-3-hydroxy-5-(oxan-4-yl)cyclohex-2-en-1-one Chemical compound C1C(=O)C(C(=NOC\C=C\Cl)CC)=C(O)CC1C1CCOCC1 IOYNQIMAUDJVEI-ZFNPBRLTSA-N 0.000 description 1
- IVDRCZNHVGQBHZ-UHFFFAOYSA-N 2-butoxyethyl 2-(3,5,6-trichloropyridin-2-yl)oxyacetate Chemical group CCCCOCCOC(=O)COC1=NC(Cl)=C(Cl)C=C1Cl IVDRCZNHVGQBHZ-UHFFFAOYSA-N 0.000 description 1
- WKGKFWXGAHXMCE-UHFFFAOYSA-N 2-butoxyethyl 2-(4-chloro-2-methylphenoxy)acetate Chemical group CCCCOCCOC(=O)COC1=CC=C(Cl)C=C1C WKGKFWXGAHXMCE-UHFFFAOYSA-N 0.000 description 1
- QEGVVEOAVNHRAA-UHFFFAOYSA-N 2-chloro-6-(4,6-dimethoxypyrimidin-2-yl)sulfanylbenzoic acid Chemical compound COC1=CC(OC)=NC(SC=2C(=C(Cl)C=CC=2)C(O)=O)=N1 QEGVVEOAVNHRAA-UHFFFAOYSA-N 0.000 description 1
- OWDLFBLNMPCXSD-UHFFFAOYSA-N 2-chloro-N-(2,6-dimethylphenyl)-N-(2-oxotetrahydrofuran-3-yl)acetamide Chemical compound CC1=CC=CC(C)=C1N(C(=O)CCl)C1C(=O)OCC1 OWDLFBLNMPCXSD-UHFFFAOYSA-N 0.000 description 1
- KZNDFYDURHAESM-UHFFFAOYSA-N 2-chloro-n-(2-ethyl-6-methylphenyl)-n-(propan-2-yloxymethyl)acetamide Chemical compound CCC1=CC=CC(C)=C1N(COC(C)C)C(=O)CCl KZNDFYDURHAESM-UHFFFAOYSA-N 0.000 description 1
- MIJLZGZLQLAQCM-UHFFFAOYSA-N 2-ethoxyethyl 2-(4-{[3-chloro-5-(trifluoromethyl)pyridin-2-yl]oxy}phenoxy)propanoate Chemical group C1=CC(OC(C)C(=O)OCCOCC)=CC=C1OC1=NC=C(C(F)(F)F)C=C1Cl MIJLZGZLQLAQCM-UHFFFAOYSA-N 0.000 description 1
- IDGRPSMONFWWEK-UHFFFAOYSA-N 2-ethylhexyl 2-(4-chloro-2-methylphenoxy)acetate Chemical group CCCCC(CC)COC(=O)COC1=CC=C(Cl)C=C1C IDGRPSMONFWWEK-UHFFFAOYSA-N 0.000 description 1
- AWSZRJQNBMEZOI-UHFFFAOYSA-N 2-methoxyethyl 2-(4-tert-butylphenyl)-2-cyano-3-oxo-3-[2-(trifluoromethyl)phenyl]propanoate Chemical compound C=1C=C(C(C)(C)C)C=CC=1C(C#N)(C(=O)OCCOC)C(=O)C1=CC=CC=C1C(F)(F)F AWSZRJQNBMEZOI-UHFFFAOYSA-N 0.000 description 1
- JAAQRPKKYYHALF-UHFFFAOYSA-N 2-methoxyethyl 2-[(5-but-3-yn-2-yloxy-4-chloro-2-fluorophenyl)-(3-fluorobenzoyl)carbamoyl]cyclohexene-1-carboxylate Chemical compound C1CCCC(C(=O)OCCOC)=C1C(=O)N(C=1C(=CC(Cl)=C(OC(C)C#C)C=1)F)C(=O)C1=CC=CC(F)=C1 JAAQRPKKYYHALF-UHFFFAOYSA-N 0.000 description 1
- 125000004200 2-methoxyethyl group Chemical group [H]C([H])([H])OC([H])([H])C([H])([H])* 0.000 description 1
- LLWADFLAOKUBDR-UHFFFAOYSA-N 2-methyl-4-chlorophenoxybutyric acid Chemical compound CC1=CC(Cl)=CC=C1OCCCC(O)=O LLWADFLAOKUBDR-UHFFFAOYSA-N 0.000 description 1
- 229940044120 2-n-octyl-4-isothiazolin-3-one Drugs 0.000 description 1
- AVGVFDSUDIUXEU-UHFFFAOYSA-N 2-octyl-1,2-thiazolidin-3-one Chemical compound CCCCCCCCN1SCCC1=O AVGVFDSUDIUXEU-UHFFFAOYSA-N 0.000 description 1
- MFUPLJQNEXUUDW-UHFFFAOYSA-N 2-phenylisoindole-1,3-dione Chemical class O=C1C2=CC=CC=C2C(=O)N1C1=CC=CC=C1 MFUPLJQNEXUUDW-UHFFFAOYSA-N 0.000 description 1
- UFAPVJDEYHLLBG-UHFFFAOYSA-N 2-{2-chloro-4-(methylsulfonyl)-3-[(tetrahydrofuran-2-ylmethoxy)methyl]benzoyl}cyclohexane-1,3-dione Chemical compound ClC1=C(COCC2OCCC2)C(S(=O)(=O)C)=CC=C1C(=O)C1C(=O)CCCC1=O UFAPVJDEYHLLBG-UHFFFAOYSA-N 0.000 description 1
- 108020005065 3' Flanking Region Proteins 0.000 description 1
- 101710087550 3-phosphoshikimate 1-carboxyvinyltransferase 1 Proteins 0.000 description 1
- SBUKOHLFHYSZNG-UHFFFAOYSA-N 4-dodecyl-2,6-dimethylmorpholine Chemical compound CCCCCCCCCCCCN1CC(C)OC(C)C1 SBUKOHLFHYSZNG-UHFFFAOYSA-N 0.000 description 1
- QQOGZMUZAZWLJH-UHFFFAOYSA-N 5-[2-chloro-6-fluoro-4-(trifluoromethyl)phenoxy]-n-ethylsulfonyl-2-nitrobenzamide Chemical compound C1=C([N+]([O-])=O)C(C(=O)NS(=O)(=O)CC)=CC(OC=2C(=CC(=CC=2F)C(F)(F)F)Cl)=C1 QQOGZMUZAZWLJH-UHFFFAOYSA-N 0.000 description 1
- PVSGXWMWNRGTKE-UHFFFAOYSA-N 5-methyl-2-[4-methyl-5-oxo-4-(propan-2-yl)-4,5-dihydro-1H-imidazol-2-yl]pyridine-3-carboxylic acid Chemical compound N1C(=O)C(C(C)C)(C)N=C1C1=NC=C(C)C=C1C(O)=O PVSGXWMWNRGTKE-UHFFFAOYSA-N 0.000 description 1
- 101150052384 50 gene Proteins 0.000 description 1
- IBSREHMXUMOFBB-JFUDTMANSA-N 5u8924t11h Chemical compound O1[C@@H](C)[C@H](O)[C@@H](OC)C[C@@H]1O[C@@H]1[C@@H](OC)C[C@H](O[C@@H]2C(=C/C[C@@H]3C[C@@H](C[C@@]4(O3)C=C[C@H](C)[C@@H](C(C)C)O4)OC(=O)[C@@H]3C=C(C)[C@@H](O)[C@H]4OC\C([C@@]34O)=C/C=C/[C@@H]2C)/C)O[C@H]1C.C1=C[C@H](C)[C@@H]([C@@H](C)CC)O[C@]11O[C@H](C\C=C(C)\[C@@H](O[C@@H]2O[C@@H](C)[C@H](O[C@@H]3O[C@@H](C)[C@H](O)[C@@H](OC)C3)[C@@H](OC)C2)[C@@H](C)\C=C\C=C/2[C@]3([C@H](C(=O)O4)C=C(C)[C@@H](O)[C@H]3OC\2)O)C[C@H]4C1 IBSREHMXUMOFBB-JFUDTMANSA-N 0.000 description 1
- HZKBYBNLTLVSPX-UHFFFAOYSA-N 6-[(6,6-dimethyl-5,7-dihydropyrrolo[2,1-c][1,2,4]thiadiazol-3-ylidene)amino]-7-fluoro-4-prop-2-ynyl-1,4-benzoxazin-3-one Chemical compound C#CCN1C(=O)COC(C=C2F)=C1C=C2N=C1SN=C2CC(C)(C)CN21 HZKBYBNLTLVSPX-UHFFFAOYSA-N 0.000 description 1
- ZUSHSDOEVHPTCU-UHFFFAOYSA-N 6-chloro-3-phenyl-1h-pyridazin-4-one Chemical compound N1C(Cl)=CC(=O)C(C=2C=CC=CC=2)=N1 ZUSHSDOEVHPTCU-UHFFFAOYSA-N 0.000 description 1
- WFPZSXYXPSUOPY-ROYWQJLOSA-N ADP alpha-D-glucoside Chemical compound C([C@H]1O[C@H]([C@@H]([C@@H]1O)O)N1C=2N=CN=C(C=2N=C1)N)OP(O)(=O)OP(O)(=O)O[C@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O WFPZSXYXPSUOPY-ROYWQJLOSA-N 0.000 description 1
- 239000005875 Acetamiprid Substances 0.000 description 1
- QTBSBXVTEAMEQO-UHFFFAOYSA-M Acetate Chemical compound CC([O-])=O QTBSBXVTEAMEQO-UHFFFAOYSA-M 0.000 description 1
- VTNQPKFIQCLBDU-UHFFFAOYSA-N Acetochlor Chemical compound CCOCN(C(=O)CCl)C1=C(C)C=CC=C1CC VTNQPKFIQCLBDU-UHFFFAOYSA-N 0.000 description 1
- 244000056974 Adansonia digitata Species 0.000 description 1
- 235000003320 Adansonia digitata Nutrition 0.000 description 1
- 235000003319 Adansonia gregorii Nutrition 0.000 description 1
- 229920000936 Agarose Polymers 0.000 description 1
- 241000589158 Agrobacterium Species 0.000 description 1
- 241001621841 Alopecurus myosuroides Species 0.000 description 1
- 235000013479 Amaranthus retroflexus Nutrition 0.000 description 1
- 235000004135 Amaranthus viridis Nutrition 0.000 description 1
- 235000003133 Ambrosia artemisiifolia Nutrition 0.000 description 1
- 239000005727 Amisulbrom Substances 0.000 description 1
- KLSJWNVTNUYHDU-UHFFFAOYSA-N Amitrole Chemical compound NC1=NC=NN1 KLSJWNVTNUYHDU-UHFFFAOYSA-N 0.000 description 1
- GEHMBYLTCISYNY-UHFFFAOYSA-N Ammonium sulfamate Chemical compound [NH4+].NS([O-])(=O)=O GEHMBYLTCISYNY-UHFFFAOYSA-N 0.000 description 1
- NXQDBZGWYSEGFL-UHFFFAOYSA-N Anilofos Chemical compound COP(=S)(OC)SCC(=O)N(C(C)C)C1=CC=C(Cl)C=C1 NXQDBZGWYSEGFL-UHFFFAOYSA-N 0.000 description 1
- 108010007730 Apyrase Proteins 0.000 description 1
- 102000007347 Apyrase Human genes 0.000 description 1
- 101100371686 Arabidopsis thaliana UBQ10 gene Proteins 0.000 description 1
- 235000005781 Avena Nutrition 0.000 description 1
- 235000007320 Avena fatua Nutrition 0.000 description 1
- 244000075850 Avena orientalis Species 0.000 description 1
- 241001647031 Avena sterilis Species 0.000 description 1
- 235000004535 Avena sterilis Nutrition 0.000 description 1
- 239000005730 Azoxystrobin Substances 0.000 description 1
- 241001645380 Bassia scoparia Species 0.000 description 1
- 239000005734 Benalaxyl Substances 0.000 description 1
- PFJJMJDEVDLPNE-UHFFFAOYSA-N Benoxacor Chemical compound C1=CC=C2N(C(=O)C(Cl)Cl)C(C)COC2=C1 PFJJMJDEVDLPNE-UHFFFAOYSA-N 0.000 description 1
- 239000005472 Bensulfuron methyl Substances 0.000 description 1
- RRNIZKPFKNDSRS-UHFFFAOYSA-N Bensulide Chemical compound CC(C)OP(=S)(OC(C)C)SCCNS(=O)(=O)C1=CC=CC=C1 RRNIZKPFKNDSRS-UHFFFAOYSA-N 0.000 description 1
- 239000005476 Bentazone Substances 0.000 description 1
- 239000005736 Benthiavalicarb Substances 0.000 description 1
- 239000005884 Beta-Cyfluthrin Substances 0.000 description 1
- 239000005653 Bifenazate Substances 0.000 description 1
- 239000005740 Boscalid Substances 0.000 description 1
- 235000011293 Brassica napus Nutrition 0.000 description 1
- 240000002791 Brassica napus Species 0.000 description 1
- XTFNPKDYCLFGPV-OMCISZLKSA-N Bromofenoxim Chemical compound C1=C(Br)C(O)=C(Br)C=C1\C=N\OC1=CC=C([N+]([O-])=O)C=C1[N+]([O-])=O XTFNPKDYCLFGPV-OMCISZLKSA-N 0.000 description 1
- 241000978501 Brunnichia Species 0.000 description 1
- 241000978499 Brunnichia ovata Species 0.000 description 1
- 239000005742 Bupirimate Substances 0.000 description 1
- ZZVVDIVWGXTDRQ-BSYVCWPDSA-N Buthiobate Chemical compound C=1C=CN=CC=1\N=C(/SCCCC)SCC1=CC=C(C(C)(C)C)C=C1 ZZVVDIVWGXTDRQ-BSYVCWPDSA-N 0.000 description 1
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 1
- 235000014552 Cassia tora Nutrition 0.000 description 1
- 208000013641 Cerebrofacial arteriovenous metameric syndrome Diseases 0.000 description 1
- 235000011498 Chenopodium album var missouriense Nutrition 0.000 description 1
- 235000013328 Chenopodium album var. album Nutrition 0.000 description 1
- 235000014052 Chenopodium album var. microphyllum Nutrition 0.000 description 1
- 235000014050 Chenopodium album var. stevensii Nutrition 0.000 description 1
- 235000013012 Chenopodium album var. striatum Nutrition 0.000 description 1
- 244000281762 Chenopodium ambrosioides Species 0.000 description 1
- 235000000509 Chenopodium ambrosioides Nutrition 0.000 description 1
- 235000005484 Chenopodium berlandieri Nutrition 0.000 description 1
- 235000005490 Chenopodium botrys Nutrition 0.000 description 1
- 235000009332 Chenopodium rubrum Nutrition 0.000 description 1
- DXXVCXKMSWHGTF-UHFFFAOYSA-N Chlomethoxyfen Chemical compound C1=C([N+]([O-])=O)C(OC)=CC(OC=2C(=CC(Cl)=CC=2)Cl)=C1 DXXVCXKMSWHGTF-UHFFFAOYSA-N 0.000 description 1
- NLYNUTMZTCLNOO-UHFFFAOYSA-N Chlorbromuron Chemical compound CON(C)C(=O)NC1=CC=C(Br)C(Cl)=C1 NLYNUTMZTCLNOO-UHFFFAOYSA-N 0.000 description 1
- 239000005493 Chloridazon (aka pyrazone) Substances 0.000 description 1
- 239000005647 Chlorpropham Substances 0.000 description 1
- 108010077544 Chromatin Proteins 0.000 description 1
- WMLPCIHUFDKWJU-UHFFFAOYSA-N Cinosulfuron Chemical compound COCCOC1=CC=CC=C1S(=O)(=O)NC(=O)NC1=NC(OC)=NC(OC)=N1 WMLPCIHUFDKWJU-UHFFFAOYSA-N 0.000 description 1
- 239000005497 Clethodim Substances 0.000 description 1
- 239000005888 Clothianidin Substances 0.000 description 1
- 241000222199 Colletotrichum Species 0.000 description 1
- 241000233839 Commelina communis Species 0.000 description 1
- 241000207894 Convolvulus arvensis Species 0.000 description 1
- 244000074881 Conyza canadensis Species 0.000 description 1
- RYGMFSIKBFXOCR-UHFFFAOYSA-N Copper Chemical compound [Cu] RYGMFSIKBFXOCR-UHFFFAOYSA-N 0.000 description 1
- JJLJMEJHUUYSSY-UHFFFAOYSA-L Copper hydroxide Chemical compound [OH-].[OH-].[Cu+2] JJLJMEJHUUYSSY-UHFFFAOYSA-L 0.000 description 1
- 239000005750 Copper hydroxide Substances 0.000 description 1
- 239000005752 Copper oxychloride Substances 0.000 description 1
- MIKUYHXYGGJMLM-GIMIYPNGSA-N Crotonoside Natural products C1=NC2=C(N)NC(=O)N=C2N1[C@H]1O[C@@H](CO)[C@H](O)[C@@H]1O MIKUYHXYGGJMLM-GIMIYPNGSA-N 0.000 description 1
- 239000005754 Cyazofamid Substances 0.000 description 1
- 229920000089 Cyclic olefin copolymer Polymers 0.000 description 1
- DFCAFRGABIXSDS-UHFFFAOYSA-N Cycloate Chemical compound CCSC(=O)N(CC)C1CCCCC1 DFCAFRGABIXSDS-UHFFFAOYSA-N 0.000 description 1
- OFSLKOLYLQSJPB-UHFFFAOYSA-N Cyclosulfamuron Chemical compound COC1=CC(OC)=NC(NC(=O)NS(=O)(=O)NC=2C(=CC=CC=2)C(=O)C2CC2)=N1 OFSLKOLYLQSJPB-UHFFFAOYSA-N 0.000 description 1
- 239000005501 Cycloxydim Substances 0.000 description 1
- 239000005755 Cyflufenamid Substances 0.000 description 1
- 239000005655 Cyflumetofen Substances 0.000 description 1
- 241000234653 Cyperus Species 0.000 description 1
- 244000285774 Cyperus esculentus Species 0.000 description 1
- 244000075634 Cyperus rotundus Species 0.000 description 1
- 102000018832 Cytochromes Human genes 0.000 description 1
- 108010052832 Cytochromes Proteins 0.000 description 1
- IGXWBGJHJZYPQS-SSDOTTSWSA-N D-Luciferin Chemical compound OC(=O)[C@H]1CSC(C=2SC3=CC=C(O)C=C3N=2)=N1 IGXWBGJHJZYPQS-SSDOTTSWSA-N 0.000 description 1
- NYHBQMYGNKIUIF-UHFFFAOYSA-N D-guanosine Natural products C1=2NC(N)=NC(=O)C=2N=CN1C1OC(CO)C(O)C1O NYHBQMYGNKIUIF-UHFFFAOYSA-N 0.000 description 1
- 102000012410 DNA Ligases Human genes 0.000 description 1
- 108010061982 DNA Ligases Proteins 0.000 description 1
- 238000007400 DNA extraction Methods 0.000 description 1
- 101710096830 DNA-3-methyladenine glycosylase Proteins 0.000 description 1
- 102100039128 DNA-3-methyladenine glycosylase Human genes 0.000 description 1
- NPOJQCVWMSKXDN-UHFFFAOYSA-N Dacthal Chemical group COC(=O)C1=C(Cl)C(Cl)=C(C(=O)OC)C(Cl)=C1Cl NPOJQCVWMSKXDN-UHFFFAOYSA-N 0.000 description 1
- NDUPDOJHUQKPAG-UHFFFAOYSA-N Dalapon Chemical compound CC(Cl)(Cl)C(O)=O NDUPDOJHUQKPAG-UHFFFAOYSA-N 0.000 description 1
- CYCGRDQQIOGCKX-UHFFFAOYSA-N Dehydro-luciferin Natural products OC(=O)C1=CSC(C=2SC3=CC(O)=CC=C3N=2)=N1 CYCGRDQQIOGCKX-UHFFFAOYSA-N 0.000 description 1
- 229940123980 Desaturase inhibitor Drugs 0.000 description 1
- PQUCIEFHOVEZAU-UHFFFAOYSA-N Diammonium sulfite Chemical compound [NH4+].[NH4+].[O-]S([O-])=O PQUCIEFHOVEZAU-UHFFFAOYSA-N 0.000 description 1
- 239000005759 Diethofencarb Substances 0.000 description 1
- 239000005893 Diflubenzuron Substances 0.000 description 1
- 235000017896 Digitaria Nutrition 0.000 description 1
- 241001303487 Digitaria <clam> Species 0.000 description 1
- 244000152970 Digitaria sanguinalis Species 0.000 description 1
- 235000010823 Digitaria sanguinalis Nutrition 0.000 description 1
- IKYICRRUVNIHPP-UHFFFAOYSA-N Dimethametryn Chemical compound CCNC1=NC(NC(C)C(C)C)=NC(SC)=N1 IKYICRRUVNIHPP-UHFFFAOYSA-N 0.000 description 1
- PHVNLLCAQHGNKU-UHFFFAOYSA-N Dimethipin Chemical compound CC1=C(C)S(=O)(=O)CCS1(=O)=O PHVNLLCAQHGNKU-UHFFFAOYSA-N 0.000 description 1
- 108010028143 Dioxygenases Proteins 0.000 description 1
- 102000016680 Dioxygenases Human genes 0.000 description 1
- QAHFOPIILNICLA-UHFFFAOYSA-N Diphenamid Chemical compound C=1C=CC=CC=1C(C(=O)N(C)C)C1=CC=CC=C1 QAHFOPIILNICLA-UHFFFAOYSA-N 0.000 description 1
- 239000005630 Diquat Substances 0.000 description 1
- 208000035240 Disease Resistance Diseases 0.000 description 1
- QXNVGIXVLWOKEQ-UHFFFAOYSA-N Disodium Chemical class [Na][Na] QXNVGIXVLWOKEQ-UHFFFAOYSA-N 0.000 description 1
- 239000005764 Dithianon Substances 0.000 description 1
- 239000005765 Dodemorph Substances 0.000 description 1
- 241000192043 Echinochloa Species 0.000 description 1
- 235000007351 Eleusine Nutrition 0.000 description 1
- 241000209215 Eleusine Species 0.000 description 1
- 102100023882 Endoribonuclease ZC3H12A Human genes 0.000 description 1
- 101710112715 Endoribonuclease ZC3H12A Proteins 0.000 description 1
- 244000148064 Enicostema verticillatum Species 0.000 description 1
- 241000044408 Eriochloa Species 0.000 description 1
- PTFJIKYUEPWBMS-UHFFFAOYSA-N Ethalfluralin Chemical compound CC(=C)CN(CC)C1=C([N+]([O-])=O)C=C(C(F)(F)F)C=C1[N+]([O-])=O PTFJIKYUEPWBMS-UHFFFAOYSA-N 0.000 description 1
- 239000005976 Ethephon Substances 0.000 description 1
- KCOCSOWTADCKOL-UHFFFAOYSA-N Ethidimuron Chemical compound CCS(=O)(=O)C1=NN=C(N(C)C(=O)NC)S1 KCOCSOWTADCKOL-UHFFFAOYSA-N 0.000 description 1
- UWVKRNOCDUPIDM-UHFFFAOYSA-N Ethoxysulfuron Chemical compound CCOC1=CC=CC=C1OS(=O)(=O)NC(=O)NC1=NC(OC)=CC(OC)=N1 UWVKRNOCDUPIDM-UHFFFAOYSA-N 0.000 description 1
- ICWUMLXQKFTJMH-UHFFFAOYSA-N Etobenzanid Chemical compound C1=CC(OCOCC)=CC=C1C(=O)NC1=CC=CC(Cl)=C1Cl ICWUMLXQKFTJMH-UHFFFAOYSA-N 0.000 description 1
- 241001473317 Eupatorium cannabinum Species 0.000 description 1
- 241001553700 Euphorbia lathyris Species 0.000 description 1
- WZKSXHQDXQKIQJ-UHFFFAOYSA-N F[C](F)F Chemical group F[C](F)F WZKSXHQDXQKIQJ-UHFFFAOYSA-N 0.000 description 1
- 244000248416 Fagopyrum cymosum Species 0.000 description 1
- 108010087894 Fatty acid desaturases Proteins 0.000 description 1
- GMBRUAIJEFRHFQ-UHFFFAOYSA-N Fenchlorazole-ethyl Chemical group N1=C(C(=O)OCC)N=C(C(Cl)(Cl)Cl)N1C1=CC=C(Cl)C=C1Cl GMBRUAIJEFRHFQ-UHFFFAOYSA-N 0.000 description 1
- HMIBKHHNXANVHR-UHFFFAOYSA-N Fenothiocarb Chemical compound CN(C)C(=O)SCCCCOC1=CC=CC=C1 HMIBKHHNXANVHR-UHFFFAOYSA-N 0.000 description 1
- PQKBPHSEKWERTG-UHFFFAOYSA-N Fenoxaprop ethyl Chemical group C1=CC(OC(C)C(=O)OCC)=CC=C1OC1=NC2=CC=C(Cl)C=C2O1 PQKBPHSEKWERTG-UHFFFAOYSA-N 0.000 description 1
- XDWSZOWLJIWERG-UHFFFAOYSA-N Fenuron-TCA Chemical compound [O-]C(=O)C(Cl)(Cl)Cl.C[NH+](C)C(=O)NC1=CC=CC=C1 XDWSZOWLJIWERG-UHFFFAOYSA-N 0.000 description 1
- 239000004606 Fillers/Extenders Substances 0.000 description 1
- BJGNCJDXODQBOB-UHFFFAOYSA-N Fivefly Luciferin Natural products OC(=O)C1CSC(C=2SC3=CC(O)=CC=C3N=2)=N1 BJGNCJDXODQBOB-UHFFFAOYSA-N 0.000 description 1
- 239000005901 Flubendiamide Substances 0.000 description 1
- FICWGWVVIRLNRB-UHFFFAOYSA-N Flucetosulfuron Chemical compound COCC(=O)OC(C(C)F)C1=NC=CC=C1S(=O)(=O)NC(=O)NC1=NC(OC)=CC(OC)=N1 FICWGWVVIRLNRB-UHFFFAOYSA-N 0.000 description 1
- PXRROZVNOOEPPZ-UHFFFAOYSA-N Flupropanate Chemical compound OC(=O)C(F)(F)C(F)F PXRROZVNOOEPPZ-UHFFFAOYSA-N 0.000 description 1
- GXAMYUGOODKVRM-UHFFFAOYSA-N Flurecol Chemical compound C1=CC=C2C(C(=O)O)(O)C3=CC=CC=C3C2=C1 GXAMYUGOODKVRM-UHFFFAOYSA-N 0.000 description 1
- 239000005535 Flurochloridone Substances 0.000 description 1
- 239000005558 Fluroxypyr Substances 0.000 description 1
- UKSLKNUCVPZQCQ-UHFFFAOYSA-N Fluxofenim Chemical compound C=1C=C(Cl)C=CC=1C(C(F)(F)F)=NOCC1OCCO1 UKSLKNUCVPZQCQ-UHFFFAOYSA-N 0.000 description 1
- 239000005791 Fuberidazole Substances 0.000 description 1
- 108700039691 Genetic Promoter Regions Proteins 0.000 description 1
- 108700007698 Genetic Terminator Regions Proteins 0.000 description 1
- 239000005980 Gibberellic acid Substances 0.000 description 1
- 235000010469 Glycine max Nutrition 0.000 description 1
- 244000068988 Glycine max Species 0.000 description 1
- 108030006517 Glyphosate oxidoreductases Proteins 0.000 description 1
- 241000208818 Helianthus Species 0.000 description 1
- 241001262556 Helianthus ciliaris Species 0.000 description 1
- 241000701443 Helicoverpa zea single nucleopolyhedrovirus Species 0.000 description 1
- GVGLGOZIDCSQPN-PVHGPHFFSA-N Heroin Chemical compound O([C@H]1[C@H](C=C[C@H]23)OC(C)=O)C4=C5[C@@]12CCN(C)[C@@H]3CC5=CC=C4OC(C)=O GVGLGOZIDCSQPN-PVHGPHFFSA-N 0.000 description 1
- 108010034791 Heterochromatin Proteins 0.000 description 1
- CAWXEEYDBZRFPE-UHFFFAOYSA-N Hexazinone Chemical compound O=C1N(C)C(N(C)C)=NC(=O)N1C1CCCCC1 CAWXEEYDBZRFPE-UHFFFAOYSA-N 0.000 description 1
- 239000005661 Hexythiazox Substances 0.000 description 1
- 240000005979 Hordeum vulgare Species 0.000 description 1
- 235000007340 Hordeum vulgare Nutrition 0.000 description 1
- 206010020649 Hyperkeratosis Diseases 0.000 description 1
- 101150062179 II gene Proteins 0.000 description 1
- PMAAYIYCDXGUAP-UHFFFAOYSA-N Indanofan Chemical compound O=C1C2=CC=CC=C2C(=O)C1(CC)CC1(C=2C=C(Cl)C=CC=2)CO1 PMAAYIYCDXGUAP-UHFFFAOYSA-N 0.000 description 1
- 108091092195 Intron Proteins 0.000 description 1
- QBEXFUOWUYCXNI-UHFFFAOYSA-N Ioxynil octanoate Chemical compound CCCCCCCC(=O)OC1=C(I)C=C(C#N)C=C1I QBEXFUOWUYCXNI-UHFFFAOYSA-N 0.000 description 1
- 239000005867 Iprodione Substances 0.000 description 1
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 description 1
- 235000009496 Juglans regia Nutrition 0.000 description 1
- 240000007049 Juglans regia Species 0.000 description 1
- 241000088861 Latua Species 0.000 description 1
- 108090001090 Lectins Proteins 0.000 description 1
- 102000004856 Lectins Human genes 0.000 description 1
- 239000005572 Lenacil Substances 0.000 description 1
- 241000209510 Liliopsida Species 0.000 description 1
- 235000003403 Limnocharis flava Nutrition 0.000 description 1
- 108060001084 Luciferase Proteins 0.000 description 1
- 239000005089 Luciferase Substances 0.000 description 1
- DDWFXDSYGUXRAY-UHFFFAOYSA-N Luciferin Natural products CCc1c(C)c(CC2NC(=O)C(=C2C=C)C)[nH]c1Cc3[nH]c4C(=C5/NC(CC(=O)O)C(C)C5CC(=O)O)CC(=O)c4c3C DDWFXDSYGUXRAY-UHFFFAOYSA-N 0.000 description 1
- KDXKERNSBIXSRK-UHFFFAOYSA-N Lysine Natural products NCCCCC(N)C(O)=O KDXKERNSBIXSRK-UHFFFAOYSA-N 0.000 description 1
- 239000004472 Lysine Substances 0.000 description 1
- 239000005574 MCPA Substances 0.000 description 1
- AZFKQCNGMSSWDS-UHFFFAOYSA-N MCPA-thioethyl Chemical group CCSC(=O)COC1=CC=C(Cl)C=C1C AZFKQCNGMSSWDS-UHFFFAOYSA-N 0.000 description 1
- 239000005575 MCPB Substances 0.000 description 1
- 101150039283 MCPB gene Proteins 0.000 description 1
- 239000005804 Mandipropamid Substances 0.000 description 1
- 240000006236 Martynia annua Species 0.000 description 1
- 239000005576 Mecoprop-P Substances 0.000 description 1
- OKIBNKKYNPBDRS-UHFFFAOYSA-N Mefluidide Chemical compound CC(=O)NC1=CC(NS(=O)(=O)C(F)(F)F)=C(C)C=C1C OKIBNKKYNPBDRS-UHFFFAOYSA-N 0.000 description 1
- 239000005805 Mepanipyrim Substances 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- BWPYBAJTDILQPY-UHFFFAOYSA-N Methoxyphenone Chemical compound C1=C(C)C(OC)=CC=C1C(=O)C1=CC=CC(C)=C1 BWPYBAJTDILQPY-UHFFFAOYSA-N 0.000 description 1
- FMINYZXVCTYSNY-UHFFFAOYSA-N Methyldymron Chemical compound C=1C=CC=CC=1N(C)C(=O)NC(C)(C)C1=CC=CC=C1 FMINYZXVCTYSNY-UHFFFAOYSA-N 0.000 description 1
- 239000005809 Metiram Substances 0.000 description 1
- 239000005583 Metribuzin Substances 0.000 description 1
- BYBLEWFAAKGYCD-UHFFFAOYSA-N Miconazole Chemical compound ClC1=CC(Cl)=CC=C1COC(C=1C(=CC(Cl)=CC=1)Cl)CN1C=NC=C1 BYBLEWFAAKGYCD-UHFFFAOYSA-N 0.000 description 1
- 102000029749 Microtubule Human genes 0.000 description 1
- 108091022875 Microtubule Proteins 0.000 description 1
- 229920000881 Modified starch Polymers 0.000 description 1
- LKJPSUCKSLORMF-UHFFFAOYSA-N Monolinuron Chemical compound CON(C)C(=O)NC1=CC=C(Cl)C=C1 LKJPSUCKSLORMF-UHFFFAOYSA-N 0.000 description 1
- 239000005811 Myclobutanil Substances 0.000 description 1
- IUOKJNROJISWRO-UHFFFAOYSA-N N-(2-cyano-3-methylbutan-2-yl)-2-(2,4-dichlorophenoxy)propanamide Chemical compound CC(C)C(C)(C#N)NC(=O)C(C)OC1=CC=C(Cl)C=C1Cl IUOKJNROJISWRO-UHFFFAOYSA-N 0.000 description 1
- NQRFDNJEBWAUBL-UHFFFAOYSA-N N-[cyano(2-thienyl)methyl]-4-ethyl-2-(ethylamino)-1,3-thiazole-5-carboxamide Chemical compound S1C(NCC)=NC(CC)=C1C(=O)NC(C#N)C1=CC=CS1 NQRFDNJEBWAUBL-UHFFFAOYSA-N 0.000 description 1
- NTBVTCXMRYKRTB-UHFFFAOYSA-N N-{2-[(4,6-dimethoxypyrimidin-2-yl)(hydroxy)methyl]-6-(methoxymethyl)phenyl}-1,1-difluoromethanesulfonamide Chemical compound COCC1=CC=CC(C(O)C=2N=C(OC)C=C(OC)N=2)=C1NS(=O)(=O)C(F)F NTBVTCXMRYKRTB-UHFFFAOYSA-N 0.000 description 1
- CCGPUGMWYLICGL-UHFFFAOYSA-N Neburon Chemical compound CCCCN(C)C(=O)NC1=CC=C(Cl)C(Cl)=C1 CCGPUGMWYLICGL-UHFFFAOYSA-N 0.000 description 1
- 108020004711 Nucleic Acid Probes Proteins 0.000 description 1
- 239000005587 Oryzalin Substances 0.000 description 1
- WFVUIONFJOAYPK-KAMYIIQDSA-N Oxabetrinil Chemical compound C=1C=CC=CC=1C(/C#N)=N\OCC1OCCO1 WFVUIONFJOAYPK-KAMYIIQDSA-N 0.000 description 1
- FCOHEOSCARXMMS-UHFFFAOYSA-N Oxaziclomefone Chemical compound C1OC(C)=C(C=2C=CC=CC=2)C(=O)N1C(C)(C)C1=CC(Cl)=CC(Cl)=C1 FCOHEOSCARXMMS-UHFFFAOYSA-N 0.000 description 1
- KYGZCKSPAKDVKC-UHFFFAOYSA-N Oxolinic acid Chemical compound C1=C2N(CC)C=C(C(O)=O)C(=O)C2=CC2=C1OCO2 KYGZCKSPAKDVKC-UHFFFAOYSA-N 0.000 description 1
- 238000010222 PCR analysis Methods 0.000 description 1
- 229910019142 PO4 Inorganic materials 0.000 description 1
- 239000005985 Paclobutrazol Substances 0.000 description 1
- 239000005643 Pelargonic acid Substances 0.000 description 1
- 239000005813 Penconazole Substances 0.000 description 1
- 239000005816 Penthiopyrad Substances 0.000 description 1
- 102000035195 Peptidases Human genes 0.000 description 1
- 108091005804 Peptidases Proteins 0.000 description 1
- WHTBVLXUSXVMEV-UHFFFAOYSA-N Perfluidone Chemical compound C1=C(NS(=O)(=O)C(F)(F)F)C(C)=CC(S(=O)(=O)C=2C=CC=CC=2)=C1 WHTBVLXUSXVMEV-UHFFFAOYSA-N 0.000 description 1
- 239000005593 Pethoxamid Substances 0.000 description 1
- 241000257649 Phalaris minor Species 0.000 description 1
- 239000005594 Phenmedipham Substances 0.000 description 1
- 241000233614 Phytophthora Species 0.000 description 1
- 239000005597 Pinoxaden Substances 0.000 description 1
- UNLYSVIDNRIVFJ-UHFFFAOYSA-N Piperophos Chemical compound CCCOP(=S)(OCCC)SCC(=O)N1CCCCC1C UNLYSVIDNRIVFJ-UHFFFAOYSA-N 0.000 description 1
- 108700001094 Plant Genes Proteins 0.000 description 1
- 229930182764 Polyoxin Natural products 0.000 description 1
- 235000015926 Proboscidea louisianica ssp. fragrans Nutrition 0.000 description 1
- 235000015925 Proboscidea louisianica subsp. louisianica Nutrition 0.000 description 1
- 235000019096 Proboscidea parviflora Nutrition 0.000 description 1
- 239000005820 Prochloraz Substances 0.000 description 1
- RSVPPPHXAASNOL-UHFFFAOYSA-N Prodiamine Chemical compound CCCN(CCC)C1=C([N+]([O-])=O)C=C(C(F)(F)F)C(N)=C1[N+]([O-])=O RSVPPPHXAASNOL-UHFFFAOYSA-N 0.000 description 1
- 239000005599 Profoxydim Substances 0.000 description 1
- IPDFPNNPBMREIF-CHWSQXEVSA-N Prohydrojasmon Chemical compound CCCCC[C@@H]1[C@@H](CC(=O)OCCC)CCC1=O IPDFPNNPBMREIF-CHWSQXEVSA-N 0.000 description 1
- 239000005823 Propineb Substances 0.000 description 1
- 239000005601 Propoxycarbazone Substances 0.000 description 1
- 239000005602 Propyzamide Substances 0.000 description 1
- 239000005603 Prosulfocarb Substances 0.000 description 1
- LTUNNEGNEKBSEH-UHFFFAOYSA-N Prosulfuron Chemical compound COC1=NC(C)=NC(NC(=O)NS(=O)(=O)C=2C(=CC=CC=2)CCC(F)(F)F)=N1 LTUNNEGNEKBSEH-UHFFFAOYSA-N 0.000 description 1
- 239000005604 Prosulfuron Substances 0.000 description 1
- 239000005925 Pymetrozine Substances 0.000 description 1
- 239000005869 Pyraclostrobin Substances 0.000 description 1
- WTKZEGDFNFYCGP-UHFFFAOYSA-O Pyrazolium Chemical compound C1=CN[NH+]=C1 WTKZEGDFNFYCGP-UHFFFAOYSA-O 0.000 description 1
- 239000005663 Pyridaben Substances 0.000 description 1
- 239000005606 Pyridate Substances 0.000 description 1
- JTZCTMAVMHRNTR-UHFFFAOYSA-N Pyridate Chemical compound CCCCCCCCSC(=O)OC1=CC(Cl)=NN=C1C1=CC=CC=C1 JTZCTMAVMHRNTR-UHFFFAOYSA-N 0.000 description 1
- RRKHIAYNPVQKEF-UHFFFAOYSA-N Pyriftalid Chemical compound COC1=CC(OC)=NC(SC=2C=3C(=O)OC(C)C=3C=CC=2)=N1 RRKHIAYNPVQKEF-UHFFFAOYSA-N 0.000 description 1
- CZPWVGJYEJSRLH-UHFFFAOYSA-N Pyrimidine Chemical compound C1=CN=CN=C1 CZPWVGJYEJSRLH-UHFFFAOYSA-N 0.000 description 1
- MWMQNVGAHVXSPE-UHFFFAOYSA-N Pyriprole Chemical compound ClC=1C=C(C(F)(F)F)C=C(Cl)C=1N1N=C(C#N)C(SC(F)F)=C1NCC1=CC=CC=N1 MWMQNVGAHVXSPE-UHFFFAOYSA-N 0.000 description 1
- CNILNQMBAHKMFS-UHFFFAOYSA-M Pyrithiobac-sodium Chemical compound [Na+].COC1=CC(OC)=NC(SC=2C(=C(Cl)C=CC=2)C([O-])=O)=N1 CNILNQMBAHKMFS-UHFFFAOYSA-M 0.000 description 1
- 239000005608 Quinmerac Substances 0.000 description 1
- 239000005615 Quizalofop-P-tefuryl Substances 0.000 description 1
- 108091034057 RNA (poly(A)) Proteins 0.000 description 1
- 108020004511 Recombinant DNA Proteins 0.000 description 1
- 235000001537 Ribes X gardonianum Nutrition 0.000 description 1
- 235000001535 Ribes X utile Nutrition 0.000 description 1
- 235000016919 Ribes petraeum Nutrition 0.000 description 1
- 244000281247 Ribes rubrum Species 0.000 description 1
- 235000002355 Ribes spicatum Nutrition 0.000 description 1
- 244000275012 Sesbania cannabina Species 0.000 description 1
- 241000533293 Sesbania emerus Species 0.000 description 1
- 235000005775 Setaria Nutrition 0.000 description 1
- 241000232088 Setaria <nematode> Species 0.000 description 1
- 235000017016 Setaria faberi Nutrition 0.000 description 1
- 241000820191 Setaria magna Species 0.000 description 1
- KEAYESYHFKHZAL-UHFFFAOYSA-N Sodium Chemical compound [Na] KEAYESYHFKHZAL-UHFFFAOYSA-N 0.000 description 1
- 241000209072 Sorghum Species 0.000 description 1
- 235000011684 Sorghum saccharatum Nutrition 0.000 description 1
- 239000005664 Spirodiclofen Substances 0.000 description 1
- 239000005665 Spiromesifen Substances 0.000 description 1
- 239000005931 Spirotetramat Substances 0.000 description 1
- 102000004523 Sulfate Adenylyltransferase Human genes 0.000 description 1
- 108010022348 Sulfate adenylyltransferase Proteins 0.000 description 1
- SAQSTQBVENFSKT-UHFFFAOYSA-M TCA-sodium Chemical compound [Na+].[O-]C(=O)C(Cl)(Cl)Cl SAQSTQBVENFSKT-UHFFFAOYSA-M 0.000 description 1
- 239000005620 Tembotrione Substances 0.000 description 1
- KDWQYMVPYJGPHS-UHFFFAOYSA-N Thenylchlor Chemical compound C1=CSC(CN(C(=O)CCl)C=2C(=CC=CC=2C)C)=C1OC KDWQYMVPYJGPHS-UHFFFAOYSA-N 0.000 description 1
- 239000005940 Thiacloprid Substances 0.000 description 1
- 239000005622 Thiencarbazone Substances 0.000 description 1
- 239000005843 Thiram Substances 0.000 description 1
- 239000005624 Tralkoxydim Substances 0.000 description 1
- WHKUVVPPKQRRBV-UHFFFAOYSA-N Trasan Chemical compound CC1=CC(Cl)=CC=C1OCC(O)=O WHKUVVPPKQRRBV-UHFFFAOYSA-N 0.000 description 1
- 239000005625 Tri-allate Substances 0.000 description 1
- MWBPRDONLNQCFV-UHFFFAOYSA-N Tri-allate Chemical compound CC(C)N(C(C)C)C(=O)SCC(Cl)=C(Cl)Cl MWBPRDONLNQCFV-UHFFFAOYSA-N 0.000 description 1
- 239000005846 Triadimenol Substances 0.000 description 1
- 239000005627 Triclopyr Substances 0.000 description 1
- IBZHOAONZVJLOB-UHFFFAOYSA-N Tridiphane Chemical compound ClC1=CC(Cl)=CC(C2(CC(Cl)(Cl)Cl)OC2)=C1 IBZHOAONZVJLOB-UHFFFAOYSA-N 0.000 description 1
- HFBWPRKWDIRYNX-UHFFFAOYSA-N Trietazine Chemical compound CCNC1=NC(Cl)=NC(N(CC)CC)=N1 HFBWPRKWDIRYNX-UHFFFAOYSA-N 0.000 description 1
- GSEJCLTVZPLZKY-UHFFFAOYSA-N Triethanolamine Chemical class OCCN(CCO)CCO GSEJCLTVZPLZKY-UHFFFAOYSA-N 0.000 description 1
- 239000005629 Tritosulfuron Substances 0.000 description 1
- 108091023045 Untranslated Region Proteins 0.000 description 1
- 208000036142 Viral infection Diseases 0.000 description 1
- 239000005870 Ziram Substances 0.000 description 1
- 239000005863 Zoxamide Substances 0.000 description 1
- FHOXQLTVOMNIOR-QZPAGEHASA-N [(1S,2R,4R,5S,7S,11S,12S,15R,16S)-2,16-dimethyl-15-[(1S)-1-[(2R,3R)-3-[(3S)-2-methylpentan-3-yl]oxiran-2-yl]ethyl]-8-oxo-4-propanoyloxy-9-oxatetracyclo[9.7.0.02,7.012,16]octadecan-5-yl] propanoate Chemical compound CC[C@@H](C(C)C)[C@H]1O[C@@H]1[C@@H](C)[C@@H]1[C@@]2(C)CC[C@@H]3[C@@]4(C)C[C@@H](OC(=O)CC)[C@@H](OC(=O)CC)C[C@@H]4C(=O)OC[C@H]3[C@@H]2CC1 FHOXQLTVOMNIOR-QZPAGEHASA-N 0.000 description 1
- QQODLKZGRKWIFG-RUTXASTPSA-N [(R)-cyano-(4-fluoro-3-phenoxyphenyl)methyl] (1S)-3-(2,2-dichloroethenyl)-2,2-dimethylcyclopropane-1-carboxylate Chemical compound CC1(C)C(C=C(Cl)Cl)[C@@H]1C(=O)O[C@@H](C#N)C1=CC=C(F)C(OC=2C=CC=CC=2)=C1 QQODLKZGRKWIFG-RUTXASTPSA-N 0.000 description 1
- KVIZNNVXXNFLMU-AIIUZBJTSA-N [2,3,5,6-tetrafluoro-4-(methoxymethyl)phenyl]methyl (1r,3r)-2,2-dimethyl-3-[(e)-prop-1-enyl]cyclopropane-1-carboxylate Chemical compound FC1=C(F)C(COC)=C(F)C(F)=C1COC(=O)[C@H]1C(C)(C)[C@@H]1\C=C\C KVIZNNVXXNFLMU-AIIUZBJTSA-N 0.000 description 1
- OOWCJRMYMAMSOH-UHFFFAOYSA-N [2,3,5,6-tetrafluoro-4-(methoxymethyl)phenyl]methyl 2,2-dimethyl-3-(2-methylprop-1-enyl)cyclopropane-1-carboxylate Chemical compound FC1=C(F)C(COC)=C(F)C(F)=C1COC(=O)C1C(C)(C)C1C=C(C)C OOWCJRMYMAMSOH-UHFFFAOYSA-N 0.000 description 1
- JLCPHMBAVCMARE-UHFFFAOYSA-N [3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-[[3-[[3-[[3-[[3-[[3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-hydroxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methyl [5-(6-aminopurin-9-yl)-2-(hydroxymethyl)oxolan-3-yl] hydrogen phosphate Polymers Cc1cn(C2CC(OP(O)(=O)OCC3OC(CC3OP(O)(=O)OCC3OC(CC3O)n3cnc4c3nc(N)[nH]c4=O)n3cnc4c3nc(N)[nH]c4=O)C(COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3CO)n3cnc4c(N)ncnc34)n3ccc(N)nc3=O)n3cnc4c(N)ncnc34)n3ccc(N)nc3=O)n3ccc(N)nc3=O)n3ccc(N)nc3=O)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cc(C)c(=O)[nH]c3=O)n3cc(C)c(=O)[nH]c3=O)n3ccc(N)nc3=O)n3cc(C)c(=O)[nH]c3=O)n3cnc4c3nc(N)[nH]c4=O)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)O2)c(=O)[nH]c1=O JLCPHMBAVCMARE-UHFFFAOYSA-N 0.000 description 1
- 229950008167 abamectin Drugs 0.000 description 1
- 150000003869 acetamides Chemical class 0.000 description 1
- OJFDKHTZOUZBOS-CITAKDKDSA-N acetoacetyl-CoA Chemical compound O[C@@H]1[C@H](OP(O)(O)=O)[C@@H](COP(O)(=O)OP(O)(=O)OCC(C)(C)[C@@H](O)C(=O)NCCC(=O)NCCSC(=O)CC(=O)C)O[C@H]1N1C2=NC=NC(N)=C2N=C1 OJFDKHTZOUZBOS-CITAKDKDSA-N 0.000 description 1
- 125000002777 acetyl group Chemical group [H]C([H])([H])C(*)=O 0.000 description 1
- 230000000397 acetylating effect Effects 0.000 description 1
- 102000005421 acetyltransferase Human genes 0.000 description 1
- 239000002535 acidifier Substances 0.000 description 1
- 229940095602 acidifiers Drugs 0.000 description 1
- NUFNQYOELLVIPL-UHFFFAOYSA-N acifluorfen Chemical compound C1=C([N+]([O-])=O)C(C(=O)O)=CC(OC=2C(=CC(=CC=2)C(F)(F)F)Cl)=C1 NUFNQYOELLVIPL-UHFFFAOYSA-N 0.000 description 1
- 239000012190 activator Substances 0.000 description 1
- 125000002015 acyclic group Chemical group 0.000 description 1
- 230000001464 adherent effect Effects 0.000 description 1
- 230000009418 agronomic effect Effects 0.000 description 1
- XCSGPAVHZFQHGE-UHFFFAOYSA-N alachlor Chemical compound CCC1=CC=CC(CC)=C1N(COC)C(=O)CCl XCSGPAVHZFQHGE-UHFFFAOYSA-N 0.000 description 1
- 125000003545 alkoxy group Chemical group 0.000 description 1
- 125000000217 alkyl group Chemical group 0.000 description 1
- 239000011717 all-trans-retinol Substances 0.000 description 1
- 235000019169 all-trans-retinol Nutrition 0.000 description 1
- DTOSIQBPPRVQHS-PDBXOOCHSA-N alpha-linolenic acid Chemical compound CC\C=C/C\C=C/C\C=C/CCCCCCCC(O)=O DTOSIQBPPRVQHS-PDBXOOCHSA-N 0.000 description 1
- 235000020661 alpha-linolenic acid Nutrition 0.000 description 1
- 230000004075 alteration Effects 0.000 description 1
- 150000001408 amides Chemical class 0.000 description 1
- 125000003277 amino group Chemical group 0.000 description 1
- BREATYVWRHIPIY-UHFFFAOYSA-N amisulbrom Chemical compound CN(C)S(=O)(=O)N1C=NC(S(=O)(=O)N2C3=CC(F)=CC=C3C(Br)=C2C)=N1 BREATYVWRHIPIY-UHFFFAOYSA-N 0.000 description 1
- 229960002587 amitraz Drugs 0.000 description 1
- QXAITBQSYVNQDR-ZIOPAAQOSA-N amitraz Chemical compound C=1C=C(C)C=C(C)C=1/N=C/N(C)\C=N\C1=CC=C(C)C=C1C QXAITBQSYVNQDR-ZIOPAAQOSA-N 0.000 description 1
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 description 1
- 229910052921 ammonium sulfate Inorganic materials 0.000 description 1
- 235000011130 ammonium sulphate Nutrition 0.000 description 1
- 230000002528 anti-freeze Effects 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 238000012209 assay specification Methods 0.000 description 1
- 208000014347 autosomal dominant hyaline body myopathy Diseases 0.000 description 1
- 230000000680 avirulence Effects 0.000 description 1
- AKNQMEBLVAMSNZ-UHFFFAOYSA-N azaconazole Chemical compound ClC1=CC(Cl)=CC=C1C1(CN2N=CN=C2)OCCO1 AKNQMEBLVAMSNZ-UHFFFAOYSA-N 0.000 description 1
- 229950000294 azaconazole Drugs 0.000 description 1
- XOEMATDHVZOBSG-UHFFFAOYSA-N azafenidin Chemical compound C1=C(OCC#C)C(Cl)=CC(Cl)=C1N1C(=O)N2CCCCC2=N1 XOEMATDHVZOBSG-UHFFFAOYSA-N 0.000 description 1
- QRSHQJLLXXEYPS-UHFFFAOYSA-N azane;5-ethyl-2-(4-methyl-5-oxo-4-propan-2-yl-1h-imidazol-2-yl)pyridine-3-carboxylic acid Chemical compound [NH4+].[O-]C(=O)C1=CC(CC)=CN=C1C1=NC(C)(C(C)C)C(=O)N1 QRSHQJLLXXEYPS-UHFFFAOYSA-N 0.000 description 1
- WFDXOXNFNRHQEC-GHRIWEEISA-N azoxystrobin Chemical compound CO\C=C(\C(=O)OC)C1=CC=CC=C1OC1=CC(OC=2C(=CC=CC=2)C#N)=NC=N1 WFDXOXNFNRHQEC-GHRIWEEISA-N 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- 230000010310 bacterial transformation Effects 0.000 description 1
- HYJSGOXICXYZGS-UHFFFAOYSA-N benazolin Chemical compound C1=CC=C2SC(=O)N(CC(=O)O)C2=C1Cl HYJSGOXICXYZGS-UHFFFAOYSA-N 0.000 description 1
- LVKBXDHACCFCTA-UHFFFAOYSA-N bencarbazone Chemical compound C1=C(C(N)=S)C(NS(=O)(=O)CC)=CC(N2C(N(C)C(=N2)C(F)(F)F)=O)=C1F LVKBXDHACCFCTA-UHFFFAOYSA-N 0.000 description 1
- XMQFTWRPUQYINF-UHFFFAOYSA-N bensulfuron-methyl Chemical group COC(=O)C1=CC=CC=C1CS(=O)(=O)NC(=O)NC1=NC(OC)=CC(OC)=N1 XMQFTWRPUQYINF-UHFFFAOYSA-N 0.000 description 1
- VVSLYIKSEBPRSN-PELKAZGASA-N benthiavalicarb Chemical compound C1=C(F)C=C2SC([C@@H](C)NC(=O)[C@@H](NC(O)=O)C(C)C)=NC2=C1 VVSLYIKSEBPRSN-PELKAZGASA-N 0.000 description 1
- USRKFGIXLGKMKU-ABAIWWIYSA-N benthiavalicarb-isopropyl Chemical compound C1=C(F)C=C2SC([C@@H](C)NC(=O)[C@H](C(C)C)NC(=O)OC(C)C)=NC2=C1 USRKFGIXLGKMKU-ABAIWWIYSA-N 0.000 description 1
- 229940054066 benzamide antipsychotics Drugs 0.000 description 1
- 150000003936 benzamides Chemical class 0.000 description 1
- CNBGNNVCVSKAQZ-UHFFFAOYSA-N benzidamine Natural products C12=CC=CC=C2C(OCCCN(C)C)=NN1CC1=CC=CC=C1 CNBGNNVCVSKAQZ-UHFFFAOYSA-N 0.000 description 1
- WPYMKLBDIGXBTP-UHFFFAOYSA-N benzoic acid Chemical compound OC(=O)C1=CC=CC=C1 WPYMKLBDIGXBTP-UHFFFAOYSA-N 0.000 description 1
- 235000010233 benzoic acid Nutrition 0.000 description 1
- 150000001559 benzoic acids Chemical class 0.000 description 1
- MKQSWTQPLLCSOB-UHFFFAOYSA-N benzyl 2-chloro-4-(trifluoromethyl)-1,3-thiazole-5-carboxylate Chemical compound N1=C(Cl)SC(C(=O)OCC=2C=CC=CC=2)=C1C(F)(F)F MKQSWTQPLLCSOB-UHFFFAOYSA-N 0.000 description 1
- VHLKTXFWDRXILV-UHFFFAOYSA-N bifenazate Chemical compound C1=C(NNC(=O)OC(C)C)C(OC)=CC=C1C1=CC=CC=C1 VHLKTXFWDRXILV-UHFFFAOYSA-N 0.000 description 1
- 229920000704 biodegradable plastic Polymers 0.000 description 1
- 230000033228 biological regulation Effects 0.000 description 1
- 230000005540 biological transmission Effects 0.000 description 1
- 235000010290 biphenyl Nutrition 0.000 description 1
- 239000004305 biphenyl Substances 0.000 description 1
- 230000000903 blocking effect Effects 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- DSKJPMWIHSOYEA-UHFFFAOYSA-N bupirimate Chemical compound CCCCC1=C(C)N=C(NCC)N=C1OS(=O)(=O)N(C)C DSKJPMWIHSOYEA-UHFFFAOYSA-N 0.000 description 1
- JEDYYFXHPAIBGR-UHFFFAOYSA-N butafenacil Chemical compound O=C1N(C)C(C(F)(F)F)=CC(=O)N1C1=CC=C(Cl)C(C(=O)OC(C)(C)C(=O)OCC=C)=C1 JEDYYFXHPAIBGR-UHFFFAOYSA-N 0.000 description 1
- VAIZTNZGPYBOGF-UHFFFAOYSA-N butyl 2-(4-{[5-(trifluoromethyl)pyridin-2-yl]oxy}phenoxy)propanoate Chemical group C1=CC(OC(C)C(=O)OCCCC)=CC=C1OC1=CC=C(C(F)(F)F)C=N1 VAIZTNZGPYBOGF-UHFFFAOYSA-N 0.000 description 1
- PSGPXWYGJGGEEG-UHFFFAOYSA-N butyl 9-hydroxyfluorene-9-carboxylate Chemical group C1=CC=C2C(C(=O)OCCCC)(O)C3=CC=CC=C3C2=C1 PSGPXWYGJGGEEG-UHFFFAOYSA-N 0.000 description 1
- 125000000484 butyl group Chemical group [H]C([*])([H])C([H])([H])C([H])([H])C([H])([H])[H] 0.000 description 1
- 239000011575 calcium Substances 0.000 description 1
- 229910052791 calcium Inorganic materials 0.000 description 1
- NLKUPINTOLSSLD-UHFFFAOYSA-L calcium;4-(1-oxidopropylidene)-3,5-dioxocyclohexane-1-carboxylate Chemical compound [Ca+2].CCC([O-])=C1C(=O)CC(C([O-])=O)CC1=O NLKUPINTOLSSLD-UHFFFAOYSA-L 0.000 description 1
- JHRWWRDRBPCWTF-OLQVQODUSA-N captafol Chemical compound C1C=CC[C@H]2C(=O)N(SC(Cl)(Cl)C(Cl)Cl)C(=O)[C@H]21 JHRWWRDRBPCWTF-OLQVQODUSA-N 0.000 description 1
- 239000004202 carbamide Substances 0.000 description 1
- 239000006013 carbendazim Substances 0.000 description 1
- 125000002915 carbonyl group Chemical group [*:2]C([*:1])=O 0.000 description 1
- IRUJZVNXZWPBMU-UHFFFAOYSA-N cartap Chemical compound NC(=O)SCC(N(C)C)CSC(N)=O IRUJZVNXZWPBMU-UHFFFAOYSA-N 0.000 description 1
- 150000001768 cations Chemical class 0.000 description 1
- 210000002421 cell wall Anatomy 0.000 description 1
- 230000001413 cellular effect Effects 0.000 description 1
- 239000001913 cellulose Substances 0.000 description 1
- 229920002678 cellulose Polymers 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 239000002738 chelating agent Substances 0.000 description 1
- PNCNFDRSHBFIDM-WOJGMQOQSA-N chembl111617 Chemical compound C=CCO\N=C(/CCC)C1=C(O)C(C(=O)OC)C(C)(C)CC1=O PNCNFDRSHBFIDM-WOJGMQOQSA-N 0.000 description 1
- GGWHBJGBERXSLL-NBVRZTHBSA-N chembl113137 Chemical compound C1C(=O)C(C(=N/OCC)/CCC)=C(O)CC1C1CSCCC1 GGWHBJGBERXSLL-NBVRZTHBSA-N 0.000 description 1
- MXZACTZQSGYANA-UHFFFAOYSA-N chembl545463 Chemical compound Cl.C1=CC(OC)=CC=C1C(N=C1)=CN2C1=NC(C)=C2O MXZACTZQSGYANA-UHFFFAOYSA-N 0.000 description 1
- 239000012707 chemical precursor Substances 0.000 description 1
- 239000003638 chemical reducing agent Substances 0.000 description 1
- 239000005081 chemiluminescent agent Substances 0.000 description 1
- 230000003559 chemosterilizing effect Effects 0.000 description 1
- UISUNVFOGSJSKD-UHFFFAOYSA-N chlorfluazuron Chemical compound FC1=CC=CC(F)=C1C(=O)NC(=O)NC(C=C1Cl)=CC(Cl)=C1OC1=NC=C(C(F)(F)F)C=C1Cl UISUNVFOGSJSKD-UHFFFAOYSA-N 0.000 description 1
- HKMOPYJWSFRURD-UHFFFAOYSA-N chloro hypochlorite;copper Chemical compound [Cu].ClOCl HKMOPYJWSFRURD-UHFFFAOYSA-N 0.000 description 1
- VXIVSQZSERGHQP-UHFFFAOYSA-N chloroacetamide Chemical class NC(=O)CCl VXIVSQZSERGHQP-UHFFFAOYSA-N 0.000 description 1
- 125000004218 chloromethyl group Chemical group [H]C([H])(Cl)* 0.000 description 1
- CWJSHJJYOPWUGX-UHFFFAOYSA-N chlorpropham Chemical compound CC(C)OC(=O)NC1=CC=CC(Cl)=C1 CWJSHJJYOPWUGX-UHFFFAOYSA-N 0.000 description 1
- 210000003483 chromatin Anatomy 0.000 description 1
- NNKKTZOEKDFTBU-YBEGLDIGSA-N cinidon ethyl Chemical compound C1=C(Cl)C(/C=C(\Cl)C(=O)OCC)=CC(N2C(C3=C(CCCC3)C2=O)=O)=C1 NNKKTZOEKDFTBU-YBEGLDIGSA-N 0.000 description 1
- 238000003776 cleavage reaction Methods 0.000 description 1
- SILSDTWXNBZOGF-JWGBMQLESA-N clethodim Chemical compound CCSC(C)CC1CC(O)=C(C(CC)=NOC\C=C\Cl)C(=O)C1 SILSDTWXNBZOGF-JWGBMQLESA-N 0.000 description 1
- JBDHZKLJNAIJNC-LLVKDONJSA-N clodinafop-propargyl Chemical group C1=CC(O[C@H](C)C(=O)OCC#C)=CC=C1OC1=NC=C(Cl)C=C1F JBDHZKLJNAIJNC-LLVKDONJSA-N 0.000 description 1
- 229960004022 clotrimazole Drugs 0.000 description 1
- VNFPBHJOKIVQEB-UHFFFAOYSA-N clotrimazole Chemical compound ClC1=CC=CC=C1C(N1C=NC=C1)(C=1C=CC=CC=1)C1=CC=CC=C1 VNFPBHJOKIVQEB-UHFFFAOYSA-N 0.000 description 1
- 239000003086 colorant Substances 0.000 description 1
- 239000013065 commercial product Substances 0.000 description 1
- 238000004590 computer program Methods 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- 239000010949 copper Substances 0.000 description 1
- 229910052802 copper Inorganic materials 0.000 description 1
- 229910001956 copper hydroxide Inorganic materials 0.000 description 1
- 229910000365 copper sulfate Inorganic materials 0.000 description 1
- ARUVKPQLZAKDPS-UHFFFAOYSA-L copper(II) sulfate Chemical compound [Cu+2].[O-][S+2]([O-])([O-])[O-] ARUVKPQLZAKDPS-UHFFFAOYSA-L 0.000 description 1
- 239000006184 cosolvent Substances 0.000 description 1
- 150000001907 coumarones Chemical class 0.000 description 1
- 238000012272 crop production Methods 0.000 description 1
- 238000004132 cross linking Methods 0.000 description 1
- 239000012297 crystallization seed Substances 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- YXKMMRDKEKCERS-UHFFFAOYSA-N cyazofamid Chemical compound CN(C)S(=O)(=O)N1C(C#N)=NC(Cl)=C1C1=CC=C(C)C=C1 YXKMMRDKEKCERS-UHFFFAOYSA-N 0.000 description 1
- ACMXQHFNODYQAT-UHFFFAOYSA-N cyflufenamid Chemical compound FC1=CC=C(C(F)(F)F)C(C(NOCC2CC2)=NC(=O)CC=2C=CC=CC=2)=C1F ACMXQHFNODYQAT-UHFFFAOYSA-N 0.000 description 1
- ZXQYGBMAQZUVMI-UNOMPAQXSA-N cyhalothrin Chemical compound CC1(C)C(\C=C(/Cl)C(F)(F)F)C1C(=O)OC(C#N)C1=CC=CC(OC=2C=CC=CC=2)=C1 ZXQYGBMAQZUVMI-UNOMPAQXSA-N 0.000 description 1
- OPTASPLRGRRNAP-UHFFFAOYSA-N cytosine Chemical class NC=1C=CNC(=O)N=1 OPTASPLRGRRNAP-UHFFFAOYSA-N 0.000 description 1
- 239000005547 deoxyribonucleotide Substances 0.000 description 1
- 125000002637 deoxyribonucleotide group Chemical group 0.000 description 1
- 230000001419 dependent effect Effects 0.000 description 1
- 230000008021 deposition Effects 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- WZJZMXBKUWKXTQ-UHFFFAOYSA-N desmedipham Chemical compound CCOC(=O)NC1=CC=CC(OC(=O)NC=2C=CC=CC=2)=C1 WZJZMXBKUWKXTQ-UHFFFAOYSA-N 0.000 description 1
- 230000000368 destabilizing effect Effects 0.000 description 1
- 239000003599 detergent Substances 0.000 description 1
- 238000001784 detoxification Methods 0.000 description 1
- WOWBFOBYOAGEEA-UHFFFAOYSA-N diafenthiuron Chemical compound CC(C)C1=C(NC(=S)NC(C)(C)C)C(C(C)C)=CC(OC=2C=CC=CC=2)=C1 WOWBFOBYOAGEEA-UHFFFAOYSA-N 0.000 description 1
- 229960002069 diamorphine Drugs 0.000 description 1
- FHIVAFMUCKRCQO-UHFFFAOYSA-N diazinon Chemical compound CCOP(=S)(OCC)OC1=CC(C)=NC(C(C)C)=N1 FHIVAFMUCKRCQO-UHFFFAOYSA-N 0.000 description 1
- UOAMTSKGCBMZTC-UHFFFAOYSA-N dicofol Chemical compound C=1C=C(Cl)C=CC=1C(C(Cl)(Cl)Cl)(O)C1=CC=C(Cl)C=C1 UOAMTSKGCBMZTC-UHFFFAOYSA-N 0.000 description 1
- LNJNFVJKDJYTEU-UHFFFAOYSA-N diethofencarb Chemical compound CCOC1=CC=C(NC(=O)OC(C)C)C=C1OCC LNJNFVJKDJYTEU-UHFFFAOYSA-N 0.000 description 1
