CN118147174A - 除草剂抗性蛋白、基因和用途 - Google Patents
除草剂抗性蛋白、基因和用途 Download PDFInfo
- Publication number
- CN118147174A CN118147174A CN202410244759.8A CN202410244759A CN118147174A CN 118147174 A CN118147174 A CN 118147174A CN 202410244759 A CN202410244759 A CN 202410244759A CN 118147174 A CN118147174 A CN 118147174A
- Authority
- CN
- China
- Prior art keywords
- seq
- plant
- herbicide
- protein
- spp
- 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
- 108090000623 proteins and genes Proteins 0.000 title claims abstract description 124
- 239000004009 herbicide Substances 0.000 title claims abstract description 88
- 102000004169 proteins and genes Human genes 0.000 title claims abstract description 61
- 230000002363 herbicidal effect Effects 0.000 title claims abstract description 58
- 238000000034 method Methods 0.000 claims abstract description 23
- 239000005556 hormone Substances 0.000 claims abstract description 12
- 229940088597 hormone Drugs 0.000 claims abstract description 12
- 239000003112 inhibitor Substances 0.000 claims abstract description 8
- 241000196324 Embryophyta Species 0.000 claims description 188
- 230000009261 transgenic effect Effects 0.000 claims description 94
- 108020004414 DNA Proteins 0.000 claims description 56
- 102000053602 DNA Human genes 0.000 claims description 46
- 230000014509 gene expression Effects 0.000 claims description 46
- 239000013598 vector Substances 0.000 claims description 40
- 108020004511 Recombinant DNA Proteins 0.000 claims description 33
- 150000007523 nucleic acids Chemical group 0.000 claims description 25
- FERIUCNNQQJTOY-UHFFFAOYSA-N Butyric acid Natural products CCCC(O)=O FERIUCNNQQJTOY-UHFFFAOYSA-N 0.000 claims description 16
- 108091028043 Nucleic acid sequence Proteins 0.000 claims description 15
- -1 aryloxy phenoxy propionic acids Chemical class 0.000 claims description 15
- FERIUCNNQQJTOY-UHFFFAOYSA-M Butyrate Chemical compound CCCC([O-])=O FERIUCNNQQJTOY-UHFFFAOYSA-M 0.000 claims description 14
- 150000001875 compounds Chemical class 0.000 claims description 14
- 239000012634 fragment Substances 0.000 claims description 14
- 102000040430 polynucleotide Human genes 0.000 claims description 12
- 108091033319 polynucleotide Proteins 0.000 claims description 12
- 239000002157 polynucleotide Substances 0.000 claims description 12
- RFOHRSIAXQACDB-UHFFFAOYSA-M sodium;2-(2,4-dichlorophenoxy)acetate Chemical compound [Na+].[O-]C(=O)COC1=CC=C(Cl)C=C1Cl RFOHRSIAXQACDB-UHFFFAOYSA-M 0.000 claims description 12
- 239000005627 Triclopyr Substances 0.000 claims description 11
- REEQLXCGVXDJSQ-UHFFFAOYSA-N trichlopyr Chemical compound OC(=O)COC1=NC(Cl)=C(Cl)C=C1Cl REEQLXCGVXDJSQ-UHFFFAOYSA-N 0.000 claims description 11
- 239000005558 Fluroxypyr Substances 0.000 claims description 10
- 230000002068 genetic effect Effects 0.000 claims description 10
- 230000000295 complement effect Effects 0.000 claims description 9
- 230000000694 effects Effects 0.000 claims description 9
- MEFQWPUMEMWTJP-UHFFFAOYSA-N fluroxypyr Chemical compound NC1=C(Cl)C(F)=NC(OCC(O)=O)=C1Cl MEFQWPUMEMWTJP-UHFFFAOYSA-N 0.000 claims description 9
- YIVXMZJTEQBPQO-UHFFFAOYSA-N 2,4-DB Chemical compound OC(=O)CCCOC1=CC=C(Cl)C=C1Cl YIVXMZJTEQBPQO-UHFFFAOYSA-N 0.000 claims description 8
- 230000001965 increasing effect Effects 0.000 claims description 8
- 239000005504 Dicamba Substances 0.000 claims description 5
- 239000005614 Quizalofop-P-ethyl Substances 0.000 claims description 5
- IWEDIXLBFLAXBO-UHFFFAOYSA-N dicamba Chemical compound COC1=C(Cl)C=CC(Cl)=C1C(O)=O IWEDIXLBFLAXBO-UHFFFAOYSA-N 0.000 claims description 5
- 238000004519 manufacturing process Methods 0.000 claims description 5
- 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 claims description 5
- 235000010233 benzoic acid Nutrition 0.000 claims description 3
- 150000001559 benzoic acids Chemical class 0.000 claims description 3
- 230000001939 inductive effect Effects 0.000 claims description 3
- QIIPQYDSKRYMFG-UHFFFAOYSA-N phenyl hydrogen carbonate Chemical class OC(=O)OC1=CC=CC=C1 QIIPQYDSKRYMFG-UHFFFAOYSA-N 0.000 claims description 3
- SIOXPEMLGUPBBT-UHFFFAOYSA-N picolinic acid Chemical class OC(=O)C1=CC=CC=N1 SIOXPEMLGUPBBT-UHFFFAOYSA-N 0.000 claims description 3
- BBPLSOGERZQYQC-UHFFFAOYSA-N 6-methylheptyl 2-(2,4-dichlorophenoxy)acetate Chemical compound CC(C)CCCCCOC(=O)COC1=CC=C(Cl)C=C1Cl BBPLSOGERZQYQC-UHFFFAOYSA-N 0.000 claims description 2
- 230000002708 enhancing effect Effects 0.000 claims description 2
- 238000010362 genome editing Methods 0.000 claims description 2
- 230000001172 regenerating effect Effects 0.000 claims description 2
- 230000001131 transforming effect Effects 0.000 claims description 2
- 125000003275 alpha amino acid group Chemical group 0.000 claims 3
- ABIPNDAVRBMCHV-UHFFFAOYSA-N 4,4-dimethyl-2,3-dihydro-1h-naphthalene Chemical compound C1=CC=C2C(C)(C)CCCC2=C1 ABIPNDAVRBMCHV-UHFFFAOYSA-N 0.000 claims 1
- 239000005631 2,4-Dichlorophenoxyacetic acid Substances 0.000 description 68
- 235000018102 proteins Nutrition 0.000 description 52
- 210000004027 cell Anatomy 0.000 description 37
- 241000219194 Arabidopsis Species 0.000 description 33
- 230000009466 transformation Effects 0.000 description 26
- 239000002609 medium Substances 0.000 description 24
- 230000015556 catabolic process Effects 0.000 description 23
- 235000010469 Glycine max Nutrition 0.000 description 22
- 244000068988 Glycine max Species 0.000 description 22
- 238000006731 degradation reaction Methods 0.000 description 22
- 238000012360 testing method Methods 0.000 description 22
- 108090000765 processed proteins & peptides Proteins 0.000 description 21
- 230000002018 overexpression Effects 0.000 description 20
- 229920001184 polypeptide Polymers 0.000 description 19
- 102000004196 processed proteins & peptides Human genes 0.000 description 19
- 238000012216 screening Methods 0.000 description 19
- 240000008042 Zea mays Species 0.000 description 18
- 235000002017 Zea mays subsp mays Nutrition 0.000 description 18
- 102000004190 Enzymes Human genes 0.000 description 15
- 108090000790 Enzymes Proteins 0.000 description 15
- 244000025254 Cannabis sativa Species 0.000 description 14
- 108700019146 Transgenes Proteins 0.000 description 14
- 150000001413 amino acids Chemical group 0.000 description 13
- 241000894006 Bacteria Species 0.000 description 12
- 239000007788 liquid Substances 0.000 description 12
- 235000007688 Lycopersicon esculentum Nutrition 0.000 description 11
- 240000003768 Solanum lycopersicum Species 0.000 description 11
- 235000020971 citrus fruits Nutrition 0.000 description 11
- 102000039446 nucleic acids Human genes 0.000 description 11
- 108020004707 nucleic acids Proteins 0.000 description 11
- 241000219195 Arabidopsis thaliana Species 0.000 description 10
- 241000588724 Escherichia coli Species 0.000 description 10
- 229930006000 Sucrose Natural products 0.000 description 10
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 10
- 235000016383 Zea mays subsp huehuetenangensis Nutrition 0.000 description 10
- 238000006243 chemical reaction Methods 0.000 description 10
- 235000009973 maize Nutrition 0.000 description 10
- 239000007921 spray Substances 0.000 description 10
- 239000005720 sucrose Substances 0.000 description 10
- 240000002791 Brassica napus Species 0.000 description 9
- 235000006008 Brassica napus var napus Nutrition 0.000 description 9
- 150000003839 salts Chemical class 0.000 description 9
- 240000007124 Brassica oleracea Species 0.000 description 8
- 235000003899 Brassica oleracea var acephala Nutrition 0.000 description 8
- 235000006618 Brassica rapa subsp oleifera Nutrition 0.000 description 8
- 235000004977 Brassica sinapistrum Nutrition 0.000 description 8
- 241000207199 Citrus Species 0.000 description 8
- 240000007594 Oryza sativa Species 0.000 description 8
- 235000007164 Oryza sativa Nutrition 0.000 description 8
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 description 8
- 235000005822 corn Nutrition 0.000 description 8
- 239000013604 expression vector Substances 0.000 description 8
- 230000001105 regulatory effect Effects 0.000 description 8
- JQXXHWHPUNPDRT-WLSIYKJHSA-N rifampicin Chemical compound O([C@](C1=O)(C)O/C=C/[C@@H]([C@H]([C@@H](OC(C)=O)[C@H](C)[C@H](O)[C@H](C)[C@@H](O)[C@@H](C)\C=C\C=C(C)/C(=O)NC=2C(O)=C3C([O-])=C4C)C)OC)C4=C1C3=C(O)C=2\C=N\N1CC[NH+](C)CC1 JQXXHWHPUNPDRT-WLSIYKJHSA-N 0.000 description 8
- 229960001225 rifampicin Drugs 0.000 description 8
- 239000000758 substrate Substances 0.000 description 8
- 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 7
- 241000589158 Agrobacterium Species 0.000 description 7
- 102000016680 Dioxygenases Human genes 0.000 description 7
- 108010028143 Dioxygenases Proteins 0.000 description 7
- 239000005562 Glyphosate Substances 0.000 description 7
- 244000062793 Sorghum vulgare Species 0.000 description 7
- XDDAORKBJWWYJS-UHFFFAOYSA-N glyphosate Chemical compound OC(=O)CNCP(O)(O)=O XDDAORKBJWWYJS-UHFFFAOYSA-N 0.000 description 7
- 229940097068 glyphosate Drugs 0.000 description 7