- 229940019503 diflubenzuron Drugs 0.000 description 1
- 235000019621 digestibility Nutrition 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 239000000539 dimer Substances 0.000 description 1
- 125000002147 dimethylamino group Chemical group [H]C([H])([H])N(*)C([H])([H])[H] 0.000 description 1
- 230000003467 diminishing effect Effects 0.000 description 1
- YKBZOVFACRVRJN-UHFFFAOYSA-N dinotefuran Chemical compound [O-][N+](=O)\N=C(/NC)NCC1CCOC1 YKBZOVFACRVRJN-UHFFFAOYSA-N 0.000 description 1
- SDIXRDNYIMOKSG-UHFFFAOYSA-L disodium methyl arsenate Chemical compound [Na+].[Na+].C[As]([O-])([O-])=O SDIXRDNYIMOKSG-UHFFFAOYSA-L 0.000 description 1
- MWEQTWJABOLLOS-UHFFFAOYSA-L disodium;[[[5-(6-aminopurin-9-yl)-3,4-dihydroxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-oxidophosphoryl] hydrogen phosphate;trihydrate Chemical compound O.O.O.[Na+].[Na+].C1=NC=2C(N)=NC=NC=2N1C1OC(COP(O)(=O)OP([O-])(=O)OP(O)([O-])=O)C(O)C1O MWEQTWJABOLLOS-UHFFFAOYSA-L 0.000 description 1
- 239000002270 dispersing agent Substances 0.000 description 1
- PYZSVQVRHDXQSL-UHFFFAOYSA-N dithianon Chemical compound S1C(C#N)=C(C#N)SC2=C1C(=O)C1=CC=CC=C1C2=O PYZSVQVRHDXQSL-UHFFFAOYSA-N 0.000 description 1
- NAGJZTKCGNOGPW-UHFFFAOYSA-N dithiophosphoric acid Chemical class OP(O)(S)=S NAGJZTKCGNOGPW-UHFFFAOYSA-N 0.000 description 1
- JMXKCYUTURMERF-UHFFFAOYSA-N dodemorph Chemical compound C1C(C)OC(C)CN1C1CCCCCCCCCCC1 JMXKCYUTURMERF-UHFFFAOYSA-N 0.000 description 1
- 239000012154 double-distilled water Substances 0.000 description 1
- 229960003913 econazole Drugs 0.000 description 1
- 235000013399 edible fruits Nutrition 0.000 description 1
- 238000001962 electrophoresis Methods 0.000 description 1
- 239000003995 emulsifying agent Substances 0.000 description 1
- DWRKFAJEBUWTQM-UHFFFAOYSA-N etaconazole Chemical compound O1C(CC)COC1(C=1C(=CC(Cl)=CC=1)Cl)CN1N=CN=C1 DWRKFAJEBUWTQM-UHFFFAOYSA-N 0.000 description 1
- ZINJLDJMHCUBIP-UHFFFAOYSA-N ethametsulfuron-methyl Chemical group CCOC1=NC(NC)=NC(NC(=O)NS(=O)(=O)C=2C(=CC=CC=2)C(=O)OC)=N1 ZINJLDJMHCUBIP-UHFFFAOYSA-N 0.000 description 1
- BBXXLROWFHWFQY-UHFFFAOYSA-N ethirimol Chemical compound CCCCC1=C(C)NC(NCC)=NC1=O BBXXLROWFHWFQY-UHFFFAOYSA-N 0.000 description 1
- OSUHJPCHFDQAIT-UHFFFAOYSA-N ethyl 2-{4-[(6-chloroquinoxalin-2-yl)oxy]phenoxy}propanoate Chemical group C1=CC(OC(C)C(=O)OCC)=CC=C1OC1=CN=C(C=C(Cl)C=C2)C2=N1 OSUHJPCHFDQAIT-UHFFFAOYSA-N 0.000 description 1
- XNKARWLGLZGMGX-UHFFFAOYSA-N ethyl 4-(4-chloro-2-methylphenoxy)butanoate Chemical group CCOC(=O)CCCOC1=CC=C(Cl)C=C1C XNKARWLGLZGMGX-UHFFFAOYSA-N 0.000 description 1
- 125000001495 ethyl group Chemical group [H]C([H])([H])C([H])([H])* 0.000 description 1
- IZBNNCFOBMGTQX-UHFFFAOYSA-N etoperidone Chemical group O=C1N(CC)C(CC)=NN1CCCN1CCN(C=2C=C(Cl)C=CC=2)CC1 IZBNNCFOBMGTQX-UHFFFAOYSA-N 0.000 description 1
- 241001233957 eudicotyledons Species 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 238000001704 evaporation Methods 0.000 description 1
- 230000008020 evaporation Effects 0.000 description 1
- QMTNOLKHSWIQBE-FGTMMUONSA-N exo-(+)-cinmethylin Chemical compound O([C@H]1[C@]2(C)CC[C@@](O2)(C1)C(C)C)CC1=CC=CC=C1C QMTNOLKHSWIQBE-FGTMMUONSA-N 0.000 description 1
- 239000000284 extract Substances 0.000 description 1
- 230000002349 favourable effect Effects 0.000 description 1
- JFSPBVWPKOEZCB-UHFFFAOYSA-N fenfuram Chemical compound O1C=CC(C(=O)NC=2C=CC=CC=2)=C1C JFSPBVWPKOEZCB-UHFFFAOYSA-N 0.000 description 1
- WDQNIWFZKXZFAY-UHFFFAOYSA-M fentin acetate Chemical compound CC([O-])=O.C1=CC=CC=C1[Sn+](C=1C=CC=CC=1)C1=CC=CC=C1 WDQNIWFZKXZFAY-UHFFFAOYSA-M 0.000 description 1
- WHDGWKAJBYRJJL-UHFFFAOYSA-K ferbam Chemical compound [Fe+3].CN(C)C([S-])=S.CN(C)C([S-])=S.CN(C)C([S-])=S WHDGWKAJBYRJJL-UHFFFAOYSA-K 0.000 description 1
- 244000037666 field crops Species 0.000 description 1
- ZGNITFSDLCMLGI-UHFFFAOYSA-N flubendiamide Chemical compound CC1=CC(C(F)(C(F)(F)F)C(F)(F)F)=CC=C1NC(=O)C1=CC=CC(I)=C1C(=O)NC(C)(C)CS(C)(=O)=O ZGNITFSDLCMLGI-UHFFFAOYSA-N 0.000 description 1
- GJEREQYJIQASAW-UHFFFAOYSA-N flufenerim Chemical compound CC(F)C1=NC=NC(NCCC=2C=CC(OC(F)(F)F)=CC=2)=C1Cl GJEREQYJIQASAW-UHFFFAOYSA-N 0.000 description 1
- WFZSZAXUALBVNX-UHFFFAOYSA-N flufenpyr Chemical compound O=C1C(C)=C(C(F)(F)F)C=NN1C1=CC(OCC(O)=O)=C(Cl)C=C1F WFZSZAXUALBVNX-UHFFFAOYSA-N 0.000 description 1
- DNUAYCRATWAJQE-UHFFFAOYSA-N flufenpyr-ethyl Chemical group C1=C(Cl)C(OCC(=O)OCC)=CC(N2C(C(C)=C(C=N2)C(F)(F)F)=O)=C1F DNUAYCRATWAJQE-UHFFFAOYSA-N 0.000 description 1
- FOUWCSDKDDHKQP-UHFFFAOYSA-N flumioxazin Chemical compound FC1=CC=2OCC(=O)N(CC#C)C=2C=C1N(C1=O)C(=O)C2=C1CCCC2 FOUWCSDKDDHKQP-UHFFFAOYSA-N 0.000 description 1
- 238000002875 fluorescence polarization Methods 0.000 description 1
- 239000007850 fluorescent dye Substances 0.000 description 1
- OQZCSNDVOWYALR-UHFFFAOYSA-N flurochloridone Chemical compound FC(F)(F)C1=CC=CC(N2C(C(Cl)C(CCl)C2)=O)=C1 OQZCSNDVOWYALR-UHFFFAOYSA-N 0.000 description 1
- MEFQWPUMEMWTJP-UHFFFAOYSA-N fluroxypyr Chemical compound NC1=C(Cl)C(F)=NC(OCC(O)=O)=C1Cl MEFQWPUMEMWTJP-UHFFFAOYSA-N 0.000 description 1
- GNVDAZSPJWCIQZ-UHFFFAOYSA-N flusulfamide Chemical compound ClC1=CC([N+](=O)[O-])=CC=C1NS(=O)(=O)C1=CC=C(Cl)C(C(F)(F)F)=C1 GNVDAZSPJWCIQZ-UHFFFAOYSA-N 0.000 description 1
- 239000006260 foam Substances 0.000 description 1
- 238000005755 formation reaction Methods 0.000 description 1
- 235000012055 fruits and vegetables Nutrition 0.000 description 1
- UYJUZNLFJAWNEZ-UHFFFAOYSA-N fuberidazole Chemical compound C1=COC(C=2NC3=CC=CC=C3N=2)=C1 UYJUZNLFJAWNEZ-UHFFFAOYSA-N 0.000 description 1
- 238000002825 functional assay Methods 0.000 description 1
- 230000000855 fungicidal effect Effects 0.000 description 1
- 230000004927 fusion Effects 0.000 description 1
- 238000010363 gene targeting Methods 0.000 description 1
- 230000035784 germination Effects 0.000 description 1
- IXORZMNAPKEEDV-UHFFFAOYSA-N gibberellic acid GA3 Natural products OC(=O)C1C2(C3)CC(=C)C3(O)CCC2C2(C=CC3O)C1C3(C)C(=O)O2 IXORZMNAPKEEDV-UHFFFAOYSA-N 0.000 description 1
- IXORZMNAPKEEDV-OBDJNFEBSA-N gibberellin A3 Chemical compound C([C@@]1(O)C(=C)C[C@@]2(C1)[C@H]1C(O)=O)C[C@H]2[C@]2(C=C[C@@H]3O)[C@H]1[C@]3(C)C(=O)O2 IXORZMNAPKEEDV-OBDJNFEBSA-N 0.000 description 1
- IAJOBQBIJHVGMQ-BYPYZUCNSA-N glufosinate-P Chemical compound CP(O)(=O)CC[C@H](N)C(O)=O IAJOBQBIJHVGMQ-BYPYZUCNSA-N 0.000 description 1
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 description 1
- 229910052737 gold Inorganic materials 0.000 description 1
- 239000010931 gold Substances 0.000 description 1
- 229940095686 granule product Drugs 0.000 description 1
- 239000003673 groundwater Substances 0.000 description 1
- 239000003630 growth substance Substances 0.000 description 1
- 229940029575 guanosine Drugs 0.000 description 1
- 229910052736 halogen Inorganic materials 0.000 description 1
- 150000002367 halogens Chemical class 0.000 description 1
- MFSWTRQUCLNFOM-SECBINFHSA-N haloxyfop-P-methyl Chemical group C1=CC(O[C@H](C)C(=O)OC)=CC=C1OC1=NC=C(C(F)(F)F)C=C1Cl MFSWTRQUCLNFOM-SECBINFHSA-N 0.000 description 1
- 238000003306 harvesting Methods 0.000 description 1
- 210000004458 heterochromatin Anatomy 0.000 description 1
- 125000000623 heterocyclic group Chemical group 0.000 description 1
- 239000003906 humectant Substances 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
- 239000001257 hydrogen Substances 0.000 description 1
- 125000004435 hydrogen atom Chemical class [H]* 0.000 description 1
- KGVPNLBXJKTABS-UHFFFAOYSA-N hymexazol Chemical compound CC1=CC(O)=NO1 KGVPNLBXJKTABS-UHFFFAOYSA-N 0.000 description 1
- AGKSTYPVMZODRV-UHFFFAOYSA-N imibenconazole Chemical compound C1=CC(Cl)=CC=C1CSC(CN1N=CN=C1)=NC1=CC=C(Cl)C=C1Cl AGKSTYPVMZODRV-UHFFFAOYSA-N 0.000 description 1
- RONFGUROBZGJKP-UHFFFAOYSA-N iminoctadine Chemical compound NC(N)=NCCCCCCCCNCCCCCCCCN=C(N)N RONFGUROBZGJKP-UHFFFAOYSA-N 0.000 description 1
- 238000003018 immunoassay Methods 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 238000011065 in-situ storage Methods 0.000 description 1
- 230000001939 inductive effect Effects 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 230000000977 initiatory effect Effects 0.000 description 1
- 230000000749 insecticidal effect Effects 0.000 description 1
- 238000007689 inspection Methods 0.000 description 1
- 238000007852 inverse PCR Methods 0.000 description 1
- ONUFESLQCSAYKA-UHFFFAOYSA-N iprodione Chemical compound O=C1N(C(=O)NC(C)C)CC(=O)N1C1=CC(Cl)=CC(Cl)=C1 ONUFESLQCSAYKA-UHFFFAOYSA-N 0.000 description 1
- 229960004849 isoconazole Drugs 0.000 description 1
- 125000001449 isopropyl group Chemical group [H]C([H])([H])C([H])(*)C([H])([H])[H] 0.000 description 1
- 150000002545 isoxazoles Chemical class 0.000 description 1
- 238000005304 joining Methods 0.000 description 1
- PVTHJAPFENJVNC-MHRBZPPQSA-N kasugamycin Chemical compound N[C@H]1C[C@H](NC(=N)C(O)=O)[C@@H](C)O[C@@H]1O[C@H]1[C@H](O)[C@@H](O)[C@@H](O)[C@H](O)[C@@H]1O PVTHJAPFENJVNC-MHRBZPPQSA-N 0.000 description 1
- 230000002147 killing effect Effects 0.000 description 1
- 238000002372 labelling Methods 0.000 description 1
- 238000011005 laboratory method Methods 0.000 description 1
- JJTUDXZGHPGLLC-UHFFFAOYSA-N lactide Chemical compound CC1OC(=O)C(C)OC1=O JJTUDXZGHPGLLC-UHFFFAOYSA-N 0.000 description 1
- 239000005910 lambda-Cyhalothrin Substances 0.000 description 1
- 239000002523 lectin Substances 0.000 description 1
- ZTMKADLOSYKWCA-UHFFFAOYSA-N lenacil Chemical compound O=C1NC=2CCCC=2C(=O)N1C1CCCCC1 ZTMKADLOSYKWCA-UHFFFAOYSA-N 0.000 description 1
- 239000003446 ligand Substances 0.000 description 1
- 229960004488 linolenic acid Drugs 0.000 description 1
- KQQKGWQCNNTQJW-UHFFFAOYSA-N linolenic acid Natural products CC=CCCC=CCC=CCCCCCCCC(O)=O KQQKGWQCNNTQJW-UHFFFAOYSA-N 0.000 description 1
- 150000002632 lipids Chemical class 0.000 description 1
- 125000000040 m-tolyl group Chemical group [H]C1=C([H])C(*)=C([H])C(=C1[H])C([H])([H])[H] 0.000 description 1
- 238000007403 mPCR Methods 0.000 description 1
- 230000014759 maintenance of location Effects 0.000 description 1
- 230000001404 mediated effect Effects 0.000 description 1
- CIFWZNRJIBNXRE-UHFFFAOYSA-N mepanipyrim Chemical compound CC#CC1=CC(C)=NC(NC=2C=CC=CC=2)=N1 CIFWZNRJIBNXRE-UHFFFAOYSA-N 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- HYVVJDQGXFXBRZ-UHFFFAOYSA-N metam Chemical compound CNC(S)=S HYVVJDQGXFXBRZ-UHFFFAOYSA-N 0.000 description 1
- AFCCDDWKHLHPDF-UHFFFAOYSA-M metam-sodium Chemical compound [Na+].CNC([S-])=S AFCCDDWKHLHPDF-UHFFFAOYSA-M 0.000 description 1
- IXJOSTZEBSTPAG-UHFFFAOYSA-N methasulfocarb Chemical compound CNC(=O)SC1=CC=C(OS(C)(=O)=O)C=C1 IXJOSTZEBSTPAG-UHFFFAOYSA-N 0.000 description 1
- MFSWTRQUCLNFOM-UHFFFAOYSA-N methyl 2-(4-{[3-chloro-5-(trifluoromethyl)pyridin-2-yl]oxy}phenoxy)propanoate Chemical group C1=CC(OC(C)C(=O)OC)=CC=C1OC1=NC=C(C(F)(F)F)C=C1Cl MFSWTRQUCLNFOM-UHFFFAOYSA-N 0.000 description 1
- RBNIGDFIUWJJEV-UHFFFAOYSA-N methyl 2-(n-benzoyl-3-chloro-4-fluoroanilino)propanoate Chemical group C=1C=C(F)C(Cl)=CC=1N(C(C)C(=O)OC)C(=O)C1=CC=CC=C1 RBNIGDFIUWJJEV-UHFFFAOYSA-N 0.000 description 1
- BACHBFVBHLGWSL-UHFFFAOYSA-N methyl 2-[4-(2,4-dichlorophenoxy)phenoxy]propanoate Chemical group C1=CC(OC(C)C(=O)OC)=CC=C1OC1=CC=C(Cl)C=C1Cl BACHBFVBHLGWSL-UHFFFAOYSA-N 0.000 description 1
- LYPWWQLKWQNQKV-UHFFFAOYSA-N methyl 2-[5-ethyl-2-[[4-[3-methyl-2,6-dioxo-4-(trifluoromethyl)pyrimidin-1-yl]phenoxy]methyl]phenoxy]propanoate Chemical compound COC(=O)C(C)OC1=CC(CC)=CC=C1COC1=CC=C(N2C(N(C)C(=CC2=O)C(F)(F)F)=O)C=C1 LYPWWQLKWQNQKV-UHFFFAOYSA-N 0.000 description 1
- ZTYVMAQSHCZXLF-UHFFFAOYSA-N methyl 2-[[4,6-bis(difluoromethoxy)pyrimidin-2-yl]carbamoylsulfamoyl]benzoate Chemical group COC(=O)C1=CC=CC=C1S(=O)(=O)NC(=O)NC1=NC(OC(F)F)=CC(OC(F)F)=N1 ZTYVMAQSHCZXLF-UHFFFAOYSA-N 0.000 description 1
- LINPVWIEWJTEEJ-UHFFFAOYSA-N methyl 2-chloro-9-hydroxyfluorene-9-carboxylate Chemical group C1=C(Cl)C=C2C(C(=O)OC)(O)C3=CC=CC=C3C2=C1 LINPVWIEWJTEEJ-UHFFFAOYSA-N 0.000 description 1
- VWGAYSCWLXQJBQ-UHFFFAOYSA-N methyl 4-iodo-2-[(4-methoxy-6-methyl-1,3,5-triazin-2-yl)carbamoylsulfamoyl]benzoate Chemical group COC(=O)C1=CC=C(I)C=C1S(=O)(=O)NC(=O)NC1=NC(C)=NC(OC)=N1 VWGAYSCWLXQJBQ-UHFFFAOYSA-N 0.000 description 1
- CJPQIRJHIZUAQP-UHFFFAOYSA-N methyl N-(2,6-dimethylphenyl)-N-(phenylacetyl)alaninate Chemical compound CC=1C=CC=C(C)C=1N(C(C)C(=O)OC)C(=O)CC1=CC=CC=C1 CJPQIRJHIZUAQP-UHFFFAOYSA-N 0.000 description 1
- 125000001570 methylene group Chemical group [H]C([H])([*:1])[*:2] 0.000 description 1
- 229920000257 metiram Polymers 0.000 description 1
- 229960002939 metizoline Drugs 0.000 description 1
- FOXFZRUHNHCZPX-UHFFFAOYSA-N metribuzin Chemical compound CSC1=NN=C(C(C)(C)C)C(=O)N1N FOXFZRUHNHCZPX-UHFFFAOYSA-N 0.000 description 1
- 229960002509 miconazole Drugs 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 210000004688 microtubule Anatomy 0.000 description 1
- 230000009526 moderate injury Effects 0.000 description 1
- 235000019426 modified starch Nutrition 0.000 description 1
- 239000003068 molecular probe Substances 0.000 description 1
- JITOKQVGRJSHHA-UHFFFAOYSA-M monosodium methyl arsenate Chemical compound [Na+].C[As](O)([O-])=O JITOKQVGRJSHHA-UHFFFAOYSA-M 0.000 description 1
- 238000002703 mutagenesis Methods 0.000 description 1
- 231100000350 mutagenesis Toxicity 0.000 description 1
- AIMMSOZBPYFASU-UHFFFAOYSA-N n-(4,6-dimethoxypyrimidin-2-yl)-n'-[3-(2,2,2-trifluoroethoxy)pyridin-1-ium-2-yl]sulfonylcarbamimidate Chemical compound COC1=CC(OC)=NC(NC(=O)NS(=O)(=O)C=2C(=CC=CN=2)OCC(F)(F)F)=N1 AIMMSOZBPYFASU-UHFFFAOYSA-N 0.000 description 1
- CHEDHKBPPDKBQF-UPONEAKYSA-N n-[5-[(6s,7ar)-6-fluoro-1,3-dioxo-5,6,7,7a-tetrahydropyrrolo[1,2-c]imidazol-2-yl]-2-chloro-4-fluorophenyl]-1-chloromethanesulfonamide Chemical compound N1([C@@H](C2=O)C[C@@H](C1)F)C(=O)N2C1=CC(NS(=O)(=O)CCl)=C(Cl)C=C1F CHEDHKBPPDKBQF-UPONEAKYSA-N 0.000 description 1
- 230000021616 negative regulation of cell division Effects 0.000 description 1
- 230000027498 negative regulation of mitosis Effects 0.000 description 1
- 229940079888 nitenpyram Drugs 0.000 description 1
- 231100000252 nontoxic Toxicity 0.000 description 1
- 230000003000 nontoxic effect Effects 0.000 description 1
- 238000007899 nucleic acid hybridization Methods 0.000 description 1
- 239000002853 nucleic acid probe Substances 0.000 description 1
- 230000031787 nutrient reservoir activity Effects 0.000 description 1
- 229950004864 olamine Drugs 0.000 description 1
- 210000000056 organ Anatomy 0.000 description 1
- 239000003960 organic solvent Substances 0.000 description 1
- 230000008520 organization Effects 0.000 description 1
- 239000003986 organophosphate insecticide Substances 0.000 description 1
- AHLBNYSZXLDEJQ-FWEHEUNISA-N orlistat Chemical compound CCCCCCCCCCC[C@H](OC(=O)[C@H](CC(C)C)NC=O)C[C@@H]1OC(=O)[C@H]1CCCCCC AHLBNYSZXLDEJQ-FWEHEUNISA-N 0.000 description 1
- UNAHYJYOSSSJHH-UHFFFAOYSA-N oryzalin Chemical compound CCCN(CCC)C1=C([N+]([O-])=O)C=C(S(N)(=O)=O)C=C1[N+]([O-])=O UNAHYJYOSSSJHH-UHFFFAOYSA-N 0.000 description 1
- 230000002018 overexpression Effects 0.000 description 1
- 150000001475 oxazolidinediones Chemical class 0.000 description 1
- AMEKQAFGQBKLKX-UHFFFAOYSA-N oxycarboxin Chemical compound O=S1(=O)CCOC(C)=C1C(=O)NC1=CC=CC=C1 AMEKQAFGQBKLKX-UHFFFAOYSA-N 0.000 description 1
- 125000004430 oxygen atom Chemical group O* 0.000 description 1
- 230000036961 partial effect Effects 0.000 description 1
- 235000015927 pasta Nutrition 0.000 description 1
- LKPLKUMXSAEKID-UHFFFAOYSA-N pentachloronitrobenzene Chemical compound [O-][N+](=O)C1=C(Cl)C(Cl)=C(Cl)C(Cl)=C1Cl LKPLKUMXSAEKID-UHFFFAOYSA-N 0.000 description 1
- 230000000737 periodic effect Effects 0.000 description 1
- 230000000361 pesticidal effect Effects 0.000 description 1
- CSWIKHNSBZVWNQ-UHFFFAOYSA-N pethoxamide Chemical compound CCOCCN(C(=O)CCl)C(=C(C)C)C1=CC=CC=C1 CSWIKHNSBZVWNQ-UHFFFAOYSA-N 0.000 description 1
- 239000003208 petroleum Substances 0.000 description 1
- IDOWTHOLJBTAFI-UHFFFAOYSA-N phenmedipham Chemical compound COC(=O)NC1=CC=CC(OC(=O)NC=2C=C(C)C=CC=2)=C1 IDOWTHOLJBTAFI-UHFFFAOYSA-N 0.000 description 1
- BSCCSDNZEIHXOK-UHFFFAOYSA-N phenyl carbamate Chemical class NC(=O)OC1=CC=CC=C1 BSCCSDNZEIHXOK-UHFFFAOYSA-N 0.000 description 1
- 150000008048 phenylpyrazoles Chemical class 0.000 description 1
- 239000003016 pheromone Substances 0.000 description 1
- 239000010452 phosphate Substances 0.000 description 1
- PTMHPRAIXMAOOB-UHFFFAOYSA-N phosphoramidic acid Chemical class NP(O)(O)=O PTMHPRAIXMAOOB-UHFFFAOYSA-N 0.000 description 1
- XNGIFLGASWRNHJ-UHFFFAOYSA-N phthalic acid Chemical class OC(=O)C1=CC=CC=C1C(O)=O XNGIFLGASWRNHJ-UHFFFAOYSA-N 0.000 description 1
- 230000000704 physical effect Effects 0.000 description 1
- 230000035790 physiological processes and functions Effects 0.000 description 1
- 230000000885 phytotoxic effect Effects 0.000 description 1
- SIOXPEMLGUPBBT-UHFFFAOYSA-N picolinic acid Chemical class OC(=O)C1=CC=CC=N1 SIOXPEMLGUPBBT-UHFFFAOYSA-N 0.000 description 1
- 239000000049 pigment Substances 0.000 description 1
- MGOHCFMYLBAPRN-UHFFFAOYSA-N pinoxaden Chemical compound CCC1=CC(C)=CC(CC)=C1C(C1=O)=C(OC(=O)C(C)(C)C)N2N1CCOCC2 MGOHCFMYLBAPRN-UHFFFAOYSA-N 0.000 description 1
- YEBIHIICWDDQOL-YBHNRIQQSA-N polyoxin Polymers O[C@@H]1[C@H](O)[C@@H](C(C=O)N)O[C@H]1N1C(=O)NC(=O)C(C(O)=O)=C1 YEBIHIICWDDQOL-YBHNRIQQSA-N 0.000 description 1
- 239000000249 polyoxyethylene sorbitan monopalmitate Substances 0.000 description 1
- XAEFZNCEHLXOMS-UHFFFAOYSA-M potassium benzoate Chemical compound [K+].[O-]C(=O)C1=CC=CC=C1 XAEFZNCEHLXOMS-UHFFFAOYSA-M 0.000 description 1
- ORHJUFUQMQEFPQ-UHFFFAOYSA-M potassium;2-(4-chloro-2-methylphenoxy)acetate Chemical compound [K+].CC1=CC(Cl)=CC=C1OCC([O-])=O ORHJUFUQMQEFPQ-UHFFFAOYSA-M 0.000 description 1
- ZRHANBBTXQZFSP-UHFFFAOYSA-M potassium;4-amino-3,5,6-trichloropyridine-2-carboxylate Chemical compound [K+].NC1=C(Cl)C(Cl)=NC(C([O-])=O)=C1Cl ZRHANBBTXQZFSP-UHFFFAOYSA-M 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 239000002243 precursor Substances 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 239000003755 preservative agent Substances 0.000 description 1
- 238000002203 pretreatment Methods 0.000 description 1
- QXJKBPAVAHBARF-BETUJISGSA-N procymidone Chemical compound O=C([C@]1(C)C[C@@]1(C1=O)C)N1C1=CC(Cl)=CC(Cl)=C1 QXJKBPAVAHBARF-BETUJISGSA-N 0.000 description 1
- 230000002035 prolonged effect Effects 0.000 description 1
- ZKWPMZVVAJSYNI-UHFFFAOYSA-N prop-2-enal Chemical compound C=CC=O.C=CC=O ZKWPMZVVAJSYNI-UHFFFAOYSA-N 0.000 description 1
- IKVXBIIHQGXQRQ-CYBMUJFWSA-N propan-2-yl (2r)-2-(n-benzoyl-3-chloro-4-fluoroanilino)propanoate Chemical group C=1C=C(F)C(Cl)=CC=1N([C@H](C)C(=O)OC(C)C)C(=O)C1=CC=CC=C1 IKVXBIIHQGXQRQ-CYBMUJFWSA-N 0.000 description 1
- WJNRPILHGGKWCK-UHFFFAOYSA-N propazine Chemical compound CC(C)NC1=NC(Cl)=NC(NC(C)C)=N1 WJNRPILHGGKWCK-UHFFFAOYSA-N 0.000 description 1
- KKMLIVYBGSAJPM-UHFFFAOYSA-L propineb Chemical compound [Zn+2].[S-]C(=S)NC(C)CNC([S-])=S KKMLIVYBGSAJPM-UHFFFAOYSA-L 0.000 description 1
- NQLVQOSNDJXLKG-UHFFFAOYSA-N prosulfocarb Chemical compound CCCN(CCC)C(=O)SCC1=CC=CC=C1 NQLVQOSNDJXLKG-UHFFFAOYSA-N 0.000 description 1
- 235000019833 protease Nutrition 0.000 description 1
- 238000002731 protein assay Methods 0.000 description 1
- 210000001938 protoplast Anatomy 0.000 description 1
- UHSGPDMIQQYNAX-UHFFFAOYSA-N protoporphyrinogen Chemical compound C1C(=C(C=2C=C)C)NC=2CC(=C(C=2CCC(O)=O)C)NC=2CC(N2)=C(CCC(O)=O)C(C)=C2CC2=C(C)C(C=C)=C1N2 UHSGPDMIQQYNAX-UHFFFAOYSA-N 0.000 description 1
- QHMTXANCGGJZRX-WUXMJOGZSA-N pymetrozine Chemical compound C1C(C)=NNC(=O)N1\N=C\C1=CC=CN=C1 QHMTXANCGGJZRX-WUXMJOGZSA-N 0.000 description 1
- HZRSNVGNWUDEFX-UHFFFAOYSA-N pyraclostrobin Chemical compound COC(=O)N(OC)C1=CC=CC=C1COC1=NN(C=2C=CC(Cl)=CC=2)C=C1 HZRSNVGNWUDEFX-UHFFFAOYSA-N 0.000 description 1
- DDIQWGKUSJOETH-UHFFFAOYSA-N pyrafluprole Chemical compound ClC=1C=C(C(F)(F)F)C=C(Cl)C=1N1N=C(C#N)C(SCF)=C1NCC1=CN=CC=N1 DDIQWGKUSJOETH-UHFFFAOYSA-N 0.000 description 1
- DWSPRBSLSXQIEJ-UHFFFAOYSA-N pyrasulfotole Chemical compound CC1=NN(C)C(O)=C1C(=O)C1=CC=C(C(F)(F)F)C=C1S(C)(=O)=O DWSPRBSLSXQIEJ-UHFFFAOYSA-N 0.000 description 1
- 150000003217 pyrazoles Chemical class 0.000 description 1
- DWFZBUWUXWZWKD-UHFFFAOYSA-N pyridaben Chemical compound C1=CC(C(C)(C)C)=CC=C1CSC1=C(Cl)C(=O)N(C(C)(C)C)N=C1 DWFZBUWUXWZWKD-UHFFFAOYSA-N 0.000 description 1
- 150000003222 pyridines Chemical class 0.000 description 1
- 125000004076 pyridyl group Chemical group 0.000 description 1
- USSIUIGPBLPCDF-KEBDBYFISA-N pyriminobac-methyl Chemical group CO\N=C(/C)C1=CC=CC(OC=2N=C(OC)C=C(OC)N=2)=C1C(=O)OC USSIUIGPBLPCDF-KEBDBYFISA-N 0.000 description 1
- XRJLAOUDSILTFT-UHFFFAOYSA-N pyroquilon Chemical compound O=C1CCC2=CC=CC3=C2N1CC3 XRJLAOUDSILTFT-UHFFFAOYSA-N 0.000 description 1
- 238000012175 pyrosequencing Methods 0.000 description 1
- 229960002132 pyrrolnitrin Drugs 0.000 description 1
- 238000007859 qualitative PCR Methods 0.000 description 1
- 238000003908 quality control method Methods 0.000 description 1
- LOAUVZALPPNFOQ-UHFFFAOYSA-N quinaldic acid Chemical class C1=CC=CC2=NC(C(=O)O)=CC=C21 LOAUVZALPPNFOQ-UHFFFAOYSA-N 0.000 description 1
- ALZOLUNSQWINIR-UHFFFAOYSA-N quinmerac Chemical compound OC(=O)C1=C(Cl)C=CC2=CC(C)=CN=C21 ALZOLUNSQWINIR-UHFFFAOYSA-N 0.000 description 1
- BBKDWPHJZANJGB-IKJXHCRLSA-N quizalofop-P-tefuryl Chemical group O=C([C@H](OC=1C=CC(OC=2N=C3C=CC(Cl)=CC3=NC=2)=CC=1)C)OCC1CCCO1 BBKDWPHJZANJGB-IKJXHCRLSA-N 0.000 description 1
- BACHBFVBHLGWSL-JTQLQIEISA-N rac-diclofop methyl Natural products C1=CC(O[C@@H](C)C(=O)OC)=CC=C1OC1=CC=C(Cl)C=C1Cl BACHBFVBHLGWSL-JTQLQIEISA-N 0.000 description 1
- 230000002285 radioactive effect Effects 0.000 description 1
- 238000003753 real-time PCR Methods 0.000 description 1
- 238000010188 recombinant method Methods 0.000 description 1
- 230000000306 recurrent effect Effects 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 230000008929 regeneration Effects 0.000 description 1
- 238000011069 regeneration method Methods 0.000 description 1
- 230000022983 regulation of cell cycle Effects 0.000 description 1
- 230000008439 repair process Effects 0.000 description 1
- 230000033458 reproduction Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 229920002477 rna polymer Polymers 0.000 description 1
- 230000007017 scission Effects 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 238000005204 segregation Methods 0.000 description 1
- 230000010153 self-pollination Effects 0.000 description 1
- 150000007659 semicarbazones Chemical class 0.000 description 1
- 239000003620 semiochemical Substances 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 238000012163 sequencing technique Methods 0.000 description 1
- 230000009528 severe injury Effects 0.000 description 1
- 238000009958 sewing Methods 0.000 description 1
- MGLWZSOBALDPEK-UHFFFAOYSA-N simetryn Chemical compound CCNC1=NC(NCC)=NC(SC)=N1 MGLWZSOBALDPEK-UHFFFAOYSA-N 0.000 description 1
- NLJMYIDDQXHKNR-UHFFFAOYSA-K sodium citrate Chemical compound O.O.[Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O NLJMYIDDQXHKNR-UHFFFAOYSA-K 0.000 description 1
- PDEFQWNXOUGDJR-UHFFFAOYSA-M sodium;2,2-dichloropropanoate Chemical compound [Na+].CC(Cl)(Cl)C([O-])=O PDEFQWNXOUGDJR-UHFFFAOYSA-M 0.000 description 1
- JUNDBKFAZXZKRA-MAFYXNADSA-M sodium;2-[(e)-n-[(3,5-difluorophenyl)carbamoylamino]-c-methylcarbonimidoyl]pyridine-3-carboxylate Chemical compound [Na+].N=1C=CC=C(C([O-])=O)C=1C(/C)=N/NC(=O)NC1=CC(F)=CC(F)=C1 JUNDBKFAZXZKRA-MAFYXNADSA-M 0.000 description 1
- AXKBOWBNOCUNJL-UHFFFAOYSA-M sodium;2-nitrophenolate Chemical compound [Na+].[O-]C1=CC=CC=C1[N+]([O-])=O AXKBOWBNOCUNJL-UHFFFAOYSA-M 0.000 description 1
- GABUSZPTCJGKGB-UHFFFAOYSA-M sodium;4-(4-chloro-2-methylphenoxy)butanoate Chemical compound [Na+].CC1=CC(Cl)=CC=C1OCCCC([O-])=O GABUSZPTCJGKGB-UHFFFAOYSA-M 0.000 description 1
- QGKPUZOFTJQTHL-UHFFFAOYSA-M sodium;4-cyano-2,6-diiodophenolate Chemical compound [Na+].[O-]C1=C(I)C=C(C#N)C=C1I QGKPUZOFTJQTHL-UHFFFAOYSA-M 0.000 description 1
- RVULBHWZFCBODE-UHFFFAOYSA-M sodium;5-[2-chloro-4-(trifluoromethyl)phenoxy]-2-nitrobenzoate Chemical compound [Na+].C1=C([N+]([O-])=O)C(C(=O)[O-])=CC(OC=2C(=CC(=CC=2)C(F)(F)F)Cl)=C1 RVULBHWZFCBODE-UHFFFAOYSA-M 0.000 description 1
- 244000000000 soil microbiome Species 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- DTDSAWVUFPGDMX-UHFFFAOYSA-N spirodiclofen Chemical compound CCC(C)(C)C(=O)OC1=C(C=2C(=CC(Cl)=CC=2)Cl)C(=O)OC11CCCCC1 DTDSAWVUFPGDMX-UHFFFAOYSA-N 0.000 description 1
- CLSVJBIHYWPGQY-GGYDESQDSA-N spirotetramat Chemical compound CCOC(=O)OC1=C(C=2C(=CC=C(C)C=2)C)C(=O)N[C@@]11CC[C@H](OC)CC1 CLSVJBIHYWPGQY-GGYDESQDSA-N 0.000 description 1
- 230000002269 spontaneous effect Effects 0.000 description 1
- 238000003153 stable transfection Methods 0.000 description 1
- 239000003206 sterilizing agent Substances 0.000 description 1
- 239000000021 stimulant Substances 0.000 description 1
- 125000005750 substituted cyclic group Chemical group 0.000 description 1
- OORLZFUTLGXMEF-UHFFFAOYSA-N sulfentrazone Chemical compound O=C1N(C(F)F)C(C)=NN1C1=CC(NS(C)(=O)=O)=C(Cl)C=C1Cl OORLZFUTLGXMEF-UHFFFAOYSA-N 0.000 description 1
- 125000000472 sulfonyl group Chemical group *S(*)(=O)=O 0.000 description 1
- 230000001629 suppression Effects 0.000 description 1
- 239000002352 surface water Substances 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 230000009885 systemic effect Effects 0.000 description 1
- ROZUQUDEWZIBHV-UHFFFAOYSA-N tecloftalam Chemical compound OC(=O)C1=C(Cl)C(Cl)=C(Cl)C(Cl)=C1C(=O)NC1=CC=CC(Cl)=C1Cl ROZUQUDEWZIBHV-UHFFFAOYSA-N 0.000 description 1
- XQTLDIFVVHJORV-UHFFFAOYSA-N tecnazene Chemical compound [O-][N+](=O)C1=C(Cl)C(Cl)=CC(Cl)=C1Cl XQTLDIFVVHJORV-UHFFFAOYSA-N 0.000 description 1
- IUQAXCIUEPFPSF-UHFFFAOYSA-N tembotrione Chemical compound ClC1=C(COCC(F)(F)F)C(S(=O)(=O)C)=CC=C1C(=O)C1C(=O)CCCC1=O IUQAXCIUEPFPSF-UHFFFAOYSA-N 0.000 description 1
- 230000002123 temporal effect Effects 0.000 description 1
- 239000004308 thiabendazole Substances 0.000 description 1
- WJCNZQLZVWNLKY-UHFFFAOYSA-N thiabendazole Chemical compound S1C=NC(C=2NC3=CC=CC=C3N=2)=C1 WJCNZQLZVWNLKY-UHFFFAOYSA-N 0.000 description 1
- 229960004546 thiabendazole Drugs 0.000 description 1
- 235000010296 thiabendazole Nutrition 0.000 description 1
- 150000004867 thiadiazoles Chemical class 0.000 description 1
- 239000002562 thickening agent Substances 0.000 description 1
- GLDAZAQRGCSFNP-UHFFFAOYSA-N thiencarbazone Chemical compound O=C1N(C)C(OC)=NN1C(=O)NS(=O)(=O)C1=C(C)SC=C1C(O)=O GLDAZAQRGCSFNP-UHFFFAOYSA-N 0.000 description 1
- 150000007970 thio esters Chemical class 0.000 description 1
- 150000003558 thiocarbamic acid derivatives Chemical class 0.000 description 1
- KUAZQDVKQLNFPE-UHFFFAOYSA-N thiram Chemical compound CN(C)C(=S)SSC(=S)N(C)C KUAZQDVKQLNFPE-UHFFFAOYSA-N 0.000 description 1
- 229960002447 thiram Drugs 0.000 description 1
- VJQYLJSMBWXGDV-UHFFFAOYSA-N tiadinil Chemical compound N1=NSC(C(=O)NC=2C=C(Cl)C(C)=CC=2)=C1C VJQYLJSMBWXGDV-UHFFFAOYSA-N 0.000 description 1
- HYVWIQDYBVKITD-UHFFFAOYSA-N tolylfluanid Chemical compound CN(C)S(=O)(=O)N(SC(F)(Cl)Cl)C1=CC=C(C)C=C1 HYVWIQDYBVKITD-UHFFFAOYSA-N 0.000 description 1
- IYMLUHWAJFXAQP-UHFFFAOYSA-N topramezone Chemical compound CC1=C(C(=O)C2=C(N(C)N=C2)O)C=CC(S(C)(=O)=O)=C1C1=NOCC1 IYMLUHWAJFXAQP-UHFFFAOYSA-N 0.000 description 1
- 108700012359 toxins Proteins 0.000 description 1
- DQFPEYARZIQXRM-LTGZKZEYSA-N tralkoxydim Chemical compound C1C(=O)C(C(/CC)=N/OCC)=C(O)CC1C1=C(C)C=C(C)C=C1C DQFPEYARZIQXRM-LTGZKZEYSA-N 0.000 description 1
- 230000005026 transcription initiation Effects 0.000 description 1
- 230000005030 transcription termination Effects 0.000 description 1
- 238000011426 transformation method Methods 0.000 description 1
- 230000007704 transition Effects 0.000 description 1
- 230000014621 translational initiation Effects 0.000 description 1
- 230000017105 transposition Effects 0.000 description 1
- BAZVSMNPJJMILC-UHFFFAOYSA-N triadimenol Chemical compound C1=NC=NN1C(C(O)C(C)(C)C)OC1=CC=C(Cl)C=C1 BAZVSMNPJJMILC-UHFFFAOYSA-N 0.000 description 1
- NKNFWVNSBIXGLL-UHFFFAOYSA-N triazamate Chemical compound CCOC(=O)CSC1=NC(C(C)(C)C)=NN1C(=O)N(C)C NKNFWVNSBIXGLL-UHFFFAOYSA-N 0.000 description 1
- 150000003918 triazines Chemical class 0.000 description 1
- 150000003852 triazoles Chemical class 0.000 description 1
- REEQLXCGVXDJSQ-UHFFFAOYSA-N trichlopyr Chemical compound OC(=O)COC1=NC(Cl)=C(Cl)C=C1Cl REEQLXCGVXDJSQ-UHFFFAOYSA-N 0.000 description 1
- RROQIUMZODEXOR-UHFFFAOYSA-N triforine Chemical compound O=CNC(C(Cl)(Cl)Cl)N1CCN(C(NC=O)C(Cl)(Cl)Cl)CC1 RROQIUMZODEXOR-UHFFFAOYSA-N 0.000 description 1
- HRXKRNGNAMMEHJ-UHFFFAOYSA-K trisodium citrate Chemical compound [Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O HRXKRNGNAMMEHJ-UHFFFAOYSA-K 0.000 description 1
- 229940038773 trisodium citrate Drugs 0.000 description 1
- KVEQCVKVIFQSGC-UHFFFAOYSA-N tritosulfuron Chemical compound FC(F)(F)C1=NC(OC)=NC(NC(=O)NS(=O)(=O)C=2C(=CC=CC=2)C(F)(F)F)=N1 KVEQCVKVIFQSGC-UHFFFAOYSA-N 0.000 description 1
- 241000701451 unidentified granulovirus Species 0.000 description 1
- 230000017260 vegetative to reproductive phase transition of meristem Effects 0.000 description 1
- 150000004669 very long chain fatty acids Chemical class 0.000 description 1
- 230000035899 viability Effects 0.000 description 1
- 230000009385 viral infection Effects 0.000 description 1
- 235000020234 walnut Nutrition 0.000 description 1
- 239000011534 wash buffer Substances 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
- 238000003911 water pollution Methods 0.000 description 1
- 239000000080 wetting agent Substances 0.000 description 1
- DUBNHZYBDBBJHD-UHFFFAOYSA-L ziram Chemical compound [Zn+2].CN(C)C([S-])=S.CN(C)C([S-])=S DUBNHZYBDBBJHD-UHFFFAOYSA-L 0.000 description 1
- FJBGIXKIXPUXBY-UHFFFAOYSA-N {2-[3-(4-chlorophenyl)propyl]-2,4,4-trimethyl-1,3-oxazolidin-3-yl}(imidazol-1-yl)methanone Chemical compound C1=CN=CN1C(=O)N1C(C)(C)COC1(C)CCCC1=CC=C(Cl)C=C1 FJBGIXKIXPUXBY-UHFFFAOYSA-N 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
- C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
- C12N15/82—Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
- C12N15/8241—Phenotypically and genetically modified plants via recombinant DNA technology
- C12N15/8261—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield
- C12N15/8271—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield for stress resistance, e.g. heavy metal resistance
- C12N15/8274—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield for stress resistance, e.g. heavy metal resistance for herbicide resistance
- C12N15/8275—Glyphosate
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N57/00—Biocides, pest repellants or attractants, or plant growth regulators containing organic phosphorus compounds
- A01N57/18—Biocides, pest repellants or attractants, or plant growth regulators containing organic phosphorus compounds having phosphorus-to-carbon bonds
- A01N57/20—Biocides, pest repellants or attractants, or plant growth regulators containing organic phosphorus compounds having phosphorus-to-carbon bonds containing acyclic or cycloaliphatic radicals
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
- C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
- C12N15/82—Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
- C12N15/8241—Phenotypically and genetically modified plants via recombinant DNA technology
- C12N15/8261—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield
- C12N15/8271—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield for stress resistance, e.g. heavy metal resistance
- C12N15/8274—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield for stress resistance, e.g. heavy metal resistance for herbicide resistance
- C12N15/8277—Phosphinotricin
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
- C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
- C12N15/82—Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
- C12N15/8241—Phenotypically and genetically modified plants via recombinant DNA technology
- C12N15/8261—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield
- C12N15/8271—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield for stress resistance, e.g. heavy metal resistance
- C12N15/8274—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield for stress resistance, e.g. heavy metal resistance for herbicide resistance
- C12N15/8278—Sulfonylurea
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
- C12Q1/6888—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms
- C12Q1/6895—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms for plants, fungi or algae
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Engineering & Computer Science (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Biotechnology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Molecular Biology (AREA)
- General Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- General Health & Medical Sciences (AREA)
- Plant Pathology (AREA)
- Biophysics (AREA)
- Physics & Mathematics (AREA)
- Biochemistry (AREA)
- Microbiology (AREA)
- Analytical Chemistry (AREA)
- Cell Biology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Mycology (AREA)
- Botany (AREA)
- Immunology (AREA)
- Agronomy & Crop Science (AREA)
- Pest Control & Pesticides (AREA)
- Dentistry (AREA)
- Environmental Sciences (AREA)
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
Compositions and methods related to transgenic glyphosate tolerant Brassica plants are provided. Specifically, the present invention provides Brassica plants having a DP-073496-4 event which imparts tolerance to glyphosate. The Brassica plant harboring the DP-073496-4 event at the recited chromosomal location comprises genomic/transgene junctions within SEQ ID NO: 2 or with genomic/transgene junctions as set forth in SEQ
ID NO: 12 and/or 13. The characterization of the genomic insertion site of the event provides for an enhanced breeding efficiency and enables the use of molecular markers to track the transgene insert in the breeding populations and progeny thereof.
Various methods and compositions for the identification, detection, and use of the event are provided.
ID NO: 12 and/or 13. The characterization of the genomic insertion site of the event provides for an enhanced breeding efficiency and enables the use of molecular markers to track the transgene insert in the breeding populations and progeny thereof.
Various methods and compositions for the identification, detection, and use of the event are provided.
Description
=
BR.4SSICA GAT EVENT DP-073406-4 AND COMPOSITIONS AND METHODS FOR
THE 11:IEN1IFICATION AND/OR DETECTION THEREOF
REFERENCE TO A SEQUENCE USTING SUBMITTED AS A TEXT FILE VIA EFS-WEB
The official copy of the sequence listing is submitted concurrently with the specification as a text file via EFS-Web, In compliance with the American Standard =
Code for Information Interchange (ASCII), with a file name of 399080seqiist.txt, a creation date of November 24, 2010, and a size of 40 Kb. The sequence listing filed = via EFS-Web is part of the specification =
FIELD OF THE INVENTION
This Invention Is in the field of molecular biology. More specifically, this Invention pertains to expression of a sequence that confers tolerance to glyphosate.
BACKGROUND OF THE INVENTION
The expression of foreign genes In plants is known to be Influenced by their location in the plant genome, perhaps due to chromatin structure (e.gõ
heterochromatin) or the proximity of transcriptional regulatory elements (e.g., enhancers) close to the Integration site (Weising, of al, (1088) Ann. Rev.
Genet 22:421-477). At the same time the presence of the fransoene at different locations in the genome Influences the overall phenotype of the plant In different ways.
For this reason, it Is often necessary to screen a large number of events In order to Wen* an event characterized by optima/ expression of an Introduced gene of Interest.
For example, it has been observed in plants and in Other Organisms that there may be a wide variation in levels of expression of an introduced gene among events.
There may also be differences In spatial or temporal patterns of expression, for example, differences In the relative expression of'a transgene in various plant tissues, that may not correspond to the patterns expected from transcriptional regulatory elements present in the introduced gene construct. It Is also Observed that the transgene Insertion can affect the endogenous gene expression.. For these reasons, It is common to produce hundreds to thousands of different events and screen those =
= =
=
Date Recue/Date Received 2022-05-20 events for a single event that has desired transgene expression levels and patterns for commercial purposes. An event that has desired levels or patterns of transgene expression is useful for introgressing the transgene into other genetic backgrounds by sexual outcrossing using conventional breeding methods. Progeny of such crosses maintain the transgene expression characteristics of the original transformant. This strategy is used to ensure reliable gene expression in a number of varieties that are well adapted to local growing conditions.
It would be advantageous to be able to detect the presence of a particular event in order to determine whether progeny of a sexual cross contain a transgene of interest. In addition, a method for detecting a particular event would be helpful for = complying with regulations requiring the pre-market approval and labeling of foods derived from recombinant crop plants or for use in environmental monitoring, monitoring traits In crops in the field or monitoring products derived from a crop harvest, as well as for use in ensuring compliance of parties subject to regulatory or contractual terms.
In the, commercial production of crops, it is desirable to easily and quickly eliminate unwanted plants (i.e., "weeds") from a field of crop plants. An ideal treatment would be one which could be applied to an entire field but which would eliminate only the unwanted plants while leaving the crop plants unharmed. One such treatment system would involve the use 'of crop plants which are tolerant to an herbicide so that when the herbicide was sprayed on a field of herbicide-tolerant crop plants, the crop plants would continue to thrive while non-herbicide-tolerant weeds were killed or severely damaged.
Due to local and regional variation in dominant weed species as well as preferred crop species, a continuing need exists for customized systems of crop protection and weed management which can be adapted to the needs of a particular region, geography, and/or locality. Method and compositions that allow for the rapid identification of events in plants that produce such qualities are needed. For example, a continuing need exists for methods of crop protection and weed management which can reduce the number of herbicide applications necessary to control weeds in a field, reduce the amount of herbicide necessary to control weeds in a field, reduce the amount of tilling necessary to produce a crop, and/or delay or prevent the development and/or appearance of herbicide-resistant weeds. A continuing need exists for methods and compositions of crop protection and weed management which allow the targeted use of a particular herbicide and for the efficient detection of such an event.
BR.4SSICA GAT EVENT DP-073406-4 AND COMPOSITIONS AND METHODS FOR
THE 11:IEN1IFICATION AND/OR DETECTION THEREOF
REFERENCE TO A SEQUENCE USTING SUBMITTED AS A TEXT FILE VIA EFS-WEB
The official copy of the sequence listing is submitted concurrently with the specification as a text file via EFS-Web, In compliance with the American Standard =
Code for Information Interchange (ASCII), with a file name of 399080seqiist.txt, a creation date of November 24, 2010, and a size of 40 Kb. The sequence listing filed = via EFS-Web is part of the specification =
FIELD OF THE INVENTION
This Invention Is in the field of molecular biology. More specifically, this Invention pertains to expression of a sequence that confers tolerance to glyphosate.
BACKGROUND OF THE INVENTION
The expression of foreign genes In plants is known to be Influenced by their location in the plant genome, perhaps due to chromatin structure (e.gõ
heterochromatin) or the proximity of transcriptional regulatory elements (e.g., enhancers) close to the Integration site (Weising, of al, (1088) Ann. Rev.
Genet 22:421-477). At the same time the presence of the fransoene at different locations in the genome Influences the overall phenotype of the plant In different ways.
For this reason, it Is often necessary to screen a large number of events In order to Wen* an event characterized by optima/ expression of an Introduced gene of Interest.
For example, it has been observed in plants and in Other Organisms that there may be a wide variation in levels of expression of an introduced gene among events.
There may also be differences In spatial or temporal patterns of expression, for example, differences In the relative expression of'a transgene in various plant tissues, that may not correspond to the patterns expected from transcriptional regulatory elements present in the introduced gene construct. It Is also Observed that the transgene Insertion can affect the endogenous gene expression.. For these reasons, It is common to produce hundreds to thousands of different events and screen those =
= =
=
Date Recue/Date Received 2022-05-20 events for a single event that has desired transgene expression levels and patterns for commercial purposes. An event that has desired levels or patterns of transgene expression is useful for introgressing the transgene into other genetic backgrounds by sexual outcrossing using conventional breeding methods. Progeny of such crosses maintain the transgene expression characteristics of the original transformant. This strategy is used to ensure reliable gene expression in a number of varieties that are well adapted to local growing conditions.
It would be advantageous to be able to detect the presence of a particular event in order to determine whether progeny of a sexual cross contain a transgene of interest. In addition, a method for detecting a particular event would be helpful for = complying with regulations requiring the pre-market approval and labeling of foods derived from recombinant crop plants or for use in environmental monitoring, monitoring traits In crops in the field or monitoring products derived from a crop harvest, as well as for use in ensuring compliance of parties subject to regulatory or contractual terms.
In the, commercial production of crops, it is desirable to easily and quickly eliminate unwanted plants (i.e., "weeds") from a field of crop plants. An ideal treatment would be one which could be applied to an entire field but which would eliminate only the unwanted plants while leaving the crop plants unharmed. One such treatment system would involve the use 'of crop plants which are tolerant to an herbicide so that when the herbicide was sprayed on a field of herbicide-tolerant crop plants, the crop plants would continue to thrive while non-herbicide-tolerant weeds were killed or severely damaged.
Due to local and regional variation in dominant weed species as well as preferred crop species, a continuing need exists for customized systems of crop protection and weed management which can be adapted to the needs of a particular region, geography, and/or locality. Method and compositions that allow for the rapid identification of events in plants that produce such qualities are needed. For example, a continuing need exists for methods of crop protection and weed management which can reduce the number of herbicide applications necessary to control weeds in a field, reduce the amount of herbicide necessary to control weeds in a field, reduce the amount of tilling necessary to produce a crop, and/or delay or prevent the development and/or appearance of herbicide-resistant weeds. A continuing need exists for methods and compositions of crop protection and weed management which allow the targeted use of a particular herbicide and for the efficient detection of such an event.
2 Date Recue/Date Received 2022-05-20 BRIEF SUMMARY OF THE INVENTION
Compositions and methods related to transgenic glyphosate-tolerant Brassica plants are provided. Specifically, the present invention provides Brassica plants containing a transgene which imparts tolerance to glyphosate. The event may be, for example, DP-073496-4. The Brassica plant harboring the transgene at the recited chromosomal location comprises unique genomic/transgene junctions having at least the ,polynucleotide sequence of SEQ ID NO: 2 or at least the polynucleotide sequence of SEQ ID NO: 12 and/or 13. Further provided are the seeds deposited as Patent Deposit Number PTA-11504 and plants, plant cells, plant parts, seed and plant products derived therefrom. Characterization of the genomic insertion site of DP-073496-4 or any other event comprising integration of the glyphosate-tolerance transgene provides for an enhanced breeding efficiency and enables the use of molecular markers to track the transgene insert in the breeding populations and progeny thereof. Various methods and compositions for the identification, detection, and use of the glyphosate-N-acetyltransferase ("GAT" or ''glyat") transformation event in Brassica are provided.
BRIEF DESCRIPTION OF THE DRAWINGS
Figure 1 shows synthesis of plasmid PHP28181. Plasmid PHP28181 was used to produce the GAT Brassica lines.
Figure 2 provides a schematic map of plasmid PHP28181.
Figure 3 provides a schematic map of insertion DNA, fragment PHP28181A.
Figure 4 provides a schematic representation of fragment A from PHP28181 (PHP28181A), specifically a schematic map of Hind III/Not I fragment (PHP28181A) containing the gat4621 gene cassette that was used for plant transformation to generate DP-073496-4 Brassica. The fragment size is 2112 bp. The construct-specific primer locations of 09-0-3290/09-0-3288 are indicated on the map.
Figure 5 Southern analysis of Construct Specific PCR of Leaf DNA From DP-073496-4 Brassica and Non-Genetically Modified Control Brassica, PCR
amplification with primer set 09-0-3290/09-0-3288 targeting the unique ubiquitin promoter and gat4621 junction present in DP-073496-4canola. Expected PCR
amplicon size is 675 bp.
Figure 6 Southern analysis of Brassica FatA gene PCR of leaf DNA from DP-073496-4 Brassica and Non-Genetically Modified Control Brassica. PCR
amplification of endogenous brassica FatA gene with primer set 09-0-2812/09-as positive control for PCR amplification. Expected PCR amplicon size is 506 bp.
Compositions and methods related to transgenic glyphosate-tolerant Brassica plants are provided. Specifically, the present invention provides Brassica plants containing a transgene which imparts tolerance to glyphosate. The event may be, for example, DP-073496-4. The Brassica plant harboring the transgene at the recited chromosomal location comprises unique genomic/transgene junctions having at least the ,polynucleotide sequence of SEQ ID NO: 2 or at least the polynucleotide sequence of SEQ ID NO: 12 and/or 13. Further provided are the seeds deposited as Patent Deposit Number PTA-11504 and plants, plant cells, plant parts, seed and plant products derived therefrom. Characterization of the genomic insertion site of DP-073496-4 or any other event comprising integration of the glyphosate-tolerance transgene provides for an enhanced breeding efficiency and enables the use of molecular markers to track the transgene insert in the breeding populations and progeny thereof. Various methods and compositions for the identification, detection, and use of the glyphosate-N-acetyltransferase ("GAT" or ''glyat") transformation event in Brassica are provided.
BRIEF DESCRIPTION OF THE DRAWINGS
Figure 1 shows synthesis of plasmid PHP28181. Plasmid PHP28181 was used to produce the GAT Brassica lines.
Figure 2 provides a schematic map of plasmid PHP28181.
Figure 3 provides a schematic map of insertion DNA, fragment PHP28181A.
Figure 4 provides a schematic representation of fragment A from PHP28181 (PHP28181A), specifically a schematic map of Hind III/Not I fragment (PHP28181A) containing the gat4621 gene cassette that was used for plant transformation to generate DP-073496-4 Brassica. The fragment size is 2112 bp. The construct-specific primer locations of 09-0-3290/09-0-3288 are indicated on the map.
Figure 5 Southern analysis of Construct Specific PCR of Leaf DNA From DP-073496-4 Brassica and Non-Genetically Modified Control Brassica, PCR
amplification with primer set 09-0-3290/09-0-3288 targeting the unique ubiquitin promoter and gat4621 junction present in DP-073496-4canola. Expected PCR
amplicon size is 675 bp.
Figure 6 Southern analysis of Brassica FatA gene PCR of leaf DNA from DP-073496-4 Brassica and Non-Genetically Modified Control Brassica. PCR
amplification of endogenous brassica FatA gene with primer set 09-0-2812/09-as positive control for PCR amplification. Expected PCR amplicon size is 506 bp.
- 3 -Date Recue/Date Received 2022-05-20 DETAILED DESCRIPTION OF THE INVENTION
The present Inventions now will be described more fully hereinafter with reference to the accompanying drawings, in which some, but not all embodiments of the inventions are shown. Indeed, these inventions may be embodied in many different forms and should not be construed as limited to the embodiments set forth herein; rather, these embodiments are provided so that this disclosure will satisfy applicable legal requirements. Like numbers refer to like elements throughout.
Many modifications and other embodiments of the inventions set forth herein will come to mind to one skilled in the art to which these Inventions pertain having the benefit of the teachings presented in the foregoing descriptions and the associated drawings. Therefore, it is to be understood that the inventions are not to be limited to the specific embodiments disclosed and that modifications and other embodiments are intended to be included within the scope of the appended claims. Although specific terms are employed herein, they are used in a generic and descriptive sense only and not for purposes of limitation.
Compositions and methods related to transgenic glyphosate-tolerant Brass/ca plants are provided. Specifically, the present invention provides Brass/ca plants having event DP-073496-4 or another event comprising PHP28181A or an operable =
fragment or variant thereof. A Brass/ca plant having event DP-073496-4, for example, has been modified by the insertion of the glyphosate acetyftransferase (glyat4621) gene derived from Bacillus licheniformis. The g1yat4621 gene was, functionally improved by a gene shuffling process to optimize the kinetics of glyphosate acetyftransferase (GLYN]) activity for acetylating the herbicide glyphosate.
The Insertion of the g1yat4621 gene in the plant confers tolerance to the herbicidal active ingredient glyphosate through the conversion of glyphosate to the non-toxic acetylated form. Thus, a Brass/ca plant having the event DP-073496-4 is tolerant to glyphosate.
The polynucleotides conferring the glyphosate tolerance are Inserted at a specific position in the Brass/ca genome and thereby produce, for example, the DP-073496-4 event. A Brass/ca plant harboring the DP-073496-4 event at a specific chromosomal location comprises genomicitransgene Junctions having a unique polynuoleotide sequence exemplified by SEQ ID NO: 2 or at least the polynucleotide sequence of SEQ ID NO: 12 and/or 13; SEQ ID NO: 14 and/or 15; or SEQ ID NO: 16 and/or 17. The characterization of the genomic insertion site of either event provides for an enhanced breeding efficiency and enables the use of molecular markers to track the transgene insert in .the breeding populations and progeny thereof. Various methods and compositions for the identification, detection, and use of the Brass/ca DP-073498-4 event are provided herein, in one embodiment, a brassica plant having
The present Inventions now will be described more fully hereinafter with reference to the accompanying drawings, in which some, but not all embodiments of the inventions are shown. Indeed, these inventions may be embodied in many different forms and should not be construed as limited to the embodiments set forth herein; rather, these embodiments are provided so that this disclosure will satisfy applicable legal requirements. Like numbers refer to like elements throughout.
Many modifications and other embodiments of the inventions set forth herein will come to mind to one skilled in the art to which these Inventions pertain having the benefit of the teachings presented in the foregoing descriptions and the associated drawings. Therefore, it is to be understood that the inventions are not to be limited to the specific embodiments disclosed and that modifications and other embodiments are intended to be included within the scope of the appended claims. Although specific terms are employed herein, they are used in a generic and descriptive sense only and not for purposes of limitation.
Compositions and methods related to transgenic glyphosate-tolerant Brass/ca plants are provided. Specifically, the present invention provides Brass/ca plants having event DP-073496-4 or another event comprising PHP28181A or an operable =
fragment or variant thereof. A Brass/ca plant having event DP-073496-4, for example, has been modified by the insertion of the glyphosate acetyftransferase (glyat4621) gene derived from Bacillus licheniformis. The g1yat4621 gene was, functionally improved by a gene shuffling process to optimize the kinetics of glyphosate acetyftransferase (GLYN]) activity for acetylating the herbicide glyphosate.
The Insertion of the g1yat4621 gene in the plant confers tolerance to the herbicidal active ingredient glyphosate through the conversion of glyphosate to the non-toxic acetylated form. Thus, a Brass/ca plant having the event DP-073496-4 is tolerant to glyphosate.
The polynucleotides conferring the glyphosate tolerance are Inserted at a specific position in the Brass/ca genome and thereby produce, for example, the DP-073496-4 event. A Brass/ca plant harboring the DP-073496-4 event at a specific chromosomal location comprises genomicitransgene Junctions having a unique polynuoleotide sequence exemplified by SEQ ID NO: 2 or at least the polynucleotide sequence of SEQ ID NO: 12 and/or 13; SEQ ID NO: 14 and/or 15; or SEQ ID NO: 16 and/or 17. The characterization of the genomic insertion site of either event provides for an enhanced breeding efficiency and enables the use of molecular markers to track the transgene insert in .the breeding populations and progeny thereof. Various methods and compositions for the identification, detection, and use of the Brass/ca DP-073498-4 event are provided herein, in one embodiment, a brassica plant having
4 Date Recue/Date Received 2022-05-20 in its genome in the following order: a polynucleotide comprising SEQ ID NO:
12, a polynucleotide encoding a glyphosate-N-acetyltransferase and a polynucleotide comprising SEQ ID NO: 13 is provided. The term "event DP-073496-4 specific"
refers to a polynucleotide sequence which is suitable for discriminatively identifying event DP-073496-4 in plants, plant material, or in products such as, but not limited to, oil produced from the seeds, or food or feed products (fresh or processed) comprising, or derived from, plant material.
Compositions further include seed deposited as Patent Deposit Numbers PTA-11504 and plants, plant cells, and seed derived therefrom. Applicant(s) have made a deposit of at least 2500 seeds of Brassica event DP-073496-4 with the American Type Culture Collection (ATCC), Manassas, VA 20110-2209 USA on November 24, 2010 and the deposit will be maintained under the terms of the Budapest Treaty on the International Recognition of the Deposit of Microorganisms for the Purposes of Patent Procedure. Deposits are made merely as a convenience for those of skill in the art and are not an admission that a deposit is required under 35 U.S.C. 112. The seeds deposited with the ATCC are taken from the deposit maintained by Pioneer Hi-Bred International, Inc., 7250 NW 62nd Avenue, Johnston, Iowa 50131-1000. Access to this deposit will be available during the pendency of the application to the Commissioner of Patents and Trademarks and persons determined by the Commissioner to be entitled thereto upon request. Upon allowance of any claims in the application, the Applicant(s) will make available to the public, pursuant to 37 C.F.R. 1.808, sample(s) of the deposit. The deposit of seed comprising Brassica event DP-073496-4 will be maintained in the ATCC depository, which is a public depository, for a period of 30 years or 5 years after the most recent request or for the enforceable life of the patent, whichever is longer and will be replaced if it becomes nonviable during that period. Additionally, Applicant(s) will have satisfied all the requirements of 37 C.F.R. 1.801 - 1.809, including providing an indication of the viability of the sample upon deposit. Applicant(s) have no authority to waive any restrictions imposed by law on the transfer of biological material or its transportation in commerce. Applicant(s) do not waive any infringement of their rights granted under this patent or rights applicable to event DP-073496-4 under the Plant Variety Protection Act (7 USC 2321, et seq.). Unauthorized seed multiplication prohibited.
The seed may be regulated.
As used herein, the term "Brassica" means any Brassica plant and includes all plant varieties that can be bred with Brassica. As used herein, the term plant includes plant cells, plant organs, plant protoplasts, plant cell tissue cultures from which plants can be regenerated, plant calli, plant clumps and plant cells that are
12, a polynucleotide encoding a glyphosate-N-acetyltransferase and a polynucleotide comprising SEQ ID NO: 13 is provided. The term "event DP-073496-4 specific"
refers to a polynucleotide sequence which is suitable for discriminatively identifying event DP-073496-4 in plants, plant material, or in products such as, but not limited to, oil produced from the seeds, or food or feed products (fresh or processed) comprising, or derived from, plant material.
Compositions further include seed deposited as Patent Deposit Numbers PTA-11504 and plants, plant cells, and seed derived therefrom. Applicant(s) have made a deposit of at least 2500 seeds of Brassica event DP-073496-4 with the American Type Culture Collection (ATCC), Manassas, VA 20110-2209 USA on November 24, 2010 and the deposit will be maintained under the terms of the Budapest Treaty on the International Recognition of the Deposit of Microorganisms for the Purposes of Patent Procedure. Deposits are made merely as a convenience for those of skill in the art and are not an admission that a deposit is required under 35 U.S.C. 112. The seeds deposited with the ATCC are taken from the deposit maintained by Pioneer Hi-Bred International, Inc., 7250 NW 62nd Avenue, Johnston, Iowa 50131-1000. Access to this deposit will be available during the pendency of the application to the Commissioner of Patents and Trademarks and persons determined by the Commissioner to be entitled thereto upon request. Upon allowance of any claims in the application, the Applicant(s) will make available to the public, pursuant to 37 C.F.R. 1.808, sample(s) of the deposit. The deposit of seed comprising Brassica event DP-073496-4 will be maintained in the ATCC depository, which is a public depository, for a period of 30 years or 5 years after the most recent request or for the enforceable life of the patent, whichever is longer and will be replaced if it becomes nonviable during that period. Additionally, Applicant(s) will have satisfied all the requirements of 37 C.F.R. 1.801 - 1.809, including providing an indication of the viability of the sample upon deposit. Applicant(s) have no authority to waive any restrictions imposed by law on the transfer of biological material or its transportation in commerce. Applicant(s) do not waive any infringement of their rights granted under this patent or rights applicable to event DP-073496-4 under the Plant Variety Protection Act (7 USC 2321, et seq.). Unauthorized seed multiplication prohibited.
The seed may be regulated.
As used herein, the term "Brassica" means any Brassica plant and includes all plant varieties that can be bred with Brassica. As used herein, the term plant includes plant cells, plant organs, plant protoplasts, plant cell tissue cultures from which plants can be regenerated, plant calli, plant clumps and plant cells that are
- 5 -Date Recue/Date Received 2022-05-20 intact in plants or parts of plants such as embryos, pollen, ovules, seeds, leaves, flowers, branches, fruit, Date Recue/Date Received 2022-05-20 stalks, roots, root tips, anthers, and the like. Mature seed produced may be used for food, feed, fuel or other commercial or industrial purposes or for purposes of growing or reproducing the species. Progeny, variants and mutants of the regenerated plants are also included within the scope of the invention, provided that these parts comprise a DP-073496-4 event.
A transgenic "event" is produced by transformation of plant cells with a heterologous DNA construct(s) including a nucleic acid expression cassette that comprises a transgene of interest, the regeneration of a population of plants from cells which each comprise the inserted transgene and selection of a particular plant characterized by insertion into a particular genome location. An event is characterized phenotypically by the expression of the transgene(s). At the genetic level, an event Is part of the genetic makeup of a plant. The term "event" also refers to progeny, produced by a sexual outcross between the transformant and another variety, that Include the heterologous DNA. Even after repeated back-crossing to a recurrent parent, the inserted DNA and flanking DNA from the transformed parent are present in the progeny of the cross at the same chromosomal location. The term "event"
also refers to DNA from the original transformant comprising the inserted DNA and flanking sequence immediately adjacent to the inserted DNA that would be expected to be transferred to a progeny as the result of a sexual cross of one parental line that includes the inserted DNA (e.g., the original transformant and progeny resulting from selfing) and a parental line that does not contain the inserted DNA.
As used herein, "insert DNA' refers to the heterologous DNA within the expression cassettes used to transform the plant material while "flanking DNA"
can comprise either genornic DNA naturally present In an organism such as a plant, or foreign (heterologous) DNA introduced via the transformation process which is extraneous to the original insert DNA molecule, e.g. fragments associated with the transformation event. A "flanking region" or tanking sequence" as used herein refers to a sequence of at least 20, 50, 100, 200, 300, 400, 1000, 1500, 2000, 2500 or 5000 base pairs or greater which Is located either immediately upstream of and contiguous with, or immediately downstream of and contiguous with, the original foreign Insert DNA molecule. Non-limiting examples of the flanking regions of the DP-073496-4 event comprise polynucieotide sequences that are set faith in SEC) ID NO: 2, 8 and/or 9 and variants and fragments thereof.
Transformation procedures leading to random integration of the foreign DNA
will result in transformants containing different flanking regions characteristic of and unique for each transformant. " When recombinant DNA is introduced into a plant through traditional crossing, its flanking regions will generally not be changed.
A transgenic "event" is produced by transformation of plant cells with a heterologous DNA construct(s) including a nucleic acid expression cassette that comprises a transgene of interest, the regeneration of a population of plants from cells which each comprise the inserted transgene and selection of a particular plant characterized by insertion into a particular genome location. An event is characterized phenotypically by the expression of the transgene(s). At the genetic level, an event Is part of the genetic makeup of a plant. The term "event" also refers to progeny, produced by a sexual outcross between the transformant and another variety, that Include the heterologous DNA. Even after repeated back-crossing to a recurrent parent, the inserted DNA and flanking DNA from the transformed parent are present in the progeny of the cross at the same chromosomal location. The term "event"
also refers to DNA from the original transformant comprising the inserted DNA and flanking sequence immediately adjacent to the inserted DNA that would be expected to be transferred to a progeny as the result of a sexual cross of one parental line that includes the inserted DNA (e.g., the original transformant and progeny resulting from selfing) and a parental line that does not contain the inserted DNA.
As used herein, "insert DNA' refers to the heterologous DNA within the expression cassettes used to transform the plant material while "flanking DNA"
can comprise either genornic DNA naturally present In an organism such as a plant, or foreign (heterologous) DNA introduced via the transformation process which is extraneous to the original insert DNA molecule, e.g. fragments associated with the transformation event. A "flanking region" or tanking sequence" as used herein refers to a sequence of at least 20, 50, 100, 200, 300, 400, 1000, 1500, 2000, 2500 or 5000 base pairs or greater which Is located either immediately upstream of and contiguous with, or immediately downstream of and contiguous with, the original foreign Insert DNA molecule. Non-limiting examples of the flanking regions of the DP-073496-4 event comprise polynucieotide sequences that are set faith in SEC) ID NO: 2, 8 and/or 9 and variants and fragments thereof.
Transformation procedures leading to random integration of the foreign DNA
will result in transformants containing different flanking regions characteristic of and unique for each transformant. " When recombinant DNA is introduced into a plant through traditional crossing, its flanking regions will generally not be changed.
6 =
Date Recue/Date Received 2022-05-20 Transformants will also contain unique junctions between a piece of heterologous insert DNA and genomic DNA or two pieces of genomic DNA or two pieces of heterologous DNA. A "junction' is a point where two specific DNA fragments join. For example, a junction exists where insert DNA joins flanking DNA. A junction point also exists in a transformed organism where two DNA fragments join together in a manner that is modified from that found in the native organism. As used herein, 'Junction DNA"
refers to DNA that comprises a junction point. Non-limiting examples of junction DNA
from the DP-073496-4 event are forth in SEO ID NO: 2, 11, 12, 13, 14, 15, 16, 17, 18, and/or 19 or variants and fragments thereof.
A DP073496-4 plant can be bred by first sexually crossing a first parental Brassica plant grown from the transgenic DP-073496-4 Brassica plant (or progeny thereof derived from transformation with the expression cassettes of the embodiments of the present Invention that confer herbicide tolerance) and a second parental Brassica plant that lacks the herbicide tolerance phenotype, thereby producing a plurality of first progeny plants and then selecting a first progeny plant that displays the desired herbicide tolerance and selfing the first progeny plant, thereby producing a plurality of second progeny plants and then selecting from the second progeny plants which display the desired herbicide tolerance. These steps can further include the back-crossing of the first herbicide tolerant progeny plant or the second herbicide tolerant progeny plant to the second parental Brassica plant or a third parental Brassica plant, thereby producing a Brassies plant that displays the desired herbicide tolerance. It is further recognized that assaying progeny for phenotype ii not required.
Various methods and compositions, as disclosed elsewhere herein, can be used to detect and/or Identify the DP073496-4 or other event.
Two different transgenio plants can also be sexually crossed to produce offspring that contain two independently-segregating exogenous genes. Seifing of =
appropriate progeny can produce plants that are homozygous for both exogenous genes. Sack-crossing to a parental plant and out-crossing with a non-transganic plant are also contemplated, as is vegetative propagation. Descriptions of other breeding methods that are commonly used for different traits and crops can be found in one of = several references, e.gõ Fehr, in Breeding Methods for Cultivar Development, VVilcos, ed., American Society of Agronomy, Madison Wis. (1987).
The term "germplasm" refers to an individual, a group of individuals or a clone representing a genotype, variety, species or culture or the genetic material thereof.
A line" or "strain" is a group of individuals of identical parentage that are generally inbred to some degree and that are generally isogonic or near isogenic.
Date Recue/Date Received 2022-05-20 Transformants will also contain unique junctions between a piece of heterologous insert DNA and genomic DNA or two pieces of genomic DNA or two pieces of heterologous DNA. A "junction' is a point where two specific DNA fragments join. For example, a junction exists where insert DNA joins flanking DNA. A junction point also exists in a transformed organism where two DNA fragments join together in a manner that is modified from that found in the native organism. As used herein, 'Junction DNA"
refers to DNA that comprises a junction point. Non-limiting examples of junction DNA
from the DP-073496-4 event are forth in SEO ID NO: 2, 11, 12, 13, 14, 15, 16, 17, 18, and/or 19 or variants and fragments thereof.
A DP073496-4 plant can be bred by first sexually crossing a first parental Brassica plant grown from the transgenic DP-073496-4 Brassica plant (or progeny thereof derived from transformation with the expression cassettes of the embodiments of the present Invention that confer herbicide tolerance) and a second parental Brassica plant that lacks the herbicide tolerance phenotype, thereby producing a plurality of first progeny plants and then selecting a first progeny plant that displays the desired herbicide tolerance and selfing the first progeny plant, thereby producing a plurality of second progeny plants and then selecting from the second progeny plants which display the desired herbicide tolerance. These steps can further include the back-crossing of the first herbicide tolerant progeny plant or the second herbicide tolerant progeny plant to the second parental Brassica plant or a third parental Brassica plant, thereby producing a Brassies plant that displays the desired herbicide tolerance. It is further recognized that assaying progeny for phenotype ii not required.
Various methods and compositions, as disclosed elsewhere herein, can be used to detect and/or Identify the DP073496-4 or other event.
Two different transgenio plants can also be sexually crossed to produce offspring that contain two independently-segregating exogenous genes. Seifing of =
appropriate progeny can produce plants that are homozygous for both exogenous genes. Sack-crossing to a parental plant and out-crossing with a non-transganic plant are also contemplated, as is vegetative propagation. Descriptions of other breeding methods that are commonly used for different traits and crops can be found in one of = several references, e.gõ Fehr, in Breeding Methods for Cultivar Development, VVilcos, ed., American Society of Agronomy, Madison Wis. (1987).
The term "germplasm" refers to an individual, a group of individuals or a clone representing a genotype, variety, species or culture or the genetic material thereof.
A line" or "strain" is a group of individuals of identical parentage that are generally inbred to some degree and that are generally isogonic or near isogenic.
7 Date Recue/Date Received 2022-05-20 Inbred lines tend to be highly homogeneous, homozygous and reproducible.
Many analytical methods are available to determine the homozygosity and phenotypic stability of inbred lines.
The phrase "hybrid plants" refers to plants which result from a cross between genetically different individuals.
The term "crossed" or "cross" in the context of this Invention means the fusion of gametes, e.g., via pollination to produce progeny (i.e., cells, seeds, or plants) in the case of plants. The term encompasses both sexual crosses (the pollination of one plant by another) and, in the case of plants, sewing (self-pollination, i.e., when the pollen and ovule are from the same plant).
The term "Introgression" refers to the transmission of a desired allele of a genetic locus from one genetic background to another. In one method, the desired alleles can be Introgressed through a sexual cross between two parents, wherein at least one of the parents has the desired allele in its genome.
In some embodiments, the polynucleotides conferring the brassica DP-073496-4 event of the invention are engineered into a molecular stack. In other embodiments, the molecular stack further comprises at least one additional polynucleotide that confers tolerance to a second. herbicide. In one embodiment, the sequence confers tolerance to glufosinate, and in a specific embodiment, the sequence comprises pat =
gene, In another embodiment, the additional polynudeotide provides tolerance to ALS-inhibitor herbicides.
In other embodiments, an event of the invention comprises one or more traits of interest, and in more specific embodiments, the plant is stacked with any combination of polynucleotide sequences of interest in order to create plants with a desired combination of traits. A trait, as used herein, refers to the phenotype derived from a particular sequence or groups of sequences. For example, herbicide-tolerance polynucleotides may be stacked with any other polynucleotides encoding poiypeptides having pesticidal andior insecticidal activity, such as Bacillus thuringiensis toxic proteins (described in US Patent Numbers 5,366,892; 5,747,450; 5,737,514;
34 5,723,756; 5,593,881; Geiser, at al., (1986) Gene 48;109; Lee, et al., (2003) App!.
Environ. Microbial. 69:4648-4657 (Vip3A); Galitzky, at at., (2001) Acta Crystallogr, D.
BM. Crystallogr. 57:1101-1109 (Cty3Bb1) and Herman, et al., (2004) J. Agrio.
Food Chem. 52:2726-2734 (CtylF)), lectins (Van Demme, at at., (1994) Plant Mal.
Biol.
24:825, pentin (described in US Patent Number 5,981,722), and the like. The combinations generated can also include multiple copies of any one of the polynucleotides of interest.
Many analytical methods are available to determine the homozygosity and phenotypic stability of inbred lines.
The phrase "hybrid plants" refers to plants which result from a cross between genetically different individuals.
The term "crossed" or "cross" in the context of this Invention means the fusion of gametes, e.g., via pollination to produce progeny (i.e., cells, seeds, or plants) in the case of plants. The term encompasses both sexual crosses (the pollination of one plant by another) and, in the case of plants, sewing (self-pollination, i.e., when the pollen and ovule are from the same plant).
The term "Introgression" refers to the transmission of a desired allele of a genetic locus from one genetic background to another. In one method, the desired alleles can be Introgressed through a sexual cross between two parents, wherein at least one of the parents has the desired allele in its genome.
In some embodiments, the polynucleotides conferring the brassica DP-073496-4 event of the invention are engineered into a molecular stack. In other embodiments, the molecular stack further comprises at least one additional polynucleotide that confers tolerance to a second. herbicide. In one embodiment, the sequence confers tolerance to glufosinate, and in a specific embodiment, the sequence comprises pat =
gene, In another embodiment, the additional polynudeotide provides tolerance to ALS-inhibitor herbicides.
In other embodiments, an event of the invention comprises one or more traits of interest, and in more specific embodiments, the plant is stacked with any combination of polynucleotide sequences of interest in order to create plants with a desired combination of traits. A trait, as used herein, refers to the phenotype derived from a particular sequence or groups of sequences. For example, herbicide-tolerance polynucleotides may be stacked with any other polynucleotides encoding poiypeptides having pesticidal andior insecticidal activity, such as Bacillus thuringiensis toxic proteins (described in US Patent Numbers 5,366,892; 5,747,450; 5,737,514;
34 5,723,756; 5,593,881; Geiser, at al., (1986) Gene 48;109; Lee, et al., (2003) App!.
Environ. Microbial. 69:4648-4657 (Vip3A); Galitzky, at at., (2001) Acta Crystallogr, D.
BM. Crystallogr. 57:1101-1109 (Cty3Bb1) and Herman, et al., (2004) J. Agrio.
Food Chem. 52:2726-2734 (CtylF)), lectins (Van Demme, at at., (1994) Plant Mal.
Biol.
24:825, pentin (described in US Patent Number 5,981,722), and the like. The combinations generated can also include multiple copies of any one of the polynucleotides of interest.
8 Date Recue/Date Received 2022-05-20 In some embodiments, an event of the invention may be stacked with other herbicide-tolerance traits to create a transgenic plant of the invention with further improved properties. Other herbicide-tolerance polynucleotides that could be used In such embodiments include those conferring tolerance to glyphosate by other modes of action, such as, for example, a gene that encodes a glyphosate oxido-reductase enzyme as described more fully in US Patent Numbers 5,776,760 and 5,463,175.
Other traits that could be combined with an event of the invention include .those derived from polynucleotides that confer on the plant the capacity to produce a higher level of 6-enolpyruvylshikimate-3-phosphate synthase (EPSPS), for example, as more fully described in US Patent Numbers 6,248,876 B1; 5,627,061; 5,804,425;
5,633,435;
5,145,783; 4,971,908; 5,312,910; 5,188,642; 4,940,835; 5,866,775; 6,225,114 13.1;
6,130,366; 5,310,667; 4,535,060; 4,769,061; 5,633,448; 5,510,471; Re. 36,449;
RE
37,287 E and 5,491,288 and International Publication Numbers WO 97/04103; WO
00/66746; WO 01/66704 and WO 00/66747. Other traits that could be combined with the an event of the invention include those conferring tolerance to sulfonylurea and/or imidazolinone, for example, as described more fully in US Patent Numbers 5,605,011;
5,013,659; 5,141,870; 5,767,361; 5,731,180; 5,304,732; 4,761,373; 5,331,107;
5,928,937 and 5,378,824 and International Publication Number WO 96/33270.
In some. embodiments, an event of the Invention may be stacked with, for example, hydroxyphenylpyruvatedloxygenases which are enzymes that catalyze the reaction in which para-hydroxyphenylpyruvate (HPP) is transformed into ' homogentisate. Molecules which inhibit this enzyme and which bind to the enzyme in order to inhibit transformation of the HPP into hornogentisate are useful as herbicides.
Traits conferring tolerance to such herbicides in plants are described in US
Patent Numbers 6,245,968 B1 ; 6,268,549 and 6,069,115 and International Publication Number WO 99/23856. Other examples of suitable herbicide-tolerance traits that could be stacked with an event of the invention Include aryloxyalkanoate dioxygenase polynucleotides (which reportedly confer tolerance to 2,4-D and other phenoxy auxin herbicides as well as to aryloxyphenoxypropionate herbicides as described, for example, in International Publication WO 05/107437) and dicamba-tolerance polynucleotides as described, for example, in Herman, et al., (2005) J. Biol.
Chem.
280:24759-24767.
Other examples of herbicide-tolerance traits that could be combined with an event disclosed herein include those conferred by polynucleotides encoding an exogenous phosphinothricin acetyltransferase, as described In US Patent Numbers 5,969,213; 5,489,520; 5,550,318; 5,874,265.; 5,919,675; 5,561,236; 5,648,477;
5,646,024; 6,177,616 and 5,879,903. Plants containing an exogenous
Other traits that could be combined with an event of the invention include .those derived from polynucleotides that confer on the plant the capacity to produce a higher level of 6-enolpyruvylshikimate-3-phosphate synthase (EPSPS), for example, as more fully described in US Patent Numbers 6,248,876 B1; 5,627,061; 5,804,425;
5,633,435;
5,145,783; 4,971,908; 5,312,910; 5,188,642; 4,940,835; 5,866,775; 6,225,114 13.1;
6,130,366; 5,310,667; 4,535,060; 4,769,061; 5,633,448; 5,510,471; Re. 36,449;
RE
37,287 E and 5,491,288 and International Publication Numbers WO 97/04103; WO
00/66746; WO 01/66704 and WO 00/66747. Other traits that could be combined with the an event of the invention include those conferring tolerance to sulfonylurea and/or imidazolinone, for example, as described more fully in US Patent Numbers 5,605,011;
5,013,659; 5,141,870; 5,767,361; 5,731,180; 5,304,732; 4,761,373; 5,331,107;
5,928,937 and 5,378,824 and International Publication Number WO 96/33270.
In some. embodiments, an event of the Invention may be stacked with, for example, hydroxyphenylpyruvatedloxygenases which are enzymes that catalyze the reaction in which para-hydroxyphenylpyruvate (HPP) is transformed into ' homogentisate. Molecules which inhibit this enzyme and which bind to the enzyme in order to inhibit transformation of the HPP into hornogentisate are useful as herbicides.
Traits conferring tolerance to such herbicides in plants are described in US
Patent Numbers 6,245,968 B1 ; 6,268,549 and 6,069,115 and International Publication Number WO 99/23856. Other examples of suitable herbicide-tolerance traits that could be stacked with an event of the invention Include aryloxyalkanoate dioxygenase polynucleotides (which reportedly confer tolerance to 2,4-D and other phenoxy auxin herbicides as well as to aryloxyphenoxypropionate herbicides as described, for example, in International Publication WO 05/107437) and dicamba-tolerance polynucleotides as described, for example, in Herman, et al., (2005) J. Biol.
Chem.
280:24759-24767.
Other examples of herbicide-tolerance traits that could be combined with an event disclosed herein include those conferred by polynucleotides encoding an exogenous phosphinothricin acetyltransferase, as described In US Patent Numbers 5,969,213; 5,489,520; 5,550,318; 5,874,265.; 5,919,675; 5,561,236; 5,648,477;
5,646,024; 6,177,616 and 5,879,903. Plants containing an exogenous
9 Date Recue/Date Received 2022-05-20 =
=
phosphinothricin acetyltransferase can exhibit improved tolerance to glufosinate = herbicides, which inhibit the enzyme glutamine synthase. Other examples of herbicide-tolerance traits that could be combined with an event disclosed herein Include those conferred by polynucleotides conferring altered protoporphydnogen = 5 coddase (protox) activfty;as described in US Patent Numbers 8,288,306 81; 6,262,837 131 and 5,707,373 and International Publication Number WO 01/12825. Plants = containing such polynucleotides can exhibit Improved tolerance to any of a variety of herbicides which target the protox enzyme (also referred to as "protox Inhibitors".
In other embodiments, an ALS inhibRokolmant trait is combined with the event =
= 10 disclosed herein. As used herein, an "ALS Inhibitor-tolerant polypeptide' comprises any polypeptide which when expressed in a plant confers tolerance to at least one ALS
Inhibitor. A variety of ALS Inhibitors are known and include, for example, sulfonyiurea, imidazolinone, triazoloprimidines, myimIclinyoxy(thlo)benzoates, and/or sulfonylamlnocarbonyltriazolinone herbicide, Additional ALS Inhibitors are known and 15 are disclosed elsewhere herein. It Is known in the art that ALS
mutations fall into different classes with regard to tolerance to sulfonylureas, imidazolinones, triazolopyrimidinea, and pyrimiglinyl(thio)benzoates. Including mutations having the following characteristics: (1) broad tolerance to all four of these groups;
(2) tolerance to imidazolinonee and pyrimidinyl(thio)benzoates; (3) tolerance to sulfonylureas and 20 triazolopyrimiclines; and (4) tolerance to sulfonylureas and imidazolinones.
Various ALS inhibitor-tolerant polypeptides can be employed. In some embodiments, the ALS Inhibitor-tolerant polynucleotides contain at least one nucleotide mutation resulting in one amino acid change In the ALS polypeptide.
In specific embodiments, the change maxis in one of seven substantially conserved 25 regions of acetotactate synthase. See, for example, Hatted et al.
(1995) Molecular = Genetics and Genomes 248;419-425; Lee et (1996) 81480 Journal 7:1241-1248;
Mazur of at. (1989) Ann. Rev. Plant Phys. 40:441-470; and U.S. Patent No.
5,505,011.
The ALS inhibitor-tolerant polypeptide can be encoded by, for example, the SuRA or SuRB locus of ALS. In 30 specific embodiments, the ALS inhibitor-tolerant polypeptlde comprises the 03 ALS
mutant, the HRA ALS mutant, the $4 mutant or the S4IHRA mutant or any combination thereof. Different mutations in ALS are known to confer tolerance to different herbicides and groups (and/or subgroups) of herbicides; see, e.g., Tranal and Wright (2002) Weed Science 50:700-712. See also, U.S. Patent No. 5,805,011, 35 5,378,824, 5,141,870, and 5,013,659 See also, SEQ ID NO:85 comprising a soybean HRA
sequence; SEQ ID N0:66 comprising a maize HRA sequence; SEC? ID NO:67 =
= 10=
=
Date Recue/Date Received 2022-05-20 comprising an ArabMoods HRA sequence; and SEQ ID N0:86 comprising an HRA
sequence used in coiton. The HRA mutation In ALS finds particular Use In one embodiment of the invention. The mutation results in the production bf an acetolactate synthase polypeptkie which Is resistant to at least one ALS inhibitor chemistry in comparison to the wad-type protein. For example, a plant expressing an ALS
Inhibitor-tolerant polypeptide may be tolerant of a dose of sulfonyfurea, Imidazolinone, triazolopyrimidines, prylmidnyloxy(thlo)benzoates, and/or sulfonylamlnocarbonyitriazolinone herbicide that is at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40, 50, 70, 80, 100, 125, 150, 200, 500, or 1000 times higher than a dose of the herbicide that would cause damage to an appropriate control plant.
In some embodiments, an ALS inhibitor-tolerant polypeptide comprises a number of mutations. Additionally, plants having an ALS inhibitorpolypeptkie can be generated through the selection of naturally occurring mutations that Impart tolerance to glyphosate.
In some embodiments, the ALS inhibitor-tolerant polypeptlde confers tolerance to sulfonylursa and Imidazolinone herbicides. Sulfonylurea and imidazolinone herbicides Inhibit growth of higher plants by blocking acetoiactate synthase (ALS), also known as, acetohydroxy acid synthase (AHAS). For example, plants containing particular mutations in ALS (e.g., the $4 and/or HRA mutations) are tolerant to sulfonyiurea herbicides. The production of suffonyiurea-tolerant plants and =
imidazollnone-tolerant plants is described more fully In U.S. Patent Nos.
5,605,011;, 5,013,659; 5,141,870; 5,767,381; 5,731,180; 5,304,732:4,781,373; 5,331,107;
5,928,937; and 5,378,824; and International publication WO 96/33270.
specific embodiments, the ALS inhibitor-tolerant polypeptide comprises a sulfonamide-tolerant acetolactate synthase (otherwise known as a sulfonamide-tolerant acetohydroxy acid synthase) or an imidaiolinone-tolerant acatolactate synihase (otherwise known as an imidazolinone-tolerant acetohydroxy acid synthase).
Other examples of herbicide-tolerance traits that could be combined with an event disclosed herein include those conferring tolerance to at least one herbicide in a plant such an, for example, a brassies plant or horseweed. Herbicide-tolerant weeds are known In the art, as are plants that vary In their tolerance to particular herbicides.
See, e.g., Green and Williams, (2004) COrrelation of Corn (Zen mays) inbred Response to Nicosutfuron and Mesotrione, poster presented at the WSSA Annual Meeting in Kansas City, Missouri, February 9-12, 2004; Green, (1998) Weed Technology 12;474-477; Green and Ulrich, (1993) Weed Science 41:508-516. The trait(s) responsible for these tolerances can be combined by breeding or via other =
Date Recue/Date Received 2022-05-20 methods with an event disclosed herein .to provide a plant of the invention as well as methods of use thereof.
An event disclosed herein can also be combined with at least one other trait to produce plants of the present invention that further comprise a variety of desired trait combinations including, but not limited to, trails desirable for animal feed such as high oil content (e.g., US Patent Number 6,232,529); balanced amino acid content (e.g., .
hordothionins (US Patent Numbers 5,990,389; 5,885,801; 5,885,802 and 5,703,409;
US Patent Number 5,850,016); barley high lysine of 8f., (1987) Ear. J.
Blochem. 165:99-108 and WO 98/20122) and high methlonirie proteins (Pedersen, of at., (1988) J. ?Wet Chem. 2131:6279; Kirlhara, at at., (1988) Gene 71:359 and Musumura, of al., (1989) Plant Md. BM. 12:123)); = increased digestibility (e.g., modified storage proteins (US Patent Application Serial Number 10/053,410, filed November 7, 2001) and thioredans (US Application Serial Number 10/005,429, fled December 3, 2001)) .
Desired trait combinations also include LLNC (low linolenic acid content see, e.g., Dyer, of at, (2002) App!. Micmblol. Biotech:lot 59:224-230) and OLCH (high oleic acid content; see, e.g., Fernandez-Moya, et al., (2005) 1, Aga . Food (Them.
53:5328-5330).
An event disclosed herein may also be combined with other desirable traits =
such as, for example, furnonisin detoxification genes (US Patent Number 5,792,931), avIrulence and disease resistance genes (Jones, of a/., (1994) Sofence 266:769;
Martin, et al., (1993) Science 262:1432; Mindrinos, et al., (1994) Cell 78:11389) and traits desirable for processing or process products .such as modified oh (e.g., fatty acid desaturase genes (US Patent Number 5,952,544; WO 94/11515)); modified starches (e.g., ADPG pyrophosPhorylases (AGPase), starch syntheses ($S), starch . branching enzymes (SSE), and starch debranching enzymes (SDEIE)) and polymers or bioplastics (e.g.. US Patent Number 5,602,321; beta-ketothlolase, potyhydroxybutyrate synthase, and acetoacetyl-CoA reductade (Schubert, of at, (1985) J. Baobab!.
170:5837-5847) facilitate expression of polyhydroxyalkanoates (PHAs)).
One could also combine herbicide-tolerant polynuctootidos with polynucleotides providing agronomic trails such as male sterility (e.g., see; US Patent Number 5:583,210), stalk strength, flowering time or transformation technology traits such as cell cycle regulation or gene targeting (e.g., WO 99/81619, WO 00/17364 and WO 99/25821) .
In another embodiment, an event disclosed herein can also be combined with the Rcg1 sequence or biologically active variant or fragment thereof. The Reg1 =
Date Recue/Date Received 2022-05-20 sequence Is an anthracnose stalk rot resistance gene in corn. See, -for example, US
Patent Application Serial Number 11/397,153, 11/397,275 and 111397,247.
These stacked combinations can be created by any method Including, but not 5, limited to, breeding plants by any conventional methodology or genetic transformation. =
If the sequences are stacked by genetically transforming the plants, the polynucleotide sequences of interest'can be combined at any time and in any order. The traits can be Introduced simultaneously in a co-transformation protocol with the polynucleotides of interest provided by any combination of transformation cassettes. For example, if two sequences will be Introduced, the two sequences can be contained in separate transformation cassettes (trans) or contained on the same transformation cassette (cis). Expression of the sequences can be driven by the same promoter or by different promoters. In certain cases, it may be desirable to introduce a transformation cassette that will suppress the expression of the polynucleotide of interest. This may be combined with any combination of other suppression cassettes or overexpression cassettes to generate the desired combination of traits in the plant It is further recognized that polynucleotide sequences can be stacked at a desired genornic location using a site-specific recombination system. See, for example, W099/25821, W099/25854, W099/25840, W099/25855 and W099/25853 As used herein, the use of the term *polynudeotide* is not Intended to limit the present invention to polynucleotides comprising DNA. Those of ordinary skill in the art Will recognize that polynucleotides, can comprise ribonucleotides and combinations of ribonucleotides and deoxyribonu=cleotides. Such deoxyribonucleotides and-ribonucleotides Include both naturally occurring molecules and synthetic analogues.
The polynucleotides of the invention also encompass all forms of sequences including.
but not limited to, single-stranded forms, double-stranded forms, hairpins, stem-and-loop structures, and the like.
= A DP-073498-4 Brassica plant comprises an expression cassette having an optimized glyphcsate acetylbensferase polynucleotide. The cassette can include 5' and 3' regulatory sequences operably finked to the Wat polynucleotides.
*Operably linked* Is intended to mean a functional linkage between two or more elements.
For example, an operable linkage between a polynucleotide of Interest and a regulatory sequence (i.e., a promoter) is functional grik that allows for the expression of the polynucleotide of Interest. Operably linked elements may be contiguous or non-contiguous. When used to refer to the Joining of two protein coding regions, by operably linked it is Intended that the coding regions are in the -same reading frame.
= 13 Date Recue/Date Received 2022-05-20 The cassette may additionally.contain at least one additional gene to be cotransformed into the organism. Alternatively, the additional gene(s) can be provided on multiple expression cassettes. Such an expression cassette. is provided with a plurality of restriction sites and/or recombination sites for insertion of the polynucleotide to be under the transcriptional regulation of the regulatory regions. The expression cassette .may additionally contain selectable marker genes.
The expression cassette can include in the 5'-3' direction of transcription, a transcriptional and translational initiation region (i.e., a promoter), a coding region and a transcriptional and translational termination region functional In plants.
"Promoter"
refers to a nucleotide sequence capable of controlling the expression Of a coding sequence or functional RNA. In general, a coding sequence is located 3' to a promoter sequence. The promoter sequence can comprise proximal and more distal upstream elements, the latter elements are often referred to as enhancers.
Accordingly, an 'enhancer" is a nucleotide sequence that can stimulate promoter activity and may be an innate element of the promoter or a heterologous element = inserted to enhance the level or tissue-specificity of a promoter.
Promoters may be derived In their entirety from a native gene, or be composed of different elements = derived from different promoters found in nature or even comprise synthetic nucleotide segments. It is understood by those skilled in the art that different promoters may direct the expression of a gene in different tissues or cell types or at different stages of development or in response to different environmental conditions. Promoters that cause a nucleic acid fragment to be expressed in most cell types at most times are commonly referred to as 'constitutive promoters". New promoters of various types useful in plant cells are constantly being discovered; numerous examples may be found in the compilation by Okamunl and Goldberg, (1989) Biochemistry of Plants 15:1-82. It is further recognized that since In most cases the exact boundaries of regulatory sequences have not been completely defined, nucleic acid fragments of different lengths may have identical promoter activity.
The expression cassettes may also contain 5' leader sequences. Such leader sequences can act to enhance translation. The regulatory regions (i.e., promoters, transcriptional regulatory regions, RNA processing or stability regions, introns, polyadenylation signals, transcriptional termination regions and translational termination regions) and/or the coding region may be native/analogous or heterologous to the host cell or to each other.
The "translation leader sequence" refers to a nucleotide sequence located between the promoter sequence of a gene, and the coding sequence. The translation leader sequence is present In the fully processed mRNA upstream of the translation Date Recue/Date Received 2022-05-20 start sequence. The translation leader sdquence may affect numerous parameters Including, processing of the primary transcript to mRNA, mRNA stability and/or translation efficiency. Examples of translation leader sequences have been described (Turner and Foster, (1995) Mc!. Blotechnot 3:225-236). The non-coding sequences" refer to nucleotide sequences located downstream of a coding sequence and include polyadenylation recognition sequences and other sequences encoding regulatory signals capable of affecting mRNA processing or gene expression.
The polyadenylation signal is usually characterized by affecting the addition of polyadenylic acid tracts to the 3` end of the mRNA precursor. The use of different 3' non-coding sequences Is exemplified by Ingelbrecht, etal., (1989) Plant Cell 1:671-680.
As used herein, "heterologous* in reference to a sequence is a sequence that originates from a foreign species, or, if from the same species, Is substantially modified from its native form in composition and/or genomic locus by deliberate human Intervention. For example, a promoter operably linked to a heterologous polynucleotide is from a species different from the species from which the polynucleotide was derived, or, if from the same/analogous species, one or both are substantially modified from their original form and/or genomic locus or the promoter is not the native promoter for the operably linked polynucleotide.
In preparing the expression cassette, the various DNA fragments may be manipulated, so as to provide for the DNA sequences in the proper orientation and, as appropriate, in the proper reading frame, Toward this end, adapters or linkers may be employed. to Join the DNA fragments or other manipulations may be involved to provide for convenient restriction sites, removal of superfluous DNA, removal of restriction sites, or the like. For this purpose, in vitro mutagenesis, primer repair, restriction, annealing, resubstitutions, e.g., transitions and transversions, may be involved. The expression cassette can also comprise a selectable marker gene for the selection of transformed cells. Selectable marker genes are utilized for the selection of transformed cells or tissues.
Isolated polynucleotides are provided that can be used in various methods for the detection and/or identification of the brassica DP-073496-4 event. An 'isolated" or "purified" polynucleotide or biologically active portion thereof, is substantially or essentially free from components that normally accompany or interact with the polynucleoticie as found in its naturally occurring environment. Thus, an isolated or purified polynudeotide is substantially free of other cellular material or culture medium when produced by recombinant techniques or substantially free of chemical precursors or other chemicals when chemically synthesized. Optimally, an 'isolated"
polynucleotide Is free of sequences (optimally protein encoding sequences) that .15 Date Recue/Date Received 2022-05-20 =
naturally flank the polynucleotide (I.e., sequences located at the 5' and 3' ends of the polynucleotide) in the genomic DNA of the organism from which the polynucleotide is derived. For example, in yarious embodiments, the isolated polynucleotide can contain less than about 5 kb, 4 kb, 3 kb, 2 kb, 1 kb, 0.5 kb or 0.1 kb of nucleotide sequence that naturally flank the polynucleotide in genomio DNA of the cell from which the polynucleotide is derived.
In specific embodiments, the polynucleotides of the Invention comprise the junction DNA sequence set forth in NO: 2, or variants and/or fragments thereof or the junction DNA sequence set forth in SEQ ID NO:12 and/or 13. In other embodiments, the polynucleotides of the invention comprise the junction DNA sequences set forth in SEQ ID NO: 14, 15, 16, 17, 18 and/or 19 or variants and fragments thereof. In specific embodiments, methods of detection described herein amplify a polynucleotide comprising a junction of the specific DP-073496-4 event. Fragments and variants of Junction DNA sequences are suitable for discriminatively identifying either event DP-073496-4. As discussed elsewhere herein, such sequences find use as primers and/or probes.
= In other embodiments, the polynucleotides of the invention comprise polynucleotides that can detect a DP-073496-4 event or a region specific to DP-073496-4. Such sequences include any polynucleotide set forth in SEQ ID NO: 2, 4, 5, 6,7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27 or variants and fragments thereof. Fragments and variants of polynucleotides that detect a DP-073496-4 event or a region specific to DP-073496-4 are suitable for discriminatively identifying event DP-073496-4, As discussed elsewhere herein, such sequences find use as primers and/or probes.
"Variants" is intended to mean substantially similar sequences. For polynucleotides, a variant comprises a polynucleotide having deletions (i.e., truncations) at the 5' and/or 3' end; deletion and/or addition of one or more nucleotides at one or more internal sites in the native polynucleotide and/or substitution of one or more nucleotides at one or more sites in the native polynucleotide.
As used heroin, a "probe" Is an Isolated polynucleotide to which is attached a = conventional detectable label or reporter molecule, e.g., a radioactive isotope, ligand, chemiluminescent agent, enzyme, etc. Such a probe is complementary to a strand of a target polynucleotide. In the case of the present invention, the probe Is complementary to a strand of isolated DNA from Brassies event DP-073496-4, whether from a Brassies plant or from a sample that includes DNA from the event.
Probes according to the present Invention Include not only deoxyribonucleic or Date Recue/Date Received 2022-05-20 = ribonucleic acids but also poiyamidee and other probe materials that can specifically detect the presence of the target DNA Sequence.
= As used herein, *primers' are isolated polynucleotides that are annealed to a complementary target DNA strand by nucleic add hybridization to form a hybrid between the primer and the target DNA strand, then extended along the target DNA
strand by a polymerase, e.g., a DNA poiymerase. Primer pairs of the invention refer to their use for amplification of a target polynucleotIde, e.g., by the polymerase chain reaction (PCR) or other conventional nucleic-acid amplification methods. 'VCR*
or 'polymerase chain reaction' is a technique used for the amplification of specific DNA
segments (see, US Patent Numbers 4.883,195 and 4,800,159).
Any combination of primers can be used such that the pair allows tor the detection of a DP-073498-4 event or a region specific to DP-073496-4.
Probes and primers are of sufficient nucleotide length to bind lo the target DNA
sequence and specifically detect ancifor identify a polynucleotide having a DP-4 event. It is recognized that the hybridization conditions or reaction conditions can be determined by the operator to 'achieve this result. This length may be of any length that is of sufficient length to be usefill in a detection method of choice.
Generally, 8, 11, 14, 18, 18, 20, 22, 24, 28; 28, 30, 40, 50, 75, 100, 200, 300, 400, 500, 600, 700 nucleotides or more or between about 11-20, 20-30, 30-40, 40-50, 50-100, 100-200, 200-300, 300-400, 400-500, 500-800, 600-700, 700-800 or more nucleotides In length are used. Such probes and primers can hybridize specifically to a target sequence under high stringency hybridization conditions. . Probes and primers according to embodiments of the present invention may have complete DNA sequence identity of contiguous nucleotides with the target sequence, although probes differing from the target DNA sequence and that retain the ability to specifically detect andfor Identify a target DNA sequence may be designed by conventional methods. Accordingly, probes and primers can share about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or greater sequence Identity or complementarily to the' target polynucleotide, or can differ from the target sequence by 1, 2, 3, 4, 5, 6 or more = 30 nucleotides. Probes Can be used as primers, but are generally designed to bind to the target DNA or RNA and are not used .in an amplification process. In one non-limiting embodiment, a probe can comprises a polynucleotide encoding the gl1et4621 sequence or any variant or fragment thereof, Specific primers can be used to amplify an integration fragment to produce an emplicon that can be used as a 'specific probe' or can herr be 4aetected for identifying event DP-073498-4 in biological samples. Alternatively, a probe of the invention can be used during the PCR reaction to allow for the detection of the amplification event Date Recue/Date Received 2022-05-20 (Le., a TaqmanTo probe or an MOB probe, so called real-time PCP). When the probe Is hybridized with the polynucleofides of a biological sample under conditions which allow for the binding of the probe to the sample, this binding can be detected and thus allow for an indication of the presence of event DP-073496-4 in the biological sample.
Such Identification of a bound probe has been described In the art. In an embodiment of the invention, the specific probe is a sequence which, under optimized conditions, hybridizes specifically to a region within the 51 or 3' flanking region of the event and also comprises a part of the foreign DNA contiguous therewith. The specific probe may comprise a sequence of at least 80%, between 80 and 85%, between 85 and 90%, between 90 and 95% and between 95 and 100% identical (or complementary) to a specific region of the DP-0734964 event.
As used herein, "amplified DNA" or "amplicon* refers to the product of polynucleotide amplification of a target polynucleotide that is part of a nucleic acid template. For example, to determine whether a Brass/ca plant resulting from a sexual cross contains the DP-073496-4 event, DNA extracted from the Brass/ca plant tissue sample may be subjected to a polynucleotide amplification method using a DNA
primer pair that includes a first primer derived from flanking sequence adjacent to the Insertion site of Inserted heterologous DNA and a second primer derived from the inserted heterologous DNA to produce an amplicon that is diagnostic for the presence of the DP-073496-4 event DNA. In specific embodiments, the amplicon comprises a DP-073495-4 junction polynucleotide (i.e., a portion of SEQ ID NO: 2 which spans the junction site, such as, for example, SEQ ID NO: 10, 11, 12, 13, 14, 15, 16, 17, 18 and/or 19 or variants and fragments thereof). By "diagnostic* for a DP-073496-event, the use of any method or assay which discriminates between the presence or the absence of a DP-073496-4 event in a biological sample is intended.
Alternatively, the second primer may be derived from the flanking sequence. In still other embodiments, primer pairs can be derived from flanking sequence on both sides of the inserted DNA so as to produce an amplicon that includes the entire insert polynucleotide of the expression construct as well as the sequence flanking the transgenic insert. See, Figure 3. The amplicon Is of a length and has a sequence that is also diagnostic for the event (Leõ has a junction DNA from a DP-073496-4 event).
The amplicon may range in length from the combined length of the primer pairs plus one nucleotide base pair to any length of amplicon producible by a DNA
amplification protocol. A member of a primer pair derived from the flanking sequence may be located a distance from the inserted DNA sequence, this distance can range from one nucleotide base pair up to the limits of the amplification reaction or about twenty Date Recue/Date Received 2022-05-20 =
=
= thousand nucleotide base pairs. The use of the term 'amplicorr specifically excludes primer dImers that may be formed in the DNA thermal amplification reaction.
Methods for preparing and using probes and primers are described, for example, in Molecular Cloning: A Laboratory Manual, 2nd ed, vol. 1-3, ed.
, 5 Sambrook, etal., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y. 1989 (hereinafter, 'Sambrook, at at, 19891; Current Protocols In Molecular Biology, ed.
ifusubei, at aL, Greene Publishing and Wiley-Intelscience, New York, 1992 (with periodic updates) (hereinafter, "Ausubel, at al., 1992") and Innis, at aL, PCR
Protocols:
A Guide to Methods and Applications, Academic Press: San Diego, 1990. PCR
primer pairs can be derived from a known sequence, for example, by using computer programs intended for that purpose such as the PCR primer analysis tool in Vector Nil version 0 (Informax Inc., Bethesda Md.); PrimerSelect (DNASTAR Inc., Madison, VVia.); and Primer (Version 0.5©, 1991, Whitehead Institute for Biomedical Research, Cambridge, Mass.). Additionally, the sequence can 'be visualy scanned IS and primers manually identified using guidelines known to one of skill in the art It Is to be understood that as used herein the term "transgenie inciudes. any cell, cell line, callus, tissue; plant part or plant, the genotype of Which has been altered =
by the presence of a heterologous nucleic acid including those transgenics initially so altered as well as those created by sexual crosses or asexual propagation from the initial transgenlc. The tenn Iranagenie as used herein does not encompass the =
alteration of the genorne (chromosomal or extra-chromosomal) by conventional plant.
breeding methods or by naturally occurring events such as random cross-fertilization, non-recombinant viral infection, = non-recombinant bacterial transformation, non-recombinant transposition or spontaneous mutation.
"Transformation' refers to the transfer of a nucleic acid fragment into the genome of a host organism, resulting in genetically stable inheritance. Host organisms = containing the transformed nucleic acid fragments are referred to as Iransgenle organisms. Examples of methods of plant transformation. include Agrobacterium-mediated transformation (De Moore, of af., (1987) Meth. Enzymol. 143:277) and .
particle-accelerated or 'gene gun' transformation technology (Klein, et el, (1907) Nature (London) 327:70-73; US Patent Number 4,845,050).
Additional transformation methods are disclosed below.
Thus, isolated polynucleotides of the Invention can be incorporated into recombinant constructs, typically DNA constructs, which are capable of introduction into and replication In a host cell: Such a construct can be a vector that includes a replication system and sequences that are capable of transcription and translation of a poiypeptide-encoding sequence in a given host cell. A number of vectors suitable for i9=
=
=
Date Recue/Date Received 2022-05-20 stable transfection of plant cells or for the establishment of transgenic plants have been described in, e.g., Pouwels, of al., (1985; Supp. 1987) Cloning Vectors:
A
Laboratory Manual, Weissbach and Weissbach, (1989) Methods for Plant Molecular Biology (Academic Press, New York) and Flevin, 81 84'., (1990) Plant Molecular Biology Manual (Kluwer Academic Publishers). Typically, plant expression vectors Include, for example, one or more cloned plant genes under the transcriptional control of 6' and 3' regulatory sequences and a dominant selectable marker. Such plant expression vectors also can contain a promoter regulatory region (e.g., a regulatory region controlling inducible or constitutive, environmentally- or developmentally-regulated or cell- or tissue-specific expression), a transcription initiation start site, a ribosome binding site, an RNA processing signal, a transcription termination site and/or a polyadenylation signal.
Various methods and compositions for Identifying event DP-073496-4 are provided. Such methods find use in identifying and/or detecting a DP-073496-4 event In any biological material. Such methods include, for example, methods to confirm seed purity and methods for screening seeds in a seed lot for a DP-073496-4 event.
In one embodiment, a method for identifying event DP-073496-4 in a biological sample is provided and comprises contacting the sample with a first and a second primer; and, amplifying a polynucleotide comprising a DP-073496-4 specific region.
A biological sample can comprise any sample in which one desires to determine if DNA having event DP-073496-4 is present. For example, a biological sample can comprise any plant material or material comprising or derived from a plant material such as, but not limited to, food or feed products. As used herein, "plant material" refers to material which is obtained or derived from a plant or plant part. In specific embodiments, the biological sample comprises a brassica tissue.
Primers and probes based on the flanking DNA and insert sequences disclosed herein can be used to confirm (and, if necessary, to correct) the disclosed sequences by conventional methods, e.g., by re-cloning and sequencing such sequences.
The polynucteotide probes and primers of the present Invention specifically detect a target DNA sequence. Any conventional nucleic acid hybridization or amplification method can be used to identify the presence of DNA from a transgenic event in a sample. By "specifically detect" it is intended that the polynucieotide can be used either as a primer to amplify a DP-073496-4 specific region or the polynuoleotide can be used as a probe that hybridizes under stringent conditions to a pOlynucleotide from a DP-073496-4 event. The level or degree of hybridization which allows for the specific detection of a DP-073496-4 event or a specific region of a DP-073496-4 event is sufficient to distinguish the polynudeotIde with the DP-073496-4 specific region from a Date Recue/Date Received 2022-05-20 polynucleotide lacking this region and thereby allow for discriminately identifying a DP-073496-4 event By "shares sufficient sequence identity or complentarity to allow for the amplification of a DP-073496-4 specific event" is intended the sequence shares at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100%
Identity or compiementarity to a fragment or across the full length of the polynucleotide from the DP-073496-4 specific region.