- 235000019713 millet Nutrition 0.000 description 7
- 239000000047 product Substances 0.000 description 7
- 239000002689 soil Substances 0.000 description 7
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 7
- 229920001817 Agar Polymers 0.000 description 6
- 244000075850 Avena orientalis Species 0.000 description 6
- 235000014698 Brassica juncea var multisecta Nutrition 0.000 description 6
- 235000011301 Brassica oleracea var capitata Nutrition 0.000 description 6
- 235000001169 Brassica oleracea var oleracea Nutrition 0.000 description 6
- 241001301148 Brassica rapa subsp. oleifera Species 0.000 description 6
- 229920000742 Cotton Polymers 0.000 description 6
- 235000003198 Cynara Nutrition 0.000 description 6
- 241000208947 Cynara Species 0.000 description 6
- 244000127993 Elaeis melanococca Species 0.000 description 6
- 239000005561 Glufosinate Substances 0.000 description 6
- 241000219146 Gossypium Species 0.000 description 6
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N Phenol Natural products OC1=CC=CC=C1 ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 description 6
- 240000006394 Sorghum bicolor Species 0.000 description 6
- 230000009471 action Effects 0.000 description 6
- 239000008272 agar Substances 0.000 description 6
- 230000001580 bacterial effect Effects 0.000 description 6
- 238000010276 construction Methods 0.000 description 6
- 108010083942 mannopine synthase Proteins 0.000 description 6
- LCPDWSOZIOUXRV-UHFFFAOYSA-N phenoxyacetic acid Chemical compound OC(=O)COC1=CC=CC=C1 LCPDWSOZIOUXRV-UHFFFAOYSA-N 0.000 description 6
- 235000009566 rice Nutrition 0.000 description 6
- 239000000243 solution Substances 0.000 description 6
- 210000001519 tissue Anatomy 0.000 description 6
- 239000011782 vitamin Substances 0.000 description 6
- 235000013343 vitamin Nutrition 0.000 description 6
- 229940088594 vitamin Drugs 0.000 description 6
- 229930003231 vitamin Natural products 0.000 description 6
- 244000105624 Arachis hypogaea Species 0.000 description 5
- 235000011293 Brassica napus Nutrition 0.000 description 5
- 239000007987 MES buffer Substances 0.000 description 5
- 244000269722 Thea sinensis Species 0.000 description 5
- 239000002253 acid Substances 0.000 description 5
- 244000193174 agave Species 0.000 description 5
- 125000000539 amino acid group Chemical group 0.000 description 5
- 238000003556 assay Methods 0.000 description 5
- 230000015572 biosynthetic process Effects 0.000 description 5
- 230000000593 degrading effect Effects 0.000 description 5
- 238000011161 development Methods 0.000 description 5
- 230000018109 developmental process Effects 0.000 description 5
- 210000002257 embryonic structure Anatomy 0.000 description 5
- 230000012010 growth Effects 0.000 description 5
- 229930027917 kanamycin Natural products 0.000 description 5
- SBUJHOSQTJFQJX-NOAMYHISSA-N kanamycin Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CN)O[C@@H]1O[C@H]1[C@H](O)[C@@H](O[C@@H]2[C@@H]([C@@H](N)[C@H](O)[C@@H](CO)O2)O)[C@H](N)C[C@@H]1N SBUJHOSQTJFQJX-NOAMYHISSA-N 0.000 description 5
- 229960000318 kanamycin Drugs 0.000 description 5
- 229930182823 kanamycin A Natural products 0.000 description 5
- 239000013642 negative control Substances 0.000 description 5
- 239000002773 nucleotide Substances 0.000 description 5
- 125000003729 nucleotide group Chemical group 0.000 description 5
- 241000894007 species Species 0.000 description 5
- 239000000126 substance Substances 0.000 description 5
- 238000003786 synthesis reaction Methods 0.000 description 5
- HFZWRUODUSTPEG-UHFFFAOYSA-N 2,4-dichlorophenol Chemical compound OC1=CC=C(Cl)C=C1Cl HFZWRUODUSTPEG-UHFFFAOYSA-N 0.000 description 4
- RTGHRDFWYQHVFW-UHFFFAOYSA-N 3-oxoadipic acid Chemical compound OC(=O)CCC(=O)CC(O)=O RTGHRDFWYQHVFW-UHFFFAOYSA-N 0.000 description 4
- RLFWWDJHLFCNIJ-UHFFFAOYSA-N 4-aminoantipyrine Chemical compound CN1C(C)=C(N)C(=O)N1C1=CC=CC=C1 RLFWWDJHLFCNIJ-UHFFFAOYSA-N 0.000 description 4
- 241001075517 Abelmoschus Species 0.000 description 4
- 241000234282 Allium Species 0.000 description 4
- 244000099147 Ananas comosus Species 0.000 description 4
- 240000007087 Apium graveolens Species 0.000 description 4
- 235000010777 Arachis hypogaea Nutrition 0.000 description 4
- 235000005340 Asparagus officinalis Nutrition 0.000 description 4
- 235000007319 Avena orientalis Nutrition 0.000 description 4
- 241000219198 Brassica Species 0.000 description 4
- 235000011299 Brassica oleracea var botrytis Nutrition 0.000 description 4
- 235000017647 Brassica oleracea var italica Nutrition 0.000 description 4
- 240000003259 Brassica oleracea var. botrytis Species 0.000 description 4
- 240000008100 Brassica rapa Species 0.000 description 4
- 240000006432 Carica papaya Species 0.000 description 4
- 235000009467 Carica papaya Nutrition 0.000 description 4
- 241000252867 Cupriavidus metallidurans Species 0.000 description 4
- 241000234653 Cyperus Species 0.000 description 4
- 244000000626 Daucus carota Species 0.000 description 4
- 235000002767 Daucus carota Nutrition 0.000 description 4
- 235000001950 Elaeis guineensis Nutrition 0.000 description 4
- 241000220223 Fragaria Species 0.000 description 4
- 235000008100 Ginkgo biloba Nutrition 0.000 description 4
- 244000194101 Ginkgo biloba Species 0.000 description 4
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 description 4
- 241000209082 Lolium Species 0.000 description 4
- 235000018330 Macadamia integrifolia Nutrition 0.000 description 4
- 240000007575 Macadamia integrifolia Species 0.000 description 4
- 241000219823 Medicago Species 0.000 description 4
- 235000017587 Medicago sativa ssp. sativa Nutrition 0.000 description 4
- 240000007817 Olea europaea Species 0.000 description 4
- 235000011684 Sorghum saccharatum Nutrition 0.000 description 4
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 description 4
- 229940024606 amino acid Drugs 0.000 description 4
- 235000001014 amino acid Nutrition 0.000 description 4
- 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 4
- AZZMGZXNTDTSME-JUZDKLSSSA-M cefotaxime sodium Chemical compound [Na+].N([C@@H]1C(N2C(=C(COC(C)=O)CS[C@@H]21)C([O-])=O)=O)C(=O)\C(=N/OC)C1=CSC(N)=N1 AZZMGZXNTDTSME-JUZDKLSSSA-M 0.000 description 4
- 230000006378 damage Effects 0.000 description 4
- 230000003054 hormonal effect Effects 0.000 description 4
- 238000005286 illumination Methods 0.000 description 4
- 239000003550 marker Substances 0.000 description 4
- 239000000463 material Substances 0.000 description 4
- 239000013612 plasmid Substances 0.000 description 4
- 238000005507 spraying Methods 0.000 description 4
- 239000006228 supernatant Substances 0.000 description 4
- XOAAWQZATWQOTB-UHFFFAOYSA-N taurine Chemical compound NCCS(O)(=O)=O XOAAWQZATWQOTB-UHFFFAOYSA-N 0.000 description 4
- 235000013616 tea Nutrition 0.000 description 4
- 238000009333 weeding Methods 0.000 description 4
- 108010053957 2,4-dichlorophenoxyacetate-alpha-ketoglutarate dioxygenase Proteins 0.000 description 3
- 244000198134 Agave sisalana Species 0.000 description 3
- 235000011624 Agave sisalana Nutrition 0.000 description 3
- 235000002732 Allium cepa var. cepa Nutrition 0.000 description 3
- 241000219318 Amaranthus Species 0.000 description 3
- 235000007119 Ananas comosus Nutrition 0.000 description 3
- 235000015849 Apium graveolens Dulce Group Nutrition 0.000 description 3
- 235000010591 Appio Nutrition 0.000 description 3
- 241000047982 Axonopus Species 0.000 description 3
- 235000003351 Brassica cretica Nutrition 0.000 description 3
- 235000012905 Brassica oleracea var viridis Nutrition 0.000 description 3
- 235000003343 Brassica rupestris Nutrition 0.000 description 3
- 235000014036 Castanea Nutrition 0.000 description 3
- 241001070941 Castanea Species 0.000 description 3
- 244000146553 Ceiba pentandra Species 0.000 description 3
- 235000003301 Ceiba pentandra Nutrition 0.000 description 3
- 235000013162 Cocos nucifera Nutrition 0.000 description 3
- 244000060011 Cocos nucifera Species 0.000 description 3
- 108020004705 Codon Proteins 0.000 description 3
- 244000241257 Cucumis melo Species 0.000 description 3
- 235000009854 Cucurbita moschata Nutrition 0.000 description 3
- 244000075634 Cyperus rotundus Species 0.000 description 3
- 235000016854 Cyperus rotundus Nutrition 0.000 description 3
- 244000281702 Dioscorea villosa Species 0.000 description 3
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 3
- 235000018060 Elaeis melanococca Nutrition 0.000 description 3
- 244000025670 Eleusine indica Species 0.000 description 3
- 235000014716 Eleusine indica Nutrition 0.000 description 3
- YQYJSBFKSSDGFO-UHFFFAOYSA-N Epihygromycin Natural products OC1C(O)C(C(=O)C)OC1OC(C(=C1)O)=CC=C1C=C(C)C(=O)NC1C(O)C(O)C2OCOC2C1O YQYJSBFKSSDGFO-UHFFFAOYSA-N 0.000 description 3
- 241000234642 Festuca Species 0.000 description 3
- 235000017317 Fortunella Nutrition 0.000 description 3
- 235000016623 Fragaria vesca Nutrition 0.000 description 3
- 235000011363 Fragaria x ananassa Nutrition 0.000 description 3
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 3