Regarding the amplification of a target polynucleotide (e.g., by PCR) using a particular amplification primer pair, "stringent conditions" are conditions that permit the primer pair to hybridize to the target polynucleotide to which one primer having the corresponding wild-type sequence (or its complement) and another primer having the corresponding DP-073496-4 inserted DNA sequence would bind and preferably to produce an identifiable amplification product (the amplicon) having a DP-specific region In a DNA thermal amplification reaction. In a PCR approach, oligonucleotide primers can be designed for use in PCR reactions to amplify a DP-073496-4 specific region. Methods for designing PCR primers and PCR cloning are generally known in the art and are disclosed In Sambrook, of al., (1989) Molecular Cloning: A Laboratory Manual (2d ed., Cold Spring Harbor Laboratory Press, Plainview, New York). See also, Innis, et al., eds. (1990) PCR Protocols: A
Guide to Methods and Applications (Academic Press, New York); Innis and Gelfand, eds.
(1995) PCR Strategies (Academic Press, New York) and Innis and Gelfand, eds.
(1999) PCR Methods Manual (Academic Press, New York). Methods of amplification are further described In US Patent Number 4,683,195, 4,683,202 and Chen, of al., (1994) PNAS 91:5695-5699. These methods as well as other methods known in the art of DNA amplification may be used in the practice of the embodiments of the present invention. It is understood that a number of parameters in a specific PCR
protocol may need to be adjusted to specific laboratory conditions and may be slightly modified and yet allow for the collection of similar results. These adjustments will be apparent to a person skilled in the art.
The amplified polynucleotide (amplicon) can be of any length that allows for the detection of the DP-073496-4 event or a DP-073496-4 specific region. For example, the amplicon can be about 10, 50, 100, 200, 300, 500, 700, 100, 2000, 3000, 4000, 5000 nucleotides in length or longer.
In specific embodiments, the specific region of the DP-073496-4 event is detected.
Any primer can be employed in the methods of the invention that allows a DP-0734964 specific region to be amplified and/or detected. For example, in specific embodiments, the first primer comprises a fragment of a polynucleotide of SEQ
ID NO:
Date Recue/Date Received 2022-05-20 2 or 3, wherein the first or the second primer shares sufficient sequence Identity or complementarity to the polynucleotlde to amplify the DP-073496-4 specific region.
-The primer pair cart comprise a fragment of SEQ ID NO: 2 or 3. in another embodiment, the primer pair comprises a fret printer comprising a fragment of SEQ ID
NO: 8 and a second primer comprising a fragment of SEQ ID NO: 9 or 10; or, alternatively, the primer pair comprises a first primer comprising a fragment of SEC ID
NO: 9 and the second primer comprises a fragment of SEQ ID NO; 8 or 10. The =
primers can be of any length sufficient to amplify a DP-073496-4 specific region including, for example, at least 8, 7, 8, 9, 10, 15, 20, 15 or 30 or about 7-
=
phosphinothricin acetyltransferase can exhibit improved tolerance to glufosinate = herbicides, which inhibit the enzyme glutamine synthase. Other examples of herbicide-tolerance traits that could be combined with an event disclosed herein Include those conferred by polynucleotides conferring altered protoporphydnogen = 5 coddase (protox) activfty;as described in US Patent Numbers 8,288,306 81; 6,262,837 131 and 5,707,373 and International Publication Number WO 01/12825. Plants = containing such polynucleotides can exhibit Improved tolerance to any of a variety of herbicides which target the protox enzyme (also referred to as "protox Inhibitors".
In other embodiments, an ALS inhibRokolmant trait is combined with the event =
= 10 disclosed herein. As used herein, an "ALS Inhibitor-tolerant polypeptide' comprises any polypeptide which when expressed in a plant confers tolerance to at least one ALS
Inhibitor. A variety of ALS Inhibitors are known and include, for example, sulfonyiurea, imidazolinone, triazoloprimidines, myimIclinyoxy(thlo)benzoates, and/or sulfonylamlnocarbonyltriazolinone herbicide, Additional ALS Inhibitors are known and 15 are disclosed elsewhere herein. It Is known in the art that ALS
mutations fall into different classes with regard to tolerance to sulfonylureas, imidazolinones, triazolopyrimidinea, and pyrimiglinyl(thio)benzoates. Including mutations having the following characteristics: (1) broad tolerance to all four of these groups;
(2) tolerance to imidazolinonee and pyrimidinyl(thio)benzoates; (3) tolerance to sulfonylureas and 20 triazolopyrimiclines; and (4) tolerance to sulfonylureas and imidazolinones.
Various ALS inhibitor-tolerant polypeptides can be employed. In some embodiments, the ALS Inhibitor-tolerant polynucleotides contain at least one nucleotide mutation resulting in one amino acid change In the ALS polypeptide.
In specific embodiments, the change maxis in one of seven substantially conserved 25 regions of acetotactate synthase. See, for example, Hatted et al.
(1995) Molecular = Genetics and Genomes 248;419-425; Lee et (1996) 81480 Journal 7:1241-1248;
Mazur of at. (1989) Ann. Rev. Plant Phys. 40:441-470; and U.S. Patent No.
5,505,011.
The ALS inhibitor-tolerant polypeptide can be encoded by, for example, the SuRA or SuRB locus of ALS. In 30 specific embodiments, the ALS inhibitor-tolerant polypeptlde comprises the 03 ALS
mutant, the HRA ALS mutant, the $4 mutant or the S4IHRA mutant or any combination thereof. Different mutations in ALS are known to confer tolerance to different herbicides and groups (and/or subgroups) of herbicides; see, e.g., Tranal and Wright (2002) Weed Science 50:700-712. See also, U.S. Patent No. 5,805,011, 35 5,378,824, 5,141,870, and 5,013,659 See also, SEQ ID NO:85 comprising a soybean HRA
sequence; SEQ ID N0:66 comprising a maize HRA sequence; SEC? ID NO:67 =
= 10=
=
Date Recue/Date Received 2022-05-20 comprising an ArabMoods HRA sequence; and SEQ ID N0:86 comprising an HRA
sequence used in coiton. The HRA mutation In ALS finds particular Use In one embodiment of the invention. The mutation results in the production bf an acetolactate synthase polypeptkie which Is resistant to at least one ALS inhibitor chemistry in comparison to the wad-type protein. For example, a plant expressing an ALS
Inhibitor-tolerant polypeptide may be tolerant of a dose of sulfonyfurea, Imidazolinone, triazolopyrimidines, prylmidnyloxy(thlo)benzoates, and/or sulfonylamlnocarbonyitriazolinone herbicide that is at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40, 50, 70, 80, 100, 125, 150, 200, 500, or 1000 times higher than a dose of the herbicide that would cause damage to an appropriate control plant.
In some embodiments, an ALS inhibitor-tolerant polypeptide comprises a number of mutations. Additionally, plants having an ALS inhibitorpolypeptkie can be generated through the selection of naturally occurring mutations that Impart tolerance to glyphosate.
In some embodiments, the ALS inhibitor-tolerant polypeptlde confers tolerance to sulfonylursa and Imidazolinone herbicides. Sulfonylurea and imidazolinone herbicides Inhibit growth of higher plants by blocking acetoiactate synthase (ALS), also known as, acetohydroxy acid synthase (AHAS). For example, plants containing particular mutations in ALS (e.g., the $4 and/or HRA mutations) are tolerant to sulfonyiurea herbicides. The production of suffonyiurea-tolerant plants and =
imidazollnone-tolerant plants is described more fully In U.S. Patent Nos.
5,605,011;, 5,013,659; 5,141,870; 5,767,381; 5,731,180; 5,304,732:4,781,373; 5,331,107;
5,928,937; and 5,378,824; and International publication WO 96/33270.
specific embodiments, the ALS inhibitor-tolerant polypeptide comprises a sulfonamide-tolerant acetolactate synthase (otherwise known as a sulfonamide-tolerant acetohydroxy acid synthase) or an imidaiolinone-tolerant acatolactate synihase (otherwise known as an imidazolinone-tolerant acetohydroxy acid synthase).
Other examples of herbicide-tolerance traits that could be combined with an event disclosed herein include those conferring tolerance to at least one herbicide in a plant such an, for example, a brassies plant or horseweed. Herbicide-tolerant weeds are known In the art, as are plants that vary In their tolerance to particular herbicides.
See, e.g., Green and Williams, (2004) COrrelation of Corn (Zen mays) inbred Response to Nicosutfuron and Mesotrione, poster presented at the WSSA Annual Meeting in Kansas City, Missouri, February 9-12, 2004; Green, (1998) Weed Technology 12;474-477; Green and Ulrich, (1993) Weed Science 41:508-516. The trait(s) responsible for these tolerances can be combined by breeding or via other =
Date Recue/Date Received 2022-05-20 methods with an event disclosed herein .to provide a plant of the invention as well as methods of use thereof.
An event disclosed herein can also be combined with at least one other trait to produce plants of the present invention that further comprise a variety of desired trait combinations including, but not limited to, trails desirable for animal feed such as high oil content (e.g., US Patent Number 6,232,529); balanced amino acid content (e.g., .
hordothionins (US Patent Numbers 5,990,389; 5,885,801; 5,885,802 and 5,703,409;
US Patent Number 5,850,016); barley high lysine of 8f., (1987) Ear. J.
Blochem. 165:99-108 and WO 98/20122) and high methlonirie proteins (Pedersen, of at., (1988) J. ?Wet Chem. 2131:6279; Kirlhara, at at., (1988) Gene 71:359 and Musumura, of al., (1989) Plant Md. BM. 12:123)); = increased digestibility (e.g., modified storage proteins (US Patent Application Serial Number 10/053,410, filed November 7, 2001) and thioredans (US Application Serial Number 10/005,429, fled December 3, 2001)) .
Desired trait combinations also include LLNC (low linolenic acid content see, e.g., Dyer, of at, (2002) App!. Micmblol. Biotech:lot 59:224-230) and OLCH (high oleic acid content; see, e.g., Fernandez-Moya, et al., (2005) 1, Aga . Food (Them.
53:5328-5330).
An event disclosed herein may also be combined with other desirable traits =
such as, for example, furnonisin detoxification genes (US Patent Number 5,792,931), avIrulence and disease resistance genes (Jones, of a/., (1994) Sofence 266:769;
Martin, et al., (1993) Science 262:1432; Mindrinos, et al., (1994) Cell 78:11389) and traits desirable for processing or process products .such as modified oh (e.g., fatty acid desaturase genes (US Patent Number 5,952,544; WO 94/11515)); modified starches (e.g., ADPG pyrophosPhorylases (AGPase), starch syntheses ($S), starch . branching enzymes (SSE), and starch debranching enzymes (SDEIE)) and polymers or bioplastics (e.g.. US Patent Number 5,602,321; beta-ketothlolase, potyhydroxybutyrate synthase, and acetoacetyl-CoA reductade (Schubert, of at, (1985) J. Baobab!.
170:5837-5847) facilitate expression of polyhydroxyalkanoates (PHAs)).
One could also combine herbicide-tolerant polynuctootidos with polynucleotides providing agronomic trails such as male sterility (e.g., see; US Patent Number 5:583,210), stalk strength, flowering time or transformation technology traits such as cell cycle regulation or gene targeting (e.g., WO 99/81619, WO 00/17364 and WO 99/25821) .
In another embodiment, an event disclosed herein can also be combined with the Rcg1 sequence or biologically active variant or fragment thereof. The Reg1 =
Date Recue/Date Received 2022-05-20 sequence Is an anthracnose stalk rot resistance gene in corn. See, -for example, US
Patent Application Serial Number 11/397,153, 11/397,275 and 111397,247.
These stacked combinations can be created by any method Including, but not 5, limited to, breeding plants by any conventional methodology or genetic transformation. =
If the sequences are stacked by genetically transforming the plants, the polynucleotide sequences of interest'can be combined at any time and in any order. The traits can be Introduced simultaneously in a co-transformation protocol with the polynucleotides of interest provided by any combination of transformation cassettes. For example, if two sequences will be Introduced, the two sequences can be contained in separate transformation cassettes (trans) or contained on the same transformation cassette (cis). Expression of the sequences can be driven by the same promoter or by different promoters. In certain cases, it may be desirable to introduce a transformation cassette that will suppress the expression of the polynucleotide of interest. This may be combined with any combination of other suppression cassettes or overexpression cassettes to generate the desired combination of traits in the plant It is further recognized that polynucleotide sequences can be stacked at a desired genornic location using a site-specific recombination system. See, for example, W099/25821, W099/25854, W099/25840, W099/25855 and W099/25853 As used herein, the use of the term *polynudeotide* is not Intended to limit the present invention to polynucleotides comprising DNA. Those of ordinary skill in the art Will recognize that polynucleotides, can comprise ribonucleotides and combinations of ribonucleotides and deoxyribonu=cleotides. Such deoxyribonucleotides and-ribonucleotides Include both naturally occurring molecules and synthetic analogues.
The polynucleotides of the invention also encompass all forms of sequences including.
but not limited to, single-stranded forms, double-stranded forms, hairpins, stem-and-loop structures, and the like.
= A DP-073498-4 Brassica plant comprises an expression cassette having an optimized glyphcsate acetylbensferase polynucleotide. The cassette can include 5' and 3' regulatory sequences operably finked to the Wat polynucleotides.
*Operably linked* Is intended to mean a functional linkage between two or more elements.
For example, an operable linkage between a polynucleotide of Interest and a regulatory sequence (i.e., a promoter) is functional grik that allows for the expression of the polynucleotide of Interest. Operably linked elements may be contiguous or non-contiguous. When used to refer to the Joining of two protein coding regions, by operably linked it is Intended that the coding regions are in the -same reading frame.
= 13 Date Recue/Date Received 2022-05-20 The cassette may additionally.contain at least one additional gene to be cotransformed into the organism. Alternatively, the additional gene(s) can be provided on multiple expression cassettes. Such an expression cassette. is provided with a plurality of restriction sites and/or recombination sites for insertion of the polynucleotide to be under the transcriptional regulation of the regulatory regions. The expression cassette .may additionally contain selectable marker genes.
The expression cassette can include in the 5'-3' direction of transcription, a transcriptional and translational initiation region (i.e., a promoter), a coding region and a transcriptional and translational termination region functional In plants.
"Promoter"
refers to a nucleotide sequence capable of controlling the expression Of a coding sequence or functional RNA. In general, a coding sequence is located 3' to a promoter sequence. The promoter sequence can comprise proximal and more distal upstream elements, the latter elements are often referred to as enhancers.
Accordingly, an 'enhancer" is a nucleotide sequence that can stimulate promoter activity and may be an innate element of the promoter or a heterologous element = inserted to enhance the level or tissue-specificity of a promoter.
Promoters may be derived In their entirety from a native gene, or be composed of different elements = derived from different promoters found in nature or even comprise synthetic nucleotide segments. It is understood by those skilled in the art that different promoters may direct the expression of a gene in different tissues or cell types or at different stages of development or in response to different environmental conditions. Promoters that cause a nucleic acid fragment to be expressed in most cell types at most times are commonly referred to as 'constitutive promoters". New promoters of various types useful in plant cells are constantly being discovered; numerous examples may be found in the compilation by Okamunl and Goldberg, (1989) Biochemistry of Plants 15:1-82. It is further recognized that since In most cases the exact boundaries of regulatory sequences have not been completely defined, nucleic acid fragments of different lengths may have identical promoter activity.
The expression cassettes may also contain 5' leader sequences. Such leader sequences can act to enhance translation. The regulatory regions (i.e., promoters, transcriptional regulatory regions, RNA processing or stability regions, introns, polyadenylation signals, transcriptional termination regions and translational termination regions) and/or the coding region may be native/analogous or heterologous to the host cell or to each other.
The "translation leader sequence" refers to a nucleotide sequence located between the promoter sequence of a gene, and the coding sequence. The translation leader sequence is present In the fully processed mRNA upstream of the translation Date Recue/Date Received 2022-05-20 start sequence. The translation leader sdquence may affect numerous parameters Including, processing of the primary transcript to mRNA, mRNA stability and/or translation efficiency. Examples of translation leader sequences have been described (Turner and Foster, (1995) Mc!. Blotechnot 3:225-236). The non-coding sequences" refer to nucleotide sequences located downstream of a coding sequence and include polyadenylation recognition sequences and other sequences encoding regulatory signals capable of affecting mRNA processing or gene expression.
The polyadenylation signal is usually characterized by affecting the addition of polyadenylic acid tracts to the 3` end of the mRNA precursor. The use of different 3' non-coding sequences Is exemplified by Ingelbrecht, etal., (1989) Plant Cell 1:671-680.
As used herein, "heterologous* in reference to a sequence is a sequence that originates from a foreign species, or, if from the same species, Is substantially modified from its native form in composition and/or genomic locus by deliberate human Intervention. For example, a promoter operably linked to a heterologous polynucleotide is from a species different from the species from which the polynucleotide was derived, or, if from the same/analogous species, one or both are substantially modified from their original form and/or genomic locus or the promoter is not the native promoter for the operably linked polynucleotide.
In preparing the expression cassette, the various DNA fragments may be manipulated, so as to provide for the DNA sequences in the proper orientation and, as appropriate, in the proper reading frame, Toward this end, adapters or linkers may be employed. to Join the DNA fragments or other manipulations may be involved to provide for convenient restriction sites, removal of superfluous DNA, removal of restriction sites, or the like. For this purpose, in vitro mutagenesis, primer repair, restriction, annealing, resubstitutions, e.g., transitions and transversions, may be involved. The expression cassette can also comprise a selectable marker gene for the selection of transformed cells. Selectable marker genes are utilized for the selection of transformed cells or tissues.
Isolated polynucleotides are provided that can be used in various methods for the detection and/or identification of the brassica DP-073496-4 event. An 'isolated" or "purified" polynucleotide or biologically active portion thereof, is substantially or essentially free from components that normally accompany or interact with the polynucleoticie as found in its naturally occurring environment. Thus, an isolated or purified polynudeotide is substantially free of other cellular material or culture medium when produced by recombinant techniques or substantially free of chemical precursors or other chemicals when chemically synthesized. Optimally, an 'isolated"
polynucleotide Is free of sequences (optimally protein encoding sequences) that .15 Date Recue/Date Received 2022-05-20 =
naturally flank the polynucleotide (I.e., sequences located at the 5' and 3' ends of the polynucleotide) in the genomic DNA of the organism from which the polynucleotide is derived. For example, in yarious embodiments, the isolated polynucleotide can contain less than about 5 kb, 4 kb, 3 kb, 2 kb, 1 kb, 0.5 kb or 0.1 kb of nucleotide sequence that naturally flank the polynucleotide in genomio DNA of the cell from which the polynucleotide is derived.
In specific embodiments, the polynucleotides of the Invention comprise the junction DNA sequence set forth in NO: 2, or variants and/or fragments thereof or the junction DNA sequence set forth in SEQ ID NO:12 and/or 13. In other embodiments, the polynucleotides of the invention comprise the junction DNA sequences set forth in SEQ ID NO: 14, 15, 16, 17, 18 and/or 19 or variants and fragments thereof. In specific embodiments, methods of detection described herein amplify a polynucleotide comprising a junction of the specific DP-073496-4 event. Fragments and variants of Junction DNA sequences are suitable for discriminatively identifying either event DP-073496-4. As discussed elsewhere herein, such sequences find use as primers and/or probes.
= In other embodiments, the polynucleotides of the invention comprise polynucleotides that can detect a DP-073496-4 event or a region specific to DP-073496-4. Such sequences include any polynucleotide set forth in SEQ ID NO: 2, 4, 5, 6,7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27 or variants and fragments thereof. Fragments and variants of polynucleotides that detect a DP-073496-4 event or a region specific to DP-073496-4 are suitable for discriminatively identifying event DP-073496-4, As discussed elsewhere herein, such sequences find use as primers and/or probes.
"Variants" is intended to mean substantially similar sequences. For polynucleotides, a variant comprises a polynucleotide having deletions (i.e., truncations) at the 5' and/or 3' end; deletion and/or addition of one or more nucleotides at one or more internal sites in the native polynucleotide and/or substitution of one or more nucleotides at one or more sites in the native polynucleotide.
As used heroin, a "probe" Is an Isolated polynucleotide to which is attached a = conventional detectable label or reporter molecule, e.g., a radioactive isotope, ligand, chemiluminescent agent, enzyme, etc. Such a probe is complementary to a strand of a target polynucleotide. In the case of the present invention, the probe Is complementary to a strand of isolated DNA from Brassies event DP-073496-4, whether from a Brassies plant or from a sample that includes DNA from the event.
Probes according to the present Invention Include not only deoxyribonucleic or Date Recue/Date Received 2022-05-20 = ribonucleic acids but also poiyamidee and other probe materials that can specifically detect the presence of the target DNA Sequence.
= As used herein, *primers' are isolated polynucleotides that are annealed to a complementary target DNA strand by nucleic add hybridization to form a hybrid between the primer and the target DNA strand, then extended along the target DNA
strand by a polymerase, e.g., a DNA poiymerase. Primer pairs of the invention refer to their use for amplification of a target polynucleotIde, e.g., by the polymerase chain reaction (PCR) or other conventional nucleic-acid amplification methods. 'VCR*
or 'polymerase chain reaction' is a technique used for the amplification of specific DNA
segments (see, US Patent Numbers 4.883,195 and 4,800,159).
Any combination of primers can be used such that the pair allows tor the detection of a DP-073498-4 event or a region specific to DP-073496-4.
Probes and primers are of sufficient nucleotide length to bind lo the target DNA
sequence and specifically detect ancifor identify a polynucleotide having a DP-4 event. It is recognized that the hybridization conditions or reaction conditions can be determined by the operator to 'achieve this result. This length may be of any length that is of sufficient length to be usefill in a detection method of choice.
Generally, 8, 11, 14, 18, 18, 20, 22, 24, 28; 28, 30, 40, 50, 75, 100, 200, 300, 400, 500, 600, 700 nucleotides or more or between about 11-20, 20-30, 30-40, 40-50, 50-100, 100-200, 200-300, 300-400, 400-500, 500-800, 600-700, 700-800 or more nucleotides In length are used. Such probes and primers can hybridize specifically to a target sequence under high stringency hybridization conditions. . Probes and primers according to embodiments of the present invention may have complete DNA sequence identity of contiguous nucleotides with the target sequence, although probes differing from the target DNA sequence and that retain the ability to specifically detect andfor Identify a target DNA sequence may be designed by conventional methods. Accordingly, probes and primers can share about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or greater sequence Identity or complementarily to the' target polynucleotide, or can differ from the target sequence by 1, 2, 3, 4, 5, 6 or more = 30 nucleotides. Probes Can be used as primers, but are generally designed to bind to the target DNA or RNA and are not used .in an amplification process. In one non-limiting embodiment, a probe can comprises a polynucleotide encoding the gl1et4621 sequence or any variant or fragment thereof, Specific primers can be used to amplify an integration fragment to produce an emplicon that can be used as a 'specific probe' or can herr be 4aetected for identifying event DP-073498-4 in biological samples. Alternatively, a probe of the invention can be used during the PCR reaction to allow for the detection of the amplification event Date Recue/Date Received 2022-05-20 (Le., a TaqmanTo probe or an MOB probe, so called real-time PCP). When the probe Is hybridized with the polynucleofides of a biological sample under conditions which allow for the binding of the probe to the sample, this binding can be detected and thus allow for an indication of the presence of event DP-073496-4 in the biological sample.
Such Identification of a bound probe has been described In the art. In an embodiment of the invention, the specific probe is a sequence which, under optimized conditions, hybridizes specifically to a region within the 51 or 3' flanking region of the event and also comprises a part of the foreign DNA contiguous therewith. The specific probe may comprise a sequence of at least 80%, between 80 and 85%, between 85 and 90%, between 90 and 95% and between 95 and 100% identical (or complementary) to a specific region of the DP-0734964 event.
As used herein, "amplified DNA" or "amplicon* refers to the product of polynucleotide amplification of a target polynucleotide that is part of a nucleic acid template. For example, to determine whether a Brass/ca plant resulting from a sexual cross contains the DP-073496-4 event, DNA extracted from the Brass/ca plant tissue sample may be subjected to a polynucleotide amplification method using a DNA
primer pair that includes a first primer derived from flanking sequence adjacent to the Insertion site of Inserted heterologous DNA and a second primer derived from the inserted heterologous DNA to produce an amplicon that is diagnostic for the presence of the DP-073496-4 event DNA. In specific embodiments, the amplicon comprises a DP-073495-4 junction polynucleotide (i.e., a portion of SEQ ID NO: 2 which spans the junction site, such as, for example, SEQ ID NO: 10, 11, 12, 13, 14, 15, 16, 17, 18 and/or 19 or variants and fragments thereof). By "diagnostic* for a DP-073496-event, the use of any method or assay which discriminates between the presence or the absence of a DP-073496-4 event in a biological sample is intended.
Alternatively, the second primer may be derived from the flanking sequence. In still other embodiments, primer pairs can be derived from flanking sequence on both sides of the inserted DNA so as to produce an amplicon that includes the entire insert polynucleotide of the expression construct as well as the sequence flanking the transgenic insert. See, Figure 3. The amplicon Is of a length and has a sequence that is also diagnostic for the event (Leõ has a junction DNA from a DP-073496-4 event).
The amplicon may range in length from the combined length of the primer pairs plus one nucleotide base pair to any length of amplicon producible by a DNA
amplification protocol. A member of a primer pair derived from the flanking sequence may be located a distance from the inserted DNA sequence, this distance can range from one nucleotide base pair up to the limits of the amplification reaction or about twenty Date Recue/Date Received 2022-05-20 =
=
= thousand nucleotide base pairs. The use of the term 'amplicorr specifically excludes primer dImers that may be formed in the DNA thermal amplification reaction.
Methods for preparing and using probes and primers are described, for example, in Molecular Cloning: A Laboratory Manual, 2nd ed, vol. 1-3, ed.
, 5 Sambrook, etal., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y. 1989 (hereinafter, 'Sambrook, at at, 19891; Current Protocols In Molecular Biology, ed.
ifusubei, at aL, Greene Publishing and Wiley-Intelscience, New York, 1992 (with periodic updates) (hereinafter, "Ausubel, at al., 1992") and Innis, at aL, PCR
Protocols:
A Guide to Methods and Applications, Academic Press: San Diego, 1990. PCR
primer pairs can be derived from a known sequence, for example, by using computer programs intended for that purpose such as the PCR primer analysis tool in Vector Nil version 0 (Informax Inc., Bethesda Md.); PrimerSelect (DNASTAR Inc., Madison, VVia.); and Primer (Version 0.5©, 1991, Whitehead Institute for Biomedical Research, Cambridge, Mass.). Additionally, the sequence can 'be visualy scanned IS and primers manually identified using guidelines known to one of skill in the art It Is to be understood that as used herein the term "transgenie inciudes. any cell, cell line, callus, tissue; plant part or plant, the genotype of Which has been altered =
by the presence of a heterologous nucleic acid including those transgenics initially so altered as well as those created by sexual crosses or asexual propagation from the initial transgenlc. The tenn Iranagenie as used herein does not encompass the =
alteration of the genorne (chromosomal or extra-chromosomal) by conventional plant.
breeding methods or by naturally occurring events such as random cross-fertilization, non-recombinant viral infection, = non-recombinant bacterial transformation, non-recombinant transposition or spontaneous mutation.
"Transformation' refers to the transfer of a nucleic acid fragment into the genome of a host organism, resulting in genetically stable inheritance. Host organisms = containing the transformed nucleic acid fragments are referred to as Iransgenle organisms. Examples of methods of plant transformation. include Agrobacterium-mediated transformation (De Moore, of af., (1987) Meth. Enzymol. 143:277) and .
particle-accelerated or 'gene gun' transformation technology (Klein, et el, (1907) Nature (London) 327:70-73; US Patent Number 4,845,050).
Additional transformation methods are disclosed below.
Thus, isolated polynucleotides of the Invention can be incorporated into recombinant constructs, typically DNA constructs, which are capable of introduction into and replication In a host cell: Such a construct can be a vector that includes a replication system and sequences that are capable of transcription and translation of a poiypeptide-encoding sequence in a given host cell. A number of vectors suitable for i9=
=
=
Date Recue/Date Received 2022-05-20 stable transfection of plant cells or for the establishment of transgenic plants have been described in, e.g., Pouwels, of al., (1985; Supp. 1987) Cloning Vectors:
A
Laboratory Manual, Weissbach and Weissbach, (1989) Methods for Plant Molecular Biology (Academic Press, New York) and Flevin, 81 84'., (1990) Plant Molecular Biology Manual (Kluwer Academic Publishers). Typically, plant expression vectors Include, for example, one or more cloned plant genes under the transcriptional control of 6' and 3' regulatory sequences and a dominant selectable marker. Such plant expression vectors also can contain a promoter regulatory region (e.g., a regulatory region controlling inducible or constitutive, environmentally- or developmentally-regulated or cell- or tissue-specific expression), a transcription initiation start site, a ribosome binding site, an RNA processing signal, a transcription termination site and/or a polyadenylation signal.
Various methods and compositions for Identifying event DP-073496-4 are provided. Such methods find use in identifying and/or detecting a DP-073496-4 event In any biological material. Such methods include, for example, methods to confirm seed purity and methods for screening seeds in a seed lot for a DP-073496-4 event.
In one embodiment, a method for identifying event DP-073496-4 in a biological sample is provided and comprises contacting the sample with a first and a second primer; and, amplifying a polynucleotide comprising a DP-073496-4 specific region.
A biological sample can comprise any sample in which one desires to determine if DNA having event DP-073496-4 is present. For example, a biological sample can comprise any plant material or material comprising or derived from a plant material such as, but not limited to, food or feed products. As used herein, "plant material" refers to material which is obtained or derived from a plant or plant part. In specific embodiments, the biological sample comprises a brassica tissue.
Primers and probes based on the flanking DNA and insert sequences disclosed herein can be used to confirm (and, if necessary, to correct) the disclosed sequences by conventional methods, e.g., by re-cloning and sequencing such sequences.
The polynucteotide probes and primers of the present Invention specifically detect a target DNA sequence. Any conventional nucleic acid hybridization or amplification method can be used to identify the presence of DNA from a transgenic event in a sample. By "specifically detect" it is intended that the polynucieotide can be used either as a primer to amplify a DP-073496-4 specific region or the polynuoleotide can be used as a probe that hybridizes under stringent conditions to a pOlynucleotide from a DP-073496-4 event. The level or degree of hybridization which allows for the specific detection of a DP-073496-4 event or a specific region of a DP-073496-4 event is sufficient to distinguish the polynudeotIde with the DP-073496-4 specific region from a Date Recue/Date Received 2022-05-20 polynucleotide lacking this region and thereby allow for discriminately identifying a DP-073496-4 event By "shares sufficient sequence identity or complentarity to allow for the amplification of a DP-073496-4 specific event" is intended the sequence shares at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100%
Identity or compiementarity to a fragment or across the full length of the polynucleotide from the DP-073496-4 specific region.
Regarding the amplification of a target polynucleotide (e.g., by PCR) using a particular amplification primer pair, "stringent conditions" are conditions that permit the primer pair to hybridize to the target polynucleotide to which one primer having the corresponding wild-type sequence (or its complement) and another primer having the corresponding DP-073496-4 inserted DNA sequence would bind and preferably to produce an identifiable amplification product (the amplicon) having a DP-specific region In a DNA thermal amplification reaction. In a PCR approach, oligonucleotide primers can be designed for use in PCR reactions to amplify a DP-073496-4 specific region. Methods for designing PCR primers and PCR cloning are generally known in the art and are disclosed In Sambrook, of al., (1989) Molecular Cloning: A Laboratory Manual (2d ed., Cold Spring Harbor Laboratory Press, Plainview, New York). See also, Innis, et al., eds. (1990) PCR Protocols: A
Guide to Methods and Applications (Academic Press, New York); Innis and Gelfand, eds.
(1995) PCR Strategies (Academic Press, New York) and Innis and Gelfand, eds.
(1999) PCR Methods Manual (Academic Press, New York). Methods of amplification are further described In US Patent Number 4,683,195, 4,683,202 and Chen, of al., (1994) PNAS 91:5695-5699. These methods as well as other methods known in the art of DNA amplification may be used in the practice of the embodiments of the present invention. It is understood that a number of parameters in a specific PCR
protocol may need to be adjusted to specific laboratory conditions and may be slightly modified and yet allow for the collection of similar results. These adjustments will be apparent to a person skilled in the art.
The amplified polynucleotide (amplicon) can be of any length that allows for the detection of the DP-073496-4 event or a DP-073496-4 specific region. For example, the amplicon can be about 10, 50, 100, 200, 300, 500, 700, 100, 2000, 3000, 4000, 5000 nucleotides in length or longer.
In specific embodiments, the specific region of the DP-073496-4 event is detected.
Any primer can be employed in the methods of the invention that allows a DP-0734964 specific region to be amplified and/or detected. For example, in specific embodiments, the first primer comprises a fragment of a polynucleotide of SEQ
ID NO:
Date Recue/Date Received 2022-05-20 2 or 3, wherein the first or the second primer shares sufficient sequence Identity or complementarity to the polynucleotlde to amplify the DP-073496-4 specific region.
-The primer pair cart comprise a fragment of SEQ ID NO: 2 or 3. in another embodiment, the primer pair comprises a fret printer comprising a fragment of SEQ ID
NO: 8 and a second primer comprising a fragment of SEQ ID NO: 9 or 10; or, alternatively, the primer pair comprises a first primer comprising a fragment of SEC ID
NO: 9 and the second primer comprises a fragment of SEQ ID NO; 8 or 10. The =
primers can be of any length sufficient to amplify a DP-073496-4 specific region including, for example, at least 8, 7, 8, 9, 10, 15, 20, 15 or 30 or about 7-
10, 10-15, 15-20, 20-25, 25-30, 30-35, 35-40, 40-46 nucleotides or longer. Additional primer are = also set forth herein In Table 11:
As discussed elsewhere herein, any method to PCR ampfify the DP-073498-4 = event or specific region can be employed, including for example, real time PCR. See, for example, Link, of at, (1995a). Oligonucleotides with fluorescent dyes at opposite = ends provide a quenched probe system for detecting PCR product and nucleic add hybridization. PCR methods and Applications. 4:357-382; US Patent Number 5,538,848; US Patent Number 5,723,591; Applied Blosystems User Bulletin No. 2, "Relative Quantitation of Gene Expression,' P/N 4303859 and Applied Blosystems User Bialetin No. 5, 'Multiplex PCR with Taqman VIC probes,' PM 4308236,.
= Thus, In specific embodiments, a method of detecting the presence of brassica event DP-073496-4 or progeny thereof in a biological sample is provided. The method comprises (a) extracting a DNA sample from the biological sample; (b) providing a pair of DNA primer molecules targeting the insert and/or junction (c) providing DNA
amplification reaction conditions; (d) perforating the DNA amplification reaction, thereby producing a DNA amplicon molecule and (e) detecting the DNA amplicon molecule, wherein the detection Of said DNA amplicon molecule In the DNA
amplification reaction indicates the presence of Brassloa event DP-0734984. In oi;der for a nucleic acid molecule to serve as a primer or probe it needs only be sufficiently complementary in sequence to be able to tom) a stable double-stranded Structure under the particular solvent and salt concentrations employed;
In hybridization techniques, all or part of a polynucleotide that selectively hybridizes to a target polynuoleotide having a DP-073496-4 specific event Is employed. By 'stringent conditions" or 'stringent hybridization conditions' when referring to a polynudeotide probe conditions under which a probe will hybridize to its target sequence to a detectably greater degree than to other sequences (e.g., at least 2-fold over background) are intended. Regarding the amplification of a -target =
= =
Date Recue/Date Received 2022-05-20 polynucleotide (e.g., by PCR) using a particular amplification primer pair, "stringent conditions" are conditions that permit the primer pair to hybridize to the target polynucleotide to which one primer having the corresponding wild-type sequence and another primer having the corresponding DP-073496-4 inserted DNA sequence.
Stringent conditions are sequence-dependent and will be variable in different circumstances. By controlling the stringency of the hybridization and/or washing conditions, target sequences that are 100% complementary to the probe can be Identified (homologous probing). Alternatively, stringency conditions can be adjusted to allow some mismatching in sequences so that lower degrees of identity are detected (heterologous probing). Generally, a probe is less than about 1000 nucleotides in length or less than 500 nucleotides in length.
As used herein, a substantially identical or complementary sequence Is a polynucleotide that will specifically hybridize to the complement of the nucleic acid molecule to which it is being compared under high stringency conditions, Appropriate stringency conditions which promote DNA hybridization, for example, 6X sodium chloride/sodium citrate (SSC) at about 45 C., followed by a wash of 2X8SC at 50 C., are known to those skilled in the art or can be found in Current Protocols in Molecular Biology, John Wiley & Sons, N.Y. (1989), 6.3.1-6,3.6. Typically, stringent conditions for hybridization and detection will be those in which the salt concentration is less than .
about 1.5 M Na ion, typically about 0.01 to 1.0 M Na ion concentration (or other salts) at pH 7.0 to 8.3 and the temperature Is at least about 30 C for short probes (e.g., 10 to 50 nucleotides) and at least about 60 C for long probes (e.g., greater than 50 nucleotides). Stringent conditions may also be achieved with the addition of destabilizing agents such as formamide. Exemplary low stringency conditions include hybridization with a buffer solution of 30 to 35% forrnamide, 1 M NaCl, 1% SDS
(sodium dodecyl sulphate) at 37 C,. and a wash in 1X to 2X SSC (20X SSC = 3.0 M
NaCl/0.3 M trisodium citrate) at 50 to 55 C. Exemplary moderate stringency conditions include hybridization in 40 to 45% formamicie, 1.0 M NaCI, 1% WS at 37 C, and a wash in 0.5X to 1X SSC at 55 to 60 C. Exemplary high stringency conditions include hybridization in 50% formamide, 1 M NaCl, 1% SDS at 37 C
and a wash in 0.1X SSC at 60 to 65 C. Optionally, wash buffers may comprise about 0.1%
to about 1% SDS. Duration of hybridization is generally less than about 24 hours, usually about 4 to about 12 hours. The duration of the wash time will be at least a length of time sufficient to reach equilibrium.
In hybridization reactions, specificity is typically the function of post-hybridization washes, the critical factors being the ionic strength and temperature of the final wash solution. For DNA-DNA hybrids, the Tri, can be approximated from the Date Recue/Date Received 2022-05-20 equation of Meinkoth and Wahl, (1984) Anal. Biochem. 138:267-284: Tm = 81.5 C
+
16.6 (log M) + 0.41 (%GC) - 0.61 (% form) - 500/L; where M is the molarity. of monovalent cations, %GC is the percentage of guanosine and cytosine nucleotides in the DNA, % form is the percentage of forniamIcie In the hybridization solution and L is the length of the hybrid in base pairs. The Tin Is the temperature (under defined ionic strength and pH) at which 50% of a complementary target sequence hybridizes to a perfectly matched probe. Tm is reduced by about 1 C for each 1% of mismatching;
thus, Tm, hybridization, and/or wash conditions can be adjusted to hybridize to sequences of the desired identity. For example, if sequences with *0% identity are sought, the Tm can be decreased 10 C. Generally, stringent conditions are selected to be about 5 C lower than the thermal melting point (Tm) for the specific sequence and its complement at a defined ionic strength and pH. However, severely stringent conditions can utilize a hybridization and/or wash at 1, 2, 3 or 4 C lower than the thermal melting point (TO; moderately stringent conditions can utilize a hybridization and/or wash at 6, 7, 8, 9 or 10 C lower than the thermal melting point (TO;
low stringency conditions can utilize a hybridization and/or wash at 11, 12, 13, 14, 16 or C lower than the thermal melting point (Tm). Using the equation, hybridization and wash compositions, and desired Tin, those of ordinary skill will understand that variations In the stringency of hybridization and/or wash solutions are inherently 20 described. If the desired degree of mismatching results in a Tin of less than 45 C
(aqueous solution) or 32 C (formamide solution), it Is optimal to increase the SSG
concentration so that a higher temperature can be used. An extensive guide to the hybridization of nucleic acids is found in Tijssen, (1993) Laboratory Techniques in Biochemistry and Molecular Biology¨Hybridization with Nucleic Acid Probes, Part I, Chapter 2 (Elsevier, New York) and Ausubel, at al., eds. (1995) Current Protocols in Molecular Biology, Chapter 2 (Greene Publishing and VViley-Interscienoe, New York).
See, Sambrook, at a/., (1089) Molecular Cloning: A Laboratory Manual (2d ed., Cold Spring Harbor Laboratory Press, Plainview, New York) and Haymes, et al., (1985) In:
Nucleic Ackl Hybridization, a Practical Approach, IRL Press, Washington, D.C.
A polynucleotide Is said to be the 'complement" of another polynucleotide if they exhibit complementarity. As used herein, molecules are said to exhibit 'complete complementarity" when every nucleotide of one of the polynucleotide molecules is complementary to a nucleotide of the other. Two molecules are said to be "minimally complementary' if they can hybridize to one another with sufficient stability to permit them to remain annealed to one another under at least conventional low-stringency' conditions. Similarly, the molecules are said to be "complementary" if they can Date Recue/Date Received 2022-05-20 hybridize to one another with sufficient stability to permit them to remain annealed to one another under conventional 'high-stringencr conditions.
Further provided are methods of detecting the presence of DNA corresponding to the DP-073496-4 event in a sample. In one embodiment, the method comprises (a) contacting the biological sample with a polynucleotide probe that hybridizes under stringent hybridization conditions with DNA from brassica event DP-073496-4 and specifically detects the DP-073496-4 event; (b) subjecting the sample and probe to stringent hybridization conditions and (c) detecting hybridization of the probe to the DNA, wherein detection of hybridization indicates the presence of the DP-event.
Various methods can be used to detect the DP-073496-4 specific region or amplicon thereof, including, but not limited to, Genetic Bit Analysis (Nikiforov, at al., (1994) Nucleic Add Res. 22:4167-4176) where a DNA oligonucleotide is designed which overlaps both the adjacent flanking DNA sequence and the inserted DNA
sequence. The oligonucleotide is immobilized in wells of a microwell plate.
Following PCR of the region of Interest (using one primer in the inserted sequence and one in the adjacent flanking sequence) a single-stranded PCR product can be annealed to the immobilized ofigonudeotide and serve as a template for a single base extension reaction using a DNA polymerase and labeled ddNTPs specific for the expected next base. Readout may be fluorescent or ELISA-based. A signal indicates presence of the insert/flanking sequence due to successful amplification, hybridization and single base extension.
Another detection method is the Pyrosequencing technique as described by =
Winge, ((2000) lnnov. Pharma. Tech, 001 8-24). In this method, an oligonucleotide is designed that overlaps the adjacent DNA and insert DNA junction. The oligonucleotide Is annealed to a single-stranded PCR product from the region of Interest (one primer In the Inserted sequence and one in the flanking sequence) and incubated in the presence of a DNA polymerase, ATP, sulfurylase, luciferase, apyrase, adenosine 5' phosphosuifate and luciferin. dNTPs are added individually and the incorporation results In a fight signal which is measured. A light signal indicates the presence of the transgene Insert/flanking sequence due to successful amplification, hybridization and single or multi-base extension.
Fluorescence Polarization as described by Chen, of al., ((1999) Gomm Res.
9:492-498) is also a method that can be used to detect an amplicon of the invention, Using this method, an oligonucleotide is designed which overlaps the flanking and Inserted DNA junction. The oligonucleotide is hybridized to a single-stranded PCR
product from the region of interest ,(one primer in the inserted DNA and one in the = 25 =
Date Recue/Date Received 2022-05-20 flanking DNA sequence) and incubated In the presence of a DNA polymerase and a fluorescent-labeled ddNTP. Single base extension results in incorporation of the ddNTP. Incorporation can be measured as a change in polarization using a fluorometer. A change in polarization indicates the presence of the transgene insert/flanking sequence due to successful amplification, hybridization and single base extension.
Taqmanek (PE Applied 13losystems, Foster City, Calif.) is described as, a method of detecting and quantifying the presence of a DNA sequence and is fully understood in the Instructions provided by the manufacturer. Briefly, a FRET
oligonucleotide probe is designed which overlaps the flanking and insert DNA
junction.
The FRET probe and PCR primers (one primer in the insert DNA sequence and one in the flanking genomic sequence) are cycled in the presence of a thermostable polymerase and dNTPs. Hybridization of the FRET probe results in cleavage and release of the fluorescent moiety away from the quenching moiety on the FRET
probe.
A fluorescent signal indicates the presence of the flankIng/transgene insert sequence due to successful amplification and hybridization.
Molecular Beacons have been described for use in sequence detection as described in Tyangi, at at., ((1996) Nature Biotech. 14:303-308). Briefly, a FRET
oligonucleotide probe is designed that overlaps the flanking and insert DNA
junction.
The unique structure of the FRET probe results In it containing secondary structure that keeps the fluorescent and quenching moieties in close proximity. The FRET
probe and PCR primers (one primer in the insert DNA sequence and one in the = flanking sequence) are cycled in the presence of a thermostable polymerase and dNTPs. Following successful PCR amplification, hybridization of the FRET probe to the target sequence results in the removal of the probe secondary structure and spatial separation of the fluorescent and quenching moieties. A fluorescent signal results. A
fluorescent signal indicates the presence of the flanking/transgene insert sequence due to successful amplification and hybridization.
A hybridization reaction using a probe specific to a sequence found within the amplicon is yet another method used to detect the amplicon produced by a PCR
reaction.
As used herein, "kit" refers to a .set of reagents for the purpose of performing the method embodiments of the invention, more particularly, the identification and/or the detection of the DP-073495-4 event in biological samples. The kit of the invention can be used and its components can be specifically adjusted, for purposes of quality control (e.g. purity of seed lots), detection of event DP-073496-4 in plant material or Date Recue/Date Received 2022-05-20 material comprising or derived from plant material, such as but not limited to food or feed products.
In specific embodiments, a kit for identifying event DP-073496-4 in a biologic sample is provided. The kit comprises a first and a second primer, wherein the first and second primer amplify a polynucleotide comprising a DP-073496-4 specific region.
In further embodiments, the kit also comprises a polynucleotide for the detection of the DP-073496-4 specific region. The kit can comprise, for example, a first primer comprising a fragment of a polynucleotide of SEQ ID NO: 2, 3, 8, 9, or 10, wherein the first or the second primer shares sufficient sequence homology or complementally and specificity to the polynucleotide to amplify said DP-073496-4 specific region. For example, in specific embodiments, the first primer comprises a fragment of a polynucleotide of SEQ ID NO: 2 or 3, wherein the first or the second primer shares sufficient sequence homology or complementarity to the polynucleotide to amplify the DP-073496-4 specific region, In other embodiments, the first primer comprises a fragment of a polynucleotide of SEQ ID NO: 8 and the second primer comprises a fragment of SEQ ID NO: 9 or 10, wherein the first or the second primer shares sufficient sequence homology or complementarity to the polynucleotide to amplify the DP061061-7 specific region. Alternatively, the first primer pair comprises SEQ
ID
NO:9 or a variant or fragment thereof and the second primer comprises SEQ ID
NO: 8 or 10 or a variant or fragment thereof. In other embodiments, the primer pair can = comprise a fragment of SEQ ID NO: 2 and a fragment of SEQ ID NO: 3. The primers can be of any length sufficient to amplify the DP-073496-4 region including, for = example, at least 6, 7, 8, 9, 10, 15, 20, 15 or 30 or about 7-10, 10-15, 15-20, 20-25, 25-30, 30-35, 35-40, 40-45 nucleotides or longer. Further provided are DNA
detection kits comprising at least one polynucleotide that can specifically detect a DP-specific region or insert DNA, wherein said polynucleotide comprises at least one DNA
molecule of a sufficient length of contiguous nucleotides homologous or complementary to SEQ ID NO: 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 16 19, 20, 21, 22, 23, 24, 25, 26, or 27.
In one embodiment, a kit for identifying event DP-073496-4 in a biological sample Is provided, The kit comprises a first and a second primer, wherein said first and said second primer amplify a polynucleotide comprising a DP-073496-4 specific region. In further embodiments, the kit further comprises a polynucleotide for the detection of the DP-073498-4 specific region. Thus, in one non-limiting embodiment, the first primer comprises a first fragment of SEQ ID NO; 11 and the second primer comprises a second fragment of SEQ ID NO:11, wherein the first and the second primer flank the DP-073496-4 specific region and share sufficient sequence homology Date Recue/Date Received 2022-05-20 or complementarity to the polynucleotide to amplify said DP-073498-4 specific region.
As such, a kit can therefore include a first primer comprising a fragment of SEQ ID
NO:8 and a second primer comprising a fragment of SEQ ID NO:9; or a first or a second primer comprising at least 8 consecutive polynucleotides of SEQ ID NO:
As discussed elsewhere herein, any method to PCR ampfify the DP-073498-4 = event or specific region can be employed, including for example, real time PCR. See, for example, Link, of at, (1995a). Oligonucleotides with fluorescent dyes at opposite = ends provide a quenched probe system for detecting PCR product and nucleic add hybridization. PCR methods and Applications. 4:357-382; US Patent Number 5,538,848; US Patent Number 5,723,591; Applied Blosystems User Bulletin No. 2, "Relative Quantitation of Gene Expression,' P/N 4303859 and Applied Blosystems User Bialetin No. 5, 'Multiplex PCR with Taqman VIC probes,' PM 4308236,.
= Thus, In specific embodiments, a method of detecting the presence of brassica event DP-073496-4 or progeny thereof in a biological sample is provided. The method comprises (a) extracting a DNA sample from the biological sample; (b) providing a pair of DNA primer molecules targeting the insert and/or junction (c) providing DNA
amplification reaction conditions; (d) perforating the DNA amplification reaction, thereby producing a DNA amplicon molecule and (e) detecting the DNA amplicon molecule, wherein the detection Of said DNA amplicon molecule In the DNA
amplification reaction indicates the presence of Brassloa event DP-0734984. In oi;der for a nucleic acid molecule to serve as a primer or probe it needs only be sufficiently complementary in sequence to be able to tom) a stable double-stranded Structure under the particular solvent and salt concentrations employed;
In hybridization techniques, all or part of a polynucleotide that selectively hybridizes to a target polynuoleotide having a DP-073496-4 specific event Is employed. By 'stringent conditions" or 'stringent hybridization conditions' when referring to a polynudeotide probe conditions under which a probe will hybridize to its target sequence to a detectably greater degree than to other sequences (e.g., at least 2-fold over background) are intended. Regarding the amplification of a -target =
= =
Date Recue/Date Received 2022-05-20 polynucleotide (e.g., by PCR) using a particular amplification primer pair, "stringent conditions" are conditions that permit the primer pair to hybridize to the target polynucleotide to which one primer having the corresponding wild-type sequence and another primer having the corresponding DP-073496-4 inserted DNA sequence.
Stringent conditions are sequence-dependent and will be variable in different circumstances. By controlling the stringency of the hybridization and/or washing conditions, target sequences that are 100% complementary to the probe can be Identified (homologous probing). Alternatively, stringency conditions can be adjusted to allow some mismatching in sequences so that lower degrees of identity are detected (heterologous probing). Generally, a probe is less than about 1000 nucleotides in length or less than 500 nucleotides in length.
As used herein, a substantially identical or complementary sequence Is a polynucleotide that will specifically hybridize to the complement of the nucleic acid molecule to which it is being compared under high stringency conditions, Appropriate stringency conditions which promote DNA hybridization, for example, 6X sodium chloride/sodium citrate (SSC) at about 45 C., followed by a wash of 2X8SC at 50 C., are known to those skilled in the art or can be found in Current Protocols in Molecular Biology, John Wiley & Sons, N.Y. (1989), 6.3.1-6,3.6. Typically, stringent conditions for hybridization and detection will be those in which the salt concentration is less than .
about 1.5 M Na ion, typically about 0.01 to 1.0 M Na ion concentration (or other salts) at pH 7.0 to 8.3 and the temperature Is at least about 30 C for short probes (e.g., 10 to 50 nucleotides) and at least about 60 C for long probes (e.g., greater than 50 nucleotides). Stringent conditions may also be achieved with the addition of destabilizing agents such as formamide. Exemplary low stringency conditions include hybridization with a buffer solution of 30 to 35% forrnamide, 1 M NaCl, 1% SDS
(sodium dodecyl sulphate) at 37 C,. and a wash in 1X to 2X SSC (20X SSC = 3.0 M
NaCl/0.3 M trisodium citrate) at 50 to 55 C. Exemplary moderate stringency conditions include hybridization in 40 to 45% formamicie, 1.0 M NaCI, 1% WS at 37 C, and a wash in 0.5X to 1X SSC at 55 to 60 C. Exemplary high stringency conditions include hybridization in 50% formamide, 1 M NaCl, 1% SDS at 37 C
and a wash in 0.1X SSC at 60 to 65 C. Optionally, wash buffers may comprise about 0.1%
to about 1% SDS. Duration of hybridization is generally less than about 24 hours, usually about 4 to about 12 hours. The duration of the wash time will be at least a length of time sufficient to reach equilibrium.
In hybridization reactions, specificity is typically the function of post-hybridization washes, the critical factors being the ionic strength and temperature of the final wash solution. For DNA-DNA hybrids, the Tri, can be approximated from the Date Recue/Date Received 2022-05-20 equation of Meinkoth and Wahl, (1984) Anal. Biochem. 138:267-284: Tm = 81.5 C
+
16.6 (log M) + 0.41 (%GC) - 0.61 (% form) - 500/L; where M is the molarity. of monovalent cations, %GC is the percentage of guanosine and cytosine nucleotides in the DNA, % form is the percentage of forniamIcie In the hybridization solution and L is the length of the hybrid in base pairs. The Tin Is the temperature (under defined ionic strength and pH) at which 50% of a complementary target sequence hybridizes to a perfectly matched probe. Tm is reduced by about 1 C for each 1% of mismatching;
thus, Tm, hybridization, and/or wash conditions can be adjusted to hybridize to sequences of the desired identity. For example, if sequences with *0% identity are sought, the Tm can be decreased 10 C. Generally, stringent conditions are selected to be about 5 C lower than the thermal melting point (Tm) for the specific sequence and its complement at a defined ionic strength and pH. However, severely stringent conditions can utilize a hybridization and/or wash at 1, 2, 3 or 4 C lower than the thermal melting point (TO; moderately stringent conditions can utilize a hybridization and/or wash at 6, 7, 8, 9 or 10 C lower than the thermal melting point (TO;
low stringency conditions can utilize a hybridization and/or wash at 11, 12, 13, 14, 16 or C lower than the thermal melting point (Tm). Using the equation, hybridization and wash compositions, and desired Tin, those of ordinary skill will understand that variations In the stringency of hybridization and/or wash solutions are inherently 20 described. If the desired degree of mismatching results in a Tin of less than 45 C
(aqueous solution) or 32 C (formamide solution), it Is optimal to increase the SSG
concentration so that a higher temperature can be used. An extensive guide to the hybridization of nucleic acids is found in Tijssen, (1993) Laboratory Techniques in Biochemistry and Molecular Biology¨Hybridization with Nucleic Acid Probes, Part I, Chapter 2 (Elsevier, New York) and Ausubel, at al., eds. (1995) Current Protocols in Molecular Biology, Chapter 2 (Greene Publishing and VViley-Interscienoe, New York).
See, Sambrook, at a/., (1089) Molecular Cloning: A Laboratory Manual (2d ed., Cold Spring Harbor Laboratory Press, Plainview, New York) and Haymes, et al., (1985) In:
Nucleic Ackl Hybridization, a Practical Approach, IRL Press, Washington, D.C.
A polynucleotide Is said to be the 'complement" of another polynucleotide if they exhibit complementarity. As used herein, molecules are said to exhibit 'complete complementarity" when every nucleotide of one of the polynucleotide molecules is complementary to a nucleotide of the other. Two molecules are said to be "minimally complementary' if they can hybridize to one another with sufficient stability to permit them to remain annealed to one another under at least conventional low-stringency' conditions. Similarly, the molecules are said to be "complementary" if they can Date Recue/Date Received 2022-05-20 hybridize to one another with sufficient stability to permit them to remain annealed to one another under conventional 'high-stringencr conditions.
Further provided are methods of detecting the presence of DNA corresponding to the DP-073496-4 event in a sample. In one embodiment, the method comprises (a) contacting the biological sample with a polynucleotide probe that hybridizes under stringent hybridization conditions with DNA from brassica event DP-073496-4 and specifically detects the DP-073496-4 event; (b) subjecting the sample and probe to stringent hybridization conditions and (c) detecting hybridization of the probe to the DNA, wherein detection of hybridization indicates the presence of the DP-event.
Various methods can be used to detect the DP-073496-4 specific region or amplicon thereof, including, but not limited to, Genetic Bit Analysis (Nikiforov, at al., (1994) Nucleic Add Res. 22:4167-4176) where a DNA oligonucleotide is designed which overlaps both the adjacent flanking DNA sequence and the inserted DNA
sequence. The oligonucleotide is immobilized in wells of a microwell plate.
Following PCR of the region of Interest (using one primer in the inserted sequence and one in the adjacent flanking sequence) a single-stranded PCR product can be annealed to the immobilized ofigonudeotide and serve as a template for a single base extension reaction using a DNA polymerase and labeled ddNTPs specific for the expected next base. Readout may be fluorescent or ELISA-based. A signal indicates presence of the insert/flanking sequence due to successful amplification, hybridization and single base extension.
Another detection method is the Pyrosequencing technique as described by =
Winge, ((2000) lnnov. Pharma. Tech, 001 8-24). In this method, an oligonucleotide is designed that overlaps the adjacent DNA and insert DNA junction. The oligonucleotide Is annealed to a single-stranded PCR product from the region of Interest (one primer In the Inserted sequence and one in the flanking sequence) and incubated in the presence of a DNA polymerase, ATP, sulfurylase, luciferase, apyrase, adenosine 5' phosphosuifate and luciferin. dNTPs are added individually and the incorporation results In a fight signal which is measured. A light signal indicates the presence of the transgene Insert/flanking sequence due to successful amplification, hybridization and single or multi-base extension.
Fluorescence Polarization as described by Chen, of al., ((1999) Gomm Res.
9:492-498) is also a method that can be used to detect an amplicon of the invention, Using this method, an oligonucleotide is designed which overlaps the flanking and Inserted DNA junction. The oligonucleotide is hybridized to a single-stranded PCR
product from the region of interest ,(one primer in the inserted DNA and one in the = 25 =
Date Recue/Date Received 2022-05-20 flanking DNA sequence) and incubated In the presence of a DNA polymerase and a fluorescent-labeled ddNTP. Single base extension results in incorporation of the ddNTP. Incorporation can be measured as a change in polarization using a fluorometer. A change in polarization indicates the presence of the transgene insert/flanking sequence due to successful amplification, hybridization and single base extension.
Taqmanek (PE Applied 13losystems, Foster City, Calif.) is described as, a method of detecting and quantifying the presence of a DNA sequence and is fully understood in the Instructions provided by the manufacturer. Briefly, a FRET
oligonucleotide probe is designed which overlaps the flanking and insert DNA
junction.
The FRET probe and PCR primers (one primer in the insert DNA sequence and one in the flanking genomic sequence) are cycled in the presence of a thermostable polymerase and dNTPs. Hybridization of the FRET probe results in cleavage and release of the fluorescent moiety away from the quenching moiety on the FRET
probe.
A fluorescent signal indicates the presence of the flankIng/transgene insert sequence due to successful amplification and hybridization.
Molecular Beacons have been described for use in sequence detection as described in Tyangi, at at., ((1996) Nature Biotech. 14:303-308). Briefly, a FRET
oligonucleotide probe is designed that overlaps the flanking and insert DNA
junction.
The unique structure of the FRET probe results In it containing secondary structure that keeps the fluorescent and quenching moieties in close proximity. The FRET
probe and PCR primers (one primer in the insert DNA sequence and one in the = flanking sequence) are cycled in the presence of a thermostable polymerase and dNTPs. Following successful PCR amplification, hybridization of the FRET probe to the target sequence results in the removal of the probe secondary structure and spatial separation of the fluorescent and quenching moieties. A fluorescent signal results. A
fluorescent signal indicates the presence of the flanking/transgene insert sequence due to successful amplification and hybridization.
A hybridization reaction using a probe specific to a sequence found within the amplicon is yet another method used to detect the amplicon produced by a PCR
reaction.
As used herein, "kit" refers to a .set of reagents for the purpose of performing the method embodiments of the invention, more particularly, the identification and/or the detection of the DP-073495-4 event in biological samples. The kit of the invention can be used and its components can be specifically adjusted, for purposes of quality control (e.g. purity of seed lots), detection of event DP-073496-4 in plant material or Date Recue/Date Received 2022-05-20 material comprising or derived from plant material, such as but not limited to food or feed products.
In specific embodiments, a kit for identifying event DP-073496-4 in a biologic sample is provided. The kit comprises a first and a second primer, wherein the first and second primer amplify a polynucleotide comprising a DP-073496-4 specific region.
In further embodiments, the kit also comprises a polynucleotide for the detection of the DP-073496-4 specific region. The kit can comprise, for example, a first primer comprising a fragment of a polynucleotide of SEQ ID NO: 2, 3, 8, 9, or 10, wherein the first or the second primer shares sufficient sequence homology or complementally and specificity to the polynucleotide to amplify said DP-073496-4 specific region. For example, in specific embodiments, the first primer comprises a fragment of a polynucleotide of SEQ ID NO: 2 or 3, wherein the first or the second primer shares sufficient sequence homology or complementarity to the polynucleotide to amplify the DP-073496-4 specific region, In other embodiments, the first primer comprises a fragment of a polynucleotide of SEQ ID NO: 8 and the second primer comprises a fragment of SEQ ID NO: 9 or 10, wherein the first or the second primer shares sufficient sequence homology or complementarity to the polynucleotide to amplify the DP061061-7 specific region. Alternatively, the first primer pair comprises SEQ
ID
NO:9 or a variant or fragment thereof and the second primer comprises SEQ ID
NO: 8 or 10 or a variant or fragment thereof. In other embodiments, the primer pair can = comprise a fragment of SEQ ID NO: 2 and a fragment of SEQ ID NO: 3. The primers can be of any length sufficient to amplify the DP-073496-4 region including, for = example, at least 6, 7, 8, 9, 10, 15, 20, 15 or 30 or about 7-10, 10-15, 15-20, 20-25, 25-30, 30-35, 35-40, 40-45 nucleotides or longer. Further provided are DNA
detection kits comprising at least one polynucleotide that can specifically detect a DP-specific region or insert DNA, wherein said polynucleotide comprises at least one DNA
molecule of a sufficient length of contiguous nucleotides homologous or complementary to SEQ ID NO: 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 16 19, 20, 21, 22, 23, 24, 25, 26, or 27.
In one embodiment, a kit for identifying event DP-073496-4 in a biological sample Is provided, The kit comprises a first and a second primer, wherein said first and said second primer amplify a polynucleotide comprising a DP-073496-4 specific region. In further embodiments, the kit further comprises a polynucleotide for the detection of the DP-073498-4 specific region. Thus, in one non-limiting embodiment, the first primer comprises a first fragment of SEQ ID NO; 11 and the second primer comprises a second fragment of SEQ ID NO:11, wherein the first and the second primer flank the DP-073496-4 specific region and share sufficient sequence homology Date Recue/Date Received 2022-05-20 or complementarity to the polynucleotide to amplify said DP-073498-4 specific region.
As such, a kit can therefore include a first primer comprising a fragment of SEQ ID
NO:8 and a second primer comprising a fragment of SEQ ID NO:9; or a first or a second primer comprising at least 8 consecutive polynucleotides of SEQ ID NO:
11; or a first or a second primer comprising at least 8 consecutive polynucleotides of SEQ ID
NO:8 or 9.
In further embodiments, methods are provided for detecting a glyphosate-N-acetyltranferase polypeptide comprising analysing brassica plant tissues using an immunoassay comprising a glyphosate-N-acetyltranferase polypeptide-specific antibody or antibodies, In other embodiments, methods for detecting the presence of a polynucleotide that encodes a glyphosate-N-acetyltranferase polypeptide are provide and comprise assaying brassica plant tissue using PCR amplification. Kits for employing such methods are further provided.
Any of the polynucleotides and fragments and variants thereof employed in the methods and compositions of the invention can share sequence identity to a region of the transgene insert of the DP-073498-4 event, a Junction sequence of the DP-073498-4 event, or a region of the insert in combination with a region of the flanking sequence of the DP-073496-4 event. Methods to determine the relationship of various sequences are known. As used herein, "reference sequence' is a defined sequence used as a basis for sequence comparison. A reference sequence may be a subset or the entirety of a specified sequence; for example, as a segment of a full-length cDNA
or gene sequence, or the complete cDNA or gene sequence. As used herein, "comparison window" makes reference to a contiguous and specified segment of a polynucleotide sequence, wherein the polynucleotide sequence in the comparison window may comprise additions or deletions gaps) compared to the reference sequence (which does not comprise additions or deletions) for optimal alignment of the two polynucleotides. Generally, the comparison window is at least 20 contiguous nucleotides in length and optionally can be 30, 40, 50, 100 or longer. Those of skill in the art understand that to avoid a high similarity to a reference sequence due to inclusion of gaps in the polynucleotide sequence a gap penalty is typically introduced and is subtracted from the number of matches.
Methods of alignment of sequences for comparison are well known in the art.
Thus, the determination of percent sequence identity between any two sequences can be accomplished using a mathematical algorithm. Non-limiting examples of such mathematical algorithms are the algorithm of Myers and Miller, (1988) CABIOS
4;11-17; the local alignment algorithm of Smith, at al., (1981) Adv. App!. Math.
2:482; the global alignment algorithm of Needleman and Wunsch, (1970) J. Md. Biol. 48:443-Date Recue/Date Received 2022-05-20 453; the search-for-local alignment method of Pearson and Uprnan, (1988) Proc.
Natl.
Acad. Set 85:2444-2448; the algorithm of Karlin and Altschul, (1990) Proc.
Nat! : Acad.
Sot USA 872264, modified as in Karlin and Altschul, (1993) Proc. Natl. Acad.
Set USA
90:5873-5877.
Computer implementations of these mathematical algorithms can be utilized for comparison of sequences to determine sequence identity. Such implementations Include, but are not limited 'to: CLUSTAL In the PC/Gene program (available from IntelligenetIcs, Mountain View, California); the AUGN program (Version 2.0) and GAP, BESTFIT, BLAST, PASTA and TFASTA in the GCG Wisconsin Genetics Software Package, Version 10 (available from Accelrys Inc., 9685 Scranton Road, San Diego, California, USA). Alignments using these programs can be performed using the default parameters. The CLUSTAL program is well described by Higgins, of al., (1988) Gene 73:237-244 (1988); Higgins, at al., (1989) CABIOS 5:151-153; Carpet, et al., (1988) Nuo/elc Acids Res. 16:10881-90; Huang, at al., (1992) CAMS 8:155-65 and Pearson, of al., (1994) Meth. Mot Biol. 24:307-331. The ALIGN program is based on the algorithm of Myers and Miller, (1988) supra. A PAM120 weight residue table, a gap length penalty of 12 and a gap penalty of 4 can be used with the ALIGN
program when comparing amino acid sequences. The BLAST programs of Altschul, at al., (1990) J. Mol. Biol. 215:403 are based on the algorithm of Karlin and Altschul, (1990) supra. BLAST nucleotide searches can be performed with the BLASTN program, score = 100, wordlength = 12, to obtain nucleotide sequences homologous to a nucleotide sequence encoding a protein of the invention. BLAST protein searches can be performed with the BLASTX program, score = 50, wordlength = 3, to obtain amino acid sequences homologous to a protein or polypeptide of the invention. To obtain gapped alignments for comparison purposes, Gapped BLAST (in BLAST 2.0) can be utilized as described in Altschul, at at, (1997) Nucleic Acids Res. 26:3389.
Alternatively, PSI-BLAST (in BLAST 2.0) can be used to perform an iterated search that detects distant relationships between molecules. See, Altschul, at a/., (1997) supra. When utilizing BLAST, Gapped BLAST, PSI-BLAST, the default parameters of the respective programs (e.g., BLASTN for nucleotide sequences, BLASTX for proteins) can be used. See www.ncbl.nlm.nih,gov. Alignment may also be performed manually by Inspection.
Unless otherwise stated, sequence Identity/similarity values provided herein refer to the value obtained using GAP Version 10 using the following parameters: %
identity and % similarity for a nucleotide sequence using GAP Weight of 50 and Length Weight of 3, and the nwsgapdna.cmp scoring matrix; % Identity and % similarity for an amino acid sequence using GAP Weight of 8 and Length Weight of 2 and the Date Recue/Date Received 2022-05-20 BLOSUfv182 scoring matrix or any equivalent program thereof. By "equivalent program" any sequence comparison program that, for any two sequences in question, = generates an alignment having Identical nucleotide or amino acid residue matches and an identical percent sequence identity when compared to the corresponding alignment generated by GAP Version 10 is intended.
GAP uses the algorithm of Needleman and Wunsch, (1970) J. Md. Biol.
48:443-453, to find the alignment of two complete sequences that maximizes the number of matches and minimizes the number of gaps. GAP considers all possible alignments and gap positions and creates the alignment with the largest number of matched bases and the fewest gaps. It allows for the provision of a gap creation penally and a gap extension penalty in units of matched bases. GAP must make a = profit of gap creation penalty number of matches for each gap it Inserts.
If a gap extension penalty greater than zero is chosen, GAP must, in addition, make a profit for each gap inserted of the length of the gap times the gap extension penalty.
Default gap creation penalty values and gap extension penalty values In Version 10 of the GCG Wisconsin Genetics Software Package for protein sequences are 8 and 2, respectively. For nucleotide sequences the default gap creation penalty is 50 while the default gap extension penalty is 3. The gap creation and gap extension penalties can be expressed as an Integer selected from the group of integers consisting of from 0 to 200. Thus, for example, the gap creation and gap extension penalties can be 0, 1, 2, 3,4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65 or greater.
GAP presents one member of the family of best alignments. There may be many members of this family, but no other member has a better quality. GAP
displays four figures of merit for alignments: Quality, Ratio, identity and Similarity.
The Quality Is the metric maximized in order to align the sequences. Ratio Is the Quality divided by the number of bases in the shorter segment. Percent Identity is the percent of the symbols that actually match. Percent Similarity Is the percent of the symbols that are similar. Symbols that are across from gaps are ignored. A similarity is scored when the scoring matrix value for a pair of symbols is greater than or equal to 0.50, the similarity threshold. The scoring matrix used in Version 10 of the GCG
Wisconsin Genetics Software Package is BLOSUM62 (see, Henikoff and Henikoff, (1989) Proc.
Natl. Acad. Sc). USA 89:10915).
As used herein, "sequence Identity' or "identity' in the context of two polynucleotides or polypeptide sequences makes reference to the residues in the two sequences that are the same when aligned for maximum correspondence over a specified comparison window. When percentage of sequence identity is used in reference to proteins it Is reco;tmlzed that residue positions which are not identical Date Recue/Date Received 2022-05-20 often differ by conservative amino acid substitutions, where amino acid residues are substituted for other amino acid residues with similar chemical properties (e.g., charge or hydrophobicity) and therefore do not change the functional properties of the molecule. When sequences differ in conservative stbstitutions, the percent sequence identity may be adjusted upwards to correct for the conservative nature of the substitution. Sequences that differ by such conservative substitutions are said to have "sequence similarity or 'similarity", Means for making this adjustment are well known to those of skill In the art. Typically this involves scoring a conservative substitution as a partial rather than a full mismatch, thereby increasing the = percentage sequence identity. Thus, for example, where an Identical amino acid is given a score of 1 and a non-conservative substitution Is given a score of zero, a conservative substitution is given a score between zero and 1. The scoring of conservative substitutions is calculated, e.g., as implemented in the program PC/GENE (Intelligenetics, Mountain View, California).
As used herein, "percentage of sequence Identity" means the value determined by comparing two optimally aligned sequences over a comparison window, wherein the portion of the polynucleotide sequence in the comparison window may comprise additions or deletions (I.e., gaps) as compared to the reference sequence (which does not comprise additions or deletions) for optimal alignment of the two sequences. The percentage is calculated by determining the number of positions at which the identical nucleic acid base or amino acid residue occurs In both sequences to yield the number of matched positions, dividing the number of matched positions by the total number of positions in the window of comparison and multiplying the result by 100 to yield the percentage of sequence identity.
The present invention provides methods for controlling weeds in an area of cultivation, preventing the development or the appearance of herbicide resistant weeds in an area of cultivation, producing a crop and Increasing .crop safety. The term *controlling," and derivations thereof, for example, as In "controlling weeds"
refers to one or more of inhibiting the growth, germination, reproduction and/or proliferation of and/or killing, removing, destroying or otherwise diminishing the occurrence and/or activity of a weed.
As used herein, an area of cultivation" comprises any region in which one desires to grow a plant. Such areas of cultivations include, but are not limited to, a field in which a plant Is cultivated (such as a crop field, a sod field, a tree field, a managed forest a field for culturing fruits and vegetables, etc), a greenhouse, a growth chamber, etc.
= 31 Date Recue/Date Received 2022-05-20 The methods of the Invention comprise planting the area of cultivation with the Brassies DP-073496-4 seeds or plants, and In specific embodiments, applying to the crop, seed, weed or area of cultivation thereof an effective amount of a herbicide of interest. It is recognized that the herbicide can be applied before or after the crop is planted in the area of cultivation. Such herbicide applications can include an application of glyphosale.
In one embodiment, the method of controlling weeds comprises planting the area with the DP-073496-4 Brassica seeds or plants and applying to the crop, crop part, seed of said crop or the area under cultivation, an effective amount of a herbicide, wherein said effective amount comprises an amount that is not tolerated by a second control crop when applied to the second crop, crop part, seed or the area of cultivation, wherein said second control crop does not express the GLYAT polynucleotide.
In another embodiment, the method of controlling weeds comprises planting the area with a DP-073496-4 Brassies crop seed or plant and applying to the crop, crop part, seed of said crop or the area under cultivation, an effective amount of a glyphosate herbicide, wherein said effective amount comprises a level that Is above the. recommended label use rate for the crop, wherein said effective amount Is tolerated when applied to the DP-073496-4 Brass/ca crop, crop part, seed or the area of cultivation thereof.
A "control" or "control plant' or 'control plant cell' provides a reference point for measuring changes In phenotype of the subject plant or plant cell, and may be any suitable plant or plant cell. A control plant or plant cell may comprise, for example: (a) a wild-type plant or cell, I.e., of the same genotype as the starting material for the genetic alteration which resulted In the subject plant or cell; (b) a plant or plant cell of the same genotype as the starting material but which has been transformed with a null construct (I.e., with a construct which has no known effect on the trait of interest, such as a construct comprising a marker gene); (c) a plant or plant cell which is a non-transformed segregant among progeny of a subject plant or plant cell; (d) a plant or plant cell which is genetically identical to the subject plant or plant cell but which is not exposed to the same treatment (e.g., herbicide treatment) as the subject plant or plant cell; (e) the subject plant or plant cell itself, under conditions in which the gene of interest is not expressed or (f) the subject plant or plant cell itself, under conditions in which it has not been exposed to a particular treatment such as, for example, a herbicide or combination of herbicides and/or other chemicals. In some instances, an appropriate control plant or control plant cell may have a different genotype from the subject plant or plant cell but may share the herbicide-sensitive characteristics of the starting material for the genetic alteration(s) which resulted in the subject plant or cell =
Date Recue/Date Received 2022-05-20 (see, e.g., Green, (1998) Weed Technology 12;474-477; Green and Ulrich, (1993) Weed Science 41:506-516. In other embodiments, the null segregant can be used as a control, as they are genetically identical to DP-073496-4 with the exception of the transgenic insert DNA.
Classification of herbicides (i.e., the grouping of herbicides into classes and subclasses) Is well-known in the art and includes classifications by HRAC
(Herbicide Resistance Action Committee) and WSSA (the Weed Science Society of America) (see also, Retzinger and Mallory-Smith, (1997) Weed Technology 11:384-393). An abbreviated version of the HRAC classification (with notes regarding the corresponding WSSA group) is set forth below in Table 1.
Herbicides can be classified by their mode of action and/or site of action and can also be classified by the time at which they are applied (e.g., preemergent or postemergent), by the method of application (e.g., foliar application or soil application) or by how they are taken up by or affect the plant. For example, thifensulfuron-methyl and tribenuron-methyl are applied to the foliage of a crop and are generally metabolized there, while rimsuifuron and chlorimuron-ethyl are generally taken up = through both the roots and foliage of a plant. "Mode of action* generally refers to the metabolic or physiological process within the plant that the herbicide inhibits or otherwise impairs, whereas 'site of action" generally refers to the physical location or biochemical site within the plant where the herbicide acts or directly Interacts.
Herbicides can be classified in various ways, including by mode of action and/or site of action (see, e.g., Table 1).
Often, a herbicide-tolerance gene that confers tolerance to a particular herbicide or other chemical on a plant expressing It will also confer tolerance to other herbicides or chemicals In the same class or subclass, for example, a class or subclass set forth in Table 1. Thus, in some embodiments of the invention, a transgenic plant of the invention is tolerant to more than one herbicide or chemical in the same class or subclass, such as, for example, an inhibitor of E'PO, a sulfonylurea or a synthetic auxin.
Typically, the plants of the present Invention can tolerate treatment with different types of herbicides (i.e., herbicides having different modes of action and/or different sites of action) as well as with higher amounts of herbicides than previously known plants, thereby permitting improved weed management strategies that are recommended in order to reduce the incidence and prevalence of herbicide-tolerant weeds. Specific herbicide combinations can be employed to effectively control weeds.
The invention thereby provides a transgenic brassica plant which can be selected for use in crop production based on the prevalence of herbicide-tolerant weed Date Recue/Date Received 2022-05-20 species In the area where the transgenic crop is to be grown. Methods are known in the art for assessing the herbicide tolerance of various weed species. Weed management techniques are also known in the art, such as for example, crop rotation using a crop that is tolerant to a herbicide to which the local weed species are not tolerant A number of entities monitor and publicly report the Incidence and characteristics of herbicide-tolerant weeds, including the Herbicide Resistance Action Committee (HRAC), the Weed Science Society of America and various state agencies (see, for example, herbicide tolerance scores for various broadleaf weeds from the 2004 Illinois Agricultural Pest Management Handbook) and one of skill in the art would be able to use this information to determine which crop and herbicide combinations should be used in a particular location.
These entities also publish advice and guidelines for preventing the development and/or appearance of and controlling the spread of herbicide tolerant weeds (see, e.g., Owen and Hartzler, (2004), 2005 Herbicide Manual for Agricultural Professionals, Pub. WC 92 Revised (Iowa State University Extension, Iowa State .
University of Science and Technology, Ames, Iowa); Weed Control for Corn, Brassicas, and Sorghum, Chapter 2 of '2004 Illinois Agricultural Pest Management Handbook" (University of Illinois Extension, University of Illinois at Urbana-Champaign, Illinois); Weed Control Guide for Field Crops, MSU Extension Bulletin E434 (Michigan State University, East Lansing, Michigan)).
Table 1: Abbreviated version of HRAC Herbicide. Classification I. ALS Inhibitors (WSSA Group 2) A. Sulfonylureas 1. Azimsulfuron 2. Chlorlmuron-ethyl 3. Metsulfuron-methyl 4. Nicosulfuron 5. Rimsulfuron 6. Sulfometuron-methyl 7. ThIfensulfuron-methyl = 8. TrIbenuron-methyl =
= = 9. Amidosulfuron 10. Bensulfuron-rnethyl = 11. Chlorsulfumn
NO:8 or 9.
In further embodiments, methods are provided for detecting a glyphosate-N-acetyltranferase polypeptide comprising analysing brassica plant tissues using an immunoassay comprising a glyphosate-N-acetyltranferase polypeptide-specific antibody or antibodies, In other embodiments, methods for detecting the presence of a polynucleotide that encodes a glyphosate-N-acetyltranferase polypeptide are provide and comprise assaying brassica plant tissue using PCR amplification. Kits for employing such methods are further provided.
Any of the polynucleotides and fragments and variants thereof employed in the methods and compositions of the invention can share sequence identity to a region of the transgene insert of the DP-073498-4 event, a Junction sequence of the DP-073498-4 event, or a region of the insert in combination with a region of the flanking sequence of the DP-073496-4 event. Methods to determine the relationship of various sequences are known. As used herein, "reference sequence' is a defined sequence used as a basis for sequence comparison. A reference sequence may be a subset or the entirety of a specified sequence; for example, as a segment of a full-length cDNA
or gene sequence, or the complete cDNA or gene sequence. As used herein, "comparison window" makes reference to a contiguous and specified segment of a polynucleotide sequence, wherein the polynucleotide sequence in the comparison window may comprise additions or deletions gaps) compared to the reference sequence (which does not comprise additions or deletions) for optimal alignment of the two polynucleotides. Generally, the comparison window is at least 20 contiguous nucleotides in length and optionally can be 30, 40, 50, 100 or longer. Those of skill in the art understand that to avoid a high similarity to a reference sequence due to inclusion of gaps in the polynucleotide sequence a gap penalty is typically introduced and is subtracted from the number of matches.
Methods of alignment of sequences for comparison are well known in the art.
Thus, the determination of percent sequence identity between any two sequences can be accomplished using a mathematical algorithm. Non-limiting examples of such mathematical algorithms are the algorithm of Myers and Miller, (1988) CABIOS
4;11-17; the local alignment algorithm of Smith, at al., (1981) Adv. App!. Math.
2:482; the global alignment algorithm of Needleman and Wunsch, (1970) J. Md. Biol. 48:443-Date Recue/Date Received 2022-05-20 453; the search-for-local alignment method of Pearson and Uprnan, (1988) Proc.
Natl.
Acad. Set 85:2444-2448; the algorithm of Karlin and Altschul, (1990) Proc.
Nat! : Acad.
Sot USA 872264, modified as in Karlin and Altschul, (1993) Proc. Natl. Acad.
Set USA
90:5873-5877.
Computer implementations of these mathematical algorithms can be utilized for comparison of sequences to determine sequence identity. Such implementations Include, but are not limited 'to: CLUSTAL In the PC/Gene program (available from IntelligenetIcs, Mountain View, California); the AUGN program (Version 2.0) and GAP, BESTFIT, BLAST, PASTA and TFASTA in the GCG Wisconsin Genetics Software Package, Version 10 (available from Accelrys Inc., 9685 Scranton Road, San Diego, California, USA). Alignments using these programs can be performed using the default parameters. The CLUSTAL program is well described by Higgins, of al., (1988) Gene 73:237-244 (1988); Higgins, at al., (1989) CABIOS 5:151-153; Carpet, et al., (1988) Nuo/elc Acids Res. 16:10881-90; Huang, at al., (1992) CAMS 8:155-65 and Pearson, of al., (1994) Meth. Mot Biol. 24:307-331. The ALIGN program is based on the algorithm of Myers and Miller, (1988) supra. A PAM120 weight residue table, a gap length penalty of 12 and a gap penalty of 4 can be used with the ALIGN
program when comparing amino acid sequences. The BLAST programs of Altschul, at al., (1990) J. Mol. Biol. 215:403 are based on the algorithm of Karlin and Altschul, (1990) supra. BLAST nucleotide searches can be performed with the BLASTN program, score = 100, wordlength = 12, to obtain nucleotide sequences homologous to a nucleotide sequence encoding a protein of the invention. BLAST protein searches can be performed with the BLASTX program, score = 50, wordlength = 3, to obtain amino acid sequences homologous to a protein or polypeptide of the invention. To obtain gapped alignments for comparison purposes, Gapped BLAST (in BLAST 2.0) can be utilized as described in Altschul, at at, (1997) Nucleic Acids Res. 26:3389.
Alternatively, PSI-BLAST (in BLAST 2.0) can be used to perform an iterated search that detects distant relationships between molecules. See, Altschul, at a/., (1997) supra. When utilizing BLAST, Gapped BLAST, PSI-BLAST, the default parameters of the respective programs (e.g., BLASTN for nucleotide sequences, BLASTX for proteins) can be used. See www.ncbl.nlm.nih,gov. Alignment may also be performed manually by Inspection.
Unless otherwise stated, sequence Identity/similarity values provided herein refer to the value obtained using GAP Version 10 using the following parameters: %
identity and % similarity for a nucleotide sequence using GAP Weight of 50 and Length Weight of 3, and the nwsgapdna.cmp scoring matrix; % Identity and % similarity for an amino acid sequence using GAP Weight of 8 and Length Weight of 2 and the Date Recue/Date Received 2022-05-20 BLOSUfv182 scoring matrix or any equivalent program thereof. By "equivalent program" any sequence comparison program that, for any two sequences in question, = generates an alignment having Identical nucleotide or amino acid residue matches and an identical percent sequence identity when compared to the corresponding alignment generated by GAP Version 10 is intended.
GAP uses the algorithm of Needleman and Wunsch, (1970) J. Md. Biol.
48:443-453, to find the alignment of two complete sequences that maximizes the number of matches and minimizes the number of gaps. GAP considers all possible alignments and gap positions and creates the alignment with the largest number of matched bases and the fewest gaps. It allows for the provision of a gap creation penally and a gap extension penalty in units of matched bases. GAP must make a = profit of gap creation penalty number of matches for each gap it Inserts.
If a gap extension penalty greater than zero is chosen, GAP must, in addition, make a profit for each gap inserted of the length of the gap times the gap extension penalty.
Default gap creation penalty values and gap extension penalty values In Version 10 of the GCG Wisconsin Genetics Software Package for protein sequences are 8 and 2, respectively. For nucleotide sequences the default gap creation penalty is 50 while the default gap extension penalty is 3. The gap creation and gap extension penalties can be expressed as an Integer selected from the group of integers consisting of from 0 to 200. Thus, for example, the gap creation and gap extension penalties can be 0, 1, 2, 3,4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65 or greater.
GAP presents one member of the family of best alignments. There may be many members of this family, but no other member has a better quality. GAP
displays four figures of merit for alignments: Quality, Ratio, identity and Similarity.
The Quality Is the metric maximized in order to align the sequences. Ratio Is the Quality divided by the number of bases in the shorter segment. Percent Identity is the percent of the symbols that actually match. Percent Similarity Is the percent of the symbols that are similar. Symbols that are across from gaps are ignored. A similarity is scored when the scoring matrix value for a pair of symbols is greater than or equal to 0.50, the similarity threshold. The scoring matrix used in Version 10 of the GCG
Wisconsin Genetics Software Package is BLOSUM62 (see, Henikoff and Henikoff, (1989) Proc.
Natl. Acad. Sc). USA 89:10915).
As used herein, "sequence Identity' or "identity' in the context of two polynucleotides or polypeptide sequences makes reference to the residues in the two sequences that are the same when aligned for maximum correspondence over a specified comparison window. When percentage of sequence identity is used in reference to proteins it Is reco;tmlzed that residue positions which are not identical Date Recue/Date Received 2022-05-20 often differ by conservative amino acid substitutions, where amino acid residues are substituted for other amino acid residues with similar chemical properties (e.g., charge or hydrophobicity) and therefore do not change the functional properties of the molecule. When sequences differ in conservative stbstitutions, the percent sequence identity may be adjusted upwards to correct for the conservative nature of the substitution. Sequences that differ by such conservative substitutions are said to have "sequence similarity or 'similarity", Means for making this adjustment are well known to those of skill In the art. Typically this involves scoring a conservative substitution as a partial rather than a full mismatch, thereby increasing the = percentage sequence identity. Thus, for example, where an Identical amino acid is given a score of 1 and a non-conservative substitution Is given a score of zero, a conservative substitution is given a score between zero and 1. The scoring of conservative substitutions is calculated, e.g., as implemented in the program PC/GENE (Intelligenetics, Mountain View, California).
As used herein, "percentage of sequence Identity" means the value determined by comparing two optimally aligned sequences over a comparison window, wherein the portion of the polynucleotide sequence in the comparison window may comprise additions or deletions (I.e., gaps) as compared to the reference sequence (which does not comprise additions or deletions) for optimal alignment of the two sequences. The percentage is calculated by determining the number of positions at which the identical nucleic acid base or amino acid residue occurs In both sequences to yield the number of matched positions, dividing the number of matched positions by the total number of positions in the window of comparison and multiplying the result by 100 to yield the percentage of sequence identity.
The present invention provides methods for controlling weeds in an area of cultivation, preventing the development or the appearance of herbicide resistant weeds in an area of cultivation, producing a crop and Increasing .crop safety. The term *controlling," and derivations thereof, for example, as In "controlling weeds"
refers to one or more of inhibiting the growth, germination, reproduction and/or proliferation of and/or killing, removing, destroying or otherwise diminishing the occurrence and/or activity of a weed.
As used herein, an area of cultivation" comprises any region in which one desires to grow a plant. Such areas of cultivations include, but are not limited to, a field in which a plant Is cultivated (such as a crop field, a sod field, a tree field, a managed forest a field for culturing fruits and vegetables, etc), a greenhouse, a growth chamber, etc.
= 31 Date Recue/Date Received 2022-05-20 The methods of the Invention comprise planting the area of cultivation with the Brassies DP-073496-4 seeds or plants, and In specific embodiments, applying to the crop, seed, weed or area of cultivation thereof an effective amount of a herbicide of interest. It is recognized that the herbicide can be applied before or after the crop is planted in the area of cultivation. Such herbicide applications can include an application of glyphosale.
In one embodiment, the method of controlling weeds comprises planting the area with the DP-073496-4 Brassica seeds or plants and applying to the crop, crop part, seed of said crop or the area under cultivation, an effective amount of a herbicide, wherein said effective amount comprises an amount that is not tolerated by a second control crop when applied to the second crop, crop part, seed or the area of cultivation, wherein said second control crop does not express the GLYAT polynucleotide.
In another embodiment, the method of controlling weeds comprises planting the area with a DP-073496-4 Brassies crop seed or plant and applying to the crop, crop part, seed of said crop or the area under cultivation, an effective amount of a glyphosate herbicide, wherein said effective amount comprises a level that Is above the. recommended label use rate for the crop, wherein said effective amount Is tolerated when applied to the DP-073496-4 Brass/ca crop, crop part, seed or the area of cultivation thereof.
A "control" or "control plant' or 'control plant cell' provides a reference point for measuring changes In phenotype of the subject plant or plant cell, and may be any suitable plant or plant cell. A control plant or plant cell may comprise, for example: (a) a wild-type plant or cell, I.e., of the same genotype as the starting material for the genetic alteration which resulted In the subject plant or cell; (b) a plant or plant cell of the same genotype as the starting material but which has been transformed with a null construct (I.e., with a construct which has no known effect on the trait of interest, such as a construct comprising a marker gene); (c) a plant or plant cell which is a non-transformed segregant among progeny of a subject plant or plant cell; (d) a plant or plant cell which is genetically identical to the subject plant or plant cell but which is not exposed to the same treatment (e.g., herbicide treatment) as the subject plant or plant cell; (e) the subject plant or plant cell itself, under conditions in which the gene of interest is not expressed or (f) the subject plant or plant cell itself, under conditions in which it has not been exposed to a particular treatment such as, for example, a herbicide or combination of herbicides and/or other chemicals. In some instances, an appropriate control plant or control plant cell may have a different genotype from the subject plant or plant cell but may share the herbicide-sensitive characteristics of the starting material for the genetic alteration(s) which resulted in the subject plant or cell =
Date Recue/Date Received 2022-05-20 (see, e.g., Green, (1998) Weed Technology 12;474-477; Green and Ulrich, (1993) Weed Science 41:506-516. In other embodiments, the null segregant can be used as a control, as they are genetically identical to DP-073496-4 with the exception of the transgenic insert DNA.
Classification of herbicides (i.e., the grouping of herbicides into classes and subclasses) Is well-known in the art and includes classifications by HRAC
(Herbicide Resistance Action Committee) and WSSA (the Weed Science Society of America) (see also, Retzinger and Mallory-Smith, (1997) Weed Technology 11:384-393). An abbreviated version of the HRAC classification (with notes regarding the corresponding WSSA group) is set forth below in Table 1.
Herbicides can be classified by their mode of action and/or site of action and can also be classified by the time at which they are applied (e.g., preemergent or postemergent), by the method of application (e.g., foliar application or soil application) or by how they are taken up by or affect the plant. For example, thifensulfuron-methyl and tribenuron-methyl are applied to the foliage of a crop and are generally metabolized there, while rimsuifuron and chlorimuron-ethyl are generally taken up = through both the roots and foliage of a plant. "Mode of action* generally refers to the metabolic or physiological process within the plant that the herbicide inhibits or otherwise impairs, whereas 'site of action" generally refers to the physical location or biochemical site within the plant where the herbicide acts or directly Interacts.
Herbicides can be classified in various ways, including by mode of action and/or site of action (see, e.g., Table 1).
Often, a herbicide-tolerance gene that confers tolerance to a particular herbicide or other chemical on a plant expressing It will also confer tolerance to other herbicides or chemicals In the same class or subclass, for example, a class or subclass set forth in Table 1. Thus, in some embodiments of the invention, a transgenic plant of the invention is tolerant to more than one herbicide or chemical in the same class or subclass, such as, for example, an inhibitor of E'PO, a sulfonylurea or a synthetic auxin.
Typically, the plants of the present Invention can tolerate treatment with different types of herbicides (i.e., herbicides having different modes of action and/or different sites of action) as well as with higher amounts of herbicides than previously known plants, thereby permitting improved weed management strategies that are recommended in order to reduce the incidence and prevalence of herbicide-tolerant weeds. Specific herbicide combinations can be employed to effectively control weeds.
The invention thereby provides a transgenic brassica plant which can be selected for use in crop production based on the prevalence of herbicide-tolerant weed Date Recue/Date Received 2022-05-20 species In the area where the transgenic crop is to be grown. Methods are known in the art for assessing the herbicide tolerance of various weed species. Weed management techniques are also known in the art, such as for example, crop rotation using a crop that is tolerant to a herbicide to which the local weed species are not tolerant A number of entities monitor and publicly report the Incidence and characteristics of herbicide-tolerant weeds, including the Herbicide Resistance Action Committee (HRAC), the Weed Science Society of America and various state agencies (see, for example, herbicide tolerance scores for various broadleaf weeds from the 2004 Illinois Agricultural Pest Management Handbook) and one of skill in the art would be able to use this information to determine which crop and herbicide combinations should be used in a particular location.
These entities also publish advice and guidelines for preventing the development and/or appearance of and controlling the spread of herbicide tolerant weeds (see, e.g., Owen and Hartzler, (2004), 2005 Herbicide Manual for Agricultural Professionals, Pub. WC 92 Revised (Iowa State University Extension, Iowa State .
University of Science and Technology, Ames, Iowa); Weed Control for Corn, Brassicas, and Sorghum, Chapter 2 of '2004 Illinois Agricultural Pest Management Handbook" (University of Illinois Extension, University of Illinois at Urbana-Champaign, Illinois); Weed Control Guide for Field Crops, MSU Extension Bulletin E434 (Michigan State University, East Lansing, Michigan)).
Table 1: Abbreviated version of HRAC Herbicide. Classification I. ALS Inhibitors (WSSA Group 2) A. Sulfonylureas 1. Azimsulfuron 2. Chlorlmuron-ethyl 3. Metsulfuron-methyl 4. Nicosulfuron 5. Rimsulfuron 6. Sulfometuron-methyl 7. ThIfensulfuron-methyl = 8. TrIbenuron-methyl =
= = 9. Amidosulfuron 10. Bensulfuron-rnethyl = 11. Chlorsulfumn
12. Clnosulfuron
13. Cyclosulfamumn
14. Ethamelsulfuron-metyl
15. Elhoxysulfuron
16. Flazasulfuron
17. Flupyrsulfuron- methyl Date Recue/Date Received 2022-05-20
18. Foramsulfuron
19. Imazosulfuron
20. loclosulfuron-methyl
21. Mesosulfuron-methyl
22. Oxasulfuron
23. Plimisulfuron-methyl
24. Proailfuron
25. Pyrazosulfuron-ethyl
26. SulfostilfUron
27. Triasulfuron
28. TrMoxysulfuron
29. Triflusulfuron-methyl
30. Tritosulfuron
31. Halosulfuron-methyl
32, Flucetosulfuron B. Sulfonylaminocarbonyitriazolinones 1. Flucarbazone 2. Procarbazone C. Triazolopyrimidines =
1. Cloransulam-methyl 2. Flumetsulam 3. Diclosulam 4. Florasulam = 5. Metosulam .
6. Penoxsulam 7. Pyroxsulam D. Pyrimklinyloxy(thio)benzoates 1. Bispyribac 2. Pyriftand = 3. Pyribenzoxim = 4. Pyrithiobac 5. Pyriminabac-methyl = E. ImIdazolinones 1. Imazapyr 2. Imazethapyr 3. Imazaquin 4. Imazaplc 5. Imazamethabenz-methyl 6. Imazamox II. Other Herbicides-Active Ingredients/
Additional Modes of Action A. Inhibitors of Acetyl CoA carboxylase (ACCase) (WSSA Group 1) 1. Aryloxyphenoxypropionates ('FOPS') a. Quizalofop-P-ethyl Date Recue/Date Received 2022-05-20 b. Diclofopmethyl o. C1odinafop-propargyl d. Fenoxaprop-P-ethyl e. FluazIfop-P-butyl f. Propaquizafop = = g. Haloxyfop-P-methyl h. Cyhalofop-butyl I. Quizalofop-P-ethyl 2. Cyclohexanedlones ('D1Ms') a. Alloxydlm b. Butroxydim c. Clethodim d. Cycloxycim e. Sethoxydim Tepraloxyd1m g. Tralkoxydlm B. Inhibitors of Photosystem 11¨H RAC
Group C1/ VVSSA Group 6 1. Triazines a. Ametryne b. Atrazine C. Cyanazine d. Desmetryne e. Dimethametryne f. Prometon g. Prometryne =
Ii. Propazlne I. Simazine J. Mehra k. Terbumeton I. Terbuthylazine m. Terbutryne n. Trietazine 2. Triazinones a. Hexazlnone b. Metribuzin o. Metamitron 3. Triazollnone a. Amicarbazone 4. Uraoils a. Bromacil b. Lenaoll c. TerbacIl 5. Pyridazinones a. Pyrazon Date Recue/Date Received 2022-05-20 6. Phenyl carbamates a- Desmedipham b. Phenmedipharn C, Inhibitors of Photosystem II¨HRAC
Group C2/WSSA Group 7 1. Ureas a. Fluometuron b. LInuron c. Chiorobromuron d. Chlorotoluron e. Chlormairon f. Dimefuron g. Diuron h. Ethidimuron I. Fenuron J. Isoproturon K. Isouron I. Methabenzthiazuron m. Metobromuron n. Metoxuron o. Monollnuron p. Neburori q. Siduron r. Tebuthiuron 2. Amides a. Propanil b. Pentanochlor D. Inhibitors of Photosystem II¨HRAC
=
Group C3/ WSSA Group 6 1. Nitriles a. Brornofenoxlm b. Bromoxynil C. loxynil 2. Benzothiadlazinone (Bentazon) a. Bentazon 3. Phenylpyrkiazines a. Pyridate b. Pyridafol E. Photosystem-i-electron diversion (Bipyrldyliums) (WSSA Group 22) 1. Piquet 2. Paraquat F. Inhibitors of PPO
(protoporphyrinogen oxides.) (WSSA
Group 14) Date Recue/Date Received 2022-05-20 1. Diphenylethers a. Acifluorfen-Na b. Blfenox c. Chlornethoxyfen d. Fluoroglycofen-ethyl e. Fornesafen f. Halosafen g. Lactofen I,. Oxyfluorfen 2. Phenylpyrazoles a. Fluazolata b. Pyraflufen-ethyl 3. N-phenylphthalimides a. Clnidon-ethyl b. Flurnioxazin C. Flumiciorac-pentyl 4. Thiadiazoles =
a. Fluthiacet-methyl b. Thidiazimin 5. Oxadiazoies a. Oxadiazon b. Oxadiargyi B. Triazolinones a. Carfentrazone-ethyl b. Suifentrazone 7. Oxazolidinediones a. Pentoxazone 8. Pyritaldindiones a, Benzfendizone b. Butafenicil 9. others a. Pyrazogyl b. Profluazol G. Bleaching: Inhibition of carotenoid biosynthesis at the phytoene desaturase step (PDS) (WSSA Group 12) 1. Pyridazinones a. Norfiurazon 2. Pyridineoarboxamides a. DiflufenIcan Date Recue/Date Received 2022-05-20 b. Plcolinafen 3. Others a, Befiubutamid b. Fluridone c. Flurochioridone d. Flurtamone H. =Bleaching: Inhibition of 4-hydroxyphenyl-pyruvate-dioxygenase (4-HPPD) (WSSA Group 28) 1. Triketones a. MesotrIone b. Sulcotrione C. topremezone d. temtorlone 2. Isoxazoles a. Isoxachlottole b. isoxaflutole 3. Pyrazoles a. Benzofenap =
b. Pyrazoxyfen c. Pyrazolynate 4. Others a. Benzobicyclon I. Bleaching: Inhibition of carotenoid biosynthesis (unknown target) (WSSA
Group 11 and 13) 1. Triazoles (WSSA Group 11) a. Amftrole 2, Isoxazoildinones (WSSA Group 13) a. Clomazone 3. Ureas a. Fluometuron 3, Diphenylether a. Aclonifen Inhibition of EPsP Synthase 1. Giycines (WSSA Group 9) a, Glyphosate b. Sulfosate K. Inhibition of glutamine sy.nthetase 1. Phosphinic Acids = a Glufosinate-ammonium b. Bialaphos Date Recue/Date Received 2022-05-20 =
L. Inhibition of DHP (dihydropte. roate) synthase (WSSA Group 18) Carbamates a. Asulam M. Microtubule Assembly Inhibition (VVSSA Group 3) 1. Dinitroanilines = a Benfluralln b. Butralin C. Dinitramine d. Ethalfluralin =
e. Oryzalln f. Pendimethalin g. Trifluralin 2. Phosphoroamidates a. Amlprophos-methyl b. Butamlphos 3. Pyridines a. Dithiopyr b. Thiazopyr 4. Baniamides a. Pronamide b. Tebutam 5. Benzenedicarboxylic acids a. Chlorthat-dImethyl N. Inhibition of mitosis/microtubuie organization VVSSA Group 23) 1. Carbamates a. Chiorpropham b. Propham c. Carbetamide O. Inhibition of cell division (Inhibition of very long chain fatty acids as proposed mechanism; WSSA Group 15) 1. Chloroacetamides a. Aoetochlor = b. Alachior = c. Butachlor d. Dimethachlor e. Dimethanamid f. Metazachlor g. Nletolachlor = h. Pethoxamid 1. Pretilachlor Propachlor Date Recue/Date Received 2022-05-20 K. Proplsochlor 1. Thenylchlor 2. Acetamides a. Diphenamid = b. Napropamide c. Naproanillde 3, Oxyacetamides a. Flufenacet b. Mefenacet 4. Tetrazolinones a. FentrazamIde 5. Others a. Ani!otos b. Cafenstrole c. Indent:Ian d. Plperophos P. Inhibition of cell wall (cellulose) synthesis 1. Nitriles (WSSA Group 20) a. Dichlobenil b. Chlorthiamld 2. BenzamIdes (isoxaben = (WSSA Group 21)) a. Isoxatxm 3. TriazolocarboxamIdes (flupoxam) a. Flupoxam O. Uncoupling (membrane disruption): (WSSA Group 24) 1. Dinitrophenols a. DNOC
b. Dinoseb a. Dinoterb R. Inhibition of Lipid Synthesis by = other than ACC inhibition 1. Thiocarbamates (WSSA
Group 8) a. Flutylate b. Cydoate c. Dimepiperate d. EPTC
e. Esprocarb f. Molinate =
g. Orbencarb .
h. Pebulate Date Recue/Date Received 2022-05-20 I. Prosutfocarb J, Benthiocarb K. Tiocarbazil I. Triallate rn. Vemoiate 2. Phosphorodithioates a. Bensullde 3. Benzofurans a. Benfuresate b. Ethofumesate 4. Halogenated alkanoic acids (WSSA Group 26) a TOA
b. Daiapon Flupropanate S. Synthetic auxins (IAA-like) (WSSA
Group 4) 1. Phenoxycarboxyllc acids a Clomeprop b. 2,4-D
c. Mecoprop 2. Benzoic acids a Dicamba b. Chloramben c. TSA
3. Pyridine carboxylic acids a. Clopyralld b. Fiuroxypyr C. Picloram d. Tricyclopyr 4. Quinoline carboxylic acids a. Duinclorec b. Ouinmerac 6. Others (benazolin-ethyl) a. Benazolln-ethyl T. Inhibition of Auxin Transport 1, Phthalamates;
semicarbazones (WSSA Group 19) a. Naptalam b. Diflufenzopyr-Na U. Other Mechanism of Action 1. Arylaminopropionic acids a. Flamprop-M-methyl Date Recue/Date Received 2022-05-20 isoProPY1 2. Pyrazolium a. Difenzoquat 3. Organoarsenicals a. DSMA
b. MSMA
4. Others a. Bromobutide b. CInmethylln c. Cumyluron d. Dazomet e. Daimuron-methyl f. Dimuron g. Etobenzanid = h. Fosamine I. Metam J. OxazIclomefone k. Oleic acid I. Pelargonlc acid m. PyrIbuticarb In certain methods, glyphosate, alone or in combination with another herbicide of interest, can be applied to the DP-073496-4 Brass/0a plants or their area of cultivation. Non-limiting examples of glyphosate formations are set forth In Table 2. In specific embodiments, the glyphosate is in the form of a salt, such as, ammonium, isopropylammonium, potassium, sodium (including sesquisodium) or trimesium (alternatively named suffocate).
=
Date Recue/Date Received 2022-05-20 Table 2. Glypbosate formulations comparisons.
Active Acid Acid Herbicide by ingredi- =peva- Apply: equiva-Registered eat per lent per fl or/
lent per Trademark Mannfactuer Salt gallon gallon acre acre = Roundup Oiigival Monsanto Isopropylamine 4 3 32 0.750 Roundup Original II Monsanto Isopropylanin. e 4 3 32 0.750 Roundup Original MAX Monsanto Potassium 55 4.5 22 0.773 Roundup UllraMax Monsanto IsoproPylaMine 5 3.68 26 0.748 Roundup UltraMax II Monsanto Potassium 55 4.5 22 0.773 Roundup Weathernutis Monsanto Potassium 5.5 4.5 22 0.773 Touchdown liYagotito Disramoniam 3.7 3 32 0.750 .
Touchdown HiTech Syngenta Potassium 6.16 5 20 0.781 Touchdown Total SYngenta Potassium 5.14 4.17 24 0.782 Durango Dow AgroSciences Isopropylamine 5.4 4 24 0.750 Glyphoutax Dow AgroSciences Isopropylanine 4 3 32 0.750 Glyphomax Plus DOW AgraSciences Isopropyissoine 4 3 32 0.750 Glyphorrtax 2017 Dow AgroSciences Isopropylanine 4 3 32 0.750 C31y Star Plus Alhaughami Star Isoprogyhunine 4 3 32 0.750 City Star 5 Albaugh/Agri Star Isoprogyiamirbe 5.4 4 24 0.750 ay Star Orininal Albaugh/Agri Star Tsopropylamine 4 3 32 0.750 Gly-Fla Micro Flo rsopropylamine 4 3 32 0.750 Credit Nufhrni Isopropylamine 4 3 32 0.750 Credit Extra Nufann Lsopropylamine 4 3 32 0.750 Credit Duo 24ufarm Isopro. + 4 3 32 0.750 monoarcun.
Credit Duo Extra Norma Isopro. + 4 3 32 0.750 = mortOOM01.
Extra Credit 5 Nora= Isopropylamine 5 3.68 26 0.748 Cornerstone Agriliance Lsopropylamine 4 3 32 0.750 Comentone Plus Agrdiance Bropropylamine 4 3 32 0.750 Olyfos Chennnova Isopropylninine 4 3 32 0.750 Glyfos X-TRA Cheminova Isopropylamine 4 3 32 0.750 Rattler Helena isopropyiamine 4 3 32 0.750 Rattler Plus Helena Isopropylamine 4 3 32 0.750 Mirage OAP Isopropylamine 4 3 32 0.750 Mirage Plus UAP Isoptopylamine 4 3 32 0.750 Glyphosate. 41% Hahn Agro USA Isopropylamine 4 3 32 0.750 Buccaneer Tenkoz Isopropylamine 4 3 32 0.750 Buccaneer Plus Tenkoz Isopropylamine 4 3 32 0.750 Honcho Monsanto Isoproylamine 4 3 32 0.750 Honcho Pins Monsanto Lsopropylanese 4 3 32 0.750 Cily..4 Univ. Crop Prot. Alli Lsopropylamine 4 3 32 0.750 C31y-4 Pins Univ. Corp Prot. Allt Lsopropylamine 4 3 32 0.750 CleasOut 41 Chemical Products Lsopropylamisie 4 3 32 0.750 Tech.
ClearOut 41 Plus Chemical Products Isopropylitmine 4 3 32 0.750 Tech.
SPittre Control Solutions Isopropylamine 4 3 32 0.750 Spitfire Phis Control Solutions Isopropylanaine 4 3 32 0.750 Glyphosate 4 FarmerSavercora Isopropylarnine , 4 3 32 0.750 FS Glyphosate Plus Growsuark Isopropylamine 4 3 32 0.750 Glyphosate Original Griffin. I.LC leopropylamine 4 3 32 0.750 =
Thus, in some embodiments, a transgenic plant of the invention Is used in a method of growing a DP-073496-4 brassica crop by the application of herbicides to which the plant Is tolerant. In this manner, treatment with a combination of one of more herbicides which include, but are not limited to: acetochlor, acifluorfen and its sodium salt, aclonifen, acrolein (2-propenal), alachlor, alloxydim, ametryn, amicarbazone, amidosulfuron, aminopyraild, amitrole, ammonium sulfamate, anilofos, asulaM, atrazIne, azimsulfuron, beflubutamld, benazolin, benazolin-ethyl, =
=
=
Date Recue/Date Received 2022-05-20 bencarbazone, benfluralln, benfuresate, bensulfuron-methyl, bensulide, bentazone, .benzobicyclon, benzofenap, blfenox, bilanafos, bispyribac and its sodium salt, bromacil, bromobutide, bromofenoxim, bromoxynil, bromoxynil octanoate, butachlor, butafenacif, butamlfos, butralin, butroxydim, bufylate, cafenstrole, carbetamide, carfentrazone-ethyl, catechln, chlomethoxyfen, chloramben, chlorbromuron, chlorflurenol-methyl, chloridazon, chlorimuron-ethyl, chlorotoluron, chlorpropham, chlorsulfuron, chlorthal-dimethyl, chlorthiamid, cinidon-ethyl, cinmethylin, cinosulfuron, dethodim, clodinafop-propargyl, clomazone, clomeprop, ciopyralid, clopyrarid-olamine, cloransulam-methyl, CUH-35 (2-methoxyethyl 2-11[4-chloro-2-fluoro-5-[(1-methyl-propynyl) oxy]pheny1X3-fluorobenzoyl)aminolcarbonyl]-1-cyclohexene-1-carboxylate), cumyluron, cyanazine, cycloate, cyclosulfamuron, cycloxydim, cyhalofop-butyl, 2,4-D
and its butotyl, butyl, 'sootyl and isopropyl esters and its dimethylammonium, diolamlne and trolamine salts, daimuron, dalapon, dalapon-sodium, dazomet, Z4-DE and its dimethylammonium, potassium and sodium salts, desmedipham, desmetryn, dicamba and its diglycolammonium, dimethylammonium, potassium and sodium salts, dichlobenil, dichlorprop, didolop-methyl, diclosulam, difenzoquat metilsuffate, diflufenican, diflufenzopyr, dimefuron, =dimepiperate, dimethachlor, dimetharnetryn, dimethenamld, dimethenamld-P, dimethipin, dlmethylarsinic acid and its sodium salt, dinitramine, dinoterb, diphenamld, diquat dibromide, dithiopyr, diuron, DNOC, endothal, EPTC, esprocarb, ethaffluralin, ethametsulfuron-methyl, ethofumesate, ethoxyfen, ethoxysulfuron, etobenzanld, fenoxaprop-ethyl, fenoxaprop-P-ethyl, fentrazamide, fenuron, fenuron-TCA, flamprop-methyl, flamprop-M-isopropyl, flamprop-M-methyl, flazasulfuron, florasulam, fluazifop-butyl, fluazifop-P-butyl, flucarbazone, fluceitosulfuron, fluchloralln, flufenacet, flufenpyr, flufenpyr-ethyl, flumetsulam, flumiclorac-pentyl, flumioxazin, fluometuron, fluoroglycofen-ethyl, flupyrsulfuron-methyl and its sodium salt, flurenol, flurenol-butyl, fluridone, flurochloridone, fluroxypyr, flurtamone, fluthiacet-methyl, fomesafen, foramsulfuron, fosamine-ammonium, glufosinate, glufoslnate-ammonium, glyphosate and its salts such as ammonium, isopropylammonium, potassium, sodium (including sesquisodium) and trimesium (alternatively named sulfosate), halosulfuron-methyl, haloxyfop-etotyl, haloxyfop-methyl, hexazinone, HOK-201 (N-(2,4-clifluoropheny1)-1,5-dihydro-IV-(1-methylethyl)-5-oxo-1-[(tetrahydro-21-1-pyran-2-y1)methyl]-4H-1,2,4-triazole-4-carboxamide), imazamethabenz-methyl, imazamox, imazapic, imazapyr, imazaquin, imazaquin-ammonium, imazethapyr, imazethapyr-ammonium, imazosulfuron, indanofan, iodosulfuron-methyl, loxynil, ioxynil octanoate, ioxynil-sodium, isoproturon, isouron, isoxaben, isoxaflutole, isoxachlortole, lactofen, lenacil, linuron, maleic =
hydrazide, MCPA and its salts (e.g., MCPA-dimethylammonium, MCPA-potassium and Date Recue/Date Received 2022-05-20 MCPA-sodlum, esters (e.g., MCPA-2-ethylhexyl, MCPA-butotyl) and thioesters (e.g., MCPA-thioethyl),.MCPB and its salts (e.g., MCPB-sodium) and esters (e.g., MCPB-ethyl), mecoprop, mecoprop-P, mefenacet, mefluidide, mesosulfuron-methyl, mesotrione, metam-sodium, metamifop, metamitron, metazachlor, methabenzthiazuron, methylarsonic acid and its calcium, monoammonium, monosodium and disodium salts, methyldymron, metobenzuron, metobromuron, metolachlor, S-metholachlor, metosulam, metoxuron, metribtrzin, metsuifuron-methyl, molinate, monolinuron, naproanllide, napropamide, naptalam, neburon, nicosulfuron, norfiurazon, orbencarb, oryzalin, oxadiargyl, oxadiazon, oxasulfuron, oxazidomefone, oxyfluorfen, paraquat dichloride, pebulate, pelargonic acid, pendimethalin, penoxsulam, pentanochlor, pentoxazone, perfluidone, pethoxyamid, phenmedipham, picloram, picloram-potassium, plcollnafen, pinoxaden, piperofos, pretilachlor, prImIsulfuron-methyl, prodiamine, profoxydim, prometon, prometryn, propachlor, propanil, propaquizafop, propazine, propham, propisochlor, propoxycarbazone, propyzamide, prosulfocarb, pros ulfuron, pyracionil, pyraflufen-ethyl, pyrasulfotole, pyrazogyl, pyrazolynate, pyrazoxyfen, pyrazosulfuron-ethyl, pyribenzoxim, pyributicarb; pyriciate, pyriftalid, pyriminobac-methyl, pyrimisulfan, pyrithlobac, pyrithiobac-sodium, pyroxsulam, quinclorac, quinmerac, quinodamine, quizalofop-ethyl, quizalofop-P-ethyl, quizalofop-P-tefuryl, rimsulfuron, sethoxydim, siduron, simazine, simetryn, suicotdone, suifentrazone, sulfometuron-methyl, sulfosuifuron, 2,3,6-TBA, TCA, TCA-sodium, tebutam, tebuthiuron, tefuryltrione, tembotrione, tepraloxydim, terbacil, terbumeton, terbuthylazine, terbutryn, thenyichlor, thiazopyr, thiencarbazone, thifensulfuron-methyl, thiobencarb, tiocarbazil topramezone, tralkoxydim, tri-aliate, triasulfuron, hiazifiam, tribenuron-methyl, triclopyr, triclopyr-butotyl, triclopyr-triethylammonium, tridiphane, trietazlne, trifloxysulfuron, trifluralin, triflusulfuron-methyl, tritosuifuron and vemolate is disclosed.
Other suitable herbicides and agricultural chemicals are known in the art, such as, for example, those described in WO 2005/041654. Other herbicides also Include bioherbiddes such as Altemaria destruens Simmons, Colletotrichum gioeosporlocles (Penz.) Penz. and Saco., Orechsiera monoceras (M1I3-051), Myrotheclum verrucarie (Albertini & Schweinitz) Ditmar: Fries, Phytophthora palmlvora (Butl.) But.
and Pucclnia thtaspeos Schub.. Combinations of various herbicides can result in a greater-than-additive (I.e., synergistic) effect on weeds and/or a less-than-additive effect (i.e., safening) on crops or other desirable plants. In certain instances, combinations of glyphosate with other herbicides having a similar spectrum of control but a different mode of action will be particularly advantageous for preventing the development of = 46 Date Recue/Date Received 2022-05-20 resistant weeds. HerbicideIly effective amounts of any particular. herbicide can be easily determined by one Skilled In the art through simple experimentation.
Herbicides may be classified into groups and/or subgroups as described herein above with reference to their mode of action, or they may be classified into groups andfor subgroups in accordance with their chemical structure.
Sulfonamide herbicides have as an essential molecular structure feature a sulfonamide moiety (¨S(0)2NH¨). As referred to herein, sulfonamide herbicides particularly comprise sulfonylurea herbicides, suffonylaminocarbonyltriazolinone herbicides and triazoiopyrimidine herbicides. In sulfonylurea herbicides the sulfonamide moiety is a component in a sulfonyiurea bridge (--S(0)2NHC(0)NH(R)¨).
In suifonylurea herbicides the sutfonyi end of the sulfonyfurea bridge Is connected either directly or by way of an oxygen atom or an optionally substituted amino or methylene group to a typically substituted cyclic or acyclic group. At the opposite end of the sulfonyiurea bridge, the amino group, which may have a substituent such as methyl (R being CH) instead of hydrogen, is connected to a heterocyclic group, typically a symmetric pyrimidine or triazine ring, having one or two substituents such as methyl, ethyl, trifluOromethyl, methoxy, ethoxy, methylamlno, dimethylamino, ethyiamino and the halogens. In suffonylaminocarbonyitrlazollnone herbicides, the sulfonamide moiety is a component of a sulfonylaminocarbonyl bridge (-S(0)2NHC(0)¨), In sulfonylaminocarbonyltriazolinone herbicides the suifonyl end of the sulfonylaminocarbonyl bridge is typically connected to substituted phenyl ring. At the opposite end of the sulfonylaminocarbonyl bridge, the carbonyl is connected to the 1-position of a triazolinone ring, which is typically substituted with groups such as alkyl and alkoxy. In triazoiopyrimidine herbicides the sulfonyi end of the sulfonamide moiety is connected to the 2-position of a substituted [1,2,41triazolopyrimidine ring system and the amino end of the sulfonamide moiety Is connected to a substituted aryl, typically phenyl, group or alternatively the amino end of the sulfonamide moiety is connected to the 2-position of a substituted (1,2,4]triazolopyrImIdlne ring system and the sulfonyl end of the sulfonamide moiety is connected to .a substituted aryl, typically pyridinyl, group.
= The. methods further comprise applying to the crop and the weeds in a field a sufficient amount of at least one herbicide .to which the crop seeds or plants are tolerant, such as, for example, glyphosate, a hydroxyphenylpyruvatedioxygenase inhibitor (e.g., mesotrione or sulcotrione), a phytoene desaturase Inhibitor (e.g., diflufenican), a pigment synthesis inhibitor, sulfonamide, imidazolinone, bialaphos, phosphinothricin, azafenidin, butafenacil, sulfosate, giufosinate, triazolopyrimidine, pyrimidinyloxy(thio)benzoate or sulonylaminocarbonyitriazolinone, an acetyl Co-A
Date Recue/Date Received 2022-05-20 =
carboxyiase inhlbfior such as quizalofop-P-ethyl, a synthetic auxin such as quInclorac, KIN-485 or a protox inhibitor to control the weeds without significantly damaging the crop plants.
Generally, the effective amount of herbicide applied to the field is sufficient to selectively control the weeds without significantly affecting the crop, "Weed"
as used herein refers to a plant which is not desirable in a particular area.
Conversely, a "crop planr as used herein refers to a plant=which is desired in a particular area, such as, for example, a Brassica plant. Thus, in some embodiments, a weed is a non-crop plant or a non-crop species, while in some embodiments, a weed is a crop species which is sought to be eliminated from a particular area, such as, for example, an inferior and/or non-transgenic Brassica plant in a field planted with Brass/ca event DP-073496-4 or a non-Brass/ca crop plant in a field planted with DP-073496-4. Weeds can be classified into two major groups: monocots and dicots.
Many plant species can be controlled (i.e., killed or damaged) by the herbicides described herein. Accordingly, the methods of the invention are useful in controlling these plant species where they are undesirable ,(i.e., where they are weeds).
These plant species include crop plants as well as species commonly considered weeds, Including but not limited to species such as: blackgrass (A(opecurus myosuroldes), giant foxtail (Setarie faben), large crabgrass (Digitaria sanguine/is), Surinam grass (Brachiaria decumbens), wild oat (Avena latua), common cocklebur (Xanthium pensylvanicum), common lambsquarters (Chenopodium album), morning glory (1pomoea coccinea), pigweed (Amaranthus spp,), velvetieaf (Abutilion theophrastr), common bamyardgrass (Echinochloa crus-galb), bermudagrass (Cynodon dactylon), downy brome (Bromus tectorum), goosegrass (Eleusine id/ca), green foxtall (Setaria virld(s), Italian ryegrass (Lolium multitIorum), Johnsongrass (Sorghum halepense), lesser canarygrass (Phateris minor), windgrass (Apera splca-venti), wooly cupgrass (Erichloa vfllosa), yellow nutsedge (Cypenrs esculentus), common chickweed (Stellaria media), common ragweed (Ambrosia artemIslifolia), Kochia scoparia, horseweed (Conyza canadensls), rigid ryegrass (Latium rigidum), goosegrass (Eleucine indica), = 30 hairy fleabane (Conyza bonariensis), buckhom plantain (Plantago lanceolate), tropical splderwort (Commelina benghatensls), field bindweed (Convoivulus antens(s), purple nutsedge (Cyperus rolundus), redvine (Brunnichia ovate), hemp sesbania (Sesbania exattata), sicklepod (Senna obtusifolia), Texas blueweed (Helianthus Nana) and Devil's claws (Proboscides touisianica). In other embodiments, the weed comprises a herbicide-resistant ryegrass, for example, a glyphosate resistant ryegrass, a paraquat resistant ryegrass, a ACCase-inhibitor resistant ryegrass and a non-selective, herbicide Date Recue/Date Received 2022-05-20 resistant ryegrass. In some embodiments, the undesired plants are proximate the crop plants.