- 240000008669 Hedera helix Species 0.000 description 3
- 244000020551 Helianthus annuus Species 0.000 description 3
- 235000003222 Helianthus annuus Nutrition 0.000 description 3
- 240000005979 Hordeum vulgare Species 0.000 description 3
- 235000007340 Hordeum vulgare Nutrition 0.000 description 3
- 241000758789 Juglans Species 0.000 description 3
- 235000003228 Lactuca sativa Nutrition 0.000 description 3
- 240000008415 Lactuca sativa Species 0.000 description 3
- 235000014647 Lens culinaris subsp culinaris Nutrition 0.000 description 3
- 241000219745 Lupinus Species 0.000 description 3
- 235000003800 Macadamia tetraphylla Nutrition 0.000 description 3
- 241000213996 Melilotus Species 0.000 description 3
- 240000000249 Morus alba Species 0.000 description 3
- 235000008708 Morus alba Nutrition 0.000 description 3
- 240000002853 Nelumbo nucifera Species 0.000 description 3
- 235000006508 Nelumbo nucifera Nutrition 0.000 description 3
- 244000025272 Persea americana Species 0.000 description 3
- 235000008673 Persea americana Nutrition 0.000 description 3
- 241000746983 Phleum pratense Species 0.000 description 3
- 231100000674 Phytotoxicity Toxicity 0.000 description 3
- 239000005595 Picloram Substances 0.000 description 3
- 235000003447 Pistacia vera Nutrition 0.000 description 3
- 240000006711 Pistacia vera Species 0.000 description 3
- 241000219843 Pisum Species 0.000 description 3
- 241000508269 Psidium Species 0.000 description 3
- 240000001987 Pyrus communis Species 0.000 description 3
- 235000014443 Pyrus communis Nutrition 0.000 description 3
- 241000219492 Quercus Species 0.000 description 3
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 3
- 235000014680 Saccharomyces cerevisiae Nutrition 0.000 description 3
- 241000209051 Saccharum Species 0.000 description 3
- 235000002595 Solanum tuberosum Nutrition 0.000 description 3
- 244000061456 Solanum tuberosum Species 0.000 description 3
- 244000098338 Triticum aestivum Species 0.000 description 3
- 108090000848 Ubiquitin Proteins 0.000 description 3
- 102000044159 Ubiquitin Human genes 0.000 description 3
- 241000219873 Vicia Species 0.000 description 3
- 125000000218 acetic acid group Chemical group C(C)(=O)* 0.000 description 3
- QKSKPIVNLNLAAV-UHFFFAOYSA-N bis(2-chloroethyl) sulfide Chemical compound ClCCSCCCl QKSKPIVNLNLAAV-UHFFFAOYSA-N 0.000 description 3
- 238000010367 cloning Methods 0.000 description 3
- 235000004879 dioscorea Nutrition 0.000 description 3
- 235000013399 edible fruits Nutrition 0.000 description 3
- 150000002148 esters Chemical class 0.000 description 3
- 235000003891 ferrous sulphate Nutrition 0.000 description 3
- 239000011790 ferrous sulphate Substances 0.000 description 3
- 230000006870 function Effects 0.000 description 3
- 239000008103 glucose Substances 0.000 description 3
- 238000003306 harvesting Methods 0.000 description 3
- 230000001976 improved effect Effects 0.000 description 3
- BAUYGSIQEAFULO-UHFFFAOYSA-L iron(2+) sulfate (anhydrous) Chemical compound [Fe+2].[O-]S([O-])(=O)=O BAUYGSIQEAFULO-UHFFFAOYSA-L 0.000 description 3
- 229910000359 iron(II) sulfate Inorganic materials 0.000 description 3
- 108020004999 messenger RNA Proteins 0.000 description 3
- 235000010460 mustard Nutrition 0.000 description 3
- NQQVFXUMIDALNH-UHFFFAOYSA-N picloram Chemical compound NC1=C(Cl)C(Cl)=NC(C(O)=O)=C1Cl NQQVFXUMIDALNH-UHFFFAOYSA-N 0.000 description 3
- 238000007747 plating Methods 0.000 description 3
- 238000000746 purification Methods 0.000 description 3
- 238000011084 recovery Methods 0.000 description 3
- 239000008223 sterile water Substances 0.000 description 3
- 229940027257 timentin Drugs 0.000 description 3
- HXKWSTRRCHTUEC-UHFFFAOYSA-N 2,4-Dichlorophenoxyaceticacid Chemical compound OC(=O)C(Cl)OC1=CC=C(Cl)C=C1 HXKWSTRRCHTUEC-UHFFFAOYSA-N 0.000 description 2
- KPGXRSRHYNQIFN-UHFFFAOYSA-L 2-oxoglutarate(2-) Chemical compound [O-]C(=O)CCC(=O)C([O-])=O KPGXRSRHYNQIFN-UHFFFAOYSA-L 0.000 description 2
- XSXYVLIPQMXCBV-UHFFFAOYSA-N 3,5-dichlorocatechol Chemical compound OC1=CC(Cl)=CC(Cl)=C1O XSXYVLIPQMXCBV-UHFFFAOYSA-N 0.000 description 2
- AJBZENLMTKDAEK-UHFFFAOYSA-N 3a,5a,5b,8,8,11a-hexamethyl-1-prop-1-en-2-yl-1,2,3,4,5,6,7,7a,9,10,11,11b,12,13,13a,13b-hexadecahydrocyclopenta[a]chrysene-4,9-diol Chemical compound CC12CCC(O)C(C)(C)C1CCC(C1(C)CC3O)(C)C2CCC1C1C3(C)CCC1C(=C)C AJBZENLMTKDAEK-UHFFFAOYSA-N 0.000 description 2
- GNKZMNRKLCTJAY-UHFFFAOYSA-N 4'-Methylacetophenone Chemical compound CC(=O)C1=CC=C(C)C=C1 GNKZMNRKLCTJAY-UHFFFAOYSA-N 0.000 description 2
- ODHCTXKNWHHXJC-VKHMYHEASA-N 5-oxo-L-proline Chemical compound OC(=O)[C@@H]1CCC(=O)N1 ODHCTXKNWHHXJC-VKHMYHEASA-N 0.000 description 2
- 241000208140 Acer Species 0.000 description 2
- 241000219068 Actinidia Species 0.000 description 2
- 108010016119 Alpha-Ketoglutarate-Dependent Dioxygenase FTO Proteins 0.000 description 2
- 244000144725 Amygdalus communis Species 0.000 description 2
- 235000011437 Amygdalus communis Nutrition 0.000 description 2
- 244000226021 Anacardium occidentale Species 0.000 description 2
- 235000017060 Arachis glabrata Nutrition 0.000 description 2
- 235000018262 Arachis monticola Nutrition 0.000 description 2
- DCXYFEDJOCDNAF-UHFFFAOYSA-N Asparagine Natural products OC(=O)C(N)CC(N)=O DCXYFEDJOCDNAF-UHFFFAOYSA-N 0.000 description 2
- 241001061264 Astragalus Species 0.000 description 2
- 235000005781 Avena Nutrition 0.000 description 2
- 235000007320 Avena fatua Nutrition 0.000 description 2
- 241000209764 Avena fatua Species 0.000 description 2
- 235000000832 Ayote Nutrition 0.000 description 2
- 244000036905 Benincasa cerifera Species 0.000 description 2
- 235000011274 Benincasa cerifera Nutrition 0.000 description 2
- 241000335053 Beta vulgaris Species 0.000 description 2
- 235000005637 Brassica campestris Nutrition 0.000 description 2
- 244000178993 Brassica juncea Species 0.000 description 2
- 235000004221 Brassica oleracea var gemmifera Nutrition 0.000 description 2
- 244000308368 Brassica oleracea var. gemmifera Species 0.000 description 2
- 235000003880 Calendula Nutrition 0.000 description 2
- 240000001432 Calendula officinalis Species 0.000 description 2
- 235000005273 Canna coccinea Nutrition 0.000 description 2
- 244000292211 Canna coccinea Species 0.000 description 2
- 235000002566 Capsicum Nutrition 0.000 description 2
- 240000008574 Capsicum frutescens Species 0.000 description 2
- 235000003255 Carthamus tinctorius Nutrition 0.000 description 2
- 244000020518 Carthamus tinctorius Species 0.000 description 2
- 241000167550 Centella Species 0.000 description 2
- 241000334161 Cercis chinensis Species 0.000 description 2
- 244000281762 Chenopodium ambrosioides Species 0.000 description 2
- KZBUYRJDOAKODT-UHFFFAOYSA-N Chlorine Chemical compound ClCl KZBUYRJDOAKODT-UHFFFAOYSA-N 0.000 description 2
- 244000241235 Citrullus lanatus Species 0.000 description 2
- 235000005979 Citrus limon Nutrition 0.000 description 2
- 244000175448 Citrus madurensis Species 0.000 description 2
- 244000131522 Citrus pyriformis Species 0.000 description 2
- 241000755729 Clivia Species 0.000 description 2
- 244000205754 Colocasia esculenta Species 0.000 description 2
- 235000006481 Colocasia esculenta Nutrition 0.000 description 2
- 244000074881 Conyza canadensis Species 0.000 description 2
- 235000004385 Conyza canadensis Nutrition 0.000 description 2
- 240000000491 Corchorus aestuans Species 0.000 description 2
- 235000011777 Corchorus aestuans Nutrition 0.000 description 2
- 235000010862 Corchorus capsularis Nutrition 0.000 description 2
- 235000002787 Coriandrum sativum Nutrition 0.000 description 2
- 244000018436 Coriandrum sativum Species 0.000 description 2
- 235000014493 Crataegus Nutrition 0.000 description 2
- 241001092040 Crataegus Species 0.000 description 2
- 241000195493 Cryptophyta Species 0.000 description 2
- 235000009847 Cucumis melo var cantalupensis Nutrition 0.000 description 2
- 240000004244 Cucurbita moschata Species 0.000 description 2
- 240000001980 Cucurbita pepo Species 0.000 description 2
- 235000009852 Cucurbita pepo Nutrition 0.000 description 2
- 244000019459 Cynara cardunculus Species 0.000 description 2
- 235000019106 Cynara scolymus Nutrition 0.000 description 2
- 235000005853 Cyperus esculentus Nutrition 0.000 description 2
- 244000285774 Cyperus esculentus Species 0.000 description 2
- 241000522190 Desmodium Species 0.000 description 2
- 101100313146 Dictyostelium discoideum tbpA gene Proteins 0.000 description 2
- 235000010823 Digitaria sanguinalis Nutrition 0.000 description 2
- 240000001008 Dimocarpus longan Species 0.000 description 2
- 235000005903 Dioscorea Nutrition 0.000 description 2
- 235000000504 Dioscorea villosa Nutrition 0.000 description 2
- 235000011511 Diospyros Nutrition 0.000 description 2
- 206010059866 Drug resistance Diseases 0.000 description 2
- 238000002965 ELISA Methods 0.000 description 2
- 235000014966 Eragrostis abyssinica Nutrition 0.000 description 2
- 244000140063 Eragrostis abyssinica Species 0.000 description 2
- 235000009008 Eriobotrya japonica Nutrition 0.000 description 2
- 244000061508 Eriobotrya japonica Species 0.000 description 2
- 235000000235 Euphoria longan Nutrition 0.000 description 2
- 235000009419 Fagopyrum esculentum Nutrition 0.000 description 2
- 240000008620 Fagopyrum esculentum Species 0.000 description 2
- 240000000731 Fagus sylvatica Species 0.000 description 2
- 235000010099 Fagus sylvatica Nutrition 0.000 description 2
- 241000234643 Festuca arundinacea Species 0.000 description 2
- 239000004471 Glycine Substances 0.000 description 2
- 235000009429 Gossypium barbadense Nutrition 0.000 description 2
- 244000299507 Gossypium hirsutum Species 0.000 description 2
- 235000005206 Hibiscus Nutrition 0.000 description 2
- 235000007185 Hibiscus lunariifolius Nutrition 0.000 description 2
- 244000284380 Hibiscus rosa sinensis Species 0.000 description 2
- 241001086809 Hordeum arizonicum Species 0.000 description 2
- 244000018434 Hydrocotyle sibthorpioides Species 0.000 description 2
- 235000002788 Hydrocotyle sibthorpioides Nutrition 0.000 description 2
- 206010020649 Hyperkeratosis Diseases 0.000 description 2
- 244000017020 Ipomoea batatas Species 0.000 description 2
- 235000002678 Ipomoea batatas Nutrition 0.000 description 2
- 235000013757 Juglans Nutrition 0.000 description 2
- DCXYFEDJOCDNAF-REOHCLBHSA-N L-asparagine Chemical compound OC(=O)[C@@H](N)CC(N)=O DCXYFEDJOCDNAF-REOHCLBHSA-N 0.000 description 2
- 240000004322 Lens culinaris Species 0.000 description 2
- 244000043158 Lens esculenta Species 0.000 description 2
- 235000004431 Linum usitatissimum Nutrition 0.000 description 2
- 240000006240 Linum usitatissimum Species 0.000 description 2
- 235000015459 Lycium barbarum Nutrition 0.000 description 2
- 241000220225 Malus Species 0.000 description 2
- 244000070406 Malus silvestris Species 0.000 description 2
- 241001481296 Malus spectabilis Species 0.000 description 2
- 235000014826 Mangifera indica Nutrition 0.000 description 2
- 240000007228 Mangifera indica Species 0.000 description 2
- 240000003183 Manihot esculenta Species 0.000 description 2
- 235000000839 Melilotus officinalis subsp suaveolens Nutrition 0.000 description 2
- 244000246386 Mentha pulegium Species 0.000 description 2
- 235000016257 Mentha pulegium Nutrition 0.000 description 2
- 235000004357 Mentha x piperita Nutrition 0.000 description 2
- 241000234295 Musa Species 0.000 description 2
- 235000006510 Nelumbo pentapetala Nutrition 0.000 description 2
- 244000183278 Nephelium litchi Species 0.000 description 2
- 235000002637 Nicotiana tabacum Nutrition 0.000 description 2
- 244000061176 Nicotiana tabacum Species 0.000 description 2
- 235000016698 Nigella sativa Nutrition 0.000 description 2
- 244000090896 Nigella sativa Species 0.000 description 2
- 240000001439 Opuntia Species 0.000 description 2
- 241000209094 Oryza Species 0.000 description 2
- 240000001516 Oryza latifolia Species 0.000 description 2
- 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 2
- 240000008114 Panicum miliaceum Species 0.000 description 2
- 235000007199 Panicum miliaceum Nutrition 0.000 description 2
- 241001520808 Panicum virgatum Species 0.000 description 2
- 241001330453 Paspalum Species 0.000 description 2
- 235000000370 Passiflora edulis Nutrition 0.000 description 2
- 244000288157 Passiflora edulis Species 0.000 description 2
- 240000004370 Pastinaca sativa Species 0.000 description 2
- 235000007195 Pennisetum typhoides Nutrition 0.000 description 2
- 240000007377 Petunia x hybrida Species 0.000 description 2
- 241000219833 Phaseolus Species 0.000 description 2
- 244000046052 Phaseolus vulgaris Species 0.000 description 2
- 244000064622 Physalis edulis Species 0.000 description 2
- 235000011611 Pinus yunnanensis Nutrition 0.000 description 2
- 241000018652 Pinus yunnanensis Species 0.000 description 2
- 241000209048 Poa Species 0.000 description 2
- 241000219000 Populus Species 0.000 description 2
- 241001494501 Prosopis <angiosperm> Species 0.000 description 2
- 108010076504 Protein Sorting Signals Proteins 0.000 description 2
- 241000220299 Prunus Species 0.000 description 2
- 235000011432 Prunus Nutrition 0.000 description 2
- 241001290151 Prunus avium subsp. avium Species 0.000 description 2
- 244000294611 Punica granatum Species 0.000 description 2
- 235000014360 Punica granatum Nutrition 0.000 description 2
- 235000004443 Ricinus communis Nutrition 0.000 description 2
- 235000007238 Secale cereale Nutrition 0.000 description 2
- 244000082988 Secale cereale Species 0.000 description 2
- 235000003434 Sesamum indicum Nutrition 0.000 description 2
- 244000000231 Sesamum indicum Species 0.000 description 2
- 244000275012 Sesbania cannabina Species 0.000 description 2
- 235000002634 Solanum Nutrition 0.000 description 2
- 241000207763 Solanum Species 0.000 description 2
- 241001327268 Sorghastrum Species 0.000 description 2
- 235000007230 Sorghum bicolor Nutrition 0.000 description 2
- 244000045719 Syzygium Species 0.000 description 2
- 235000016639 Syzygium aromaticum Nutrition 0.000 description 2
- 235000012096 Syzygium samarangense Nutrition 0.000 description 2
- 235000012308 Tagetes Nutrition 0.000 description 2
- 241000736851 Tagetes Species 0.000 description 2
- 235000006468 Thea sinensis Nutrition 0.000 description 2
- 244000152045 Themeda triandra Species 0.000 description 2
- 244000299461 Theobroma cacao Species 0.000 description 2
- 235000009470 Theobroma cacao Nutrition 0.000 description 2
- 108091036066 Three prime untranslated region Proteins 0.000 description 2
- IQFYYKKMVGJFEH-XLPZGREQSA-N Thymidine Chemical compound O=C1NC(=O)C(C)=CN1[C@@H]1O[C@H](CO)[C@@H](O)C1 IQFYYKKMVGJFEH-XLPZGREQSA-N 0.000 description 2
- 241000219793 Trifolium Species 0.000 description 2
- 235000021307 Triticum Nutrition 0.000 description 2
- 241001312894 Trollius chinensis Species 0.000 description 2
- 241000722921 Tulipa gesneriana Species 0.000 description 2
- 235000017537 Vaccinium myrtillus Nutrition 0.000 description 2
- 244000078534 Vaccinium myrtillus Species 0.000 description 2
- 235000010749 Vicia faba Nutrition 0.000 description 2
- 240000006677 Vicia faba Species 0.000 description 2
- 235000002098 Vicia faba var. major Nutrition 0.000 description 2
- 241000219977 Vigna Species 0.000 description 2
- 241001247821 Ziziphus Species 0.000 description 2
- 241000981595 Zoysia japonica Species 0.000 description 2
- 240000001102 Zoysia matrella Species 0.000 description 2
- OJOBTAOGJIWAGB-UHFFFAOYSA-N acetosyringone Chemical compound COC1=CC(C(C)=O)=CC(OC)=C1O OJOBTAOGJIWAGB-UHFFFAOYSA-N 0.000 description 2
- ODHCTXKNWHHXJC-UHFFFAOYSA-N acide pyroglutamique Natural products OC(=O)C1CCC(=O)N1 ODHCTXKNWHHXJC-UHFFFAOYSA-N 0.000 description 2
- 239000004480 active ingredient Substances 0.000 description 2
- 235000016520 artichoke thistle Nutrition 0.000 description 2
- 229960001230 asparagine Drugs 0.000 description 2
- 235000009582 asparagine Nutrition 0.000 description 2
- 235000006533 astragalus Nutrition 0.000 description 2
- 230000004071 biological effect Effects 0.000 description 2
- 239000001390 capsicum minimum Substances 0.000 description 2
- 239000004202 carbamide Substances 0.000 description 2
- FPPNZSSZRUTDAP-UWFZAAFLSA-N carbenicillin Chemical compound N([C@H]1[C@H]2SC([C@@H](N2C1=O)C(O)=O)(C)C)C(=O)C(C(O)=O)C1=CC=CC=C1 FPPNZSSZRUTDAP-UWFZAAFLSA-N 0.000 description 2
- 229960003669 carbenicillin Drugs 0.000 description 2
- 230000003197 catalytic effect Effects 0.000 description 2
- 238000006555 catalytic reaction Methods 0.000 description 2
- 229960004261 cefotaxime Drugs 0.000 description 2
- 229960002727 cefotaxime sodium Drugs 0.000 description 2
- 238000005119 centrifugation Methods 0.000 description 2
- 235000019693 cherries Nutrition 0.000 description 2
- 238000003776 cleavage reaction Methods 0.000 description 2
- 239000012881 co-culture medium Substances 0.000 description 2
- 244000038559 crop plants Species 0.000 description 2
- 238000012364 cultivation method Methods 0.000 description 2
- 238000005520 cutting process Methods 0.000 description 2
- 230000001419 dependent effect Effects 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 210000005069 ears Anatomy 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 125000001495 ethyl group Chemical group [H]C([H])([H])C([H])([H])* 0.000 description 2
- IAJOBQBIJHVGMQ-BYPYZUCNSA-N glufosinate-P Chemical compound CP(O)(=O)CC[C@H](N)C(O)=O IAJOBQBIJHVGMQ-BYPYZUCNSA-N 0.000 description 2
- 230000009036 growth inhibition Effects 0.000 description 2
- 235000001050 hortel pimenta Nutrition 0.000 description 2
- 238000009396 hybridization Methods 0.000 description 2
- JTEDVYBZBROSJT-UHFFFAOYSA-N indole-3-butyric acid Chemical compound C1=CC=C2C(CCCC(=O)O)=CNC2=C1 JTEDVYBZBROSJT-UHFFFAOYSA-N 0.000 description 2
- 230000006698 induction Effects 0.000 description 2
- 208000015181 infectious disease Diseases 0.000 description 2
- BPHPUYQFMNQIOC-NXRLNHOXSA-N isopropyl beta-D-thiogalactopyranoside Chemical compound CC(C)S[C@@H]1O[C@H](CO)[C@H](O)[C@H](O)[C@H]1O BPHPUYQFMNQIOC-NXRLNHOXSA-N 0.000 description 2