As used herein, by "selectively controlled" it is intended that the majority of weeds in an area of cultivation are significantly damaged or killed, while If crop plants are also present In the field, the majority of the crop plants are not significantly damaged. Thus, a method is considered to selectively control weeds when at least 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or more of the weeds are significantly damaged or killed, while If crop plants are also present In the field, less than 45%, 40%, 35%, 30%, 25%, 20%, 15%, 10%, 5% or 1% of the crop plants are significantly damaged or killed.
In some embodiments, a Brassica DP-073496-4 plant of the invention is not significantly damaged by treatment with a particular herbicide applied to that plant at a dose equivalent to a rate of at least 0.5, 1,2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, =
20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 110, 120, 160, 170, 200, 300, 400, 500, 600, 700, 800, 800, 1000, 2000, 3000, 4000, 5000 or more grams or ounces (1 ounce = 29.57m1) of active ingredient or commercial product or herbicide formulation per acre or per hectare, whereas an appropriate control plant is significantly damaged by the same treatment..
In specific embodiments, an effective amount of an ALS inhibitor herbicide comprises at least about 0.1, 1, 5, 10, 25,50, 75, 100, 150, 200, 250, 300, 350, 400, 450, 500, 600, 700, 750, 800, 850, 900, 950,1000, 2000, 3000, 4000, 5000 or more grams or ounces (1 ounce = 29.57m1) of active Ingredient per hectare. In other embodiments, an effective amount of an ALS inhibitor comprises at least about 0.1-50, about 25-75, about 50-100, about 100-110, about 110-120, about 120-130, about 140, about 140-150, about 150-200, about 200-500, about 500-600, about 600-800, about. 800-1000 or greater grams or ounces (1 ounce = 29.57m1) of active ingredient per hectare. Any ALS inhibitor, for example, those listed in Table 1 can be applied at these levels.
In other embodiments, an effective amount of a suifonylurea comprises at least 0.1, 1, 5, 10, 25, 50, 75, 100, 150, 200, 250, 300, 350, 400, 450, 500, 600, 700, 800, 900, 1000, 5000 or more grams or ounces (1 ounce = 29.57m1) of active Ingredient per hectare. In other embodiments, an effective amount of a sulfonylures comprises at least about 0,1-50, about 25-75, about 50-100, about 100-110, about 110-120, about 120-130, about 130-140, about 140-150, about 150-160, about 160-170, about 170-180, about 190-200, about 200-250, about 250-300, about 300-350, about 350-400, about 400-450, about 450-500, about 500-550, about 550-600, about 600-850, about 650-700, about 700-800, about 800-900, about 900-1000,. about 1000-2000 or more Date Recue/Date Received 2022-05-20 grams=or ounces (1 ounce = 29.57m1) of active Ingredient per hectare.
Representative sutfonylureas that can be applied at this level are set forth in Table 1, In other embodiments, an effective amount of a sulfonylaminocarbonylirlazollnones, triazolopyrimidines, pyrimidinyloxy(thlo)benzoates, and imidazolinones can comprise at least about 0.1, 1, 5, 10, 26, 60, 75, 100, 150, 200, 250, 300, 350, 400, 460, 500, 550, 600, 650,700, 750, 800, 850, 900, 950, 1000, 1050, 1100, 1150, 1200, 1250, 1300, 1350, 1400, 1500, 1550, 1600, 1050, 1700, 1800. 1850, 1900, 1950, 2000, 2500, 3500, 4000, 4500, 5000 or greater grams or ounces (1 ounce = 29.57i1l) active Ingredient per hectare. In other embodiments, an effective amount of a sulfonyiuminocarbonyttrlazolines, biezolopyrImidines, pyrimidinyloxy(thlo)benzoates or imldazolinones comprises at least about 01-50, about 25-75, about 50-100, about 100-110, about 110-120, about 120-130, about 140, about 140-150, about 150-160, about 160-170, about 170-180, about 190-200.
about 200-250, about 250-300, about 300-350, about 350-400, about 400-450, about 450.500, about 500-550, about 550-600, about 600-550, about 650-700, about 700-800, about 800-900, about 900-1000, about 1000-2000 or more grams or ounces (1 ounce = 29.57 ml) active Ingredient per hectare.
Additional ranges of the effective amounts of herbicides can be found, for example, in various publications from University Extension services. Sea, for example, Bernards, et at., (2006) Gulde for Weed Management in Nebraska (www.ianrpubs.uri.adu/sendlt/ec130); Regher, of al., (2005) Chemical Weed Control for Fields Crops, Pastures, Rangeldnd, and Noncropland. Kansas State University Agricultural Extension Station and Corporate Extension Service; Zollinger, el ed., (2006) North Dakota Weed Control Guide, North Dakota Extension Service and the Iowa State University Extension at wviwweeds.lastate.eduõ
In some embodiments of the Invention, glyphosate is applied to an area of cultivation andtor to at least one plant In an area of cultivation at rates between 8 'OM
= 32 ounces of acid equivalent per acre, or at rates between 10, 12, 14, 16, 18, 20, 22, 24, 26, 28 and 30 ounces of acid equivalent per acre at the lower end of the range of application and between 12, 14, 18, 18, 20, 22, 24, 28, 28, 30 and 32 ounces of acid equivalent per acre at the higher end of the range of application (1 ounce =
29.57 ml).
In other embodiments, glyphosate is applied at least at 1, 5,= 10, 20, 30, 40, 50, 60, 70, 80, 90 or greater ounce of active ingredient per hectare (1 ounce = 29.57 ml).
in some embodiments of the Invention, a sutfonylorea herbicide is applied to a *field andfor to at least one plant in a held at rates between 0,04 and 1.0 ounces of active Ingredient per acre, or at rates between 0.1, 0.2,0,4, 0.6 and 0.8 ounces of active Ingredient per acre 50 = =
=
= =
Date Recue/Date Received 2022-05-20 at the lower end of the range of application and between 0.2, 0.4, 0.6, 0.8 and 1.0 ounces of active Ingredient per acre at the higher end of the range of application. (1 ounce = 29.57 m1).
As Is known in the art, glyphosate herbicides as a class contain the same active ingredient, but the active ingredient is present as one of a number of different salts and/or formulations. However, herbicides known to inhibit ALS vary in their active ingredient as well as their chemical formulations. One of skill in the art is faniillar with the determination of the amount of active ingredient and/or acid equivalent present in a particular volume and/or weight of herbicide preparation.
In some embodiments, an ALS inhibitor herbicide is employed. Rates at which the ALS inhibitor herbicide is applied to the crop, crop part, seed or area of cultivation can be any of the rates disclosed herein. In specific embodiments, the rate for the ALS inhibitor herbicide is about 0.1 to about 5000 g al/hectare, about 0.5 to about 300 g al/hectare or about 1 to about 150 g al/hectare.
. Generally, a particular herbicide is applied to a particular field (and any plants growing in it) no more than 1, 2, 3, 4, 5, 8, 7 or 8 times a year, or no more than 1, 2, 3, 4 or 5 times per growing season.
By 'treated with a combination of or "applying a combination of herbicides to a crop, area of cultivation or field' it is intended that a particular field, crop or weed is treated with each of the herbicides and/or chemicals indicated to be part of the = combination so that a desired effect is achieved, i.e., so that weeds are selectively controlled while the crop is not significantly damaged. In some embodiments, weeds which are susceptible to each of the herbicides exhibit damage from treatment with each of the herbicides which Is additive or synergistic. The application of each herbicide and/or chemical may be simultaneous or the applications may be at different times, so long as the desired effect is achieved. Furthermore, the application can occur prior to the planting of the crop.
The proportions of herbicides used In the methods of the invention with other herbicidal active ingredients in herbicidal compositions .are generally in the ratio Of 5000:1 to 1:5000, 1000:1 to 1:1000, 100:1 to 1:100, 10:1 to 1:10 or 5:1 to 1:5 by weight. The optimum ratios can be easily determined by those skilled in the art based on the weed control spectrum desired. Moreover, any combinations of ranges of the various herbicides disclosed in Table 1 can also be applied in the methods of the Invention.
Thus, in some embodiments, the Invention provides improved methods for selectively controlling weeds in a field wherein the total herbicide application may be less than 90%, 85%, 80%, 75%, 70%, 65%, 60%, 55%, 50%, 45%, 40%, 35%, 30%, Date Recue/Date Received 2022-05-20 25%, 20%, 15%, 10%, 5% or 1% of that used in other methods. Similarly, in some embodiments, the amount of a particular herbicide used for selectively controlling weeds in a field may be less than 90%, 85%, 80%, 75%, 70%, 65%, 60%, 55%, 50%, 45%, 40%, 35%, 30%, 25%, 20%, 15%, 10%, 5% or 1% of the amount of that particular herbicide that would be used in other methods, i.e., methods not utilizing a plant of the invention.
in some embodirrionts, a DP-073496-4 Brass/ca plant of the invention benefits from a synergistic effect, wherein the herbicide tolerance conferred by the GLYAT
polypeptide and that conferred by a polypeptide providing tolerance to another herbicide ii greater than expected from simply combining the herbicide tolerance conferred by each gene separately. See, e.g., McCutchen, of al., (1997) J.
Econ.
Entotnot. 90:1170-1180; Priesler, etal., (1999) J. Econ. Entomot 92:598-603.
As used . herein, the terms 'synergy,' "synergistic; "synergistically" and derivations thereof, .such as in a "synergistic effect" or a "synergistic herbicide combination" or a "synergistic herbicide composition" refer to circumstances under which the biological activity of a combination of herbicides, such as at least a first herbicide and a second herbicide, is greater than the sum of the biological activities of the individual herbicides. Synergy, expressed in terms of a "Synergy Index (SI)," generally can be determined by the method described by Kull, eta!,, (1961) Applied Microbiology 9:533.
= 20 See also, Colby, (1967) Weeds 15:20-22.
In other instances, the herbicide tolerance conferred On a DP-073496-4 plant of the invention is additive; that is, the herbicide tolerance profile conferred by the herbicide tolerance genes is what would be expected from simply combining the herbicide tolerance conferred by each gene separately to a transgenic plant containing them individually. Additive and/or synergistic activity for two or more herbicides against key weed species will increase the overall effectiveness and/or reduce the actual amount of active ingredient(s) needed to control said weeds. Where such synergy is observed, the plant of the invention may display tolerance to a higher dose or rate of herbicide and/or the plant may display tolerance to additional herbicides or other chemicals beyond those to which it would be expected to display tolerance. For example, a DP-073496-4 Brasslca plant may show tolerance to organophosphate compounds such as insecticides and/or Inhibitors of 4-hydroxyphenylpyruvate dioxygenase.
Thus, for example, the DP-073496-4 Brassica plants of the invention, when further comprising genes conferring tolerance to other herbicides, can exhibit greater than expected tolerance to various herbicides, Including but not limited to glyphosate, ALS inhibitor chemistries and sulfonylurea herbicides. The DP-073496-4 Brassica 52 =
Date Recue/Date Received 2022-05-20 plant plants of the invention may show tolerance to a particular herbicide or herbicide combination that Is at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 12%, 16%, 17%, 20%, 22%, 25%, 27%, 30%, 35%, 40%, 46%, 50%, 55%, 60%, 65%, 70%, 80%, 90%, 100%, 125%, 150%, 175%, 200%, 300%, 400% or 500% or more higher than the tolerance of an appropriate control plant that contains only a single herbicide tolerance gene which confers tolerance to the same herbicide or herbicide combination. Thus, DP-073496-4 Brassica plants may show decreased damage from the same dose of herbicide in comparison to an appropriate control plant, or they may show the same degree of damage in response to a much higher dose of herbicide than the control plant. Accordingly, in specific embodiments, a particular herbicide used for selectively containing weeds in a field is more than 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 60%, 60%, 70%, 80%, 90%, 100% or greater than the amount of that particular herbicide that wodd be used in other methods, i.e., methods not utilizing a plant of the invention.
In the same manner, in some embodiments, a DP-073496-4 Brassica plant of the invention shows improved tolerance to a particular formulation of a herbicide active Ingredient in comparison to an appropriate control plant, Herbicides are sold commercially as formulations which typically include other ingredients in addition to the herbicide active ingredient; these ingredients are often intended to enhance the efficacy of the active ingredient. Such other ingredients can include, for example, safeners and adjuvants (see, e.g., Green and Foy, (2003) "Adjuvants: Tools for Enhancing Herbicide Performance," In Weed Biology and Management, ed. Inderjit (Kluwer Academic Publishers, The Netherlands)). Thus, a DP-073496-4 Brassica plant of the invention can show tolerance to a particular formulation of a herbicide (e.g., a particular commercially available herbicide product) that is at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 12%, 15%, 17%, 20%, 22%, 26%, 27%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 80%, 90%, 100%, 125%, 150%, 175%, 200%, 300%, 400%, 500%, 600%, 700%, 800%, 900%, 1000%, 1100%, 1200%, 1300%, 1400%, 1500%, 1600%, 1700%, 1800%, 1900% or 2000% or more higher than the tolerance of an appropriate control plant that contains only a single herbicide tolerance gene which confers tolerance to the same herbicide formulation.
In some embodiments, a DP-073496-4 Bressica plant of the invention shows improved tolerance to a herbicide or herbicide class to which at least one other herbicide tolerance gene confers tolerance as well as Improved tolerance to at least one other herbicide or chemical which has a different mechanism or basis of action than either glyphosate or the herbicide corresponding to said at least one other herbicide tolerance gene. This surprising benefit of the invention finds use in methods Date Recue/Date Received 2022-05-20 of growing crops that comprise treatment with various combinations of chemicals, Including, for example, other chemicals used for growing crops. Thus, for example, a DP-073496-4 Brassica plant may also show improved tolerance to chlorpyrifos, a systemic organophosphate insecticide. Thus, the invention also provides a DP-073496-4 Brassica plant that confers tolerance to giyphosate (i.e., a GLYAT
gene) which shows improved tolerance to chemicals which affect the cytochrome P4.50 gene, and methods of use thereof. In some embodiments, the DP-073496-4 Bressica plants also show improved tolerance to dicamba. In these embodiments, the improved tolerance to dicamba may be evident in the presence of giyphosate and a sulfonyiurea herbicide.
In other methods, a herbicide combination is applied over a DP-073496-4 Brassica plant, where the herbicide combination produces either an additive or a synergistic effect for controlling weeds. Such combinations of herbicides can allow the application rate to be reduced, a broader spectrum of undesired vegetation to be controlled, improved control of the undesired vegetation with fewer applications, more rapid onset of the herbicidal activity or more prolonged herbicidal activity.
An 'additive herbicidal composition" has a herbicidal activity that is about equal to the observed activities of the individual components. A "synergistic herbicidal combination" has a herbicidal activity higher than what can be expected based on the observed activities of the Individual components when used alone. Accordingly, the presently disclosed subject matter provides a synergistic herbicide combination, wherein the degree of weed control of the mixture exceeds the sum of control of the individual herbicides. In some embodiments, the degree of weed control of the mixture exceeds the sum of control of the individual herbicides by any statistically significant amount including, for example, about 1% to 5%, about 5% to about 10%, about 10% to about 20%, about 20% to about 30%, about 30% to 40%, about 40% to about 50%, about 50% to about 60%, about 60% to about 70%, about 70% to about 80%, about 80% to about 90%. about 90% to about 100%, about 100% to 120% or greater.
Further, a "synergistically effective amount" of a herbicide refers to the amount of one herbicide necessary to elicit a synergistic effect in another herbicide present in the herbicide composition. Thus, the term "synergist," and derivations thereof, refer to a substance that enhances the activity of an active ingredient (al), i.e., a substance in a formulation from which a biological effect Is obtained, for example, a herbicide.
Accordingly, in some embodiments, the presently disclosed subject matter provides a method for controlling weeds in an area of cultivation. In some embodiments, the method comprises: (a) planting the area with a DP-0734964 crop seeds or crop plants which also comprise polynucleotides conferring ALS-inhibitor Date Recue/Date Received 2022-05-20 tolerance; and (b) applying to the weed, the crop plants, a crop part, the area of cultivation or a combination thereof, an effective amount of a herbicide composition comprising at least one of a synergistically effective amount of glyphosate and a synergistically effective amount of an ALS inhibitor (for example, but not limited to, a sulfonylurea herbicide) or agriculturally suitable salts thereof, wherein at least one of:
(i) the synergistically effective amount of the glyphosate is lower than an amount of glyphosate required to control the weeds in the absence of the sulfonylurea herbicide;
(II) the synergistically effective amount of the ALS inhibitor herbicide is lower than an amount of the ALS inhibitor required to control the weeds in the absence of glyphosate and (iii) combinations thereof and wherein the effective amount of the herbicide composition is tolerated by the crop seeds or crop plants and controls the weeds in the area of cultivation.
In some embodiments, the herbicide composition used in the presently disclosed method for controlling weeds comprises a synergistically effective amount of glyphosate and a sulfonylurea herbicide, in further embodiments, the presently disclosed synergistic herbicide composition comprises glyphosate and a sulfonylurea herbicide selected from the group consisting of metsulfuron-methyl, chlorsulfuron and triasulfuron.
In particular embodiments, the synergistic herbicide combination further comprises an adjuvant such as, for example, an ammonium Sulfate-based adjuvant, .e.g., ADD-UPe (Wenkem., Halfway House, Midrand, South Africa). In additional embodiments, the presently disclosed synergistic herbicide compositions comprise an additional herbicide, for example, an effective amount of a pyrimidinyloxy(thio)benzoate herbicide. In some embodiments, the pyrimidinyloxy(thlo)benzoate herbicide comprises bispyribac, e.g., (VELOCITY , Valent U.S.A. Corp., Walnut Creek, California, United States of America) or an agriculturally suitable salt thereof.
= In some embodiments of the presently disclosed method for controlling undesired plants, the glyphosate is applied pre-emergence, post-emergence or pre-and post-emergence to the undesired plants or plant crops and/or the ALS
inhibitor herbicide (i.e., the sulfonylurea herbicide) is applied pre-emergence, post-emergence or pre- and post-emergence to the undesired plants or plant crops. In other embodiments, the glyphosate and/or the ALS inhibitor herbicide (i.e., the sulfonylurea herbicide) are applied together or are applied separately. In yet other embodiments, the synergistic herbicide composition is applied, e.g., step (b) above, at least once prior to planting the crop(s) of interest, e.g., step (a) above.
Date Recue/Date Received 2022-05-20 Weeds that can be difficult to control with glyphosate alone In fields where a crop is grown (such as, for example, a brassica crop) include but are not limited to the following: horserred (e.g., Conyza canadensis); rigid ryegrass (e.g., Lolium rigidum);
goosegrass (e.g., Eleuslne Indica); Italian ryegrass (e.g., Lolium multifiorum); hairy fleabane (e.g., Conyza bonarfensis); buckhom plantain (e.g., Plantago lanceolate);
common ragweed (e.g., Ambrosia artemisifolia); morning glory (e.g., /pomoea spp.);
waterhemp (e.g., Amaranthus spp.); field bindweed (e.g., Convolvulus arvensis);
yellow nutsedge (e.g., Cyperus esculentus); common Iambsquarters (e.g., Chenopodium album); wild buckwheat (e.g,, Polygon/urn convolvulus); velvetleaf (e.g., Abutilon theophrasti); kochia (e.g., Kochia scoparia) and Asiatic dayflower (e.g., Commelina spp.). In areas where such weeds are found, Brassic,a plants comprising the DP-073496-4 event and tolerance to another herbicide are particularly useful in allowing the treatment of a field (and therefore any crop growing in the field) with combinations of herbicides that would cause unacceptable damage to crop plants that = 15 did not contain both of these polynucleotldes. Plants of the invention that are tolerant to glyphosate and other herbicides such as, for example, sulfonylurea, imidazolinone, triazolopyrimidine, pyrimidinyl(thio)benzoate and/or sulfonylaminocarbonyltriazolinone herbicides in addition to being tolerant to at least one other herbicide with a different mode of action or site of action are particularly useful in situations where weeds are tolerant to at least two of the same herbicides to which the plants are tolerant. In this manner, plants of the invention make possible Improved control of weeds that are tolerant to more than one herbicide.
For example, some commonly used treatments for weed control in fields where current commercial crops (Including, for example, Brassicas) are grown include glyphosate and, optionally, 2,4-D; this combination, however, has some disadvantages. Particularly, there are weed species that it does not control well and it =
also does not work well for weed control in cold weather. Another commonly used treatment for weed control in brassica fields is the sulfonylurea herbicide dilorimuron-ethyl, which has significant residual activity in the soil and thus maintains selective pressure on all later-emerging weed species, creating a favorable environment for the growth and spread of sulfonylurea-resistant weeds. Fields may be be treated with sulfonylurea, imidazollnone, trlazolopyrimidines, pyrimidiny(thlo)benzoates and/or suifonylaminocarbonyitriazonlInone such as the sulfonylurea chlorimuron-ethyl, either alone or in combination with other herbicides, such as a combination of glyphosate and tribenuron-methyl (available commercially as express ). This combination has several advantages for weed control under some circumstances, Including the use of herbicides with different modes of action and the use of herbicides having a relatively Date Recue/Date Received 2022-05-20 short period of residual activity in the soil. A herbicide having a relatively short period of residual activity is desirable, for example, in situations where it is important to reduce selective pressure that would favor the growth of herbicide-tolerant weeds. Of course, in any particular situation where weed control is required, other considerations may be more important, such as, for example, the need to prevent the development of and/or appearance of weeds in a field prior to planting a crop by using a herbicide with a relatively long period of residual activity. Treatments that include both tribenuron-methyl and thifensulfuron-methyl may be particularly useful.
Other commonly used treatments for weed control in fields where current commercial varieties of crops (including, for example, Brasslcas) are grown include the sulfonyiurea herbicide thifensulfuron-methyl (available commercially as Harmony GTO). However, one disadvantage of thifensulfuron-methyl is that the higher application rates required for consistent weed control often cause injury to e crop growing in the same field. DP-073496-4 BrassIca plants comprising additional tolerance can be treated with a combination of glyphosate and thifensuifuron-methyl, which has the advantage of using herbicides with different modes of action.
Thus, weeds that are resistant to either herbicide alone are controlled by the combination of the two herbicides, and the improved DP-073496-4 Bressice plants would not be significantly damaged by the treatment, . Other herbicides which are used for weed control in fields where curient commercial varieties of crops (Including, for example, BrassIces) are grown are the triazolopyrimidine herbicide cloransuiam-rnethyl (available commercially as FirstRate8) and the imidazolinone herbicide imazaquin (available commercially as Sceptore). When these herbicides are used Individually they may provide only marginal control of weeds. However, may be treated, for example, with a combination of glyphosate (e.g., Roundup (glyphosate isopropyiamine salt)), imazapyr (currently available commercially as Arsenal ), chlorimuron-ethyl (currently available commercially as Classic ), quizatofop-P-ethyl (currently available commercially as Assure Ile) and fomesafen (currently available commercially as Flexstap1D).
This combination has the advantage of using herbicides with different modes of action.
Thus, weeds that are tolerant to just one or several of theses herbicides are controlled by the combination of the five herbicides. This combination provides an extremely broad spectrum of protection against the type of herbicide-tolerant weeds that might be expected to arise and spread under current weed control practices.
= Fields containing the DP-073496-4 Brassica plants with additional herbicide tolerance may also be treated, for example, with a combination of herbicides including giyphosate, rimsulfuron, and dicamba or mesotrione. This combination may be = 57 Date Recue/Date Received 2022-05-20 particularly useful in controlling weeds which have developed some tolerance to herbicides which inhibit ALS. Another combination of herbicides which may be particularly useful for weed control includes glyphosate and at least one of the following: metsulfuron-methyl (commercially available as Ally ), imazapyr (commercially available as Arsenal ), imazethapyr, imazaquin and sulfentrazone. it is understood that any of the combinations discussed above or elsewhere herein may also be used to treat areas in combination with any other herbicide or agricultural chemical.
Some commonly-used treatments for weed control in fields where current commercial crops (including, for example, Brassica) are grown include glyphosate (currently available commercially as Roundup ), rimsulfuron (currently available commercially as Resolve or Matrix ), dicamba (commercially available as Clarity ), atrazine arid mesotrione (commercially available as Cailisto0). These herbicides are sometimes used individually due to poor crop tolerance to multiple herbicides.
Unfortunately, when used individually, each of these herbicides has significant disadvantages. Particularly, the incidence of weeds that are tolerant to individual herbicides continues to increase, rendering glyphosate less effective than desired In some situations. Rimsulfuron provides better weed control at high doses which can cause injury to a crop, and alternatives such as dicamba are often more expensive than other commonly-used herbicides Some commonly-used treatments for weed control in fields where currant commercial crops are grown include glyphosate (currently available commercially as Roundup ), chlorimuron-ethyl, trlbenuron-methyl, rimsuffuron (currently available commercially as Resolve or Matrix ), imazethapyr, imazapyr and imazaquin, Unfortunately, when used individually, each of these herbicides has significant disadvantages. Particularly, the Incidence of weeds that are tolerant to individual herbicides continues to increase, rendering each individual herbicide less effective than desired in some situations. However, DP-0734964 BrassIca with an additional -herbicide tolerance trait can be treated with a combination of herbicides that would cause unacceptable damage to standard plant varieties, Including combinations of herblddes that Include at least one of those mentioned above.
In the methods of the invention, a herbicide may be formulated and applied to an area of interest such as, for example, a field or area of cultivation, in any suitable manner. A herbicide may be applied to a field In any form, such as, for example, in a liquid spray or as solid powder or granules. In specific embodiments, the herbicide or combination of herbicides that are employed in the methods comprise a tankmix or a premix. A herbicide may also be formulated, for example, as a 'homogenous granule Date Recue/Date Received 2022-05-20 blend' produced using blends technology (see, e.g., US Patent Number 6,022,552, entitled "Uniform Mixtures of Pesticide Granules"). The blends technology of US
Patent Number 6,022,552 produces a nonsegregating blend (i.e., a "homogenous granule blend") of formulated crop protection chemicals In a dry granule form that enables delivery of customized mixtures designed to solve specific problems. A
homogenous granule blend can be shipped, handled, subsampled and applied in the same manner as traditional premix products where multiple active ingredients are formulated into the same granule.
Briefly, a "homogenous granule blend' is prepared by mixing together at least two extruded formulated granule products. In some embodiments, each granule product comprises a registered formulation containing a single active ingredient which is, for example, a herbicide, a fungicide and/or an insecticide. The uniformity (homogeneity) of a "homogenous granule blend" can be optimized by controlling the relative sizes and size distributions of the granules used in the blend. The diameter of extruded granules is controlled by the size of the holes in the extruder die and a centrifugal sifting process may be used to obtain a population of extruded granules with a desired length distribution (see, e.g., US Patent Number 6,270,025).
A homogenous granule blend Is considered to be "homogenous' when it can be subsampled into appropriately sized aliquots and the composition of each aliquot will meet the required assay specifications. To demonstrate homogeneity, a large sample of the homogenous granule blend is prepared and is then subsampled into aliquots of greater than the minimum statistical sample size.Blends also afford the ability to add other agrochemicals at normal, labeled use rates such as additional herbicides (a 3"3/4th mechanism of action), fungicides, insecticides, plant growth regulators and the like thereby saving costs associated with additional applications.
Any herbicide formulation applied over the DP-073496-4 Bra ssica plant can be prepared as a "tank-mix" composition. In such embodiments, each Ingredient or a combination of ingredients can be stored separately from one another. The ingredients can then be mixed with one another prior to application.
Typically, such mixing occurs shortly before appfication. In a tank-mix process, each Ingredient, before mixing, typically is present In water or a suitable organic solvent.
For additional guidance regarding the art of formulation, see, Woods, "The Formulator's Toolbox¨
Product Forms for Modern Agriculture' Pesticide Chemistry and Bioscience, The Food-Environment Challenge, Brooks and Roberts, Eds., Proceedings of the 9th International Congress on Pesticide Chemistry, The Royal Society of Chemistry, Cambridge, 1999, pp, 120-133. See also, US Patent Number 3,235,361, Column 5, line 16 through Column 7, line 19 and Examples 1041; US Patent Number 3,309,192, Date Recue/Date Received 2022-05-20 Column 5, line 43 through Column 7, Ursa 62 and Examples 8, 12, 15, 39, 41, 52, 53, 58, 132, 138-140, 162-164, 188, 167 and 169-182; US Patent Number 2,891,855, Column 3, the 66 through Column 5, line 17 and Examples 1-4; Klingman, Weed Control as a Science, John Wiley and Sons, Inc., New York, 1961, pp 81-96 and = 5 Hance, at at, Weed Control Handbook, 8th Ed., Blackwell Scientific Publications, = Oxford, 1989 .
= The methods of the invention further allow for the development of herbicide combinations to be used with the DP-073498-4 Brassies plants. In such methods, the environmental conditions in an area of cultivation are evaluated.
Environmental condiffons that can be evaluated Include, but are not Unified to, ground and surface water pollution concerns, intended use of. the crop, crop tolerance, soil residuals, weeds present in area of cultivation, soil texture, pH of soil, amount of organic matter in soli, application equipment and tillage practices. Upon the evaluation of the environmental conditions, an effective amount of a combination of herbicides can be applied to the crop, crop part, seed of the crop or area of cultivation.
In some embodiments, the herbicide applied to the DP-073496-4 Bressica plants of the invention serves to prevent the initiation of growth of susceptible weeds and/or serve to cause damage to weeds that are growing in the area of interest in some embodiments, the herbicide or herbicide mixture exert these effects on weeds affecting crops that are subsequently planted in the area of interest (1.e., field or area of cultivation). In the methods of the invention, the application of the herbicide combination need not occur at the same time. So long as the field in which the crop is planted contains detectable amounts of the first herbicide and the second herbicide is applied at some time during the period in which the crop is in the area of cultivation, the crop is considered to have been treated with a mixture of herbicides according to ' the invention. Thus, methods of the invention encompass applications of herbicide which are preemergent,"postemergent, =preplant incorporation' and/or which Involve seed treatment prior to planting.
In one embodiment, methods are provided for coating seeds. The methods comprise coating a seed with an effective amount of a herbicide or a combination. of herbicides (as disclosed elsewhere herein). The seeds can then be planted in an area of cultivation. Further provided are seeds having a coating comprising an effective .
amount of a herbicide or a combination of herbicides (as disclosed elsewhere herein).
Treemergenr refers to a herbicide which is applied to an area of interest (e.g., .35 a field or area of cultivation) before a plant emerges visibly from the soil.
*Postemergenr refers to a herbicide which is applied to an area after a plant emerges visibly from the soil. In some Instances, the terms !preemergent- and "posternergenr Date Recue/Date Received 2022-05-20 are used with reference to a weed in an area of Interest; and in some instances these terms are used with reference to a crop plant in an area of interest When used with reference to a weed, these terms may apply to only a particular type of weed or species of weed that Is present or believed to be present in the area of interest While any herbicide may be applied in a preemergent and/or postemergent treatment, some herbicides are known to be more effective In controlling a weed or weeds when applied either preemergence or postemergence. For example, rimsulfuron has both preemergence and postemergence activity, while other herbicides have predominately preemergence (metolachlor) or postemergence (glyphosate) activity. These properties of particular herbicides are known in the art and are readily determined by one of skill in the art. Further, one of skill in the art would readily be able to select appropriate herbicides and application times for use with the transgenic plants of the Invention and/or on areas in which transgenic plants of the invention are to be planted.
'Preplent incorporation" involves the incorporation of compounds into the soil prior to planting.
' Thus, the invention provides Improved methods of growing a crop and/or controlling weeds such as, for example, "pre-planting bum down," wherein an area Is treated with herbicides prior to planting the crop of Interest in order to better control weeds. The invention also provides methods of growing a crop and/or controlling = 20 weeds which are "no-till" or "low-till" (also referred to as 'reduced tillage"). In such methods, the soli is not cultivated or is cultivated less frequently during the growing cycle In comparison to traditional methods; these methods can save costs that would otherwise be incurred due to additional Cultivation, including labor and fuel costs.
The methods of the invention encompass the use of simultaneous and/or sequential applications of multiple classes of herbicides. In some embodiments, the methods of the invention Involve treating a plant of the invention and/or an area of interest (e.g., a field or area of cultivation) and/or weed with just one herbicide or other chemical such as, for example, a sulfonylurea herbicide.
The time at which a herbicide is applied to an area of interest (and any plants therein) may be important In optimizing weed control. The time at which a herbicide is applied may be determined with reference to the size of plants and/or the stage of growth and/or development of plants in the area of interest, e.g., crop plants or weeds growing In the area. The stages of growth and/or development of plants are known in the art. Thus, for example, the time at which a herbicide or other chemical Is applied to an area of interest in which plants are growing may be the time at which some or all of the plants in a particular area have reached at least a particular size and/or stage of growth and/or development, or the time at which some or all of the plants In a Date Recue/Date Received 2022-05-20 = particular area have not yet reached a particular size and/or stage of growth and/or development.
In some embodiments, the DP-073498-4 Brassica plants of the invention show improved tolerance to postemergence herbicide treatments. For example, plants of the invention may be tolerant to higher doses of herbicide, tolerant to a broader range of herbicides, and/or may be tolerant to doses of herbicide applied at earlier or later times of development in comparison to an appropriate control plant. For example, In some embodiments, the DP-073496-4 Brassica plants of the invention show an increased resistance to morphological defects that are known to result from treatment at particular stages of development. Thus, the glyphosate-tolerant plants of the invention find use in methods involving herbicide treatments at later stages of development than were previously feasible. Thus, plants of the invention may be treated with a particular herbicide that causes morphological defects in a control plant treated at the same stage of development, but the glyphosate-tolerant plants of the invention will not be significantly damaged by the same treatment.
Different chemicals such as herbicides have different "residual' effects, i.e., different amounts of time for which treatment with the chemical or herbicide continues to have an effect on plants growing in the treated area. Such effects may be desirable or undesirable, depending on the desired future purpose of the treated area (e.g., field or area of Cultivation). Thus, a crop rotation scheme may be chosen based on residual effects from treatments that will be used for each crop and their effect on the crop that will subsequently be grown In the same area. One of skill in the art is familiar with techniques that can be used to evaluate the residual effect of a herbicide;
for example, generally, glyphosate has very little or no soli residual actMty, while herbicides that act to inhibit ALS vary in their residual activity levels. Residual activities for various herbicides are known in the art, and are also known to vary with various environmental factors such as, for example, soil moisture levels, temperature, pH and soil composition (texture and organic matter).
= Moreover, the transgenic plants of the invention provide improved tolerance to treatment with additional chemicals commonly used on crops in conjunction with herbicide treatments, such as safeners, adjuvants such as ammonium sulfonate and crop oV concentrate, and the like. The term "safener refers to a substance that when added to a herbicide formulation eliminates or reduces the phytotoxic effects of the herbicide to certain crops. One of ordinary skill in the art would appreciate that the choice of safener depends, in part, on the crop plant of interest and the particular herbicide or combination of herbicides included in the synergistic herbicide composition. Exemplary safeners suitable for use with the presently disclosed Date Recue/Date Received 2022-05-20 herbicide compositions include, but are not limited to, those disclosed In US
Patent Numbers 4,808,208; 5,502,025; 8,124,240 and US Patent Application Publication Numbers 2008/0148847; 2006/0030485; 2005/0233904; 2005/0049145;
2004/0224849; 2004/0224848; 2004/0224844; 2004/0157737; 2004/0018940;
2003/0171220; 2003/0130120; 200310078167.
The methods of the invention can Involve the use of herbicides in combination with herbicide safeners such as benoxacor, BCS (1-bromo-44(chloromethyl) suffonyabenzene), cioquintocet-mexyl, cyemetrInil, dichiormid, 2.-(dichloromettlyi)-2-methyl-1,3-diexolane (MG 191), fenchlorazole-ethyl, fenciorim, flurazole, fluxofenim, furilazole, isoxaclifen-ethyl, mafenpyr-diethyl, methoxyphenone 04-methoxy-3-rnethylphenyi)(3-methylphenyl)-methanone), naphthalic anhydride (1,8-naphthalic anhydride) and oxabetrinil to Increase crop safety. Antidotally effective amounts of the herbicide safeners can be applied at the same time as the compounds of this invention, or applied as seed treatments. Therefore an aspect of the present Invention relates to the use of a = mixture comprising glyphosate, at least one other herbicide and an antidotirily effective amount of a herbicide serener.
Seed treatment is particularly useful for selective weed control, because it physically restricts antidoting to the crop plants. Therefore a particularly useful embedment of the present Invention is a method for selectively controlling the growth of weeds in a field comprising treating the seed from which the crop Is grown with an antidotally effective amount of safener and treating the field with an effective amount of herbicide to control weeds. Antidotally effective amounts of safeners can be easily determined by one skilled in the art through simple experimentation. An antidotally effective. amount of a serener is present where a desired plant is treated with the safener so that the effect of a herbicide on the plant is decreased in comparison to the effect of the herbicide on a plant that was not treated with the safener, generally, an antidotally effective amount of safener prevents damage or severe damage to the plant treated with the safener. One of skill In the art is capable of determining whether the use of a serener is appropriate and determining the dose at which a safener should be administered to a crop.
In specific embodiments, the herbicide or herbicide combination applied to the plant of the invention acts as a safener. In this embodiment, a first herbicide or a herbicide mixture Is applied at an antidotally effect amount to the plant.
Accordingly, a method for controlling weeds In an area of cultivation is provided. The method comprises planting the area with crop seeds or plants which compose a first polynucleotide encoding a polypedide that can confer tolerance to giyphosate 63, =
=
Date Recue/Date Received 2022-05-20 operably linked to a promoter active in a plant; and, a second polynucleotIde encoding an ALS inhibitor-tolerant polypeptlde operably linked to a promoter active in a plant. A
combination of herbicides comprising at least an effective amount of a first and a second herbicide is applied to the crop, crop part, weed or area of cultivation thereof.
The effective amount of the herbicide combination controls weeds; and, the effective = amount or the first herbicide is not tolerated by the crop when applied alone when compared to a control crop that has not been exposed to the first herbicide;
and, the effective amount of the second herbicide is sufficient to produce a safening effect, wherein the safening effect provides an increase in crop tolerance upon the application of the first and the second herbicide when compared to the crop tolerance when the first herbicide is applied alone.
In specific embodiments, the combination of safening herbicides comprises a first ALS inhibitor and a second ALS inhibitor. In other embodiments, the safening effect is achieved by applying an effective amount of a combination of glyphosate and at least one ALS inhibitor chemistry. Such mixtures provides increased crop tolerance (i.e., a decrease In herbicidal injury). This method allows for increased application rates of the chemistries post or pre-treatment. Such methods find use for increased control of unwanted or undesired vegetation. In still other embodiments, a safening affect is achieved when the DP-073496-4 brassica crops, crop part, crop seed, weed or area of cultivation is treated with at least one herbicide from the sulfonylurea family of chemistry in combination with at least one herbicide from the imidazolinone family.
This method provides increased crop tolerance (i.e., a decrease In herbicidal injury).
In specific embodiments, the sulfonylurea comprises rimsulfuron and the imidazolinone comprises imazethapyr. In other embodiments, glyphosate Is also applied to the crop, crop part or area of cultivation.
As used herein, an 'adjuvant" Is any material added to a spray solution or formulation to modify the action of an agricultural chemical or the physical properties of the spray solution. See, for example, Green and Foy, .(2003) "Adjuvants: Tools for Enhancing Herbicide Performance," in Weed Biology and Management, ed. lnderjit (Kluwer Academic Publishers, The Netherlands). Adjuvants can be categorized or subclassified. as activators, acidifiers, buffers, additives, adherents, antiflocculants, antifoamers, defoamers, antifreezes, attractants, baste blends, chelating agents, cleaners, colorants or dyes, compatibility agents, cosolvents, couplers, crop oil concentrates, deposition agents, detergents, dispersants, drift control agents, emulsifiers, evaporation reducers, extenders, fertilizers, foam markers, formulants, inerts, humectants, methylated seed oils, high load COCs, polymers, modified vegetable oils, penetrators, repellents, petroleum oil concentrates, preservatives, Date Recue/Date Received 2022-05-20 rainfast agents, retention aids, solubillzers, surfactants, spreaders, stickers, spreader stickers, synergists, thickeners, translocatlan aids, uv protectants, vegetable oils, water conditioners and wetting agents.
In addition, methods of the invention can comprise the use of a herbicide or a mixture of herbicides, as well as, one or more other insecticides, fungicides, nematocides, bactericides, acaricides, growth regulators, chemosterilants, semiochemicals, repellents, attractants, pheromones, feeding stimulants or other biologically active compounds or entomopathogenic bacteria, virus or fungi to form a multi-component mixture gMng an even broader spectrum of agricultural protection.
Examples of such agricultural protectants which can be used in methods of the Invention include: Insecticides such as abamectin, acephate, acetamiprid, amidoflumet (8-1955), avermectin, azadirachtin, azinphos-methyl, bifenthrin, bifenazate, buprofezin, carbofuran, cartap, ohlorrenapyr, chlorfkaazuron, chlorpyrifos, chlorpyrifos-methyl, chromafenozide, ciothianidin, cyflumetofen, cyfluthrin, beta-cyfluthrin, cyhaiothrin, lambda-cyfialothrin, cypermethrin, cyromazine, deltamethrin, diafenthiuron, dlazinon, dieidrin, diflubenzuron, dimefluthrin, dimethoate, dinotefuran, diofenolan, emamectin, endosulfan, esfenvalerate, ethiprole, fenothiocarb, fenoxycarb, fenpropathrin, fenvalerate, flpronil, flonicamid, flubendiamide, flucythrinate, tau-fluvalinate, flufenerlm (UR-50701), flufenoxuron, fonophos, halofenozide, hexaflumuron, hydramethyinon, imidacloprid, indoxacarb, isofenphos, lufenuron, malathion, metaflurplzone, metaldehyde, methamidophos, methidathion, methomyl, methoprene, methoxychlor, metofluthrin, monocrotophos, methoxyfenozide, nitenpyram, nithiazine, novaluron, noviflumuron (XDE-007), oxamyl, parathion, parathion-methyl, permethrin, phorate, phosalone, phosmet, phosphamidon, pirimicarb, profenofos, profluthrin, pymetrozine, pyrafluprole, pyrethrtn, pyridalyi, pyriprole, pyriproxyfen, rotenone, ryanddine, spinosad, spirodiclofen, spiromesifen (BSN 2060), spirotetramat, suiprofos, tebufenozide, teflubenzuron, tefluthrin, terbufos, tetrachlorvinphos, thiacloprid, thiamethoxam, thiodicarb, thiosultap-sodium, tralomethrin, triazamate, trichlorfon and triflumuron; fungicides such as acibenzolar, aldimorph, amisulbrom, azaconazole, azoxystrobln, benalaxyl, benomyl, benthiavalicarb, benthiavalicarb-isopropyl, binomial, biphenyl, bitertanol, blasticidin-S, Bordeaux mixture (Tribasic copper sulfate), boscalidinicobifen, bromuconatole, bupirimate, buthiobate, carboxln, carpropamid, oaptafol, captan, carbendatim, chloroneb, chlorothalonil, chiozolinate, clotrimazole, copper oxychlorlde, copper salts such as copper sulfate and copper hydroxide, cyazofamid, cyflunamid, cymoxanil, cyproconazole, cyprodln11, dichlofluanld, diclocymet, diclomezine, dicloran, diethofencarb, difenoconazole,. dimethomorph, dimoxystrobin, diniconazole, Date Recue/Date Received 2022-05-20 cilniconazole-M, dinocap, discostrobin, dithianon, dodemorph, dodine, econazole, etaconazole, edifenphos, epoxiconazole, ethaboxam, ethirimol, ethridiazole, famoxadone, fenamidone, fenarimol, fenbuconazole, fencaramid, fenfuram, fenhexamicle, fenoxanil, fenpiclonil, fenpropidin, fenpropimorph, fentln acetate, fentin hydroxide, ferbam, ferfurazoate, feiimzone, fluazinam, fludioxonil, flumetover, fluoplcollde, fluoxastrobin, fluquinconazole, fluquinconazole, flusilazole, flusulfamide, flutolanil, flutriafol, folpet, fosetyl-aluminum, fuberidazole, furalaxyl, furametapyr, hexaconazoie, hymexazole, guazatine, hrnazalil, imibenconazole, iminoctadlne, iodicarb, ipconazole, iprobenfos, tprodione, iprovalioarb, isoconazole, isoprothiolane, kasugamycln, kresoxim-methyl, mancozeb, mandipropamid, maneb, mapanipyrin, = mefenoxam, mepronil, metalaxyl, metconazole, methasulfocarb, metiram, metominostrobinflenominostrobin, mepanipyrim, metrafenone, miconazole, myclobutanil, neo-asozin (ferric methanearsonate), nuarimol, octhilinone, ofurace, orysastrobin, oxadixyl, oxolinic add, oxpoconazole, oxycarboxin, paclobutrazol, penconazole, pencycuron, penthiopyrad, perfurazoate, phoSphonlc acid, phthalide, picobenzamid, plcoxystrobin, polyoxin, probenazole, brochloraz, procymidone, propamocarb, propamocarb-hydrochlorlde, propiconazole, propineb, proquinazid, prothioconazole, pyraclostrobin, pryazophos, pyrifenox, pyrimethanil, pyrifenox, pyrrolnitrine, pyroquilon, quInc,onazole, quinoxyfen, quintozene, silthlofam, simeconazole, spiroxamine, streptomycin, sulfur, tebuconazole, techrazene, tecloftalam, tecnazene, tetraconazole, thlabendazole, thifluzamIde, thlophanate, thiophanate-methyl, thlrarrt, tiadinil, toiclofos-methyl, tolyfluanid, triadimefon, triadimenol, trlarimol, trlazoxide, tridemorph, trimoprhamide tricyclazole, trifloxystrobin, triforine, triticonazole, uniconazole, validamycin, vindozolln, zineb, ziram, and zoxamide; nematocides such as aldicarb, oxamyl and fenamiphos; bactericides such as streptomycin; acaricides such as amitraz, chinomethionat, chlorobenzilate, cyhexatin, dicofot, dienochlor, etoxazole, fenazaquin, fenbutalln oxide, fenpropathrin, fenpyroximate, hexythiazox, propargite, pyridaben and tebufenpyrad and biological agents including entomopathogenic bacteria, such as Bacillus thuringlensis subsp.
Aizawal, Bacillus ihuringlensis subsp. Kurstaki, and the encapsulated delta-endotoxins of Bacillus thuringlensis (e.g., Cellcap, MPV, MPVII); entomopathogenic fungi, such as green muscardine fungus; and entomopathogenic virus including baculovirus, nucleopolyhedro virus (NPV) such as HzNPV, AfNPV; and granuiosis virus (GV) such as CpGV. The weight ratios of these various mixing partners to other compositions (e.g., herbicides) used in the methods of the invention typically are between 100:1 and 1:100, or between 30:1 and 1:30, between 10;1 and 1:10, or between 4:1 and 1:4.
Date Recue/Date Received 2022-05-20 =
The present invention also pertains to a composition comprising a biologically effective amount of a herbicide of Interest or a mixture of herbicides, and an effective amount of at least one additional biologically active compound or agent and can further comprise at least one of a surfactant, a solid diluent or a liquid diluent.
Examples of such biologically active compounds or agents are: insecticides such as abameotin, acephate, acetamiprld, amidoflumet (S-1955), avermectin, azadirachtin, azinphos-methyl, bifenthrin, binfenazate, buprofezin, carbofuran, chlorferiapyr, chlorfluazuron, chlorpyrifos, chlorpyrifos-methyl, chromafenozide, clothianidin, cyfluthrin, beta-cyfiuthrin, cyhalothrin, lambda-cyhalothrin, cypermethrin, cyromazine, deltamethrin, diafenthluron, diazinon, dfflubenzuron, dimethoate, diofenolan, emamectin, endosulfan, esfenvalerate, ethiprole, fenothicarb, fenoxycarb, fenpropathrin, fenvalerate, flpronil, flonicamid, flucythrinate, tau-fluvalinate, flufenerim (UR-50701), flufenoxuron, fonophos, halofenozide, hexaflumuron, imidacloprid, indoxacarb, Isofenphos, lufenuron, malathion, metaldehyde, methamidophos, methidathion, methomyl, methoprene., methoxychlor, monocrotophos, methoxyfenozide, nithiazin, novaluron, noviflumuron (XDE-007), oxamyl, parathion, parathion-methyl, permethrin, phorate, phosalone, phosmet, phosphamidon, pirimicarb, profenofos, pymetrozlne, pyrIdalyi, pyriproxyfen, rotenone, spinosad, splromesifin (BSN 2060), sulrxofos, tebufenozide, teflubenzuron, tefluthrin, terbufos, tetrachlorvinphos, thlacloprid, thiamethoxam, thiodicarb, thiosultap-sodium, tralomethrin, trichlorfon and triflumuron;
fungicides such as acibenzolar, azoxystrobin, benomyl, blasticidin-S, Bordeaux mixture (tribasic copper sulfate), bromuconazola, carpropamid,..captafol, captan, carbendazim, chloroneb, chlorothalonil, copper oxychloride, copper salts, cyflufenamid, cymoxanil, cyproconazole, cyprodinii, (S)-3,5-dichloro-N-(3-chloro-1-ethyl-1-methyl-2-oxopropy1)-4-methylbenzamide (RH 7281), diclocymet (5-2900), diclomezine, dicloran, difenoconazole, (S)-3,5-dihydro-5-methy1-2-(methylthio)-phenyl-3-(phenyl-amino)-4H-imidazol-4-one (RP 407213), dimethomorph, dimoxystrobin, dinlconazole, diniconazole-M, dodine, edifenphos, epoxiconazole, famoxadone, fenamidone, fenarimol, fenbuconazole, fencaramid (5ZX0722), fenpiclonil, fenpropidin, fenpropimorph, fentin acetate, fentin hydroxide, fluazinam, fludioxonil, flumetover (RPA 403397), flumorf/flumorlin (SYP-L190), fluoxastrobin (HEC
6725), fluquinconazole, flusilazole, flutolanil, flutriafol, folpet, fosetyl-aluminum, furalaxyl, furametapyr (3-82658), hexaconazole, ipconazole, iprobenfos, iprodione, = isoprothiolane, kasugamycin, kresoxim-methyl, mancozeb, maneb, mefenoxam, mepronil, metalaxyl, metconazole, metomlno-strobinffenominostrobin (SSF-126), metrafenone (AC375839), myclobutanll, neo-asozin (ferric methane-arsonate), nicobifen (BAS 510), orysastrobin, oxadixyl, penconazote, pencycuron, probenazole, Date Recue/Date Received 2022-05-20 prochloraz. propamocarb, propiconazole, proquinazId (DPX-KQ926), prothioconazole (JAU 6476), pyrifenox, pyraclostrobln,. pyrimethanil, pyrowilon, quinoxyfen, = spiroxamine, sulfur, tebuconazole, tetraconazole, thiabendazole, thifluzamide, thiophanate-methyl, thiram, tiadlnil, trladlmefon, trladimenol, tricyclazole, trifloxystrobin, triticonazole, validamycin and vinciozolin; nematocides such as aldicarb, oxamyl and fenamiphos; bactericides such as streptomycin; acaricides such .as amftraz, chinomethionat chlorobenzilate, cyhexatin, dicofol, dienochlor, etoxazole, fenazaquin, fenbutatin oxide, fenpropathrin, fenpyroximate, hexythlazox, propargite, pyriciaben and tebufenpyrad; and biological agents including entomopathogenic bacteria, such as Bacillus thuringlensis subsp. Aizawal, Bacillus thutingiensis subsp. Kurstaki, and the encapsulated delta-endotoxins of Bacillus thuringiensis (e.g., Cellcap, MPV, MPVII);
entomopathogenic fungi, such as green muscardine fungus; and entomopathogenic virus including baculovirus, nucleopolyhedro virus (NPV) such as lizNPV, AfNPV
and granulosis virus (GV) such as CpGV. Methods of the invention may also comprise the use of plants genetically transformed to express proteins toxic to invertebrate pests (such as Bacillus thuringiensis delta-endotoxins). In such embodiments, the effect of exogenously applied invertebrate pest control compounds may be synergistic with the expressed toxin proteins.
General references for these agricultural protectants .include The Pesticide Manual, 13th Edition, Tomlin, Ed., British Crop Protection Council, Farnham, Surrey, U.K., 2003 and The BloPesticide Manual, rt Edition, Copping, Ed., British Crop Protection Council, Farnham, Surrey, U.K., 2001.
In certain instances, combinations with other invertebrate pest control compounds or agents having a similar spectrum of control but a different mode of action will be particularly advantageous for resistance management. Thus, compositions of the present invention can further comprise a biologically effective amount of at least one additional invertebrate pest control compound or agent having a similar spectrum of control but a different mode of action. Contacting a plant genetically modified to express a plant protection compound (e.g., protein) or the locus of the plant with a biologically effective amount of a compound of this invention can also provide a broader spectrum of plant protection and be advantageous for resistance management.
= Thus, methods of the invention employ a herbicide or herbicide combination and may further comprise the use of insecticides and/or fungicides, and/or other agricultural chemicals such as fertilizers. The use of such combined treatments of the Invention can broaden the spectrum of activity against additional weed species and suppress the proliferation of any resistant biotypes.
Date Recue/Date Received 2022-05-20 Methods of the invention can further comprise the use of plant growth regulators such as aviglycine, N-(phenylmethyl)-1H-putin-6-amine, ethephon, epocholeone, gibberellic acid, gibbereMn A4 and A7, herein protein, mepiquat chloride, prohexadione calcium, prohydrojasmon, sodium nitrophenolate and trinexapac-methyl and plant growth modifying organisms such as Bacillus cams strain SP01.
Embodiments of the present invention are further defined in the following Examples. It should be understood that these Examples are given by way of Illustration only. From the above discussion and these Examples, one skilled in the art can ascertain the essential characteristics of this Invention, and without departing from the spirit and scope thereof, can make various changes and modifications of the embodiments of the invention to adapt it to various usages and conditions.
Thus, various modifications of the embodiments of the invention, in addition to those shown and described herein, will be apparent to those skilled in the art from the foregoing description. Such modifications are also intended to fall within the scope of the appended claims.
EXPERIMENTAL
The following abbreviations are used in describing the present invention.
ALS acetolaotate synthase protein bp base pair g1ya14621 glyphosate acetyltransferase gene GLYAT4621 glyphosate acetyltransferase protein zrn-als ' wild type acetolactate synthase gene from brassica zrn-hra modified version of acetolactate synthase gene from brassica kb kilobase PCR polymerase chain reaction UTR untranslated region Example 1. Insert and Flanking Border Sequence Characterization of Brassica Event OP0-73496-4 Brassica (Brassica napus L.) has been modified by the Insertion of the glyphosate acetyltransferase gene (g1yat4621) derived from Bacillus licheniformis and optimized by gene shuffling. Plasmld PHP28181 contains an expression cassette as further described hereafter.
Date Recue/Date Received 2022-05-20 DNA construct PHP28181 was made by cloning the GAT4621:PINII TERM
fragment excised from DNA construct pZSL149 with BamHI and Mfel double digestion downstream to the AT-UBC110 promoter of DNA construct Q0272 in the same BamHI
and Mfel sites using T4 DNA ligase (New England Lab). The resulting PHP28181 DNA contains the expression cassette: AT-UBQ10 (DUPONT) PRO:GAT4621:131N111 TERM. See, Figure 1 and Figure 2.
The 2112 bp PHP28181A DNA fragment was prepared from plasmid PHP28181 with HindiII and Noti restriction enzyme double digestion. The digested plasmid DNA was resolved in a 1% agarose gel by electrophoresis. The DNA band of the correct size was excised and DNA fragment was extracted using a Qiagen DNA
fragment extraction kit (Qiagen). DNA fragment purity was checked by PCR with a series of dilutions of amp+ positive control DNA since the PHP28181 plasmid contains an amp+ gene in its backbone. DNA fragment concentration was measured spectrophotornetrically and confirmed by comparing to DNA low mass markers (InVitrogen) in an agarose gel.
Transformation was accomplished essentially as described in Chen and Tulsieram, US Patent Application Publication Number 2007/0107077. Buds were collected from donor line N81822BC and sterilized. Buds were then homogenized, filtered, and washed to collect the microspores. The resultant microspore suspension was adjusted to a specified density and cultured for 2 days. Embryogenic microspores were then isolated via gradient centrifugation and cultured.
Gold particles coated with the PI-11328181A DNA fragment were used for transformation. Biolistic transformation was carried out using the PDS-1000/He Particle Delivery System (Bio-Fiad, Hercules, CA) as described by Klein, et al., (1987) Nature 327:70-73. Transformed embryogenic microspores were cultured in fresh medium In dark conditions for 10-12 days, then under dim light for 1-3 weeks.
Green embryos were transferred to fresh medium and cultured for two weeks to select for glyphosate tolerance. Germinated shoots and/or plants were transferred to growth medium supplemented with glyphosate.
The g/yat4621 gene was derived from the soil bacterium Bacillus licheniformis and was synthesized by a gene shuffling process to optimize the acetyitransferase activity of the GLYAT4621 enzyme (Castle, et at, (2004) gc(ence 304:1161-1164).
The inserted fragment (Figure 3) from this plasmid contains the g1yet4621 gene =
cassette. The expressien of the glyat4621 gene is controlled by the UBQ10 regulatory region from Arabldopsis and the grill terminator (see, Table 4). A summary of the transformation fragment of plasmid PHP28181 is shown in Table 4. The genetic =
=
Date Recue/Date Received 2022-05-20 elements of plasmid PHP28181 used In the creation of DP-073496-4 are shown in Table 3.
Table 3: Description of Genetic Elements in Plaantid P11P28181 ,=1.--"..;';-:r.'zL =
- = ' - 7' -."
- f=, 4 ;
Traa*rmation = -'1F,L"..7',e.4 'f.R.Pr-4-41V 1 to 2112 See Table 4 for information on ?rapidest ¨ PH-p281 the elements in this region .
:AN,rt=
git'srAss;
includes DNA from various sources for Plasmid 2113 to 4770 elements 2658 plasmid construction and Construct below plasraid replication ,=`===7=,- = ,==-=*:=,;"::,w,",..41,:',-57: 0-lactamase gene coding for =
amoicalin resistance from E eIi 2736 to 3596 bla (ApR) 861 (Sutriliffe, 1978) ((amsch-Perron, 1441 153'4! at, 1984) ,g,74 Hae II fragment containing bacterial f. = ,:=,2.41-' -µ
'I 4170 to 4539 colE1 on370 origin of replication region (colE1 V 7 , -tt = derived) (Tomizawa et id, 1977) =
=
Date Recue/Date Received 2022-05-20 Table 4: Description of Genetic Elements in the Tranaformation Fragment *WI M.1.1NiniVit-VarigiMfeitir:06,ANWW.,02AN, talW
. = -:
AP* = 1:14:6010a-afrAliPtiMil7-70. F.";A:14.At0,711VF:41-;;ZPi4 ' TP;r4"1:44P*'- 4!;'41142tjagg;TAZIA7N0-#4.farrr,"e;:'4-itAg.
A'rc.fAkr-fk.-*'ga'tiS¶'Pr4Sr!Ftk,kk,:,..LdtftA.zdiarr=':V:*Afaaa'i:k:"g'gre6t'X'!SP':Pr';:.1 .:::V:i'5C':';?
Polyiinker - I to 7 7 Region required for cloning genetic elements Region Version of the promoter region from Arabidopeis to 1312 UB,Q1 0 UBal 0 polythiquifin gene (Norris et eV, Promoter 1993) developed by the P... T. &Pont de Nemours and Company Polylinker 1313 to 1335 23 Region required for cloning genetic elements Region 1336 to 1779 4062f Gene Synthetic glyphosate N-acetyltransferase gene (Castle et al., 2004; Siehl et al., 2007) linker 1780 to 1796 Poly 17 Region required for cloning genetie elements Region 1797 t 2106 310 Terminator region from Sokoloff tuberosum proteinase o Terminator inhibitor II gene (Keil et ed, 1986; An et al, 1989) linker 2107 to 2112 Poly 6 Region required for cloning genetic elements Region The nucleotide sequence of the inserted DNA in the DP-0734964 event has been determined. PCR amplification of the unique junctions spanning the introduced genetic elements can distinguish DP-073496-4 plants from their non-genetically-modified counterparts and can .be used to screen for the presence of the inserted DNA, even at very low concentrations. Described below is a construct-specific poiymerase chain reaction (PCR) assay on genomic DNA from DP-073496-4 Brassica.
Specifically, genomic DNA from the test substance (plant material of event DP-0764954) and the control substance (plant material of a non-genetically modified Brassica with a genetic background representative of the event background) is isolated and subjected to qualitative PCR amplification using a construct-specific primer pair.