- 235000014571 nuts Nutrition 0.000 description 2
- 239000003921 oil Substances 0.000 description 2
- 230000003287 optical effect Effects 0.000 description 2
- 238000005457 optimization Methods 0.000 description 2
- 235000020232 peanut Nutrition 0.000 description 2
- 235000020233 pistachio Nutrition 0.000 description 2
- 229920000642 polymer Polymers 0.000 description 2
- 239000013641 positive control Substances 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 230000007017 scission Effects 0.000 description 2
- 238000007423 screening assay Methods 0.000 description 2
- 230000001954 sterilising effect Effects 0.000 description 2
- 229960003080 taurine Drugs 0.000 description 2
- 101150090317 tfdA gene Proteins 0.000 description 2
- 231100000331 toxic Toxicity 0.000 description 2
- 230000002588 toxic effect Effects 0.000 description 2
- 238000011426 transformation method Methods 0.000 description 2
- 238000012250 transgenic expression Methods 0.000 description 2
- ANJTVLIZGCUXLD-BDAKNGLRSA-N (-)-Cytisine Natural products C1NC[C@@H]2CN3C(=O)C=CC=C3[C@H]1C2 ANJTVLIZGCUXLD-BDAKNGLRSA-N 0.000 description 1
- SODPIMGUZLOIPE-UHFFFAOYSA-N (4-chlorophenoxy)acetic acid Chemical compound OC(=O)COC1=CC=C(Cl)C=C1 SODPIMGUZLOIPE-UHFFFAOYSA-N 0.000 description 1
- 101710128758 (S)-phenoxypropionate/alpha-ketoglutarate-dioxygenase Proteins 0.000 description 1
- 108091032973 (ribonucleotides)n+m Proteins 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
- ZPSJGADGUYYRKE-UHFFFAOYSA-N 2H-pyran-2-one Chemical compound O=C1C=CC=CO1 ZPSJGADGUYYRKE-UHFFFAOYSA-N 0.000 description 1
- WMKYKQWKLILFBM-UHFFFAOYSA-N 2h-pyridine-1-carboxylic acid Chemical class OC(=O)N1CC=CC=C1 WMKYKQWKLILFBM-UHFFFAOYSA-N 0.000 description 1
- 108010020183 3-phosphoshikimate 1-carboxyvinyltransferase Proteins 0.000 description 1
- WWOBYPKUYODHDG-UHFFFAOYSA-N 4-chlorocatechol Chemical compound OC1=CC=C(Cl)C=C1O WWOBYPKUYODHDG-UHFFFAOYSA-N 0.000 description 1
- WXNZTHHGJRFXKQ-UHFFFAOYSA-N 4-chlorophenol Chemical compound OC1=CC=C(Cl)C=C1 WXNZTHHGJRFXKQ-UHFFFAOYSA-N 0.000 description 1
- GOSWTRUMMSCNCW-HNNGNKQASA-N 9-ribosyl-trans-zeatin Chemical compound C1=NC=2C(NC\C=C(CO)/C)=NC=NC=2N1[C@@H]1O[C@H](CO)[C@@H](O)[C@H]1O GOSWTRUMMSCNCW-HNNGNKQASA-N 0.000 description 1
- 241001290610 Abildgaardia Species 0.000 description 1
- 102000000452 Acetyl-CoA carboxylase Human genes 0.000 description 1
- 108010016219 Acetyl-CoA carboxylase Proteins 0.000 description 1
- 235000009434 Actinidia chinensis Nutrition 0.000 description 1
- 244000298697 Actinidia deliciosa Species 0.000 description 1
- 235000009436 Actinidia deliciosa Nutrition 0.000 description 1
- 241000499929 Agapanthus Species 0.000 description 1
- 241000209136 Agropyron Species 0.000 description 1
- 241000743339 Agrostis Species 0.000 description 1
- 240000007241 Agrostis stolonifera Species 0.000 description 1
- 241001093951 Ailanthus altissima Species 0.000 description 1
- 235000005254 Allium ampeloprasum Nutrition 0.000 description 1
- 240000006108 Allium ampeloprasum Species 0.000 description 1
- 102000000383 Alpha-Ketoglutarate-Dependent Dioxygenase FTO Human genes 0.000 description 1
- 101710139722 Alpha-ketoglutarate-dependent taurine dioxygenase Proteins 0.000 description 1
- 235000013480 Amaranthus spinosus Nutrition 0.000 description 1
- 244000237958 Amaranthus spinosus Species 0.000 description 1
- 235000003129 Ambrosia artemisiifolia var elatior Nutrition 0.000 description 1
- QGZKDVFQNNGYKY-UHFFFAOYSA-O Ammonium Chemical compound [NH4+] QGZKDVFQNNGYKY-UHFFFAOYSA-O 0.000 description 1
- 241000380131 Ammophila arenaria Species 0.000 description 1
- 235000004047 Amorpha fruticosa Nutrition 0.000 description 1
- 240000002066 Amorpha fruticosa Species 0.000 description 1
- 235000003840 Amygdalus nana Nutrition 0.000 description 1
- 244000144730 Amygdalus persica Species 0.000 description 1
- 235000001274 Anacardium occidentale Nutrition 0.000 description 1
- 241001327399 Andropogon gerardii Species 0.000 description 1
- 241000382455 Angelica sinensis Species 0.000 description 1
- 235000007755 Annona Nutrition 0.000 description 1
- 240000006199 Annona purpurea Species 0.000 description 1
- 235000011518 Annona purpurea Nutrition 0.000 description 1
- 235000003276 Apios tuberosa Nutrition 0.000 description 1
- 235000002764 Apium graveolens Nutrition 0.000 description 1
- 101000768857 Arabidopsis thaliana 3-phosphoshikimate 1-carboxyvinyltransferase, chloroplastic Proteins 0.000 description 1
- 101100478623 Arabidopsis thaliana S-ACP-DES1 gene Proteins 0.000 description 1
- 235000003911 Arachis Nutrition 0.000 description 1
- 235000010744 Arachis villosulicarpa Nutrition 0.000 description 1
- 235000011330 Armoracia rusticana Nutrition 0.000 description 1
- 240000003291 Armoracia rusticana Species 0.000 description 1
- 244000018217 Artocarpus elasticus Species 0.000 description 1
- 241000228212 Aspergillus Species 0.000 description 1
- 235000010110 Astragalus glycyphyllos Nutrition 0.000 description 1
- 241000132012 Atractylodes Species 0.000 description 1
- 235000009393 Avena byzantina Nutrition 0.000 description 1
- 235000010082 Averrhoa carambola Nutrition 0.000 description 1
- 240000006063 Averrhoa carambola Species 0.000 description 1
- 241001532704 Azolla Species 0.000 description 1
- 241000589151 Azotobacter Species 0.000 description 1
- 241000589152 Azotobacter chroococcum Species 0.000 description 1
- 241000209128 Bambusa Species 0.000 description 1
- 235000017166 Bambusa arundinacea Nutrition 0.000 description 1
- 235000017491 Bambusa tulda Nutrition 0.000 description 1
- 235000009759 Bauhinia variegata Nutrition 0.000 description 1
- 244000259585 Bauhinia variegata Species 0.000 description 1
- 241000218993 Begonia Species 0.000 description 1
- 235000016068 Berberis vulgaris Nutrition 0.000 description 1
- 241000219164 Bertholletia Species 0.000 description 1
- 235000021533 Beta vulgaris Nutrition 0.000 description 1
- DWRXFEITVBNRMK-UHFFFAOYSA-N Beta-D-1-Arabinofuranosylthymine Natural products O=C1NC(=O)C(C)=CN1C1C(O)C(O)C(CO)O1 DWRXFEITVBNRMK-UHFFFAOYSA-N 0.000 description 1
- 235000018185 Betula X alpestris Nutrition 0.000 description 1
- 235000018212 Betula X uliginosa Nutrition 0.000 description 1
- 241000219495 Betulaceae Species 0.000 description 1
- 108010018763 Biotin carboxylase Proteins 0.000 description 1
- 240000008564 Boehmeria nivea Species 0.000 description 1
- 235000004480 Bombax malabaricum Nutrition 0.000 description 1
- 241000232312 Bouteloua Species 0.000 description 1
- 241000320720 Bouteloua dactyloides Species 0.000 description 1
- 241000339490 Brachyachne Species 0.000 description 1
- 241000743774 Brachypodium Species 0.000 description 1
- 235000006463 Brassica alba Nutrition 0.000 description 1
- 244000257790 Brassica carinata Species 0.000 description 1
- 235000011371 Brassica hirta Nutrition 0.000 description 1
- 235000011332 Brassica juncea Nutrition 0.000 description 1
- 235000014700 Brassica juncea var napiformis Nutrition 0.000 description 1
- 235000011291 Brassica nigra Nutrition 0.000 description 1
- 244000180419 Brassica nigra Species 0.000 description 1
- 244000064816 Brassica oleracea var. acephala Species 0.000 description 1
- 235000011292 Brassica rapa Nutrition 0.000 description 1
- 235000000540 Brassica rapa subsp rapa Nutrition 0.000 description 1
- 235000004936 Bromus mango Nutrition 0.000 description 1
- 241000510654 Bupleurum chinense Species 0.000 description 1
- GSCCOQHTKVBAGO-UHFFFAOYSA-N CC(C(C(C(=O)O)(Cl)Cl)(Cl)Cl)C Chemical compound CC(C(C(C(=O)O)(Cl)Cl)(Cl)Cl)C GSCCOQHTKVBAGO-UHFFFAOYSA-N 0.000 description 1
- 235000008635 Cadaba farinosa Nutrition 0.000 description 1
- 241000628166 Cadaba farinosa Species 0.000 description 1
- 241000189662 Calla Species 0.000 description 1
- 235000017595 Callicarpa japonica Nutrition 0.000 description 1
- 240000003690 Callicarpa japonica Species 0.000 description 1
- 240000001548 Camellia japonica Species 0.000 description 1
- 235000008697 Cannabis sativa Nutrition 0.000 description 1
- 235000012766 Cannabis sativa ssp. sativa var. sativa Nutrition 0.000 description 1
- 235000012765 Cannabis sativa ssp. sativa var. spontanea Nutrition 0.000 description 1
- 240000004160 Capsicum annuum Species 0.000 description 1
- 235000008534 Capsicum annuum var annuum Nutrition 0.000 description 1
- 241000722731 Carex Species 0.000 description 1
- 241000973255 Carex elata Species 0.000 description 1
- 235000014653 Carica parviflora Nutrition 0.000 description 1
- 244000132059 Carica parviflora Species 0.000 description 1
- 240000004927 Carissa macrocarpa Species 0.000 description 1
- 235000001479 Carissa macrocarpa Nutrition 0.000 description 1
- 241000723418 Carya Species 0.000 description 1
- 235000009025 Carya illinoensis Nutrition 0.000 description 1
- 244000068645 Carya illinoensis Species 0.000 description 1
- 235000009024 Ceanothus sanguineus Nutrition 0.000 description 1
- 241001148660 Cenchrus sp. Species 0.000 description 1
- 241001633683 Centipeda <firmicute> Species 0.000 description 1
- 229930186147 Cephalosporin Natural products 0.000 description 1
- 235000000509 Chenopodium ambrosioides Nutrition 0.000 description 1
- 235000005490 Chenopodium botrys Nutrition 0.000 description 1
- 241000258920 Chilopoda Species 0.000 description 1
- 241000700112 Chinchilla Species 0.000 description 1
- 241001219477 Chroococcus Species 0.000 description 1
- 235000005633 Chrysanthemum balsamita Nutrition 0.000 description 1
- 244000260524 Chrysanthemum balsamita Species 0.000 description 1
- 235000010523 Cicer arietinum Nutrition 0.000 description 1
- 244000045195 Cicer arietinum Species 0.000 description 1
- 240000006740 Cichorium endivia Species 0.000 description 1
- 235000018536 Cichorium endivia Nutrition 0.000 description 1
- 235000007542 Cichorium intybus Nutrition 0.000 description 1
- 244000298479 Cichorium intybus Species 0.000 description 1
- 235000021513 Cinchona Nutrition 0.000 description 1
- 241000157855 Cinchona Species 0.000 description 1
- 241000723347 Cinnamomum Species 0.000 description 1
- 108091062157 Cis-regulatory element Proteins 0.000 description 1
- 235000002548 Cistus Nutrition 0.000 description 1
- 241000984090 Cistus Species 0.000 description 1
- 235000009831 Citrullus lanatus Nutrition 0.000 description 1
- 235000012828 Citrullus lanatus var citroides Nutrition 0.000 description 1
- 241001672694 Citrus reticulata Species 0.000 description 1
- 239000005497 Clethodim Substances 0.000 description 1
- 239000005500 Clopyralid Substances 0.000 description 1
- 241000737241 Cocos Species 0.000 description 1
- 108700010070 Codon Usage Proteins 0.000 description 1
- 241000218631 Coniferophyta Species 0.000 description 1
- 108091035707 Consensus sequence Proteins 0.000 description 1
- 235000007706 Corchorus sp Nutrition 0.000 description 1
- 241000723382 Corylus Species 0.000 description 1
- 235000004237 Crocus Nutrition 0.000 description 1
- 241000596148 Crocus Species 0.000 description 1
- 235000015655 Crocus sativus Nutrition 0.000 description 1
- 244000124209 Crocus sativus Species 0.000 description 1
- 235000015510 Cucumis melo subsp melo Nutrition 0.000 description 1
- 240000008067 Cucumis sativus Species 0.000 description 1
- 235000010799 Cucumis sativus var sativus Nutrition 0.000 description 1
- 235000003949 Cucurbita mixta Nutrition 0.000 description 1
- 235000009804 Cucurbita pepo subsp pepo Nutrition 0.000 description 1
- 101100424929 Cupriavidus pinatubonensis (strain JMP 134 / LMG 1197) tfdDI gene Proteins 0.000 description 1
- 101100152788 Cupriavidus pinatubonensis (strain JMP 134 / LMG 1197) tfdEI gene Proteins 0.000 description 1
- 101100152790 Cupriavidus pinatubonensis (strain JMP 134 / LMG 1197) tfdFI gene Proteins 0.000 description 1
- 241000612153 Cyclamen Species 0.000 description 1
- 239000005502 Cyhalofop-butyl Substances 0.000 description 1
- 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 1
- 241000732800 Cymbidium Species 0.000 description 1
- 244000052363 Cynodon dactylon Species 0.000 description 1
- 241000817004 Cyperus amuricus Species 0.000 description 1
- 102100028717 Cytosolic 5'-nucleotidase 3A Human genes 0.000 description 1
- 238000001712 DNA sequencing Methods 0.000 description 1
- 240000006541 Dactyloctenium aegyptium Species 0.000 description 1
- 241000934856 Daphne Species 0.000 description 1
- 241000208296 Datura Species 0.000 description 1
- 241001600125 Delftia acidovorans Species 0.000 description 1
- 101100152792 Delftia acidovorans tfdF gene Proteins 0.000 description 1
- 241000202296 Delphinium Species 0.000 description 1
- 241001523681 Dendrobium Species 0.000 description 1
- 235000009355 Dianthus caryophyllus Nutrition 0.000 description 1
- 240000006497 Dianthus caryophyllus Species 0.000 description 1
- 235000001602 Digitaria X umfolozi Nutrition 0.000 description 1
- 235000017898 Digitaria ciliaris Nutrition 0.000 description 1
- 235000005476 Digitaria cruciata Nutrition 0.000 description 1
- 235000006830 Digitaria didactyla Nutrition 0.000 description 1
- 235000005804 Digitaria eriantha ssp. eriantha Nutrition 0.000 description 1
- 244000152970 Digitaria sanguinalis Species 0.000 description 1
- ROSDSFDQCJNGOL-UHFFFAOYSA-N Dimethylamine Chemical class CNC ROSDSFDQCJNGOL-UHFFFAOYSA-N 0.000 description 1
- 235000000525 Dimocarpus longan Nutrition 0.000 description 1
- 235000002723 Dioscorea alata Nutrition 0.000 description 1
- 235000007056 Dioscorea composita Nutrition 0.000 description 1
- 235000009723 Dioscorea convolvulacea Nutrition 0.000 description 1
- 235000005362 Dioscorea floribunda Nutrition 0.000 description 1
- 235000004868 Dioscorea macrostachya Nutrition 0.000 description 1
- 235000005361 Dioscorea nummularia Nutrition 0.000 description 1
- 235000005360 Dioscorea spiculiflora Nutrition 0.000 description 1
- 241000723267 Diospyros Species 0.000 description 1
- 244000236655 Diospyros kaki Species 0.000 description 1
- 235000006025 Durio zibethinus Nutrition 0.000 description 1
- 240000000716 Durio zibethinus Species 0.000 description 1
- 241000192043 Echinochloa Species 0.000 description 1
- 244000058871 Echinochloa crus-galli Species 0.000 description 1
- 240000003826 Eichhornia crassipes Species 0.000 description 1
- 241000512897 Elaeis Species 0.000 description 1
- 235000001942 Elaeis Nutrition 0.000 description 1
- 241001608549 Elaeis oleifera Species 0.000 description 1
- 235000007349 Eleusine coracana Nutrition 0.000 description 1
- 244000078127 Eleusine coracana Species 0.000 description 1
- 241000490050 Eleutherococcus Species 0.000 description 1
- 241000025852 Eremochloa ophiuroides Species 0.000 description 1
- 241000132521 Erigeron Species 0.000 description 1
- 241001013934 Erigeron breviscapus Species 0.000 description 1
- 241001198387 Escherichia coli BL21(DE3) Species 0.000 description 1
- 244000004281 Eucalyptus maculata Species 0.000 description 1
- 241000266331 Eugenia Species 0.000 description 1
- 244000061408 Eugenia caryophyllata Species 0.000 description 1
- 241000221079 Euphorbia <genus> Species 0.000 description 1
- 241001555026 Euphorbia humifusa Species 0.000 description 1
- 241001070947 Fagus Species 0.000 description 1
- 241001289529 Fallopia multiflora Species 0.000 description 1
- 235000008730 Ficus carica Nutrition 0.000 description 1
- 244000025361 Ficus carica Species 0.000 description 1
- 241000233866 Fungi Species 0.000 description 1
- 241000735598 Galbulimima belgraveana Species 0.000 description 1
- 235000014820 Galium aparine Nutrition 0.000 description 1
- 229930191978 Gibberellin Natural products 0.000 description 1
- 235000011201 Ginkgo Nutrition 0.000 description 1
- 108010063907 Glutathione Reductase Proteins 0.000 description 1
- 102100036442 Glutathione reductase, mitochondrial Human genes 0.000 description 1
- 235000000554 Glycyrrhiza uralensis Nutrition 0.000 description 1
- 240000008917 Glycyrrhiza uralensis Species 0.000 description 1
- 240000000047 Gossypium barbadense Species 0.000 description 1
- 235000009432 Gossypium hirsutum Nutrition 0.000 description 1
- 241000208818 Helianthus Species 0.000 description 1
- 241000756137 Hemerocallis Species 0.000 description 1
- 235000002941 Hemerocallis fulva Nutrition 0.000 description 1
- 240000009206 Hemerocallis fulva Species 0.000 description 1
- 244000043261 Hevea brasiliensis Species 0.000 description 1
- 241000238631 Hexapoda Species 0.000 description 1
- 240000000797 Hibiscus cannabinus Species 0.000 description 1
- 241000209219 Hordeum Species 0.000 description 1
- 235000008694 Humulus lupulus Nutrition 0.000 description 1
- 241001632576 Hyacinthus Species 0.000 description 1
- 235000003332 Ilex aquifolium Nutrition 0.000 description 1
- 235000002296 Ilex sandwicensis Nutrition 0.000 description 1
- 235000002294 Ilex volkensiana Nutrition 0.000 description 1
- 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 1
- 235000006350 Ipomoea batatas var. batatas Nutrition 0.000 description 1
- 240000001549 Ipomoea eriocarpa Species 0.000 description 1
- 235000005146 Ipomoea eriocarpa Nutrition 0.000 description 1
- 241000334160 Isatis Species 0.000 description 1
- 235000010254 Jasminum officinale Nutrition 0.000 description 1
- 240000005385 Jasminum sambac Species 0.000 description 1
- 235000013740 Juglans nigra Nutrition 0.000 description 1
- 244000184861 Juglans nigra Species 0.000 description 1
- 235000009496 Juglans regia Nutrition 0.000 description 1
- 241001166994 Kummerowia striata Species 0.000 description 1
- 244000157344 Kyllinga brevifolia Species 0.000 description 1
- 240000000161 Lagerstroemia indica Species 0.000 description 1
- 235000000283 Lagerstroemia parviflora Nutrition 0.000 description 1
- 241000219729 Lathyrus Species 0.000 description 1
- 108091026898 Leader sequence (mRNA) Proteins 0.000 description 1
- 235000010666 Lens esculenta Nutrition 0.000 description 1
- 240000003553 Leptospermum scoparium Species 0.000 description 1
- 241000244365 Ligusticum sinense Species 0.000 description 1