The PCR products are separated on 1.6% or 2% agarose gels to confirm the presence of the inserted construct in the genomic DNA generated from the test substance, and absence in the genomic DNA generated from the control substance. A reference = standard (100 base pair DNA Ladder; invitrogen Corporation Catalog #
10380-012) is used to determine the PCR product size.
Test and control samples are harvested from plant's. Genomic DNA extraction from the test and control tissues is performed using a standard urea extraction protocol, if leaf tissue. Genomic DNA from the test and control samples is isolated using Wizards Magnetic 96 DNA Plant System (Promega Corporation Catalog #
FF3760), if seed tissue. GenomIc DNA Is quantified on a spectrofiuorometer using .72 Date Recue/Date Received 2022-05-20 PicoGreene reagent (Molecular Probes, Inc., Eugene, OR)= and/or visualized on an agarose gel to confirm quantitation values and to determine the DNA quality.
Genomic DNA isolated from plant material .of event DP-073496-4 and control samples is subjected to PCR amplification (PCR Master Mix Catalog #7505 from Promega Corporation) utilizing a construct-specific primer pair which spans at least a portion of the glyat4621 coding region, and allows for the unique identification of maize event DP-073496-4. A second primer set is used to amplify an endogenous gene as a positive control for PCR amplification. The PCR target site and size of the expected PCR product for each primer set are compared to the observed results.
Example 2. Characterization of Event DP-073496-4 by Southern Blot Southern blot analyses (Southern, 1975) are performed to investigate the number of sites of insertion of the transforming DNA, the copy number and functional integrity of the genetic elements and the absence of plasmid backbone sequences.
The method used is described generally as follows. Genomic DNA is extracted from lyophilized tissue sampled from DP-073496-4 Brassica and non-genetically-modified control plants. Genomic DNA is digested with restriction endonuclease enzymes and size-separated on an agarose. gel. A molecular weight marker is run alongside samples for size estimation purposes. DNA fragments separated on agarose gel are depurinated, denatured and neutralized in situ and transferred to a , nylon membrane. Following transfer to the membrane, the DNA is bound to the membrane by UV crosslinking. Fragments homologous to the glyat4621 gene are generated by PCR from plasmid PHP28181, separated on an agarose gel by size, exsized and purified using a gel extraction kit, Labeled probe is hybridized to the = 25 target DNA on the .nylon membranes for detection of the specific fragments. Washes after hybridization are carried out at high stringency. Blots are exposed to X-ray film for one or more time points to detect hybridizing fragments and visualize molecular weight markers.
Example 3. Exaression of the insert Expression of the G1YA14621 protein is evaluated using leaf tissue collected from transgenic plants. For example, four fresh leaf punches may be collected and ground in sample extraction buffer using a GenoGrinder (Spex Certiprep). Total Extractable Protein (TEP) can be determined using the Blo-Rad Protein assay, which is based on the Bradford dye-binding procedure. Sample extracts may be diluted in sample extraction buffer for ELISA analysis.
=
Date Recue/Date Received 2022-05-20 The levels of expression of the GAT4621 protein in DP-073496-4 Brassies can be determined by quantitative enzyme linked immunosorbent assay (ELISA) of samples obtained from multiple field trial locations. Replicate seed samples (three replicates) may be obtained from DP-073496-4 plants treated with the maximum recommended label rate of Touchdown Total glyphosate herbicide (500 g/I
glyphosate as potassium salt; 0.60-1.35 I/ha), applied at the cotyledon to 8-leaf stage, as this represents a likely commercial cultivation scenario.
Another way to verify, the expression of the insert in DP-073496-4 E3rass1ca plants is to evaluate the transformed plants tolerance to glyphosate.
Muitigenerational stability and Within-generation segregation of the herbicide tolerant trait conferred by expression of the GAT4621 enzyme will be confirmed using a functional assay for herbicide tolerance. Tests are conducted on at least three generations of plant material. Herbicide injury may be scored as described in Table 5.
15Table 5: The 0 to 100 crop response rating system for herbicide injury 1-`
1.4 0 No Effect No crop reduction or injury 10 Slight crop discoloration or stunting Slight Effect Some crop discoloration, stunting, or stunt loss Crop injury more pronounced, but not lasting Moderate injury, drop usually recovers Moderate Effect Crop injury more lasting, recovery doubtful Lasting crop injury, no recovery Heavy crop injury and stand loss Severe Effect Crop nearly destroyed - A few surviving plants Only occasional live crop plants left 100 Complete Effect Complete crop destruction Example 4. Construct Specific PCR Analysis of Brassica Event DP-073496-4 Genomic DNA isolated from leaf of DP-073496-4 canola (12F2 generation) and 20 control vandal (non-genetically modified) was subjected to PCR
amplification (Roche High Fidelity PCR Master Kit, Roche Catalog # 12140314001) utilizing the construct-specific primer pair (09-0-3290/09-0-3288) which spans the ubiquitin promoter and the 98t4621 gene cassette (Figure 4). A second primer set (09-0-2812/09-0-2813) was used to amplify the endogenous canola FatA gene as a positive control for PCR
25 amplification. The PCR target site and size of the expected PCR
product for each primer sets are shown in Table 8. PCR reagents and reaction conditions are shown In Table 9. The primer sequences used in this study are listed in Table 10. In this study, 100 ng of leaf genomic DNA was used in all PCR reactions.
Date Recue/Date Received 2022-05-20 A PCR product of approximately 600 bp in size amplified by the construct-specific primer set 09-0-3290/09-0-3288 was observed in PCR reactions using plasmid PHP28181 (10 rig) as a template and three DP-073496-4 canola plants, but absent in three control canola plants and the no-template control (Figure 5), Samples were loaded as shown in Table 6.
= Table 6 Lane Sample = 1 Low Mass Molecular Weight Marker 2 Blank 3 Non-Genetically Modified canals C1 4 Non-Genetically Modified canola C2 5 = Non-Genetically Modified canola C3 6 DP-073496-4 canola T1 7 DP-073496-4 canola T2 8 DP-073496-4 canola T3 9 NT Control 10 _______________ Plasmic! PHP28181 11 Blank 12 Low Mass Molecular Weight Marker These results correspond with the expected PCR product size (675 bp) for samples containing DP-073496-4 canola genomic DNA. A PCR product approximately 450 bp in size was observed for both DP-073496-4 canola and control mole plants following PCR reaction with the primer set 09-0-2812/09-0-2813 for detection of the endogenous FatA gene (Figure 6): Samples were loaded as shown in Table 7.
Table 7 Lane Sample 1 Low Mass Molecular Weight Marker 2 Blank 3 Non-Genetically Modified canola C1 4 Non-Genetically Modified canola C2 5 Non-Genetically Modified canola C3 6 DP-073496-4 canola T1 7 DP-073496-4 canola T2 DP-073496-4 canola T3 NT Control 10 Plasmid PHP28181 11 Blank 12 Low Mass Molecule Weight Marker These results correspond with the expected PCR product size (506 bp) for genomic DNA samples containing, the canola endogenous FatA gene, The endogenous target band was not observed in the no-template control.
. , Date Recue/Date Received 2022-05-20 Table 8; PCR Genomic DNA Target Site and Expected Size of PCR Products Primer Set Target Site Expected Size of PCR Product (bp) 09-0-3290/09-0-3288 Construct-Specific ubiquitin promoter and get4621 Endogenous oanola FatA
gene' PCR: POLYMERASE CHAIN REACTION =
BP: BASE PAIRS
1. Genbank accession number for FatA gene is X87842.1. This sequence was used to design PCR primers.
Table 9: PCR Rees:lents and Reaction Conditions PCR Reagents = PCR Reaction Conditions ' =
Volume Cycle Temp Time Reagent # Cycles (pL) Element ( C) (sec) Template DNA initial (100 ng/p1) Denaturation Primer 1 (10 pM) 0.75 Denaturation 94 10 Primer 2 (10 pM) 0,75 Annealing 65 20 35 PCR Master Mix* 12.5 Elongation 72 45 ddH20 10 Final Elongation 72 180 1 Until Hold Cycle 4 analysis PCR: POLYMERASE CHAIN REACTION
DDH20: DOUBLE-DISTILLED WATER
* Roche High Fidelity Master Mix Table 10: = List of Primer Seauences Used in PM Reactions Primer Sequence 5' ¨3' Target Sequence Name SEQ ID NO 4:
09-0-3290 Ubiquitin Promoter AOCTATTGCTICACCGCCTTAGC
SEQ ID NO: 5 09-0-3288 get4821 GCTCAGCTTGGTGGAATGAAGCCAD
SEQ ID NO: 6 Canola Endogenous 09-0-2812 GACACAAGGCGGCTTCAAAGAGTTACAGATG FatA
SEQ IC NO 7: Canola Endogenous 09-0-2813 ACAATGTCATCTTGCTGGCATTCTCTTCTG FatA
Example 5. Further Insert and Flankino Border Sequence Characterization of Brass/ca Event DP-073496-4 =
To characterize the integrity of the inserted DNA and the = genomic insertion site, the flanking genomic DNA border regions of the DP-073498-4 event were = = =
Date Recue/Date Received 2022-05-20 determined. Flanking genomic sequence of DP-073496-4 Is included within SEQ ID
NO: 2. PCR amplification from the insert and border sequences confirms that the border regions are of Brassies origin and that the junction regions can be used for identification- of DP-073496-4 Brassies, Overall, characterization of the insert and genomic border sequences, along with Southern blot data, indicate a single insertion of the DNA fragment present in the Brassica genome. Various molecular techniques are then used to specifically characterize the integration site.
In the Initial characterization, the flanking genomic border regions are cloned and sequenced using the OenomeVValker end inverse PCR methods. Using information from the flanking border sequence, PCR is performed on DP-073496-4 genomic DNA and unmodified control genomic DNA, Those skilled in the art will also Include a control PCR using an endogenous gene to verify that the isolated genomic DNA Is suitable for PCR amplification.
Date Recue/Date Received 2022-05-20 CD
CD
CD
CD
CD
Table 11.
0_ PCR-based o event-specific detection methods o PCR
Primer 1 Primer 2 Probe ________________________________ Assay =
event type Mune Sequence Name Sequence Name Sequence 5 label Quencher 10-0-3514 GG Ls...toGGGC 1515 TTATOCGGICCTAG
DP-073496- Gel- SEO. ID NO:
4 based .20 GCCG 140:23 CAAACCTOC
09-0-2824 GTTaTaaTC 2325 CAAACCTCCATAG TTAGTTAGATC
DP473496. Real- SEQ ID
ATAGar.ATTAC S!QJP AGTTCAACATOTA 09-0P83 AGGATATTCTT
oe 4 time NO:21 Maim P1024 A
SEO. ID NO=26 . II FAM 4.469 FatA A- Real- SEQ ID TCGGGACGAGT 5E010 = spedfic time N022 AC F10:25 ACAiCa IAAATAT 5E0 ID N0:17 TCATGCTTC FAM MOB
Table 12. Summary Table of SEQ id NOS
SEQ ID NO Description 1 0A14621 protein 2 DP-073496-4 insert and flanking sequence Primer 09-0-3290 (SEQ ID NO: 4 4 AGCTATTGCTICACCGCCTTAGC) Target ¨ Ubiquitin Promoter Primer 09-0-3288 (SEQ ID NO: 5 GCTCAGCTTGGTGGAATGAAGCCAC) Target g8t4621 Primer 09-0-2812 (SEQ ID NO: 6 6 GACACAAGGCGGCTTCAAAGAGTTACAGATG) Target Canala Endogenous FatA
Primer 09-0-2813 (SEQ ID NO:
7 ACAATGTCATMGCTGGCATTCTCTTCTG ) Canola Endogenous FatA
8 Right border genomIc sequence 9 Left border genomic sequence Complete Internal transgene 11 Complete flanking and Internal transgene 12 Right flanking genomic/right border transgene (10 nt/10nt) 13 Left flanking genomicfleft border transgene (10 ntll Ont) 14 Right flanking genomiaright border transgene (20 nt/20nt) Left flanking genomic/left border transgene (20 nt/20nt) 16 Right flanking genomic/right border transgene (30 nt/30nt) 17 Left flanking genomialeft border transgene (30 nt/30nt) 18 Right flanking genomic/complete transgene 19 Left flanking genomic/complete transgene Primer 10-0-3514 21 Primer 09-0-2824 22 Primer 09-0-3249 23 Primer 10-0-3515 24 Primer 09-0-2825 Primer 09-0-3251 26 Primer 09-QP83 Date Recue/Date Received 2022-05-20 SEQ ID NO Description 27 Primer 09-QP87 The article a and wan' are used herein to refer to one or more than one (Le., to at least one) of the grammatical object of the article. By way of example, an element' means one or more element All publications and patent applications mentioned in the specification are indicative of the level of those skilled in the art to 'which this invention pertains.
The scope of the claims should not be limited by the preferred embodiments set forth in the examples, but should be given the broadest interpretation consistent with the description as a whole.
=
=
. =
=
Date Recue/Date Received 2022-05-20 Applicant's or agent's international applicathon No.
Me reference 35718/39908 PCT/US2010/
INDICATIONS RELATING TO DEPOSITED MICROORGANISM
OR OTHER BIOLOGICAL MATERIAL
(PCT Rule 13b1s) A. The irsications made below relate to the deposited microorganism or other blobglcal material referred to in the description on page 4:1, line 5 B. IDENTIFICATION OF DEPOSIT Further deposits are kientfled on an additional sheet 0 Name of depository Institution American Type Culture Collection Adiess of depositary institution (Including postal code and country) 10801 University Blvd.
Manassas, Virginia 20110-2/09 USA
=
Date of deposit Accession Number = 24 November 2010 PTA-C. ADDITIONAL INDICATIONS (leave blank if not epplicable) This Intimation is continued on an additional sheet El page 3, line 9; page 5, line 8; page 82, line 8; page 82, line 19; page 84, line 33; page 85, line 15 and page 85, line 21 =
D. DESIGNATED STATES FOR WHICH INDICATIONS ARE MADE (If the Indicators are not fore! designated Woks) E. SEPARATE FURNIs KING OF INDICATIONS (leave blank if not applicable) The Indications listed below will be submitted to the International Bureau later (specify the general nature of the Indications e.g. , 'Accession ivumber of cfrepostr) For receivkig Office use only For International Bureau use only 0 This sheet was raceme yinm me mternerional application CI This sheet no sow mu mu; pm nualeawirm vutood on:
Authorized officer Authorized officer =
Forrn PCT/R0/134 (July 1998) "
Date Recue/Date Received 2022-05-20
1. Cloransulam-methyl 2. Flumetsulam 3. Diclosulam 4. Florasulam = 5. Metosulam .
6. Penoxsulam 7. Pyroxsulam D. Pyrimklinyloxy(thio)benzoates 1. Bispyribac 2. Pyriftand = 3. Pyribenzoxim = 4. Pyrithiobac 5. Pyriminabac-methyl = E. ImIdazolinones 1. Imazapyr 2. Imazethapyr 3. Imazaquin 4. Imazaplc 5. Imazamethabenz-methyl 6. Imazamox II. Other Herbicides-Active Ingredients/
Additional Modes of Action A. Inhibitors of Acetyl CoA carboxylase (ACCase) (WSSA Group 1) 1. Aryloxyphenoxypropionates ('FOPS') a. Quizalofop-P-ethyl Date Recue/Date Received 2022-05-20 b. Diclofopmethyl o. C1odinafop-propargyl d. Fenoxaprop-P-ethyl e. FluazIfop-P-butyl f. Propaquizafop = = g. Haloxyfop-P-methyl h. Cyhalofop-butyl I. Quizalofop-P-ethyl 2. Cyclohexanedlones ('D1Ms') a. Alloxydlm b. Butroxydim c. Clethodim d. Cycloxycim e. Sethoxydim Tepraloxyd1m g. Tralkoxydlm B. Inhibitors of Photosystem 11¨H RAC
Group C1/ VVSSA Group 6 1. Triazines a. Ametryne b. Atrazine C. Cyanazine d. Desmetryne e. Dimethametryne f. Prometon g. Prometryne =
Ii. Propazlne I. Simazine J. Mehra k. Terbumeton I. Terbuthylazine m. Terbutryne n. Trietazine 2. Triazinones a. Hexazlnone b. Metribuzin o. Metamitron 3. Triazollnone a. Amicarbazone 4. Uraoils a. Bromacil b. Lenaoll c. TerbacIl 5. Pyridazinones a. Pyrazon Date Recue/Date Received 2022-05-20 6. Phenyl carbamates a- Desmedipham b. Phenmedipharn C, Inhibitors of Photosystem II¨HRAC
Group C2/WSSA Group 7 1. Ureas a. Fluometuron b. LInuron c. Chiorobromuron d. Chlorotoluron e. Chlormairon f. Dimefuron g. Diuron h. Ethidimuron I. Fenuron J. Isoproturon K. Isouron I. Methabenzthiazuron m. Metobromuron n. Metoxuron o. Monollnuron p. Neburori q. Siduron r. Tebuthiuron 2. Amides a. Propanil b. Pentanochlor D. Inhibitors of Photosystem II¨HRAC
=
Group C3/ WSSA Group 6 1. Nitriles a. Brornofenoxlm b. Bromoxynil C. loxynil 2. Benzothiadlazinone (Bentazon) a. Bentazon 3. Phenylpyrkiazines a. Pyridate b. Pyridafol E. Photosystem-i-electron diversion (Bipyrldyliums) (WSSA Group 22) 1. Piquet 2. Paraquat F. Inhibitors of PPO
(protoporphyrinogen oxides.) (WSSA
Group 14) Date Recue/Date Received 2022-05-20 1. Diphenylethers a. Acifluorfen-Na b. Blfenox c. Chlornethoxyfen d. Fluoroglycofen-ethyl e. Fornesafen f. Halosafen g. Lactofen I,. Oxyfluorfen 2. Phenylpyrazoles a. Fluazolata b. Pyraflufen-ethyl 3. N-phenylphthalimides a. Clnidon-ethyl b. Flurnioxazin C. Flumiciorac-pentyl 4. Thiadiazoles =
a. Fluthiacet-methyl b. Thidiazimin 5. Oxadiazoies a. Oxadiazon b. Oxadiargyi B. Triazolinones a. Carfentrazone-ethyl b. Suifentrazone 7. Oxazolidinediones a. Pentoxazone 8. Pyritaldindiones a, Benzfendizone b. Butafenicil 9. others a. Pyrazogyl b. Profluazol G. Bleaching: Inhibition of carotenoid biosynthesis at the phytoene desaturase step (PDS) (WSSA Group 12) 1. Pyridazinones a. Norfiurazon 2. Pyridineoarboxamides a. DiflufenIcan Date Recue/Date Received 2022-05-20 b. Plcolinafen 3. Others a, Befiubutamid b. Fluridone c. Flurochioridone d. Flurtamone H. =Bleaching: Inhibition of 4-hydroxyphenyl-pyruvate-dioxygenase (4-HPPD) (WSSA Group 28) 1. Triketones a. MesotrIone b. Sulcotrione C. topremezone d. temtorlone 2. Isoxazoles a. Isoxachlottole b. isoxaflutole 3. Pyrazoles a. Benzofenap =
b. Pyrazoxyfen c. Pyrazolynate 4. Others a. Benzobicyclon I. Bleaching: Inhibition of carotenoid biosynthesis (unknown target) (WSSA
Group 11 and 13) 1. Triazoles (WSSA Group 11) a. Amftrole 2, Isoxazoildinones (WSSA Group 13) a. Clomazone 3. Ureas a. Fluometuron 3, Diphenylether a. Aclonifen Inhibition of EPsP Synthase 1. Giycines (WSSA Group 9) a, Glyphosate b. Sulfosate K. Inhibition of glutamine sy.nthetase 1. Phosphinic Acids = a Glufosinate-ammonium b. Bialaphos Date Recue/Date Received 2022-05-20 =
L. Inhibition of DHP (dihydropte. roate) synthase (WSSA Group 18) Carbamates a. Asulam M. Microtubule Assembly Inhibition (VVSSA Group 3) 1. Dinitroanilines = a Benfluralln b. Butralin C. Dinitramine d. Ethalfluralin =
e. Oryzalln f. Pendimethalin g. Trifluralin 2. Phosphoroamidates a. Amlprophos-methyl b. Butamlphos 3. Pyridines a. Dithiopyr b. Thiazopyr 4. Baniamides a. Pronamide b. Tebutam 5. Benzenedicarboxylic acids a. Chlorthat-dImethyl N. Inhibition of mitosis/microtubuie organization VVSSA Group 23) 1. Carbamates a. Chiorpropham b. Propham c. Carbetamide O. Inhibition of cell division (Inhibition of very long chain fatty acids as proposed mechanism; WSSA Group 15) 1. Chloroacetamides a. Aoetochlor = b. Alachior = c. Butachlor d. Dimethachlor e. Dimethanamid f. Metazachlor g. Nletolachlor = h. Pethoxamid 1. Pretilachlor Propachlor Date Recue/Date Received 2022-05-20 K. Proplsochlor 1. Thenylchlor 2. Acetamides a. Diphenamid = b. Napropamide c. Naproanillde 3, Oxyacetamides a. Flufenacet b. Mefenacet 4. Tetrazolinones a. FentrazamIde 5. Others a. Ani!otos b. Cafenstrole c. Indent:Ian d. Plperophos P. Inhibition of cell wall (cellulose) synthesis 1. Nitriles (WSSA Group 20) a. Dichlobenil b. Chlorthiamld 2. BenzamIdes (isoxaben = (WSSA Group 21)) a. Isoxatxm 3. TriazolocarboxamIdes (flupoxam) a. Flupoxam O. Uncoupling (membrane disruption): (WSSA Group 24) 1. Dinitrophenols a. DNOC
b. Dinoseb a. Dinoterb R. Inhibition of Lipid Synthesis by = other than ACC inhibition 1. Thiocarbamates (WSSA
Group 8) a. Flutylate b. Cydoate c. Dimepiperate d. EPTC
e. Esprocarb f. Molinate =
g. Orbencarb .
h. Pebulate Date Recue/Date Received 2022-05-20 I. Prosutfocarb J, Benthiocarb K. Tiocarbazil I. Triallate rn. Vemoiate 2. Phosphorodithioates a. Bensullde 3. Benzofurans a. Benfuresate b. Ethofumesate 4. Halogenated alkanoic acids (WSSA Group 26) a TOA
b. Daiapon Flupropanate S. Synthetic auxins (IAA-like) (WSSA
Group 4) 1. Phenoxycarboxyllc acids a Clomeprop b. 2,4-D
c. Mecoprop 2. Benzoic acids a Dicamba b. Chloramben c. TSA
3. Pyridine carboxylic acids a. Clopyralld b. Fiuroxypyr C. Picloram d. Tricyclopyr 4. Quinoline carboxylic acids a. Duinclorec b. Ouinmerac 6. Others (benazolin-ethyl) a. Benazolln-ethyl T. Inhibition of Auxin Transport 1, Phthalamates;
semicarbazones (WSSA Group 19) a. Naptalam b. Diflufenzopyr-Na U. Other Mechanism of Action 1. Arylaminopropionic acids a. Flamprop-M-methyl Date Recue/Date Received 2022-05-20 isoProPY1 2. Pyrazolium a. Difenzoquat 3. Organoarsenicals a. DSMA
b. MSMA
4. Others a. Bromobutide b. CInmethylln c. Cumyluron d. Dazomet e. Daimuron-methyl f. Dimuron g. Etobenzanid = h. Fosamine I. Metam J. OxazIclomefone k. Oleic acid I. Pelargonlc acid m. PyrIbuticarb In certain methods, glyphosate, alone or in combination with another herbicide of interest, can be applied to the DP-073496-4 Brass/0a plants or their area of cultivation. Non-limiting examples of glyphosate formations are set forth In Table 2. In specific embodiments, the glyphosate is in the form of a salt, such as, ammonium, isopropylammonium, potassium, sodium (including sesquisodium) or trimesium (alternatively named suffocate).
=
Date Recue/Date Received 2022-05-20 Table 2. Glypbosate formulations comparisons.
Active Acid Acid Herbicide by ingredi- =peva- Apply: equiva-Registered eat per lent per fl or/
lent per Trademark Mannfactuer Salt gallon gallon acre acre = Roundup Oiigival Monsanto Isopropylamine 4 3 32 0.750 Roundup Original II Monsanto Isopropylanin. e 4 3 32 0.750 Roundup Original MAX Monsanto Potassium 55 4.5 22 0.773 Roundup UllraMax Monsanto IsoproPylaMine 5 3.68 26 0.748 Roundup UltraMax II Monsanto Potassium 55 4.5 22 0.773 Roundup Weathernutis Monsanto Potassium 5.5 4.5 22 0.773 Touchdown liYagotito Disramoniam 3.7 3 32 0.750 .
Touchdown HiTech Syngenta Potassium 6.16 5 20 0.781 Touchdown Total SYngenta Potassium 5.14 4.17 24 0.782 Durango Dow AgroSciences Isopropylamine 5.4 4 24 0.750 Glyphoutax Dow AgroSciences Isopropylanine 4 3 32 0.750 Glyphomax Plus DOW AgraSciences Isopropyissoine 4 3 32 0.750 Glyphorrtax 2017 Dow AgroSciences Isopropylanine 4 3 32 0.750 C31y Star Plus Alhaughami Star Isoprogyhunine 4 3 32 0.750 City Star 5 Albaugh/Agri Star Isoprogyiamirbe 5.4 4 24 0.750 ay Star Orininal Albaugh/Agri Star Tsopropylamine 4 3 32 0.750 Gly-Fla Micro Flo rsopropylamine 4 3 32 0.750 Credit Nufhrni Isopropylamine 4 3 32 0.750 Credit Extra Nufann Lsopropylamine 4 3 32 0.750 Credit Duo 24ufarm Isopro. + 4 3 32 0.750 monoarcun.
Credit Duo Extra Norma Isopro. + 4 3 32 0.750 = mortOOM01.
Extra Credit 5 Nora= Isopropylamine 5 3.68 26 0.748 Cornerstone Agriliance Lsopropylamine 4 3 32 0.750 Comentone Plus Agrdiance Bropropylamine 4 3 32 0.750 Olyfos Chennnova Isopropylninine 4 3 32 0.750 Glyfos X-TRA Cheminova Isopropylamine 4 3 32 0.750 Rattler Helena isopropyiamine 4 3 32 0.750 Rattler Plus Helena Isopropylamine 4 3 32 0.750 Mirage OAP Isopropylamine 4 3 32 0.750 Mirage Plus UAP Isoptopylamine 4 3 32 0.750 Glyphosate. 41% Hahn Agro USA Isopropylamine 4 3 32 0.750 Buccaneer Tenkoz Isopropylamine 4 3 32 0.750 Buccaneer Plus Tenkoz Isopropylamine 4 3 32 0.750 Honcho Monsanto Isoproylamine 4 3 32 0.750 Honcho Pins Monsanto Lsopropylanese 4 3 32 0.750 Cily..4 Univ. Crop Prot. Alli Lsopropylamine 4 3 32 0.750 C31y-4 Pins Univ. Corp Prot. Allt Lsopropylamine 4 3 32 0.750 CleasOut 41 Chemical Products Lsopropylamisie 4 3 32 0.750 Tech.
ClearOut 41 Plus Chemical Products Isopropylitmine 4 3 32 0.750 Tech.
SPittre Control Solutions Isopropylamine 4 3 32 0.750 Spitfire Phis Control Solutions Isopropylanaine 4 3 32 0.750 Glyphosate 4 FarmerSavercora Isopropylarnine , 4 3 32 0.750 FS Glyphosate Plus Growsuark Isopropylamine 4 3 32 0.750 Glyphosate Original Griffin. I.LC leopropylamine 4 3 32 0.750 =
Thus, in some embodiments, a transgenic plant of the invention Is used in a method of growing a DP-073496-4 brassica crop by the application of herbicides to which the plant Is tolerant. In this manner, treatment with a combination of one of more herbicides which include, but are not limited to: acetochlor, acifluorfen and its sodium salt, aclonifen, acrolein (2-propenal), alachlor, alloxydim, ametryn, amicarbazone, amidosulfuron, aminopyraild, amitrole, ammonium sulfamate, anilofos, asulaM, atrazIne, azimsulfuron, beflubutamld, benazolin, benazolin-ethyl, =
=
=
Date Recue/Date Received 2022-05-20 bencarbazone, benfluralln, benfuresate, bensulfuron-methyl, bensulide, bentazone, .benzobicyclon, benzofenap, blfenox, bilanafos, bispyribac and its sodium salt, bromacil, bromobutide, bromofenoxim, bromoxynil, bromoxynil octanoate, butachlor, butafenacif, butamlfos, butralin, butroxydim, bufylate, cafenstrole, carbetamide, carfentrazone-ethyl, catechln, chlomethoxyfen, chloramben, chlorbromuron, chlorflurenol-methyl, chloridazon, chlorimuron-ethyl, chlorotoluron, chlorpropham, chlorsulfuron, chlorthal-dimethyl, chlorthiamid, cinidon-ethyl, cinmethylin, cinosulfuron, dethodim, clodinafop-propargyl, clomazone, clomeprop, ciopyralid, clopyrarid-olamine, cloransulam-methyl, CUH-35 (2-methoxyethyl 2-11[4-chloro-2-fluoro-5-[(1-methyl-propynyl) oxy]pheny1X3-fluorobenzoyl)aminolcarbonyl]-1-cyclohexene-1-carboxylate), cumyluron, cyanazine, cycloate, cyclosulfamuron, cycloxydim, cyhalofop-butyl, 2,4-D
and its butotyl, butyl, 'sootyl and isopropyl esters and its dimethylammonium, diolamlne and trolamine salts, daimuron, dalapon, dalapon-sodium, dazomet, Z4-DE and its dimethylammonium, potassium and sodium salts, desmedipham, desmetryn, dicamba and its diglycolammonium, dimethylammonium, potassium and sodium salts, dichlobenil, dichlorprop, didolop-methyl, diclosulam, difenzoquat metilsuffate, diflufenican, diflufenzopyr, dimefuron, =dimepiperate, dimethachlor, dimetharnetryn, dimethenamld, dimethenamld-P, dimethipin, dlmethylarsinic acid and its sodium salt, dinitramine, dinoterb, diphenamld, diquat dibromide, dithiopyr, diuron, DNOC, endothal, EPTC, esprocarb, ethaffluralin, ethametsulfuron-methyl, ethofumesate, ethoxyfen, ethoxysulfuron, etobenzanld, fenoxaprop-ethyl, fenoxaprop-P-ethyl, fentrazamide, fenuron, fenuron-TCA, flamprop-methyl, flamprop-M-isopropyl, flamprop-M-methyl, flazasulfuron, florasulam, fluazifop-butyl, fluazifop-P-butyl, flucarbazone, fluceitosulfuron, fluchloralln, flufenacet, flufenpyr, flufenpyr-ethyl, flumetsulam, flumiclorac-pentyl, flumioxazin, fluometuron, fluoroglycofen-ethyl, flupyrsulfuron-methyl and its sodium salt, flurenol, flurenol-butyl, fluridone, flurochloridone, fluroxypyr, flurtamone, fluthiacet-methyl, fomesafen, foramsulfuron, fosamine-ammonium, glufosinate, glufoslnate-ammonium, glyphosate and its salts such as ammonium, isopropylammonium, potassium, sodium (including sesquisodium) and trimesium (alternatively named sulfosate), halosulfuron-methyl, haloxyfop-etotyl, haloxyfop-methyl, hexazinone, HOK-201 (N-(2,4-clifluoropheny1)-1,5-dihydro-IV-(1-methylethyl)-5-oxo-1-[(tetrahydro-21-1-pyran-2-y1)methyl]-4H-1,2,4-triazole-4-carboxamide), imazamethabenz-methyl, imazamox, imazapic, imazapyr, imazaquin, imazaquin-ammonium, imazethapyr, imazethapyr-ammonium, imazosulfuron, indanofan, iodosulfuron-methyl, loxynil, ioxynil octanoate, ioxynil-sodium, isoproturon, isouron, isoxaben, isoxaflutole, isoxachlortole, lactofen, lenacil, linuron, maleic =
hydrazide, MCPA and its salts (e.g., MCPA-dimethylammonium, MCPA-potassium and Date Recue/Date Received 2022-05-20 MCPA-sodlum, esters (e.g., MCPA-2-ethylhexyl, MCPA-butotyl) and thioesters (e.g., MCPA-thioethyl),.MCPB and its salts (e.g., MCPB-sodium) and esters (e.g., MCPB-ethyl), mecoprop, mecoprop-P, mefenacet, mefluidide, mesosulfuron-methyl, mesotrione, metam-sodium, metamifop, metamitron, metazachlor, methabenzthiazuron, methylarsonic acid and its calcium, monoammonium, monosodium and disodium salts, methyldymron, metobenzuron, metobromuron, metolachlor, S-metholachlor, metosulam, metoxuron, metribtrzin, metsuifuron-methyl, molinate, monolinuron, naproanllide, napropamide, naptalam, neburon, nicosulfuron, norfiurazon, orbencarb, oryzalin, oxadiargyl, oxadiazon, oxasulfuron, oxazidomefone, oxyfluorfen, paraquat dichloride, pebulate, pelargonic acid, pendimethalin, penoxsulam, pentanochlor, pentoxazone, perfluidone, pethoxyamid, phenmedipham, picloram, picloram-potassium, plcollnafen, pinoxaden, piperofos, pretilachlor, prImIsulfuron-methyl, prodiamine, profoxydim, prometon, prometryn, propachlor, propanil, propaquizafop, propazine, propham, propisochlor, propoxycarbazone, propyzamide, prosulfocarb, pros ulfuron, pyracionil, pyraflufen-ethyl, pyrasulfotole, pyrazogyl, pyrazolynate, pyrazoxyfen, pyrazosulfuron-ethyl, pyribenzoxim, pyributicarb; pyriciate, pyriftalid, pyriminobac-methyl, pyrimisulfan, pyrithlobac, pyrithiobac-sodium, pyroxsulam, quinclorac, quinmerac, quinodamine, quizalofop-ethyl, quizalofop-P-ethyl, quizalofop-P-tefuryl, rimsulfuron, sethoxydim, siduron, simazine, simetryn, suicotdone, suifentrazone, sulfometuron-methyl, sulfosuifuron, 2,3,6-TBA, TCA, TCA-sodium, tebutam, tebuthiuron, tefuryltrione, tembotrione, tepraloxydim, terbacil, terbumeton, terbuthylazine, terbutryn, thenyichlor, thiazopyr, thiencarbazone, thifensulfuron-methyl, thiobencarb, tiocarbazil topramezone, tralkoxydim, tri-aliate, triasulfuron, hiazifiam, tribenuron-methyl, triclopyr, triclopyr-butotyl, triclopyr-triethylammonium, tridiphane, trietazlne, trifloxysulfuron, trifluralin, triflusulfuron-methyl, tritosuifuron and vemolate is disclosed.
Other suitable herbicides and agricultural chemicals are known in the art, such as, for example, those described in WO 2005/041654. Other herbicides also Include bioherbiddes such as Altemaria destruens Simmons, Colletotrichum gioeosporlocles (Penz.) Penz. and Saco., Orechsiera monoceras (M1I3-051), Myrotheclum verrucarie (Albertini & Schweinitz) Ditmar: Fries, Phytophthora palmlvora (Butl.) But.
and Pucclnia thtaspeos Schub.. Combinations of various herbicides can result in a greater-than-additive (I.e., synergistic) effect on weeds and/or a less-than-additive effect (i.e., safening) on crops or other desirable plants. In certain instances, combinations of glyphosate with other herbicides having a similar spectrum of control but a different mode of action will be particularly advantageous for preventing the development of = 46 Date Recue/Date Received 2022-05-20 resistant weeds. HerbicideIly effective amounts of any particular. herbicide can be easily determined by one Skilled In the art through simple experimentation.
Herbicides may be classified into groups and/or subgroups as described herein above with reference to their mode of action, or they may be classified into groups andfor subgroups in accordance with their chemical structure.
Sulfonamide herbicides have as an essential molecular structure feature a sulfonamide moiety (¨S(0)2NH¨). As referred to herein, sulfonamide herbicides particularly comprise sulfonylurea herbicides, suffonylaminocarbonyltriazolinone herbicides and triazoiopyrimidine herbicides. In sulfonylurea herbicides the sulfonamide moiety is a component in a sulfonyiurea bridge (--S(0)2NHC(0)NH(R)¨).
In suifonylurea herbicides the sutfonyi end of the sulfonyfurea bridge Is connected either directly or by way of an oxygen atom or an optionally substituted amino or methylene group to a typically substituted cyclic or acyclic group. At the opposite end of the sulfonyiurea bridge, the amino group, which may have a substituent such as methyl (R being CH) instead of hydrogen, is connected to a heterocyclic group, typically a symmetric pyrimidine or triazine ring, having one or two substituents such as methyl, ethyl, trifluOromethyl, methoxy, ethoxy, methylamlno, dimethylamino, ethyiamino and the halogens. In suffonylaminocarbonyitrlazollnone herbicides, the sulfonamide moiety is a component of a sulfonylaminocarbonyl bridge (-S(0)2NHC(0)¨), In sulfonylaminocarbonyltriazolinone herbicides the suifonyl end of the sulfonylaminocarbonyl bridge is typically connected to substituted phenyl ring. At the opposite end of the sulfonylaminocarbonyl bridge, the carbonyl is connected to the 1-position of a triazolinone ring, which is typically substituted with groups such as alkyl and alkoxy. In triazoiopyrimidine herbicides the sulfonyi end of the sulfonamide moiety is connected to the 2-position of a substituted [1,2,41triazolopyrimidine ring system and the amino end of the sulfonamide moiety Is connected to a substituted aryl, typically phenyl, group or alternatively the amino end of the sulfonamide moiety is connected to the 2-position of a substituted (1,2,4]triazolopyrImIdlne ring system and the sulfonyl end of the sulfonamide moiety is connected to .a substituted aryl, typically pyridinyl, group.
= The. methods further comprise applying to the crop and the weeds in a field a sufficient amount of at least one herbicide .to which the crop seeds or plants are tolerant, such as, for example, glyphosate, a hydroxyphenylpyruvatedioxygenase inhibitor (e.g., mesotrione or sulcotrione), a phytoene desaturase Inhibitor (e.g., diflufenican), a pigment synthesis inhibitor, sulfonamide, imidazolinone, bialaphos, phosphinothricin, azafenidin, butafenacil, sulfosate, giufosinate, triazolopyrimidine, pyrimidinyloxy(thio)benzoate or sulonylaminocarbonyitriazolinone, an acetyl Co-A
Date Recue/Date Received 2022-05-20 =
carboxyiase inhlbfior such as quizalofop-P-ethyl, a synthetic auxin such as quInclorac, KIN-485 or a protox inhibitor to control the weeds without significantly damaging the crop plants.
Generally, the effective amount of herbicide applied to the field is sufficient to selectively control the weeds without significantly affecting the crop, "Weed"
as used herein refers to a plant which is not desirable in a particular area.
Conversely, a "crop planr as used herein refers to a plant=which is desired in a particular area, such as, for example, a Brassica plant. Thus, in some embodiments, a weed is a non-crop plant or a non-crop species, while in some embodiments, a weed is a crop species which is sought to be eliminated from a particular area, such as, for example, an inferior and/or non-transgenic Brassica plant in a field planted with Brass/ca event DP-073496-4 or a non-Brass/ca crop plant in a field planted with DP-073496-4. Weeds can be classified into two major groups: monocots and dicots.
Many plant species can be controlled (i.e., killed or damaged) by the herbicides described herein. Accordingly, the methods of the invention are useful in controlling these plant species where they are undesirable ,(i.e., where they are weeds).
These plant species include crop plants as well as species commonly considered weeds, Including but not limited to species such as: blackgrass (A(opecurus myosuroldes), giant foxtail (Setarie faben), large crabgrass (Digitaria sanguine/is), Surinam grass (Brachiaria decumbens), wild oat (Avena latua), common cocklebur (Xanthium pensylvanicum), common lambsquarters (Chenopodium album), morning glory (1pomoea coccinea), pigweed (Amaranthus spp,), velvetieaf (Abutilion theophrastr), common bamyardgrass (Echinochloa crus-galb), bermudagrass (Cynodon dactylon), downy brome (Bromus tectorum), goosegrass (Eleusine id/ca), green foxtall (Setaria virld(s), Italian ryegrass (Lolium multitIorum), Johnsongrass (Sorghum halepense), lesser canarygrass (Phateris minor), windgrass (Apera splca-venti), wooly cupgrass (Erichloa vfllosa), yellow nutsedge (Cypenrs esculentus), common chickweed (Stellaria media), common ragweed (Ambrosia artemIslifolia), Kochia scoparia, horseweed (Conyza canadensls), rigid ryegrass (Latium rigidum), goosegrass (Eleucine indica), = 30 hairy fleabane (Conyza bonariensis), buckhom plantain (Plantago lanceolate), tropical splderwort (Commelina benghatensls), field bindweed (Convoivulus antens(s), purple nutsedge (Cyperus rolundus), redvine (Brunnichia ovate), hemp sesbania (Sesbania exattata), sicklepod (Senna obtusifolia), Texas blueweed (Helianthus Nana) and Devil's claws (Proboscides touisianica). In other embodiments, the weed comprises a herbicide-resistant ryegrass, for example, a glyphosate resistant ryegrass, a paraquat resistant ryegrass, a ACCase-inhibitor resistant ryegrass and a non-selective, herbicide Date Recue/Date Received 2022-05-20 resistant ryegrass. In some embodiments, the undesired plants are proximate the crop plants.
As used herein, by "selectively controlled" it is intended that the majority of weeds in an area of cultivation are significantly damaged or killed, while If crop plants are also present In the field, the majority of the crop plants are not significantly damaged. Thus, a method is considered to selectively control weeds when at least 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or more of the weeds are significantly damaged or killed, while If crop plants are also present In the field, less than 45%, 40%, 35%, 30%, 25%, 20%, 15%, 10%, 5% or 1% of the crop plants are significantly damaged or killed.
In some embodiments, a Brassica DP-073496-4 plant of the invention is not significantly damaged by treatment with a particular herbicide applied to that plant at a dose equivalent to a rate of at least 0.5, 1,2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, =
20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 110, 120, 160, 170, 200, 300, 400, 500, 600, 700, 800, 800, 1000, 2000, 3000, 4000, 5000 or more grams or ounces (1 ounce = 29.57m1) of active ingredient or commercial product or herbicide formulation per acre or per hectare, whereas an appropriate control plant is significantly damaged by the same treatment..
In specific embodiments, an effective amount of an ALS inhibitor herbicide comprises at least about 0.1, 1, 5, 10, 25,50, 75, 100, 150, 200, 250, 300, 350, 400, 450, 500, 600, 700, 750, 800, 850, 900, 950,1000, 2000, 3000, 4000, 5000 or more grams or ounces (1 ounce = 29.57m1) of active Ingredient per hectare. In other embodiments, an effective amount of an ALS inhibitor comprises at least about 0.1-50, about 25-75, about 50-100, about 100-110, about 110-120, about 120-130, about 140, about 140-150, about 150-200, about 200-500, about 500-600, about 600-800, about. 800-1000 or greater grams or ounces (1 ounce = 29.57m1) of active ingredient per hectare. Any ALS inhibitor, for example, those listed in Table 1 can be applied at these levels.
In other embodiments, an effective amount of a suifonylurea comprises at least 0.1, 1, 5, 10, 25, 50, 75, 100, 150, 200, 250, 300, 350, 400, 450, 500, 600, 700, 800, 900, 1000, 5000 or more grams or ounces (1 ounce = 29.57m1) of active Ingredient per hectare. In other embodiments, an effective amount of a sulfonylures comprises at least about 0,1-50, about 25-75, about 50-100, about 100-110, about 110-120, about 120-130, about 130-140, about 140-150, about 150-160, about 160-170, about 170-180, about 190-200, about 200-250, about 250-300, about 300-350, about 350-400, about 400-450, about 450-500, about 500-550, about 550-600, about 600-850, about 650-700, about 700-800, about 800-900, about 900-1000,. about 1000-2000 or more Date Recue/Date Received 2022-05-20 grams=or ounces (1 ounce = 29.57m1) of active Ingredient per hectare.
Representative sutfonylureas that can be applied at this level are set forth in Table 1, In other embodiments, an effective amount of a sulfonylaminocarbonylirlazollnones, triazolopyrimidines, pyrimidinyloxy(thlo)benzoates, and imidazolinones can comprise at least about 0.1, 1, 5, 10, 26, 60, 75, 100, 150, 200, 250, 300, 350, 400, 460, 500, 550, 600, 650,700, 750, 800, 850, 900, 950, 1000, 1050, 1100, 1150, 1200, 1250, 1300, 1350, 1400, 1500, 1550, 1600, 1050, 1700, 1800. 1850, 1900, 1950, 2000, 2500, 3500, 4000, 4500, 5000 or greater grams or ounces (1 ounce = 29.57i1l) active Ingredient per hectare. In other embodiments, an effective amount of a sulfonyiuminocarbonyttrlazolines, biezolopyrImidines, pyrimidinyloxy(thlo)benzoates or imldazolinones comprises at least about 01-50, about 25-75, about 50-100, about 100-110, about 110-120, about 120-130, about 140, about 140-150, about 150-160, about 160-170, about 170-180, about 190-200.
about 200-250, about 250-300, about 300-350, about 350-400, about 400-450, about 450.500, about 500-550, about 550-600, about 600-550, about 650-700, about 700-800, about 800-900, about 900-1000, about 1000-2000 or more grams or ounces (1 ounce = 29.57 ml) active Ingredient per hectare.
Additional ranges of the effective amounts of herbicides can be found, for example, in various publications from University Extension services. Sea, for example, Bernards, et at., (2006) Gulde for Weed Management in Nebraska (www.ianrpubs.uri.adu/sendlt/ec130); Regher, of al., (2005) Chemical Weed Control for Fields Crops, Pastures, Rangeldnd, and Noncropland. Kansas State University Agricultural Extension Station and Corporate Extension Service; Zollinger, el ed., (2006) North Dakota Weed Control Guide, North Dakota Extension Service and the Iowa State University Extension at wviwweeds.lastate.eduõ
In some embodiments of the Invention, glyphosate is applied to an area of cultivation andtor to at least one plant In an area of cultivation at rates between 8 'OM
= 32 ounces of acid equivalent per acre, or at rates between 10, 12, 14, 16, 18, 20, 22, 24, 26, 28 and 30 ounces of acid equivalent per acre at the lower end of the range of application and between 12, 14, 18, 18, 20, 22, 24, 28, 28, 30 and 32 ounces of acid equivalent per acre at the higher end of the range of application (1 ounce =
29.57 ml).
In other embodiments, glyphosate is applied at least at 1, 5,= 10, 20, 30, 40, 50, 60, 70, 80, 90 or greater ounce of active ingredient per hectare (1 ounce = 29.57 ml).
in some embodiments of the Invention, a sutfonylorea herbicide is applied to a *field andfor to at least one plant in a held at rates between 0,04 and 1.0 ounces of active Ingredient per acre, or at rates between 0.1, 0.2,0,4, 0.6 and 0.8 ounces of active Ingredient per acre 50 = =
=
= =
Date Recue/Date Received 2022-05-20 at the lower end of the range of application and between 0.2, 0.4, 0.6, 0.8 and 1.0 ounces of active Ingredient per acre at the higher end of the range of application. (1 ounce = 29.57 m1).
As Is known in the art, glyphosate herbicides as a class contain the same active ingredient, but the active ingredient is present as one of a number of different salts and/or formulations. However, herbicides known to inhibit ALS vary in their active ingredient as well as their chemical formulations. One of skill in the art is faniillar with the determination of the amount of active ingredient and/or acid equivalent present in a particular volume and/or weight of herbicide preparation.
In some embodiments, an ALS inhibitor herbicide is employed. Rates at which the ALS inhibitor herbicide is applied to the crop, crop part, seed or area of cultivation can be any of the rates disclosed herein. In specific embodiments, the rate for the ALS inhibitor herbicide is about 0.1 to about 5000 g al/hectare, about 0.5 to about 300 g al/hectare or about 1 to about 150 g al/hectare.
. Generally, a particular herbicide is applied to a particular field (and any plants growing in it) no more than 1, 2, 3, 4, 5, 8, 7 or 8 times a year, or no more than 1, 2, 3, 4 or 5 times per growing season.
By 'treated with a combination of or "applying a combination of herbicides to a crop, area of cultivation or field' it is intended that a particular field, crop or weed is treated with each of the herbicides and/or chemicals indicated to be part of the = combination so that a desired effect is achieved, i.e., so that weeds are selectively controlled while the crop is not significantly damaged. In some embodiments, weeds which are susceptible to each of the herbicides exhibit damage from treatment with each of the herbicides which Is additive or synergistic. The application of each herbicide and/or chemical may be simultaneous or the applications may be at different times, so long as the desired effect is achieved. Furthermore, the application can occur prior to the planting of the crop.
The proportions of herbicides used In the methods of the invention with other herbicidal active ingredients in herbicidal compositions .are generally in the ratio Of 5000:1 to 1:5000, 1000:1 to 1:1000, 100:1 to 1:100, 10:1 to 1:10 or 5:1 to 1:5 by weight. The optimum ratios can be easily determined by those skilled in the art based on the weed control spectrum desired. Moreover, any combinations of ranges of the various herbicides disclosed in Table 1 can also be applied in the methods of the Invention.
Thus, in some embodiments, the Invention provides improved methods for selectively controlling weeds in a field wherein the total herbicide application may be less than 90%, 85%, 80%, 75%, 70%, 65%, 60%, 55%, 50%, 45%, 40%, 35%, 30%, Date Recue/Date Received 2022-05-20 25%, 20%, 15%, 10%, 5% or 1% of that used in other methods. Similarly, in some embodiments, the amount of a particular herbicide used for selectively controlling weeds in a field may be less than 90%, 85%, 80%, 75%, 70%, 65%, 60%, 55%, 50%, 45%, 40%, 35%, 30%, 25%, 20%, 15%, 10%, 5% or 1% of the amount of that particular herbicide that would be used in other methods, i.e., methods not utilizing a plant of the invention.
in some embodirrionts, a DP-073496-4 Brass/ca plant of the invention benefits from a synergistic effect, wherein the herbicide tolerance conferred by the GLYAT
polypeptide and that conferred by a polypeptide providing tolerance to another herbicide ii greater than expected from simply combining the herbicide tolerance conferred by each gene separately. See, e.g., McCutchen, of al., (1997) J.
Econ.
Entotnot. 90:1170-1180; Priesler, etal., (1999) J. Econ. Entomot 92:598-603.
As used . herein, the terms 'synergy,' "synergistic; "synergistically" and derivations thereof, .such as in a "synergistic effect" or a "synergistic herbicide combination" or a "synergistic herbicide composition" refer to circumstances under which the biological activity of a combination of herbicides, such as at least a first herbicide and a second herbicide, is greater than the sum of the biological activities of the individual herbicides. Synergy, expressed in terms of a "Synergy Index (SI)," generally can be determined by the method described by Kull, eta!,, (1961) Applied Microbiology 9:533.
= 20 See also, Colby, (1967) Weeds 15:20-22.
In other instances, the herbicide tolerance conferred On a DP-073496-4 plant of the invention is additive; that is, the herbicide tolerance profile conferred by the herbicide tolerance genes is what would be expected from simply combining the herbicide tolerance conferred by each gene separately to a transgenic plant containing them individually. Additive and/or synergistic activity for two or more herbicides against key weed species will increase the overall effectiveness and/or reduce the actual amount of active ingredient(s) needed to control said weeds. Where such synergy is observed, the plant of the invention may display tolerance to a higher dose or rate of herbicide and/or the plant may display tolerance to additional herbicides or other chemicals beyond those to which it would be expected to display tolerance. For example, a DP-073496-4 Brasslca plant may show tolerance to organophosphate compounds such as insecticides and/or Inhibitors of 4-hydroxyphenylpyruvate dioxygenase.
Thus, for example, the DP-073496-4 Brassica plants of the invention, when further comprising genes conferring tolerance to other herbicides, can exhibit greater than expected tolerance to various herbicides, Including but not limited to glyphosate, ALS inhibitor chemistries and sulfonylurea herbicides. The DP-073496-4 Brassica 52 =
Date Recue/Date Received 2022-05-20 plant plants of the invention may show tolerance to a particular herbicide or herbicide combination that Is at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 12%, 16%, 17%, 20%, 22%, 25%, 27%, 30%, 35%, 40%, 46%, 50%, 55%, 60%, 65%, 70%, 80%, 90%, 100%, 125%, 150%, 175%, 200%, 300%, 400% or 500% or more higher than the tolerance of an appropriate control plant that contains only a single herbicide tolerance gene which confers tolerance to the same herbicide or herbicide combination. Thus, DP-073496-4 Brassica plants may show decreased damage from the same dose of herbicide in comparison to an appropriate control plant, or they may show the same degree of damage in response to a much higher dose of herbicide than the control plant. Accordingly, in specific embodiments, a particular herbicide used for selectively containing weeds in a field is more than 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 60%, 60%, 70%, 80%, 90%, 100% or greater than the amount of that particular herbicide that wodd be used in other methods, i.e., methods not utilizing a plant of the invention.
In the same manner, in some embodiments, a DP-073496-4 Brassica plant of the invention shows improved tolerance to a particular formulation of a herbicide active Ingredient in comparison to an appropriate control plant, Herbicides are sold commercially as formulations which typically include other ingredients in addition to the herbicide active ingredient; these ingredients are often intended to enhance the efficacy of the active ingredient. Such other ingredients can include, for example, safeners and adjuvants (see, e.g., Green and Foy, (2003) "Adjuvants: Tools for Enhancing Herbicide Performance," In Weed Biology and Management, ed. Inderjit (Kluwer Academic Publishers, The Netherlands)). Thus, a DP-073496-4 Brassica plant of the invention can show tolerance to a particular formulation of a herbicide (e.g., a particular commercially available herbicide product) that is at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 12%, 15%, 17%, 20%, 22%, 26%, 27%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 80%, 90%, 100%, 125%, 150%, 175%, 200%, 300%, 400%, 500%, 600%, 700%, 800%, 900%, 1000%, 1100%, 1200%, 1300%, 1400%, 1500%, 1600%, 1700%, 1800%, 1900% or 2000% or more higher than the tolerance of an appropriate control plant that contains only a single herbicide tolerance gene which confers tolerance to the same herbicide formulation.
In some embodiments, a DP-073496-4 Bressica plant of the invention shows improved tolerance to a herbicide or herbicide class to which at least one other herbicide tolerance gene confers tolerance as well as Improved tolerance to at least one other herbicide or chemical which has a different mechanism or basis of action than either glyphosate or the herbicide corresponding to said at least one other herbicide tolerance gene. This surprising benefit of the invention finds use in methods Date Recue/Date Received 2022-05-20 of growing crops that comprise treatment with various combinations of chemicals, Including, for example, other chemicals used for growing crops. Thus, for example, a DP-073496-4 Brassica plant may also show improved tolerance to chlorpyrifos, a systemic organophosphate insecticide. Thus, the invention also provides a DP-073496-4 Brassica plant that confers tolerance to giyphosate (i.e., a GLYAT
gene) which shows improved tolerance to chemicals which affect the cytochrome P4.50 gene, and methods of use thereof. In some embodiments, the DP-073496-4 Bressica plants also show improved tolerance to dicamba. In these embodiments, the improved tolerance to dicamba may be evident in the presence of giyphosate and a sulfonyiurea herbicide.
In other methods, a herbicide combination is applied over a DP-073496-4 Brassica plant, where the herbicide combination produces either an additive or a synergistic effect for controlling weeds. Such combinations of herbicides can allow the application rate to be reduced, a broader spectrum of undesired vegetation to be controlled, improved control of the undesired vegetation with fewer applications, more rapid onset of the herbicidal activity or more prolonged herbicidal activity.
An 'additive herbicidal composition" has a herbicidal activity that is about equal to the observed activities of the individual components. A "synergistic herbicidal combination" has a herbicidal activity higher than what can be expected based on the observed activities of the Individual components when used alone. Accordingly, the presently disclosed subject matter provides a synergistic herbicide combination, wherein the degree of weed control of the mixture exceeds the sum of control of the individual herbicides. In some embodiments, the degree of weed control of the mixture exceeds the sum of control of the individual herbicides by any statistically significant amount including, for example, about 1% to 5%, about 5% to about 10%, about 10% to about 20%, about 20% to about 30%, about 30% to 40%, about 40% to about 50%, about 50% to about 60%, about 60% to about 70%, about 70% to about 80%, about 80% to about 90%. about 90% to about 100%, about 100% to 120% or greater.
Further, a "synergistically effective amount" of a herbicide refers to the amount of one herbicide necessary to elicit a synergistic effect in another herbicide present in the herbicide composition. Thus, the term "synergist," and derivations thereof, refer to a substance that enhances the activity of an active ingredient (al), i.e., a substance in a formulation from which a biological effect Is obtained, for example, a herbicide.
Accordingly, in some embodiments, the presently disclosed subject matter provides a method for controlling weeds in an area of cultivation. In some embodiments, the method comprises: (a) planting the area with a DP-0734964 crop seeds or crop plants which also comprise polynucleotides conferring ALS-inhibitor Date Recue/Date Received 2022-05-20 tolerance; and (b) applying to the weed, the crop plants, a crop part, the area of cultivation or a combination thereof, an effective amount of a herbicide composition comprising at least one of a synergistically effective amount of glyphosate and a synergistically effective amount of an ALS inhibitor (for example, but not limited to, a sulfonylurea herbicide) or agriculturally suitable salts thereof, wherein at least one of:
(i) the synergistically effective amount of the glyphosate is lower than an amount of glyphosate required to control the weeds in the absence of the sulfonylurea herbicide;
(II) the synergistically effective amount of the ALS inhibitor herbicide is lower than an amount of the ALS inhibitor required to control the weeds in the absence of glyphosate and (iii) combinations thereof and wherein the effective amount of the herbicide composition is tolerated by the crop seeds or crop plants and controls the weeds in the area of cultivation.
In some embodiments, the herbicide composition used in the presently disclosed method for controlling weeds comprises a synergistically effective amount of glyphosate and a sulfonylurea herbicide, in further embodiments, the presently disclosed synergistic herbicide composition comprises glyphosate and a sulfonylurea herbicide selected from the group consisting of metsulfuron-methyl, chlorsulfuron and triasulfuron.
In particular embodiments, the synergistic herbicide combination further comprises an adjuvant such as, for example, an ammonium Sulfate-based adjuvant, .e.g., ADD-UPe (Wenkem., Halfway House, Midrand, South Africa). In additional embodiments, the presently disclosed synergistic herbicide compositions comprise an additional herbicide, for example, an effective amount of a pyrimidinyloxy(thio)benzoate herbicide. In some embodiments, the pyrimidinyloxy(thlo)benzoate herbicide comprises bispyribac, e.g., (VELOCITY , Valent U.S.A. Corp., Walnut Creek, California, United States of America) or an agriculturally suitable salt thereof.
= In some embodiments of the presently disclosed method for controlling undesired plants, the glyphosate is applied pre-emergence, post-emergence or pre-and post-emergence to the undesired plants or plant crops and/or the ALS
inhibitor herbicide (i.e., the sulfonylurea herbicide) is applied pre-emergence, post-emergence or pre- and post-emergence to the undesired plants or plant crops. In other embodiments, the glyphosate and/or the ALS inhibitor herbicide (i.e., the sulfonylurea herbicide) are applied together or are applied separately. In yet other embodiments, the synergistic herbicide composition is applied, e.g., step (b) above, at least once prior to planting the crop(s) of interest, e.g., step (a) above.
Date Recue/Date Received 2022-05-20 Weeds that can be difficult to control with glyphosate alone In fields where a crop is grown (such as, for example, a brassica crop) include but are not limited to the following: horserred (e.g., Conyza canadensis); rigid ryegrass (e.g., Lolium rigidum);
goosegrass (e.g., Eleuslne Indica); Italian ryegrass (e.g., Lolium multifiorum); hairy fleabane (e.g., Conyza bonarfensis); buckhom plantain (e.g., Plantago lanceolate);
common ragweed (e.g., Ambrosia artemisifolia); morning glory (e.g., /pomoea spp.);
waterhemp (e.g., Amaranthus spp.); field bindweed (e.g., Convolvulus arvensis);
yellow nutsedge (e.g., Cyperus esculentus); common Iambsquarters (e.g., Chenopodium album); wild buckwheat (e.g,, Polygon/urn convolvulus); velvetleaf (e.g., Abutilon theophrasti); kochia (e.g., Kochia scoparia) and Asiatic dayflower (e.g., Commelina spp.). In areas where such weeds are found, Brassic,a plants comprising the DP-073496-4 event and tolerance to another herbicide are particularly useful in allowing the treatment of a field (and therefore any crop growing in the field) with combinations of herbicides that would cause unacceptable damage to crop plants that = 15 did not contain both of these polynucleotldes. Plants of the invention that are tolerant to glyphosate and other herbicides such as, for example, sulfonylurea, imidazolinone, triazolopyrimidine, pyrimidinyl(thio)benzoate and/or sulfonylaminocarbonyltriazolinone herbicides in addition to being tolerant to at least one other herbicide with a different mode of action or site of action are particularly useful in situations where weeds are tolerant to at least two of the same herbicides to which the plants are tolerant. In this manner, plants of the invention make possible Improved control of weeds that are tolerant to more than one herbicide.