- 241000830535 Ligustrum lucidum Species 0.000 description 1
- 241000234435 Lilium Species 0.000 description 1
- 235000003956 Luffa Nutrition 0.000 description 1
- 241000219138 Luffa Species 0.000 description 1
- 235000018780 Luffa acutangula Nutrition 0.000 description 1
- 244000280244 Luffa acutangula Species 0.000 description 1
- 241000605547 Luzula sylvatica Species 0.000 description 1
- 244000241838 Lycium barbarum Species 0.000 description 1
- 241000227653 Lycopersicon Species 0.000 description 1
- 235000002262 Lycopersicon Nutrition 0.000 description 1
- 244000276497 Lycopersicon esculentum Species 0.000 description 1
- 241000219816 Macrotyloma Species 0.000 description 1
- 241000218378 Magnolia Species 0.000 description 1
- 235000016094 Magnolia denudata Nutrition 0.000 description 1
- 240000005819 Magnolia denudata Species 0.000 description 1
- 235000014837 Malpighia glabra Nutrition 0.000 description 1
- 240000003394 Malpighia glabra Species 0.000 description 1
- 235000000889 Mammea americana Nutrition 0.000 description 1
- 240000005984 Mammea americana Species 0.000 description 1
- 235000016735 Manihot esculenta subsp esculenta Nutrition 0.000 description 1
- 235000011339 Manilkara zapota Nutrition 0.000 description 1
- 240000001794 Manilkara zapota Species 0.000 description 1
- 240000004658 Medicago sativa Species 0.000 description 1
- 235000010624 Medicago sativa Nutrition 0.000 description 1
- 235000014435 Mentha Nutrition 0.000 description 1
- 241001072983 Mentha Species 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 244000174681 Michelia champaca Species 0.000 description 1
- 235000016462 Mimosa pudica Nutrition 0.000 description 1
- 240000001140 Mimosa pudica Species 0.000 description 1
- 240000003433 Miscanthus floridulus Species 0.000 description 1
- 241000878006 Miscanthus sinensis Species 0.000 description 1
- 108010074633 Mixed Function Oxygenases Proteins 0.000 description 1
- 102000008109 Mixed Function Oxygenases Human genes 0.000 description 1
- 235000009815 Momordica Nutrition 0.000 description 1
- 241000218984 Momordica Species 0.000 description 1
- 235000009811 Momordica charantia Nutrition 0.000 description 1
- 235000009812 Momordica cochinchinensis Nutrition 0.000 description 1
- 240000001740 Momordica dioica Species 0.000 description 1
- 235000018365 Momordica dioica Nutrition 0.000 description 1
- 241000122904 Mucuna Species 0.000 description 1
- 240000005561 Musa balbisiana Species 0.000 description 1
- 240000000907 Musa textilis Species 0.000 description 1
- 235000018290 Musa x paradisiaca Nutrition 0.000 description 1
- 108010021466 Mutant Proteins Proteins 0.000 description 1
- 102000008300 Mutant Proteins Human genes 0.000 description 1
- 244000132436 Myrica rubra Species 0.000 description 1
- 241000234479 Narcissus Species 0.000 description 1
- 241001230286 Narenga Species 0.000 description 1
- 235000015742 Nephelium litchi Nutrition 0.000 description 1
- 241000208125 Nicotiana Species 0.000 description 1
- 235000007413 Nigella arvensis Nutrition 0.000 description 1
- 238000000636 Northern blotting Methods 0.000 description 1
- 241000795633 Olea <sea slug> Species 0.000 description 1
- 235000002725 Olea europaea Nutrition 0.000 description 1
- 241000233855 Orchidaceae Species 0.000 description 1
- 235000011203 Origanum Nutrition 0.000 description 1
- 240000000783 Origanum majorana Species 0.000 description 1
- 241001446528 Ornithopus Species 0.000 description 1
- 241000238814 Orthoptera Species 0.000 description 1
- 240000008346 Oryza glaberrima Species 0.000 description 1
- 239000005588 Oxadiazon Substances 0.000 description 1
- 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 1
- 102000004020 Oxygenases Human genes 0.000 description 1
- 108090000417 Oxygenases Proteins 0.000 description 1
- 235000006484 Paeonia officinalis Nutrition 0.000 description 1
- 244000170916 Paeonia officinalis Species 0.000 description 1
- 240000004371 Panax ginseng Species 0.000 description 1
- 235000002789 Panax ginseng Nutrition 0.000 description 1
- 241000180649 Panax notoginseng Species 0.000 description 1
- 235000003143 Panax notoginseng Nutrition 0.000 description 1
- 235000016496 Panda oleosa Nutrition 0.000 description 1
- 240000000220 Panda oleosa Species 0.000 description 1
- 241000282376 Panthera tigris Species 0.000 description 1
- 235000002769 Pastinaca sativa Nutrition 0.000 description 1
- 235000017769 Pastinaca sativa subsp sativa Nutrition 0.000 description 1
- 241000208181 Pelargonium Species 0.000 description 1
- 241000209046 Pennisetum Species 0.000 description 1
- 244000038248 Pennisetum spicatum Species 0.000 description 1
- 244000115721 Pennisetum typhoides Species 0.000 description 1
- 241000288936 Perodicticus potto Species 0.000 description 1
- 241000218196 Persea Species 0.000 description 1
- 240000009164 Petroselinum crispum Species 0.000 description 1
- 235000002770 Petroselinum crispum Nutrition 0.000 description 1
- 244000062780 Petroselinum sativum Species 0.000 description 1
- 241000745991 Phalaris Species 0.000 description 1
- 244000081757 Phalaris arundinacea Species 0.000 description 1
- 235000010627 Phaseolus vulgaris Nutrition 0.000 description 1
- 235000010659 Phoenix dactylifera Nutrition 0.000 description 1
- 244000104275 Phoenix dactylifera Species 0.000 description 1
- 235000014676 Phragmites communis Nutrition 0.000 description 1
- 244000273256 Phragmites communis Species 0.000 description 1
- 244000082204 Phyllostachys viridis Species 0.000 description 1
- 235000015334 Phyllostachys viridis Nutrition 0.000 description 1
- 239000005597 Pinoxaden Substances 0.000 description 1
- 235000005205 Pinus Nutrition 0.000 description 1
- 241000218602 Pinus <genus> Species 0.000 description 1
- 235000008331 Pinus X rigitaeda Nutrition 0.000 description 1
- 235000011613 Pinus brutia Nutrition 0.000 description 1
- 241000018646 Pinus brutia Species 0.000 description 1
- 235000011615 Pinus koraiensis Nutrition 0.000 description 1
- 240000007263 Pinus koraiensis Species 0.000 description 1
- 235000011609 Pinus massoniana Nutrition 0.000 description 1
- 241000018650 Pinus massoniana Species 0.000 description 1
- 241000722363 Piper Species 0.000 description 1
- 235000006440 Pistia stratiotes Nutrition 0.000 description 1
- 244000207867 Pistia stratiotes Species 0.000 description 1
- 241000209049 Poa pratensis Species 0.000 description 1
- 235000011751 Pogostemon cablin Nutrition 0.000 description 1
- 240000002505 Pogostemon cablin Species 0.000 description 1
- 241001633680 Polygonatum odoratum Species 0.000 description 1
- 241000037831 Polygonatum sibiricum Species 0.000 description 1
- 229920001213 Polysorbate 20 Polymers 0.000 description 1
- 241000183024 Populus tremula Species 0.000 description 1
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 description 1
- 235000000497 Primula Nutrition 0.000 description 1
- 241000245063 Primula Species 0.000 description 1
- ONIBWKKTOPOVIA-UHFFFAOYSA-N Proline Natural products OC(=O)C1CCCN1 ONIBWKKTOPOVIA-UHFFFAOYSA-N 0.000 description 1
- 235000001560 Prosopis chilensis Nutrition 0.000 description 1
- 235000014460 Prosopis juliflora var juliflora Nutrition 0.000 description 1
- 235000009827 Prunus armeniaca Nutrition 0.000 description 1
- 244000018633 Prunus armeniaca Species 0.000 description 1
- 235000006029 Prunus persica var nucipersica Nutrition 0.000 description 1
- 235000006040 Prunus persica var persica Nutrition 0.000 description 1
- 244000017714 Prunus persica var. nucipersica Species 0.000 description 1
- 240000001679 Psidium guajava Species 0.000 description 1
- 235000013929 Psidium pyriferum Nutrition 0.000 description 1
- 240000002292 Psychopsis papilio Species 0.000 description 1
- 244000088415 Raphanus sativus Species 0.000 description 1
- 235000006140 Raphanus sativus var sativus Nutrition 0.000 description 1
- 102000007056 Recombinant Fusion Proteins Human genes 0.000 description 1
- 108010008281 Recombinant Fusion Proteins Proteins 0.000 description 1
- 241000219061 Rheum Species 0.000 description 1
- 235000009411 Rheum rhabarbarum Nutrition 0.000 description 1
- 244000193032 Rheum rhaponticum Species 0.000 description 1
- 241000589180 Rhizobium Species 0.000 description 1
- 241000208422 Rhododendron Species 0.000 description 1
- 235000011483 Ribes Nutrition 0.000 description 1
- 241000220483 Ribes Species 0.000 description 1
- 240000001890 Ribes hudsonianum Species 0.000 description 1
- 235000016954 Ribes hudsonianum Nutrition 0.000 description 1
- 235000001466 Ribes nigrum Nutrition 0.000 description 1
- 235000003846 Ricinus Nutrition 0.000 description 1
- 241000322381 Ricinus <louse> Species 0.000 description 1
- 241000220317 Rosa Species 0.000 description 1
- 241001092459 Rubus Species 0.000 description 1
- 235000011034 Rubus glaucus Nutrition 0.000 description 1
- 244000235659 Rubus idaeus Species 0.000 description 1
- 235000009122 Rubus idaeus Nutrition 0.000 description 1
- 241000219053 Rumex Species 0.000 description 1