For example, some commonly used treatments for weed control in fields where current commercial crops (Including, for example, Brassicas) are grown include glyphosate and, optionally, 2,4-D; this combination, however, has some disadvantages. Particularly, there are weed species that it does not control well and it =
also does not work well for weed control in cold weather. Another commonly used treatment for weed control in brassica fields is the sulfonylurea herbicide dilorimuron-ethyl, which has significant residual activity in the soil and thus maintains selective pressure on all later-emerging weed species, creating a favorable environment for the growth and spread of sulfonylurea-resistant weeds. Fields may be be treated with sulfonylurea, imidazollnone, trlazolopyrimidines, pyrimidiny(thlo)benzoates and/or suifonylaminocarbonyitriazonlInone such as the sulfonylurea chlorimuron-ethyl, either alone or in combination with other herbicides, such as a combination of glyphosate and tribenuron-methyl (available commercially as express ). This combination has several advantages for weed control under some circumstances, Including the use of herbicides with different modes of action and the use of herbicides having a relatively Date Recue/Date Received 2022-05-20 short period of residual activity in the soil. A herbicide having a relatively short period of residual activity is desirable, for example, in situations where it is important to reduce selective pressure that would favor the growth of herbicide-tolerant weeds. Of course, in any particular situation where weed control is required, other considerations may be more important, such as, for example, the need to prevent the development of and/or appearance of weeds in a field prior to planting a crop by using a herbicide with a relatively long period of residual activity. Treatments that include both tribenuron-methyl and thifensulfuron-methyl may be particularly useful.
Other commonly used treatments for weed control in fields where current commercial varieties of crops (including, for example, Brasslcas) are grown include the sulfonyiurea herbicide thifensulfuron-methyl (available commercially as Harmony GTO). However, one disadvantage of thifensulfuron-methyl is that the higher application rates required for consistent weed control often cause injury to e crop growing in the same field. DP-073496-4 BrassIca plants comprising additional tolerance can be treated with a combination of glyphosate and thifensuifuron-methyl, which has the advantage of using herbicides with different modes of action.
Thus, weeds that are resistant to either herbicide alone are controlled by the combination of the two herbicides, and the improved DP-073496-4 Bressice plants would not be significantly damaged by the treatment, . Other herbicides which are used for weed control in fields where curient commercial varieties of crops (Including, for example, BrassIces) are grown are the triazolopyrimidine herbicide cloransuiam-rnethyl (available commercially as FirstRate8) and the imidazolinone herbicide imazaquin (available commercially as Sceptore). When these herbicides are used Individually they may provide only marginal control of weeds. However, may be treated, for example, with a combination of glyphosate (e.g., Roundup (glyphosate isopropyiamine salt)), imazapyr (currently available commercially as Arsenal ), chlorimuron-ethyl (currently available commercially as Classic ), quizatofop-P-ethyl (currently available commercially as Assure Ile) and fomesafen (currently available commercially as Flexstap1D).
This combination has the advantage of using herbicides with different modes of action.
Thus, weeds that are tolerant to just one or several of theses herbicides are controlled by the combination of the five herbicides. This combination provides an extremely broad spectrum of protection against the type of herbicide-tolerant weeds that might be expected to arise and spread under current weed control practices.
= Fields containing the DP-073496-4 Brassica plants with additional herbicide tolerance may also be treated, for example, with a combination of herbicides including giyphosate, rimsulfuron, and dicamba or mesotrione. This combination may be = 57 Date Recue/Date Received 2022-05-20 particularly useful in controlling weeds which have developed some tolerance to herbicides which inhibit ALS. Another combination of herbicides which may be particularly useful for weed control includes glyphosate and at least one of the following: metsulfuron-methyl (commercially available as Ally ), imazapyr (commercially available as Arsenal ), imazethapyr, imazaquin and sulfentrazone. it is understood that any of the combinations discussed above or elsewhere herein may also be used to treat areas in combination with any other herbicide or agricultural chemical.
Some commonly-used treatments for weed control in fields where current commercial crops (including, for example, Brassica) are grown include glyphosate (currently available commercially as Roundup ), rimsulfuron (currently available commercially as Resolve or Matrix ), dicamba (commercially available as Clarity ), atrazine arid mesotrione (commercially available as Cailisto0). These herbicides are sometimes used individually due to poor crop tolerance to multiple herbicides.
Unfortunately, when used individually, each of these herbicides has significant disadvantages. Particularly, the incidence of weeds that are tolerant to individual herbicides continues to increase, rendering glyphosate less effective than desired In some situations. Rimsulfuron provides better weed control at high doses which can cause injury to a crop, and alternatives such as dicamba are often more expensive than other commonly-used herbicides Some commonly-used treatments for weed control in fields where currant commercial crops are grown include glyphosate (currently available commercially as Roundup ), chlorimuron-ethyl, trlbenuron-methyl, rimsuffuron (currently available commercially as Resolve or Matrix ), imazethapyr, imazapyr and imazaquin, Unfortunately, when used individually, each of these herbicides has significant disadvantages. Particularly, the Incidence of weeds that are tolerant to individual herbicides continues to increase, rendering each individual herbicide less effective than desired in some situations. However, DP-0734964 BrassIca with an additional -herbicide tolerance trait can be treated with a combination of herbicides that would cause unacceptable damage to standard plant varieties, Including combinations of herblddes that Include at least one of those mentioned above.
In the methods of the invention, a herbicide may be formulated and applied to an area of interest such as, for example, a field or area of cultivation, in any suitable manner. A herbicide may be applied to a field In any form, such as, for example, in a liquid spray or as solid powder or granules. In specific embodiments, the herbicide or combination of herbicides that are employed in the methods comprise a tankmix or a premix. A herbicide may also be formulated, for example, as a 'homogenous granule Date Recue/Date Received 2022-05-20 blend' produced using blends technology (see, e.g., US Patent Number 6,022,552, entitled "Uniform Mixtures of Pesticide Granules"). The blends technology of US
Patent Number 6,022,552 produces a nonsegregating blend (i.e., a "homogenous granule blend") of formulated crop protection chemicals In a dry granule form that enables delivery of customized mixtures designed to solve specific problems. A
homogenous granule blend can be shipped, handled, subsampled and applied in the same manner as traditional premix products where multiple active ingredients are formulated into the same granule.
Briefly, a "homogenous granule blend' is prepared by mixing together at least two extruded formulated granule products. In some embodiments, each granule product comprises a registered formulation containing a single active ingredient which is, for example, a herbicide, a fungicide and/or an insecticide. The uniformity (homogeneity) of a "homogenous granule blend" can be optimized by controlling the relative sizes and size distributions of the granules used in the blend. The diameter of extruded granules is controlled by the size of the holes in the extruder die and a centrifugal sifting process may be used to obtain a population of extruded granules with a desired length distribution (see, e.g., US Patent Number 6,270,025).
A homogenous granule blend Is considered to be "homogenous' when it can be subsampled into appropriately sized aliquots and the composition of each aliquot will meet the required assay specifications. To demonstrate homogeneity, a large sample of the homogenous granule blend is prepared and is then subsampled into aliquots of greater than the minimum statistical sample size.Blends also afford the ability to add other agrochemicals at normal, labeled use rates such as additional herbicides (a 3"3/4th mechanism of action), fungicides, insecticides, plant growth regulators and the like thereby saving costs associated with additional applications.
Any herbicide formulation applied over the DP-073496-4 Bra ssica plant can be prepared as a "tank-mix" composition. In such embodiments, each Ingredient or a combination of ingredients can be stored separately from one another. The ingredients can then be mixed with one another prior to application.
Typically, such mixing occurs shortly before appfication. In a tank-mix process, each Ingredient, before mixing, typically is present In water or a suitable organic solvent.
For additional guidance regarding the art of formulation, see, Woods, "The Formulator's Toolbox¨
Product Forms for Modern Agriculture' Pesticide Chemistry and Bioscience, The Food-Environment Challenge, Brooks and Roberts, Eds., Proceedings of the 9th International Congress on Pesticide Chemistry, The Royal Society of Chemistry, Cambridge, 1999, pp, 120-133. See also, US Patent Number 3,235,361, Column 5, line 16 through Column 7, line 19 and Examples 1041; US Patent Number 3,309,192, Date Recue/Date Received 2022-05-20 Column 5, line 43 through Column 7, Ursa 62 and Examples 8, 12, 15, 39, 41, 52, 53, 58, 132, 138-140, 162-164, 188, 167 and 169-182; US Patent Number 2,891,855, Column 3, the 66 through Column 5, line 17 and Examples 1-4; Klingman, Weed Control as a Science, John Wiley and Sons, Inc., New York, 1961, pp 81-96 and = 5 Hance, at at, Weed Control Handbook, 8th Ed., Blackwell Scientific Publications, = Oxford, 1989 .
= The methods of the invention further allow for the development of herbicide combinations to be used with the DP-073498-4 Brassies plants. In such methods, the environmental conditions in an area of cultivation are evaluated.
Environmental condiffons that can be evaluated Include, but are not Unified to, ground and surface water pollution concerns, intended use of. the crop, crop tolerance, soil residuals, weeds present in area of cultivation, soil texture, pH of soil, amount of organic matter in soli, application equipment and tillage practices. Upon the evaluation of the environmental conditions, an effective amount of a combination of herbicides can be applied to the crop, crop part, seed of the crop or area of cultivation.
In some embodiments, the herbicide applied to the DP-073496-4 Bressica plants of the invention serves to prevent the initiation of growth of susceptible weeds and/or serve to cause damage to weeds that are growing in the area of interest in some embodiments, the herbicide or herbicide mixture exert these effects on weeds affecting crops that are subsequently planted in the area of interest (1.e., field or area of cultivation). In the methods of the invention, the application of the herbicide combination need not occur at the same time. So long as the field in which the crop is planted contains detectable amounts of the first herbicide and the second herbicide is applied at some time during the period in which the crop is in the area of cultivation, the crop is considered to have been treated with a mixture of herbicides according to ' the invention. Thus, methods of the invention encompass applications of herbicide which are preemergent,"postemergent, =preplant incorporation' and/or which Involve seed treatment prior to planting.
In one embodiment, methods are provided for coating seeds. The methods comprise coating a seed with an effective amount of a herbicide or a combination. of herbicides (as disclosed elsewhere herein). The seeds can then be planted in an area of cultivation. Further provided are seeds having a coating comprising an effective .
amount of a herbicide or a combination of herbicides (as disclosed elsewhere herein).
Treemergenr refers to a herbicide which is applied to an area of interest (e.g., .35 a field or area of cultivation) before a plant emerges visibly from the soil.
*Postemergenr refers to a herbicide which is applied to an area after a plant emerges visibly from the soil. In some Instances, the terms !preemergent- and "posternergenr Date Recue/Date Received 2022-05-20 are used with reference to a weed in an area of Interest; and in some instances these terms are used with reference to a crop plant in an area of interest When used with reference to a weed, these terms may apply to only a particular type of weed or species of weed that Is present or believed to be present in the area of interest While any herbicide may be applied in a preemergent and/or postemergent treatment, some herbicides are known to be more effective In controlling a weed or weeds when applied either preemergence or postemergence. For example, rimsulfuron has both preemergence and postemergence activity, while other herbicides have predominately preemergence (metolachlor) or postemergence (glyphosate) activity. These properties of particular herbicides are known in the art and are readily determined by one of skill in the art. Further, one of skill in the art would readily be able to select appropriate herbicides and application times for use with the transgenic plants of the Invention and/or on areas in which transgenic plants of the invention are to be planted.
'Preplent incorporation" involves the incorporation of compounds into the soil prior to planting.
' Thus, the invention provides Improved methods of growing a crop and/or controlling weeds such as, for example, "pre-planting bum down," wherein an area Is treated with herbicides prior to planting the crop of Interest in order to better control weeds. The invention also provides methods of growing a crop and/or controlling = 20 weeds which are "no-till" or "low-till" (also referred to as 'reduced tillage"). In such methods, the soli is not cultivated or is cultivated less frequently during the growing cycle In comparison to traditional methods; these methods can save costs that would otherwise be incurred due to additional Cultivation, including labor and fuel costs.
The methods of the invention encompass the use of simultaneous and/or sequential applications of multiple classes of herbicides. In some embodiments, the methods of the invention Involve treating a plant of the invention and/or an area of interest (e.g., a field or area of cultivation) and/or weed with just one herbicide or other chemical such as, for example, a sulfonylurea herbicide.
The time at which a herbicide is applied to an area of interest (and any plants therein) may be important In optimizing weed control. The time at which a herbicide is applied may be determined with reference to the size of plants and/or the stage of growth and/or development of plants in the area of interest, e.g., crop plants or weeds growing In the area. The stages of growth and/or development of plants are known in the art. Thus, for example, the time at which a herbicide or other chemical Is applied to an area of interest in which plants are growing may be the time at which some or all of the plants in a particular area have reached at least a particular size and/or stage of growth and/or development, or the time at which some or all of the plants In a Date Recue/Date Received 2022-05-20 = particular area have not yet reached a particular size and/or stage of growth and/or development.
In some embodiments, the DP-073498-4 Brassica plants of the invention show improved tolerance to postemergence herbicide treatments. For example, plants of the invention may be tolerant to higher doses of herbicide, tolerant to a broader range of herbicides, and/or may be tolerant to doses of herbicide applied at earlier or later times of development in comparison to an appropriate control plant. For example, In some embodiments, the DP-073496-4 Brassica plants of the invention show an increased resistance to morphological defects that are known to result from treatment at particular stages of development. Thus, the glyphosate-tolerant plants of the invention find use in methods involving herbicide treatments at later stages of development than were previously feasible. Thus, plants of the invention may be treated with a particular herbicide that causes morphological defects in a control plant treated at the same stage of development, but the glyphosate-tolerant plants of the invention will not be significantly damaged by the same treatment.
Different chemicals such as herbicides have different "residual' effects, i.e., different amounts of time for which treatment with the chemical or herbicide continues to have an effect on plants growing in the treated area. Such effects may be desirable or undesirable, depending on the desired future purpose of the treated area (e.g., field or area of Cultivation). Thus, a crop rotation scheme may be chosen based on residual effects from treatments that will be used for each crop and their effect on the crop that will subsequently be grown In the same area. One of skill in the art is familiar with techniques that can be used to evaluate the residual effect of a herbicide;
for example, generally, glyphosate has very little or no soli residual actMty, while herbicides that act to inhibit ALS vary in their residual activity levels. Residual activities for various herbicides are known in the art, and are also known to vary with various environmental factors such as, for example, soil moisture levels, temperature, pH and soil composition (texture and organic matter).
= Moreover, the transgenic plants of the invention provide improved tolerance to treatment with additional chemicals commonly used on crops in conjunction with herbicide treatments, such as safeners, adjuvants such as ammonium sulfonate and crop oV concentrate, and the like. The term "safener refers to a substance that when added to a herbicide formulation eliminates or reduces the phytotoxic effects of the herbicide to certain crops. One of ordinary skill in the art would appreciate that the choice of safener depends, in part, on the crop plant of interest and the particular herbicide or combination of herbicides included in the synergistic herbicide composition. Exemplary safeners suitable for use with the presently disclosed Date Recue/Date Received 2022-05-20 herbicide compositions include, but are not limited to, those disclosed In US
Patent Numbers 4,808,208; 5,502,025; 8,124,240 and US Patent Application Publication Numbers 2008/0148847; 2006/0030485; 2005/0233904; 2005/0049145;
2004/0224849; 2004/0224848; 2004/0224844; 2004/0157737; 2004/0018940;
2003/0171220; 2003/0130120; 200310078167.
The methods of the invention can Involve the use of herbicides in combination with herbicide safeners such as benoxacor, BCS (1-bromo-44(chloromethyl) suffonyabenzene), cioquintocet-mexyl, cyemetrInil, dichiormid, 2.-(dichloromettlyi)-2-methyl-1,3-diexolane (MG 191), fenchlorazole-ethyl, fenciorim, flurazole, fluxofenim, furilazole, isoxaclifen-ethyl, mafenpyr-diethyl, methoxyphenone 04-methoxy-3-rnethylphenyi)(3-methylphenyl)-methanone), naphthalic anhydride (1,8-naphthalic anhydride) and oxabetrinil to Increase crop safety. Antidotally effective amounts of the herbicide safeners can be applied at the same time as the compounds of this invention, or applied as seed treatments. Therefore an aspect of the present Invention relates to the use of a = mixture comprising glyphosate, at least one other herbicide and an antidotirily effective amount of a herbicide serener.
Seed treatment is particularly useful for selective weed control, because it physically restricts antidoting to the crop plants. Therefore a particularly useful embedment of the present Invention is a method for selectively controlling the growth of weeds in a field comprising treating the seed from which the crop Is grown with an antidotally effective amount of safener and treating the field with an effective amount of herbicide to control weeds. Antidotally effective amounts of safeners can be easily determined by one skilled in the art through simple experimentation. An antidotally effective. amount of a serener is present where a desired plant is treated with the safener so that the effect of a herbicide on the plant is decreased in comparison to the effect of the herbicide on a plant that was not treated with the safener, generally, an antidotally effective amount of safener prevents damage or severe damage to the plant treated with the safener. One of skill In the art is capable of determining whether the use of a serener is appropriate and determining the dose at which a safener should be administered to a crop.
In specific embodiments, the herbicide or herbicide combination applied to the plant of the invention acts as a safener. In this embodiment, a first herbicide or a herbicide mixture Is applied at an antidotally effect amount to the plant.
Accordingly, a method for controlling weeds In an area of cultivation is provided. The method comprises planting the area with crop seeds or plants which compose a first polynucleotide encoding a polypedide that can confer tolerance to giyphosate 63, =
=
Date Recue/Date Received 2022-05-20 operably linked to a promoter active in a plant; and, a second polynucleotIde encoding an ALS inhibitor-tolerant polypeptlde operably linked to a promoter active in a plant. A
combination of herbicides comprising at least an effective amount of a first and a second herbicide is applied to the crop, crop part, weed or area of cultivation thereof.
The effective amount of the herbicide combination controls weeds; and, the effective = amount or the first herbicide is not tolerated by the crop when applied alone when compared to a control crop that has not been exposed to the first herbicide;
and, the effective amount of the second herbicide is sufficient to produce a safening effect, wherein the safening effect provides an increase in crop tolerance upon the application of the first and the second herbicide when compared to the crop tolerance when the first herbicide is applied alone.
In specific embodiments, the combination of safening herbicides comprises a first ALS inhibitor and a second ALS inhibitor. In other embodiments, the safening effect is achieved by applying an effective amount of a combination of glyphosate and at least one ALS inhibitor chemistry. Such mixtures provides increased crop tolerance (i.e., a decrease In herbicidal injury). This method allows for increased application rates of the chemistries post or pre-treatment. Such methods find use for increased control of unwanted or undesired vegetation. In still other embodiments, a safening affect is achieved when the DP-073496-4 brassica crops, crop part, crop seed, weed or area of cultivation is treated with at least one herbicide from the sulfonylurea family of chemistry in combination with at least one herbicide from the imidazolinone family.
This method provides increased crop tolerance (i.e., a decrease In herbicidal injury).
In specific embodiments, the sulfonylurea comprises rimsulfuron and the imidazolinone comprises imazethapyr. In other embodiments, glyphosate Is also applied to the crop, crop part or area of cultivation.
As used herein, an 'adjuvant" Is any material added to a spray solution or formulation to modify the action of an agricultural chemical or the physical properties of the spray solution. See, for example, Green and Foy, .(2003) "Adjuvants: Tools for Enhancing Herbicide Performance," in Weed Biology and Management, ed. lnderjit (Kluwer Academic Publishers, The Netherlands). Adjuvants can be categorized or subclassified. as activators, acidifiers, buffers, additives, adherents, antiflocculants, antifoamers, defoamers, antifreezes, attractants, baste blends, chelating agents, cleaners, colorants or dyes, compatibility agents, cosolvents, couplers, crop oil concentrates, deposition agents, detergents, dispersants, drift control agents, emulsifiers, evaporation reducers, extenders, fertilizers, foam markers, formulants, inerts, humectants, methylated seed oils, high load COCs, polymers, modified vegetable oils, penetrators, repellents, petroleum oil concentrates, preservatives, Date Recue/Date Received 2022-05-20 rainfast agents, retention aids, solubillzers, surfactants, spreaders, stickers, spreader stickers, synergists, thickeners, translocatlan aids, uv protectants, vegetable oils, water conditioners and wetting agents.
In addition, methods of the invention can comprise the use of a herbicide or a mixture of herbicides, as well as, one or more other insecticides, fungicides, nematocides, bactericides, acaricides, growth regulators, chemosterilants, semiochemicals, repellents, attractants, pheromones, feeding stimulants or other biologically active compounds or entomopathogenic bacteria, virus or fungi to form a multi-component mixture gMng an even broader spectrum of agricultural protection.
Examples of such agricultural protectants which can be used in methods of the Invention include: Insecticides such as abamectin, acephate, acetamiprid, amidoflumet (8-1955), avermectin, azadirachtin, azinphos-methyl, bifenthrin, bifenazate, buprofezin, carbofuran, cartap, ohlorrenapyr, chlorfkaazuron, chlorpyrifos, chlorpyrifos-methyl, chromafenozide, ciothianidin, cyflumetofen, cyfluthrin, beta-cyfluthrin, cyhaiothrin, lambda-cyfialothrin, cypermethrin, cyromazine, deltamethrin, diafenthiuron, dlazinon, dieidrin, diflubenzuron, dimefluthrin, dimethoate, dinotefuran, diofenolan, emamectin, endosulfan, esfenvalerate, ethiprole, fenothiocarb, fenoxycarb, fenpropathrin, fenvalerate, flpronil, flonicamid, flubendiamide, flucythrinate, tau-fluvalinate, flufenerlm (UR-50701), flufenoxuron, fonophos, halofenozide, hexaflumuron, hydramethyinon, imidacloprid, indoxacarb, isofenphos, lufenuron, malathion, metaflurplzone, metaldehyde, methamidophos, methidathion, methomyl, methoprene, methoxychlor, metofluthrin, monocrotophos, methoxyfenozide, nitenpyram, nithiazine, novaluron, noviflumuron (XDE-007), oxamyl, parathion, parathion-methyl, permethrin, phorate, phosalone, phosmet, phosphamidon, pirimicarb, profenofos, profluthrin, pymetrozine, pyrafluprole, pyrethrtn, pyridalyi, pyriprole, pyriproxyfen, rotenone, ryanddine, spinosad, spirodiclofen, spiromesifen (BSN 2060), spirotetramat, suiprofos, tebufenozide, teflubenzuron, tefluthrin, terbufos, tetrachlorvinphos, thiacloprid, thiamethoxam, thiodicarb, thiosultap-sodium, tralomethrin, triazamate, trichlorfon and triflumuron; fungicides such as acibenzolar, aldimorph, amisulbrom, azaconazole, azoxystrobln, benalaxyl, benomyl, benthiavalicarb, benthiavalicarb-isopropyl, binomial, biphenyl, bitertanol, blasticidin-S, Bordeaux mixture (Tribasic copper sulfate), boscalidinicobifen, bromuconatole, bupirimate, buthiobate, carboxln, carpropamid, oaptafol, captan, carbendatim, chloroneb, chlorothalonil, chiozolinate, clotrimazole, copper oxychlorlde, copper salts such as copper sulfate and copper hydroxide, cyazofamid, cyflunamid, cymoxanil, cyproconazole, cyprodln11, dichlofluanld, diclocymet, diclomezine, dicloran, diethofencarb, difenoconazole,. dimethomorph, dimoxystrobin, diniconazole, Date Recue/Date Received 2022-05-20 cilniconazole-M, dinocap, discostrobin, dithianon, dodemorph, dodine, econazole, etaconazole, edifenphos, epoxiconazole, ethaboxam, ethirimol, ethridiazole, famoxadone, fenamidone, fenarimol, fenbuconazole, fencaramid, fenfuram, fenhexamicle, fenoxanil, fenpiclonil, fenpropidin, fenpropimorph, fentln acetate, fentin hydroxide, ferbam, ferfurazoate, feiimzone, fluazinam, fludioxonil, flumetover, fluoplcollde, fluoxastrobin, fluquinconazole, fluquinconazole, flusilazole, flusulfamide, flutolanil, flutriafol, folpet, fosetyl-aluminum, fuberidazole, furalaxyl, furametapyr, hexaconazoie, hymexazole, guazatine, hrnazalil, imibenconazole, iminoctadlne, iodicarb, ipconazole, iprobenfos, tprodione, iprovalioarb, isoconazole, isoprothiolane, kasugamycln, kresoxim-methyl, mancozeb, mandipropamid, maneb, mapanipyrin, = mefenoxam, mepronil, metalaxyl, metconazole, methasulfocarb, metiram, metominostrobinflenominostrobin, mepanipyrim, metrafenone, miconazole, myclobutanil, neo-asozin (ferric methanearsonate), nuarimol, octhilinone, ofurace, orysastrobin, oxadixyl, oxolinic add, oxpoconazole, oxycarboxin, paclobutrazol, penconazole, pencycuron, penthiopyrad, perfurazoate, phoSphonlc acid, phthalide, picobenzamid, plcoxystrobin, polyoxin, probenazole, brochloraz, procymidone, propamocarb, propamocarb-hydrochlorlde, propiconazole, propineb, proquinazid, prothioconazole, pyraclostrobin, pryazophos, pyrifenox, pyrimethanil, pyrifenox, pyrrolnitrine, pyroquilon, quInc,onazole, quinoxyfen, quintozene, silthlofam, simeconazole, spiroxamine, streptomycin, sulfur, tebuconazole, techrazene, tecloftalam, tecnazene, tetraconazole, thlabendazole, thifluzamIde, thlophanate, thiophanate-methyl, thlrarrt, tiadinil, toiclofos-methyl, tolyfluanid, triadimefon, triadimenol, trlarimol, trlazoxide, tridemorph, trimoprhamide tricyclazole, trifloxystrobin, triforine, triticonazole, uniconazole, validamycin, vindozolln, zineb, ziram, and zoxamide; nematocides such as aldicarb, oxamyl and fenamiphos; bactericides such as streptomycin; acaricides such as amitraz, chinomethionat, chlorobenzilate, cyhexatin, dicofot, dienochlor, etoxazole, fenazaquin, fenbutalln oxide, fenpropathrin, fenpyroximate, hexythiazox, propargite, pyridaben and tebufenpyrad and biological agents including entomopathogenic bacteria, such as Bacillus thuringlensis subsp.
Aizawal, Bacillus ihuringlensis subsp. Kurstaki, and the encapsulated delta-endotoxins of Bacillus thuringlensis (e.g., Cellcap, MPV, MPVII); entomopathogenic fungi, such as green muscardine fungus; and entomopathogenic virus including baculovirus, nucleopolyhedro virus (NPV) such as HzNPV, AfNPV; and granuiosis virus (GV) such as CpGV. The weight ratios of these various mixing partners to other compositions (e.g., herbicides) used in the methods of the invention typically are between 100:1 and 1:100, or between 30:1 and 1:30, between 10;1 and 1:10, or between 4:1 and 1:4.
Date Recue/Date Received 2022-05-20 =
The present invention also pertains to a composition comprising a biologically effective amount of a herbicide of Interest or a mixture of herbicides, and an effective amount of at least one additional biologically active compound or agent and can further comprise at least one of a surfactant, a solid diluent or a liquid diluent.
Examples of such biologically active compounds or agents are: insecticides such as abameotin, acephate, acetamiprld, amidoflumet (S-1955), avermectin, azadirachtin, azinphos-methyl, bifenthrin, binfenazate, buprofezin, carbofuran, chlorferiapyr, chlorfluazuron, chlorpyrifos, chlorpyrifos-methyl, chromafenozide, clothianidin, cyfluthrin, beta-cyfiuthrin, cyhalothrin, lambda-cyhalothrin, cypermethrin, cyromazine, deltamethrin, diafenthluron, diazinon, dfflubenzuron, dimethoate, diofenolan, emamectin, endosulfan, esfenvalerate, ethiprole, fenothicarb, fenoxycarb, fenpropathrin, fenvalerate, flpronil, flonicamid, flucythrinate, tau-fluvalinate, flufenerim (UR-50701), flufenoxuron, fonophos, halofenozide, hexaflumuron, imidacloprid, indoxacarb, Isofenphos, lufenuron, malathion, metaldehyde, methamidophos, methidathion, methomyl, methoprene., methoxychlor, monocrotophos, methoxyfenozide, nithiazin, novaluron, noviflumuron (XDE-007), oxamyl, parathion, parathion-methyl, permethrin, phorate, phosalone, phosmet, phosphamidon, pirimicarb, profenofos, pymetrozlne, pyrIdalyi, pyriproxyfen, rotenone, spinosad, splromesifin (BSN 2060), sulrxofos, tebufenozide, teflubenzuron, tefluthrin, terbufos, tetrachlorvinphos, thlacloprid, thiamethoxam, thiodicarb, thiosultap-sodium, tralomethrin, trichlorfon and triflumuron;
fungicides such as acibenzolar, azoxystrobin, benomyl, blasticidin-S, Bordeaux mixture (tribasic copper sulfate), bromuconazola, carpropamid,..captafol, captan, carbendazim, chloroneb, chlorothalonil, copper oxychloride, copper salts, cyflufenamid, cymoxanil, cyproconazole, cyprodinii, (S)-3,5-dichloro-N-(3-chloro-1-ethyl-1-methyl-2-oxopropy1)-4-methylbenzamide (RH 7281), diclocymet (5-2900), diclomezine, dicloran, difenoconazole, (S)-3,5-dihydro-5-methy1-2-(methylthio)-phenyl-3-(phenyl-amino)-4H-imidazol-4-one (RP 407213), dimethomorph, dimoxystrobin, dinlconazole, diniconazole-M, dodine, edifenphos, epoxiconazole, famoxadone, fenamidone, fenarimol, fenbuconazole, fencaramid (5ZX0722), fenpiclonil, fenpropidin, fenpropimorph, fentin acetate, fentin hydroxide, fluazinam, fludioxonil, flumetover (RPA 403397), flumorf/flumorlin (SYP-L190), fluoxastrobin (HEC
6725), fluquinconazole, flusilazole, flutolanil, flutriafol, folpet, fosetyl-aluminum, furalaxyl, furametapyr (3-82658), hexaconazole, ipconazole, iprobenfos, iprodione, = isoprothiolane, kasugamycin, kresoxim-methyl, mancozeb, maneb, mefenoxam, mepronil, metalaxyl, metconazole, metomlno-strobinffenominostrobin (SSF-126), metrafenone (AC375839), myclobutanll, neo-asozin (ferric methane-arsonate), nicobifen (BAS 510), orysastrobin, oxadixyl, penconazote, pencycuron, probenazole, Date Recue/Date Received 2022-05-20 prochloraz. propamocarb, propiconazole, proquinazId (DPX-KQ926), prothioconazole (JAU 6476), pyrifenox, pyraclostrobln,. pyrimethanil, pyrowilon, quinoxyfen, = spiroxamine, sulfur, tebuconazole, tetraconazole, thiabendazole, thifluzamide, thiophanate-methyl, thiram, tiadlnil, trladlmefon, trladimenol, tricyclazole, trifloxystrobin, triticonazole, validamycin and vinciozolin; nematocides such as aldicarb, oxamyl and fenamiphos; bactericides such as streptomycin; acaricides such .as amftraz, chinomethionat chlorobenzilate, cyhexatin, dicofol, dienochlor, etoxazole, fenazaquin, fenbutatin oxide, fenpropathrin, fenpyroximate, hexythlazox, propargite, pyriciaben and tebufenpyrad; and biological agents including entomopathogenic bacteria, such as Bacillus thuringlensis subsp. Aizawal, Bacillus thutingiensis subsp. Kurstaki, and the encapsulated delta-endotoxins of Bacillus thuringiensis (e.g., Cellcap, MPV, MPVII);
entomopathogenic fungi, such as green muscardine fungus; and entomopathogenic virus including baculovirus, nucleopolyhedro virus (NPV) such as lizNPV, AfNPV
and granulosis virus (GV) such as CpGV. Methods of the invention may also comprise the use of plants genetically transformed to express proteins toxic to invertebrate pests (such as Bacillus thuringiensis delta-endotoxins). In such embodiments, the effect of exogenously applied invertebrate pest control compounds may be synergistic with the expressed toxin proteins.
General references for these agricultural protectants .include The Pesticide Manual, 13th Edition, Tomlin, Ed., British Crop Protection Council, Farnham, Surrey, U.K., 2003 and The BloPesticide Manual, rt Edition, Copping, Ed., British Crop Protection Council, Farnham, Surrey, U.K., 2001.
In certain instances, combinations with other invertebrate pest control compounds or agents having a similar spectrum of control but a different mode of action will be particularly advantageous for resistance management. Thus, compositions of the present invention can further comprise a biologically effective amount of at least one additional invertebrate pest control compound or agent having a similar spectrum of control but a different mode of action. Contacting a plant genetically modified to express a plant protection compound (e.g., protein) or the locus of the plant with a biologically effective amount of a compound of this invention can also provide a broader spectrum of plant protection and be advantageous for resistance management.
= Thus, methods of the invention employ a herbicide or herbicide combination and may further comprise the use of insecticides and/or fungicides, and/or other agricultural chemicals such as fertilizers. The use of such combined treatments of the Invention can broaden the spectrum of activity against additional weed species and suppress the proliferation of any resistant biotypes.
Date Recue/Date Received 2022-05-20 Methods of the invention can further comprise the use of plant growth regulators such as aviglycine, N-(phenylmethyl)-1H-putin-6-amine, ethephon, epocholeone, gibberellic acid, gibbereMn A4 and A7, herein protein, mepiquat chloride, prohexadione calcium, prohydrojasmon, sodium nitrophenolate and trinexapac-methyl and plant growth modifying organisms such as Bacillus cams strain SP01.
Embodiments of the present invention are further defined in the following Examples. It should be understood that these Examples are given by way of Illustration only. From the above discussion and these Examples, one skilled in the art can ascertain the essential characteristics of this Invention, and without departing from the spirit and scope thereof, can make various changes and modifications of the embodiments of the invention to adapt it to various usages and conditions.
Thus, various modifications of the embodiments of the invention, in addition to those shown and described herein, will be apparent to those skilled in the art from the foregoing description. Such modifications are also intended to fall within the scope of the appended claims.
EXPERIMENTAL
The following abbreviations are used in describing the present invention.
ALS acetolaotate synthase protein bp base pair g1ya14621 glyphosate acetyltransferase gene GLYAT4621 glyphosate acetyltransferase protein zrn-als ' wild type acetolactate synthase gene from brassica zrn-hra modified version of acetolactate synthase gene from brassica kb kilobase PCR polymerase chain reaction UTR untranslated region Example 1. Insert and Flanking Border Sequence Characterization of Brassica Event OP0-73496-4 Brassica (Brassica napus L.) has been modified by the Insertion of the glyphosate acetyltransferase gene (g1yat4621) derived from Bacillus licheniformis and optimized by gene shuffling. Plasmld PHP28181 contains an expression cassette as further described hereafter.
Date Recue/Date Received 2022-05-20 DNA construct PHP28181 was made by cloning the GAT4621:PINII TERM
fragment excised from DNA construct pZSL149 with BamHI and Mfel double digestion downstream to the AT-UBC110 promoter of DNA construct Q0272 in the same BamHI
and Mfel sites using T4 DNA ligase (New England Lab). The resulting PHP28181 DNA contains the expression cassette: AT-UBQ10 (DUPONT) PRO:GAT4621:131N111 TERM. See, Figure 1 and Figure 2.
The 2112 bp PHP28181A DNA fragment was prepared from plasmid PHP28181 with HindiII and Noti restriction enzyme double digestion. The digested plasmid DNA was resolved in a 1% agarose gel by electrophoresis. The DNA band of the correct size was excised and DNA fragment was extracted using a Qiagen DNA
fragment extraction kit (Qiagen). DNA fragment purity was checked by PCR with a series of dilutions of amp+ positive control DNA since the PHP28181 plasmid contains an amp+ gene in its backbone. DNA fragment concentration was measured spectrophotornetrically and confirmed by comparing to DNA low mass markers (InVitrogen) in an agarose gel.
Transformation was accomplished essentially as described in Chen and Tulsieram, US Patent Application Publication Number 2007/0107077. Buds were collected from donor line N81822BC and sterilized. Buds were then homogenized, filtered, and washed to collect the microspores. The resultant microspore suspension was adjusted to a specified density and cultured for 2 days. Embryogenic microspores were then isolated via gradient centrifugation and cultured.
Gold particles coated with the PI-11328181A DNA fragment were used for transformation. Biolistic transformation was carried out using the PDS-1000/He Particle Delivery System (Bio-Fiad, Hercules, CA) as described by Klein, et al., (1987) Nature 327:70-73. Transformed embryogenic microspores were cultured in fresh medium In dark conditions for 10-12 days, then under dim light for 1-3 weeks.
Green embryos were transferred to fresh medium and cultured for two weeks to select for glyphosate tolerance. Germinated shoots and/or plants were transferred to growth medium supplemented with glyphosate.
The g/yat4621 gene was derived from the soil bacterium Bacillus licheniformis and was synthesized by a gene shuffling process to optimize the acetyitransferase activity of the GLYAT4621 enzyme (Castle, et at, (2004) gc(ence 304:1161-1164).
The inserted fragment (Figure 3) from this plasmid contains the g1yet4621 gene =
cassette. The expressien of the glyat4621 gene is controlled by the UBQ10 regulatory region from Arabldopsis and the grill terminator (see, Table 4). A summary of the transformation fragment of plasmid PHP28181 is shown in Table 4. The genetic =
=
Date Recue/Date Received 2022-05-20 elements of plasmid PHP28181 used In the creation of DP-073496-4 are shown in Table 3.
Table 3: Description of Genetic Elements in Plaantid P11P28181 ,=1.--"..;';-:r.'zL =
- = ' - 7' -."
- f=, 4 ;
Traa*rmation = -'1F,L"..7',e.4 'f.R.Pr-4-41V 1 to 2112 See Table 4 for information on ?rapidest ¨ PH-p281 the elements in this region .
:AN,rt=
git'srAss;
includes DNA from various sources for Plasmid 2113 to 4770 elements 2658 plasmid construction and Construct below plasraid replication ,=`===7=,- = ,==-=*:=,;"::,w,",..41,:',-57: 0-lactamase gene coding for =
amoicalin resistance from E eIi 2736 to 3596 bla (ApR) 861 (Sutriliffe, 1978) ((amsch-Perron, 1441 153'4! at, 1984) ,g,74 Hae II fragment containing bacterial f. = ,:=,2.41-' -µ
'I 4170 to 4539 colE1 on370 origin of replication region (colE1 V 7 , -tt = derived) (Tomizawa et id, 1977) =
=
Date Recue/Date Received 2022-05-20 Table 4: Description of Genetic Elements in the Tranaformation Fragment *WI M.1.1NiniVit-VarigiMfeitir:06,ANWW.,02AN, talW
. = -:
AP* = 1:14:6010a-afrAliPtiMil7-70. F.";A:14.At0,711VF:41-;;ZPi4 ' TP;r4"1:44P*'- 4!;'41142tjagg;TAZIA7N0-#4.farrr,"e;:'4-itAg.
A'rc.fAkr-fk.-*'ga'tiS¶'Pr4Sr!Ftk,kk,:,..LdtftA.zdiarr=':V:*Afaaa'i:k:"g'gre6t'X'!SP':Pr';:.1 .:::V:i'5C':';?
Polyiinker - I to 7 7 Region required for cloning genetic elements Region Version of the promoter region from Arabidopeis to 1312 UB,Q1 0 UBal 0 polythiquifin gene (Norris et eV, Promoter 1993) developed by the P... T. &Pont de Nemours and Company Polylinker 1313 to 1335 23 Region required for cloning genetic elements Region 1336 to 1779 4062f Gene Synthetic glyphosate N-acetyltransferase gene (Castle et al., 2004; Siehl et al., 2007) linker 1780 to 1796 Poly 17 Region required for cloning genetie elements Region 1797 t 2106 310 Terminator region from Sokoloff tuberosum proteinase o Terminator inhibitor II gene (Keil et ed, 1986; An et al, 1989) linker 2107 to 2112 Poly 6 Region required for cloning genetic elements Region The nucleotide sequence of the inserted DNA in the DP-0734964 event has been determined. PCR amplification of the unique junctions spanning the introduced genetic elements can distinguish DP-073496-4 plants from their non-genetically-modified counterparts and can .be used to screen for the presence of the inserted DNA, even at very low concentrations. Described below is a construct-specific poiymerase chain reaction (PCR) assay on genomic DNA from DP-073496-4 Brassica.
Specifically, genomic DNA from the test substance (plant material of event DP-0764954) and the control substance (plant material of a non-genetically modified Brassica with a genetic background representative of the event background) is isolated and subjected to qualitative PCR amplification using a construct-specific primer pair.
The PCR products are separated on 1.6% or 2% agarose gels to confirm the presence of the inserted construct in the genomic DNA generated from the test substance, and absence in the genomic DNA generated from the control substance. A reference = standard (100 base pair DNA Ladder; invitrogen Corporation Catalog #
10380-012) is used to determine the PCR product size.
Test and control samples are harvested from plant's. Genomic DNA extraction from the test and control tissues is performed using a standard urea extraction protocol, if leaf tissue. Genomic DNA from the test and control samples is isolated using Wizards Magnetic 96 DNA Plant System (Promega Corporation Catalog #
FF3760), if seed tissue. GenomIc DNA Is quantified on a spectrofiuorometer using .72 Date Recue/Date Received 2022-05-20 PicoGreene reagent (Molecular Probes, Inc., Eugene, OR)= and/or visualized on an agarose gel to confirm quantitation values and to determine the DNA quality.
Genomic DNA isolated from plant material .of event DP-073496-4 and control samples is subjected to PCR amplification (PCR Master Mix Catalog #7505 from Promega Corporation) utilizing a construct-specific primer pair which spans at least a portion of the glyat4621 coding region, and allows for the unique identification of maize event DP-073496-4. A second primer set is used to amplify an endogenous gene as a positive control for PCR amplification. The PCR target site and size of the expected PCR product for each primer set are compared to the observed results.
Example 2. Characterization of Event DP-073496-4 by Southern Blot Southern blot analyses (Southern, 1975) are performed to investigate the number of sites of insertion of the transforming DNA, the copy number and functional integrity of the genetic elements and the absence of plasmid backbone sequences.
The method used is described generally as follows. Genomic DNA is extracted from lyophilized tissue sampled from DP-073496-4 Brassica and non-genetically-modified control plants. Genomic DNA is digested with restriction endonuclease enzymes and size-separated on an agarose. gel. A molecular weight marker is run alongside samples for size estimation purposes. DNA fragments separated on agarose gel are depurinated, denatured and neutralized in situ and transferred to a , nylon membrane. Following transfer to the membrane, the DNA is bound to the membrane by UV crosslinking. Fragments homologous to the glyat4621 gene are generated by PCR from plasmid PHP28181, separated on an agarose gel by size, exsized and purified using a gel extraction kit, Labeled probe is hybridized to the = 25 target DNA on the .nylon membranes for detection of the specific fragments. Washes after hybridization are carried out at high stringency. Blots are exposed to X-ray film for one or more time points to detect hybridizing fragments and visualize molecular weight markers.
Example 3. Exaression of the insert Expression of the G1YA14621 protein is evaluated using leaf tissue collected from transgenic plants. For example, four fresh leaf punches may be collected and ground in sample extraction buffer using a GenoGrinder (Spex Certiprep). Total Extractable Protein (TEP) can be determined using the Blo-Rad Protein assay, which is based on the Bradford dye-binding procedure. Sample extracts may be diluted in sample extraction buffer for ELISA analysis.
=
Date Recue/Date Received 2022-05-20 The levels of expression of the GAT4621 protein in DP-073496-4 Brassies can be determined by quantitative enzyme linked immunosorbent assay (ELISA) of samples obtained from multiple field trial locations. Replicate seed samples (three replicates) may be obtained from DP-073496-4 plants treated with the maximum recommended label rate of Touchdown Total glyphosate herbicide (500 g/I
glyphosate as potassium salt; 0.60-1.35 I/ha), applied at the cotyledon to 8-leaf stage, as this represents a likely commercial cultivation scenario.
Another way to verify, the expression of the insert in DP-073496-4 E3rass1ca plants is to evaluate the transformed plants tolerance to glyphosate.
Muitigenerational stability and Within-generation segregation of the herbicide tolerant trait conferred by expression of the GAT4621 enzyme will be confirmed using a functional assay for herbicide tolerance. Tests are conducted on at least three generations of plant material. Herbicide injury may be scored as described in Table 5.
15Table 5: The 0 to 100 crop response rating system for herbicide injury 1-`
1.4 0 No Effect No crop reduction or injury 10 Slight crop discoloration or stunting Slight Effect Some crop discoloration, stunting, or stunt loss Crop injury more pronounced, but not lasting Moderate injury, drop usually recovers Moderate Effect Crop injury more lasting, recovery doubtful Lasting crop injury, no recovery Heavy crop injury and stand loss Severe Effect Crop nearly destroyed - A few surviving plants Only occasional live crop plants left 100 Complete Effect Complete crop destruction Example 4. Construct Specific PCR Analysis of Brassica Event DP-073496-4 Genomic DNA isolated from leaf of DP-073496-4 canola (12F2 generation) and 20 control vandal (non-genetically modified) was subjected to PCR
amplification (Roche High Fidelity PCR Master Kit, Roche Catalog # 12140314001) utilizing the construct-specific primer pair (09-0-3290/09-0-3288) which spans the ubiquitin promoter and the 98t4621 gene cassette (Figure 4). A second primer set (09-0-2812/09-0-2813) was used to amplify the endogenous canola FatA gene as a positive control for PCR
25 amplification. The PCR target site and size of the expected PCR
product for each primer sets are shown in Table 8. PCR reagents and reaction conditions are shown In Table 9. The primer sequences used in this study are listed in Table 10. In this study, 100 ng of leaf genomic DNA was used in all PCR reactions.
Date Recue/Date Received 2022-05-20 A PCR product of approximately 600 bp in size amplified by the construct-specific primer set 09-0-3290/09-0-3288 was observed in PCR reactions using plasmid PHP28181 (10 rig) as a template and three DP-073496-4 canola plants, but absent in three control canola plants and the no-template control (Figure 5), Samples were loaded as shown in Table 6.
= Table 6 Lane Sample = 1 Low Mass Molecular Weight Marker 2 Blank 3 Non-Genetically Modified canals C1 4 Non-Genetically Modified canola C2 5 = Non-Genetically Modified canola C3 6 DP-073496-4 canola T1 7 DP-073496-4 canola T2 8 DP-073496-4 canola T3 9 NT Control 10 _______________ Plasmic! PHP28181 11 Blank 12 Low Mass Molecular Weight Marker These results correspond with the expected PCR product size (675 bp) for samples containing DP-073496-4 canola genomic DNA. A PCR product approximately 450 bp in size was observed for both DP-073496-4 canola and control mole plants following PCR reaction with the primer set 09-0-2812/09-0-2813 for detection of the endogenous FatA gene (Figure 6): Samples were loaded as shown in Table 7.
Table 7 Lane Sample 1 Low Mass Molecular Weight Marker 2 Blank 3 Non-Genetically Modified canola C1 4 Non-Genetically Modified canola C2 5 Non-Genetically Modified canola C3 6 DP-073496-4 canola T1 7 DP-073496-4 canola T2 DP-073496-4 canola T3 NT Control 10 Plasmid PHP28181 11 Blank 12 Low Mass Molecule Weight Marker These results correspond with the expected PCR product size (506 bp) for genomic DNA samples containing, the canola endogenous FatA gene, The endogenous target band was not observed in the no-template control.
. , Date Recue/Date Received 2022-05-20 Table 8; PCR Genomic DNA Target Site and Expected Size of PCR Products Primer Set Target Site Expected Size of PCR Product (bp) 09-0-3290/09-0-3288 Construct-Specific ubiquitin promoter and get4621 Endogenous oanola FatA
gene' PCR: POLYMERASE CHAIN REACTION =
BP: BASE PAIRS
1. Genbank accession number for FatA gene is X87842.1. This sequence was used to design PCR primers.
Table 9: PCR Rees:lents and Reaction Conditions PCR Reagents = PCR Reaction Conditions ' =
Volume Cycle Temp Time Reagent # Cycles (pL) Element ( C) (sec) Template DNA initial (100 ng/p1) Denaturation Primer 1 (10 pM) 0.75 Denaturation 94 10 Primer 2 (10 pM) 0,75 Annealing 65 20 35 PCR Master Mix* 12.5 Elongation 72 45 ddH20 10 Final Elongation 72 180 1 Until Hold Cycle 4 analysis PCR: POLYMERASE CHAIN REACTION
DDH20: DOUBLE-DISTILLED WATER
* Roche High Fidelity Master Mix Table 10: = List of Primer Seauences Used in PM Reactions Primer Sequence 5' ¨3' Target Sequence Name SEQ ID NO 4:
09-0-3290 Ubiquitin Promoter AOCTATTGCTICACCGCCTTAGC
SEQ ID NO: 5 09-0-3288 get4821 GCTCAGCTTGGTGGAATGAAGCCAD
SEQ ID NO: 6 Canola Endogenous 09-0-2812 GACACAAGGCGGCTTCAAAGAGTTACAGATG FatA
SEQ IC NO 7: Canola Endogenous 09-0-2813 ACAATGTCATCTTGCTGGCATTCTCTTCTG FatA
Example 5. Further Insert and Flankino Border Sequence Characterization of Brass/ca Event DP-073496-4 =
To characterize the integrity of the inserted DNA and the = genomic insertion site, the flanking genomic DNA border regions of the DP-073498-4 event were = = =
Date Recue/Date Received 2022-05-20 determined. Flanking genomic sequence of DP-073496-4 Is included within SEQ ID
NO: 2. PCR amplification from the insert and border sequences confirms that the border regions are of Brassies origin and that the junction regions can be used for identification- of DP-073496-4 Brassies, Overall, characterization of the insert and genomic border sequences, along with Southern blot data, indicate a single insertion of the DNA fragment present in the Brassica genome. Various molecular techniques are then used to specifically characterize the integration site.
In the Initial characterization, the flanking genomic border regions are cloned and sequenced using the OenomeVValker end inverse PCR methods. Using information from the flanking border sequence, PCR is performed on DP-073496-4 genomic DNA and unmodified control genomic DNA, Those skilled in the art will also Include a control PCR using an endogenous gene to verify that the isolated genomic DNA Is suitable for PCR amplification.
Date Recue/Date Received 2022-05-20 CD
CD
CD
CD
CD
Table 11.
0_ PCR-based o event-specific detection methods o PCR
Primer 1 Primer 2 Probe ________________________________ Assay =
event type Mune Sequence Name Sequence Name Sequence 5 label Quencher 10-0-3514 GG Ls...toGGGC 1515 TTATOCGGICCTAG
DP-073496- Gel- SEO. ID NO:
4 based .20 GCCG 140:23 CAAACCTOC
09-0-2824 GTTaTaaTC 2325 CAAACCTCCATAG TTAGTTAGATC
DP473496. Real- SEQ ID
ATAGar.ATTAC S!QJP AGTTCAACATOTA 09-0P83 AGGATATTCTT
oe 4 time NO:21 Maim P1024 A
SEO. ID NO=26 . II FAM 4.469 FatA A- Real- SEQ ID TCGGGACGAGT 5E010 = spedfic time N022 AC F10:25 ACAiCa IAAATAT 5E0 ID N0:17 TCATGCTTC FAM MOB
Table 12. Summary Table of SEQ id NOS
SEQ ID NO Description 1 0A14621 protein 2 DP-073496-4 insert and flanking sequence Primer 09-0-3290 (SEQ ID NO: 4 4 AGCTATTGCTICACCGCCTTAGC) Target ¨ Ubiquitin Promoter Primer 09-0-3288 (SEQ ID NO: 5 GCTCAGCTTGGTGGAATGAAGCCAC) Target g8t4621 Primer 09-0-2812 (SEQ ID NO: 6 6 GACACAAGGCGGCTTCAAAGAGTTACAGATG) Target Canala Endogenous FatA
Primer 09-0-2813 (SEQ ID NO:
7 ACAATGTCATMGCTGGCATTCTCTTCTG ) Canola Endogenous FatA
8 Right border genomIc sequence 9 Left border genomic sequence Complete Internal transgene 11 Complete flanking and Internal transgene 12 Right flanking genomic/right border transgene (10 nt/10nt) 13 Left flanking genomicfleft border transgene (10 ntll Ont) 14 Right flanking genomiaright border transgene (20 nt/20nt) Left flanking genomic/left border transgene (20 nt/20nt) 16 Right flanking genomic/right border transgene (30 nt/30nt) 17 Left flanking genomialeft border transgene (30 nt/30nt) 18 Right flanking genomic/complete transgene 19 Left flanking genomic/complete transgene Primer 10-0-3514 21 Primer 09-0-2824 22 Primer 09-0-3249 23 Primer 10-0-3515 24 Primer 09-0-2825 Primer 09-0-3251 26 Primer 09-QP83 Date Recue/Date Received 2022-05-20 SEQ ID NO Description 27 Primer 09-QP87 The article a and wan' are used herein to refer to one or more than one (Le., to at least one) of the grammatical object of the article. By way of example, an element' means one or more element All publications and patent applications mentioned in the specification are indicative of the level of those skilled in the art to 'which this invention pertains.
The scope of the claims should not be limited by the preferred embodiments set forth in the examples, but should be given the broadest interpretation consistent with the description as a whole.
=
=
. =
=
Date Recue/Date Received 2022-05-20 Applicant's or agent's international applicathon No.
Me reference 35718/39908 PCT/US2010/
INDICATIONS RELATING TO DEPOSITED MICROORGANISM
OR OTHER BIOLOGICAL MATERIAL
(PCT Rule 13b1s) A. The irsications made below relate to the deposited microorganism or other blobglcal material referred to in the description on page 4:1, line 5 B. IDENTIFICATION OF DEPOSIT Further deposits are kientfled on an additional sheet 0 Name of depository Institution American Type Culture Collection Adiess of depositary institution (Including postal code and country) 10801 University Blvd.
Manassas, Virginia 20110-2/09 USA
=
Date of deposit Accession Number = 24 November 2010 PTA-C. ADDITIONAL INDICATIONS (leave blank if not epplicable) This Intimation is continued on an additional sheet El page 3, line 9; page 5, line 8; page 82, line 8; page 82, line 19; page 84, line 33; page 85, line 15 and page 85, line 21 =
D. DESIGNATED STATES FOR WHICH INDICATIONS ARE MADE (If the Indicators are not fore! designated Woks) E. SEPARATE FURNIs KING OF INDICATIONS (leave blank if not applicable) The Indications listed below will be submitted to the International Bureau later (specify the general nature of the Indications e.g. , 'Accession ivumber of cfrepostr) For receivkig Office use only For International Bureau use only 0 This sheet was raceme yinm me mternerional application CI This sheet no sow mu mu; pm nualeawirm vutood on:
Authorized officer Authorized officer =
Forrn PCT/R0/134 (July 1998) "
Date Recue/Date Received 2022-05-20
Claims (4)
1. A glyphosate tolerant Brassica plant having in its genome a DP-073496-4 event and further comprising at least one additional polynucleotide that confers male sterility in the plant compared to a corresponding plant that does not comprise the additional polynucleotide, wherein the DP-073496-4 event comprises in the following order: a polynucleotide comprising SEQ ID
NO: 12, a polynucleotide encoding a glyphosate-N-acetyltransferase and a polynucleotide comprising SEQ ID NO: 13.
NO: 12, a polynucleotide encoding a glyphosate-N-acetyltransferase and a polynucleotide comprising SEQ ID NO: 13.
2. The Brassica plant of claim 1, wherein the additional polynucleotide is a recombinant polynucleotide.
3. The Brassica plant of claim 2, wherein the additional polynucleotide was introduced into a Brassica plant cell by genetic transformation.
4. A method of growing a glyphosate tolerant Brassica plant comprising:
(a) planting in an area of cultivation a Brassica plant or seed having in its genome a DP-073496-4 event and a polynucleotide conferring male sterility; and (b) applying glyphosate to the area of cultivation in an amount that is effective to prevent growth of plants that are susceptible to glyphosate;
wherein the DP-073496-4 event comprises in the following order: a polynucleotide comprising SEQ ID NO: 12, a polynucleotide encoding a glyphosate-N-acetyltransferase and a polynucleotide comprising SEQ ID NO: 13.
Date Recue/Date Received 2022-05-20
(a) planting in an area of cultivation a Brassica plant or seed having in its genome a DP-073496-4 event and a polynucleotide conferring male sterility; and (b) applying glyphosate to the area of cultivation in an amount that is effective to prevent growth of plants that are susceptible to glyphosate;
wherein the DP-073496-4 event comprises in the following order: a polynucleotide comprising SEQ ID NO: 12, a polynucleotide encoding a glyphosate-N-acetyltransferase and a polynucleotide comprising SEQ ID NO: 13.
Date Recue/Date Received 2022-05-20
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CA3159670A CA3159670A1 (en) | 2010-11-24 | 2010-11-24 | Brassica gat event dp-073496-4 and compositions and methods for the identification and/or detection thereof |
Applications Claiming Priority (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CA3069427A CA3069427A1 (en) | 2010-11-24 | 2010-11-24 | Brassica gat event dp-073496-4 and compositions and methods for the identification and/or detection thereof |
CA3159670A CA3159670A1 (en) | 2010-11-24 | 2010-11-24 | Brassica gat event dp-073496-4 and compositions and methods for the identification and/or detection thereof |
CA2810180A CA2810180C (en) | 2010-11-24 | 2010-11-24 | Brassica gat event dp-073496-4 and compositions and methods for the identification and/or detection thereof |
Related Parent Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CA3069427A Division CA3069427A1 (en) | 2010-11-24 | 2010-11-24 | Brassica gat event dp-073496-4 and compositions and methods for the identification and/or detection thereof |
Publications (1)
Publication Number | Publication Date |
---|---|
CA3159670A1 true CA3159670A1 (en) | 2012-05-31 |
Family
ID=53199751
Family Applications (4)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CA2995972A Abandoned CA2995972A1 (en) | 2010-11-24 | 2010-11-24 | Brassica gat event dp-073496-4 and compositions and methods for the identification and/or detection thereof |
CA3069427A Abandoned CA3069427A1 (en) | 2010-11-24 | 2010-11-24 | Brassica gat event dp-073496-4 and compositions and methods for the identification and/or detection thereof |
CA3159670A Pending CA3159670A1 (en) | 2010-11-24 | 2010-11-24 | Brassica gat event dp-073496-4 and compositions and methods for the identification and/or detection thereof |
CA2891153A Active CA2891153C (en) | 2010-11-24 | 2010-11-24 | Brassica gat event dp-073496-4 and compositions and methods for the identification and/or detection thereof |
Family Applications Before (2)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CA2995972A Abandoned CA2995972A1 (en) | 2010-11-24 | 2010-11-24 | Brassica gat event dp-073496-4 and compositions and methods for the identification and/or detection thereof |
CA3069427A Abandoned CA3069427A1 (en) | 2010-11-24 | 2010-11-24 | Brassica gat event dp-073496-4 and compositions and methods for the identification and/or detection thereof |
Family Applications After (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CA2891153A Active CA2891153C (en) | 2010-11-24 | 2010-11-24 | Brassica gat event dp-073496-4 and compositions and methods for the identification and/or detection thereof |
Country Status (1)
Country | Link |
---|---|
CA (4) | CA2995972A1 (en) |
-
2010
- 2010-11-24 CA CA2995972A patent/CA2995972A1/en not_active Abandoned
- 2010-11-24 CA CA3069427A patent/CA3069427A1/en not_active Abandoned
- 2010-11-24 CA CA3159670A patent/CA3159670A1/en active Pending
- 2010-11-24 CA CA2891153A patent/CA2891153C/en active Active
Also Published As
Publication number | Publication date |
---|---|
CA2891153A1 (en) | 2012-05-31 |
CA3069427A1 (en) | 2012-05-31 |
CA2995972A1 (en) | 2012-05-31 |
CA2891153C (en) | 2018-04-03 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
US20230287521A1 (en) | Brassica gat event and compositions and methods for the identification and/or detection thereof | |
US7951995B2 (en) | Soybean event 3560.4.3.5 and compositions and methods for the identification and detection thereof | |
US7897846B2 (en) | Maize event DP-098140-6 and compositions and methods for the identification and/or detection thereof | |
US7928296B2 (en) | Maize event DP-098140-6 and compositions and methods for the identification and/or detection thereof | |
US8324457B2 (en) | Soybean event 3560.4.3.5 and compositions and methods for the identification and/or detection thereof | |
AU2010364322B2 (en) | Brassica GAT event DP-073496-4 and compositions and methods for the identification and/or detection thereof | |
WO2012071039A1 (en) | Brassica gat event dp-061061-7 and compositions and methods for the identification and/or detection thereof | |
US8575431B2 (en) | Brassica GAT event DP-061061-7 and compositions and methods for the identification and/or detection thereof | |
AU2015268681B2 (en) | Brassica gat event dp-073496-4 and compositions and methods for the identification and/or detection thereof | |
AU2013206424B2 (en) | Brassica gat event dp-073496-4 and compositions and methods for the identification and/or detection thereof | |
CA2891153C (en) | Brassica gat event dp-073496-4 and compositions and methods for the identification and/or detection thereof |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
EEER | Examination request |
Effective date: 20220804 |
|
EEER | Examination request |
Effective date: 20220804 |
|
EEER | Examination request |
Effective date: 20220804 |
|
EEER | Examination request |
Effective date: 20220804 |
|
EEER | Examination request |
Effective date: 20220804 |