- 240000000111 Saccharum officinarum Species 0.000 description 1
- 235000007201 Saccharum officinarum Nutrition 0.000 description 1
- 241000124033 Salix Species 0.000 description 1
- 241000921305 Salix sp. Species 0.000 description 1
- 244000124765 Salsola kali Species 0.000 description 1
- 235000007658 Salsola kali Nutrition 0.000 description 1
- 241000208829 Sambucus Species 0.000 description 1
- 235000005010 Scirpus paludosus Nutrition 0.000 description 1
- 241001558929 Sclerotium <basidiomycota> Species 0.000 description 1
- 235000017089 Scutellaria baicalensis Nutrition 0.000 description 1
- 240000004534 Scutellaria baicalensis Species 0.000 description 1
- 241000228160 Secale cereale x Triticum aestivum Species 0.000 description 1
- 235000009367 Sesamum alatum Nutrition 0.000 description 1
- 244000200882 Setaria barbata Species 0.000 description 1
- 235000001561 Setaria barbata Nutrition 0.000 description 1
- 235000008515 Setaria glauca Nutrition 0.000 description 1
- 235000007226 Setaria italica Nutrition 0.000 description 1
- 240000005498 Setaria italica Species 0.000 description 1
- 235000002248 Setaria viridis Nutrition 0.000 description 1
- 240000003461 Setaria viridis Species 0.000 description 1
- 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 1
- 241000220261 Sinapis Species 0.000 description 1
- 241001183351 Sisymbrium volgense Species 0.000 description 1
- 235000003205 Smilax rotundifolia Nutrition 0.000 description 1
- 240000009022 Smilax rotundifolia Species 0.000 description 1
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 1
- 239000005708 Sodium hypochlorite Substances 0.000 description 1
- 241001291279 Solanum galapagense Species 0.000 description 1
- 244000244100 Solanum integrifolium Species 0.000 description 1
- 235000000099 Solanum integrifolium Nutrition 0.000 description 1
- 235000002560 Solanum lycopersicum Nutrition 0.000 description 1
- 235000002597 Solanum melongena Nutrition 0.000 description 1
- 244000061458 Solanum melongena Species 0.000 description 1
- 235000015505 Sorghum bicolor subsp. bicolor Nutrition 0.000 description 1
- 238000002105 Southern blotting Methods 0.000 description 1
- 241000219315 Spinacia Species 0.000 description 1
- 235000009337 Spinacia oleracea Nutrition 0.000 description 1
- 244000300264 Spinacia oleracea Species 0.000 description 1
- 235000009184 Spondias indica Nutrition 0.000 description 1
- 108091081024 Start codon Proteins 0.000 description 1
- 241000044578 Stenotaphrum secundatum Species 0.000 description 1
- 241001626020 Syringa pubescens Species 0.000 description 1
- 244000223014 Syzygium aromaticum Species 0.000 description 1
- 235000004298 Tamarindus indica Nutrition 0.000 description 1
- 240000004584 Tamarindus indica Species 0.000 description 1
- 235000010185 Tamarix canariensis Nutrition 0.000 description 1
- 235000014265 Tamarix gallica Nutrition 0.000 description 1
- 240000001869 Tamarix ramosissima Species 0.000 description 1
- 235000010154 Tamarix ramosissima Nutrition 0.000 description 1
- 240000004460 Tanacetum coccineum Species 0.000 description 1
- 108020005038 Terminator Codon Proteins 0.000 description 1
- 241000223257 Thermomyces Species 0.000 description 1
- 244000044283 Toxicodendron succedaneum Species 0.000 description 1
- 241001521901 Tribulus lanuginosus Species 0.000 description 1
- GSEJCLTVZPLZKY-UHFFFAOYSA-N Triethanolamine Chemical class OCCN(CCO)CCO GSEJCLTVZPLZKY-UHFFFAOYSA-N 0.000 description 1
- 235000007218 Tripsacum dactyloides Nutrition 0.000 description 1
- 244000082267 Tripsacum dactyloides Species 0.000 description 1
- 235000019714 Triticale Nutrition 0.000 description 1
- 241000209140 Triticum Species 0.000 description 1
- 235000007264 Triticum durum Nutrition 0.000 description 1
- 241000209143 Triticum turgidum subsp. durum Species 0.000 description 1
- 235000004424 Tropaeolum majus Nutrition 0.000 description 1
- 240000001260 Tropaeolum majus Species 0.000 description 1
- 235000018946 Tropaeolum minus Nutrition 0.000 description 1
- 240000008573 Tropaeolum minus Species 0.000 description 1
- 235000003095 Vaccinium corymbosum Nutrition 0.000 description 1
- 240000001866 Vernicia fordii Species 0.000 description 1
- 235000002096 Vicia faba var. equina Nutrition 0.000 description 1
- 240000004922 Vigna radiata Species 0.000 description 1
- 235000010721 Vigna radiata var radiata Nutrition 0.000 description 1
- 235000011469 Vigna radiata var sublobata Nutrition 0.000 description 1
- 235000011453 Vigna umbellata Nutrition 0.000 description 1
- 240000001417 Vigna umbellata Species 0.000 description 1
- 241000405217 Viola <butterfly> Species 0.000 description 1
- 241000394440 Viola arvensis Species 0.000 description 1
- 240000009038 Viola odorata Species 0.000 description 1
- 235000013487 Viola odorata Nutrition 0.000 description 1
- 244000172533 Viola sororia Species 0.000 description 1
- 244000047670 Viola x wittrockiana Species 0.000 description 1
- 235000004031 Viola x wittrockiana Nutrition 0.000 description 1
- 235000009392 Vitis Nutrition 0.000 description 1
- 241000219095 Vitis Species 0.000 description 1
- 235000009754 Vitis X bourquina Nutrition 0.000 description 1
- 235000012333 Vitis X labruscana Nutrition 0.000 description 1
- 240000006365 Vitis vinifera Species 0.000 description 1
- 235000014787 Vitis vinifera Nutrition 0.000 description 1
- 208000027418 Wounds and injury Diseases 0.000 description 1
- 241000746966 Zizania Species 0.000 description 1
- 235000002636 Zizania aquatica Nutrition 0.000 description 1
- 241001478412 Zizania palustris Species 0.000 description 1
- ROVGZAWFACYCSP-MQBLHHJJSA-N [2-methyl-4-oxo-3-[(2z)-penta-2,4-dienyl]cyclopent-2-en-1-yl] (1r,3r)-2,2-dimethyl-3-(2-methylprop-1-enyl)cyclopropane-1-carboxylate Chemical compound CC1(C)[C@H](C=C(C)C)[C@H]1C(=O)OC1C(C)=C(C\C=C/C=C)C(=O)C1 ROVGZAWFACYCSP-MQBLHHJJSA-N 0.000 description 1
- FJJCIZWZNKZHII-UHFFFAOYSA-N [4,6-bis(cyanoamino)-1,3,5-triazin-2-yl]cyanamide Chemical compound N#CNC1=NC(NC#N)=NC(NC#N)=N1 FJJCIZWZNKZHII-UHFFFAOYSA-N 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- 150000007513 acids Chemical class 0.000 description 1
- 239000000654 additive Substances 0.000 description 1
- 235000020224 almond Nutrition 0.000 description 1
- 230000003698 anagen phase Effects 0.000 description 1
- 235000003484 annual ragweed Nutrition 0.000 description 1
- 239000001387 apium graveolens Substances 0.000 description 1
- 239000011425 bamboo Substances 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
- 230000008901 benefit Effects 0.000 description 1
- IQFYYKKMVGJFEH-UHFFFAOYSA-N beta-L-thymidine Natural products O=C1NC(=O)C(C)=CN1C1OC(CO)C(O)C1 IQFYYKKMVGJFEH-UHFFFAOYSA-N 0.000 description 1
- 235000013361 beverage Nutrition 0.000 description 1
- 238000004166 bioassay Methods 0.000 description 1
- 238000010256 biochemical assay Methods 0.000 description 1
- 230000003115 biocidal effect Effects 0.000 description 1
- 235000021014 blueberries Nutrition 0.000 description 1
- KGBXLFKZBHKPEV-UHFFFAOYSA-N boric acid Chemical compound OB(O)O KGBXLFKZBHKPEV-UHFFFAOYSA-N 0.000 description 1
- 239000004327 boric acid Substances 0.000 description 1
- 230000028446 budding cell bud growth Effects 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- 235000006263 bur ragweed Nutrition 0.000 description 1
- YHSAMQAMZAWZPE-UHFFFAOYSA-N butyl 2-[4-[3-chloro-5-(trifluoromethyl)pyridin-2-yl]oxyphenoxy]propanoate Chemical group C1=CC(OC(C)C(=O)OCCCC)=CC=C1OC1=NC=C(C(F)(F)F)C=C1Cl YHSAMQAMZAWZPE-UHFFFAOYSA-N 0.000 description 1
- 235000009120 camo Nutrition 0.000 description 1
- 239000005018 casein Substances 0.000 description 1
- BECPQYXYKAMYBN-UHFFFAOYSA-N casein, tech. Chemical compound NCCCCC(C(O)=O)N=C(O)C(CC(O)=O)N=C(O)C(CCC(O)=N)N=C(O)C(CC(C)C)N=C(O)C(CCC(O)=O)N=C(O)C(CC(O)=O)N=C(O)C(CCC(O)=O)N=C(O)C(C(C)O)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=O)N=C(O)C(CCC(O)=O)N=C(O)C(COP(O)(O)=O)N=C(O)C(CCC(O)=N)N=C(O)C(N)CC1=CC=CC=C1 BECPQYXYKAMYBN-UHFFFAOYSA-N 0.000 description 1
- 235000021240 caseins Nutrition 0.000 description 1
- 235000020226 cashew nut Nutrition 0.000 description 1
- 230000001925 catabolic effect Effects 0.000 description 1
- 230000001413 cellular effect Effects 0.000 description 1
- 230000030570 cellular localization Effects 0.000 description 1
- 229940124587 cephalosporin Drugs 0.000 description 1
- 150000001780 cephalosporins Chemical class 0.000 description 1
- 235000013339 cereals Nutrition 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 235000005607 chanvre indien Nutrition 0.000 description 1
- 238000012512 characterization method Methods 0.000 description 1
- 239000012295 chemical reaction liquid Substances 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 229910052801 chlorine Inorganic materials 0.000 description 1
- 125000001309 chloro group Chemical group Cl* 0.000 description 1
- 210000003763 chloroplast Anatomy 0.000 description 1
- OEYIOHPDSNJKLS-UHFFFAOYSA-N choline Chemical compound C[N+](C)(C)CCO OEYIOHPDSNJKLS-UHFFFAOYSA-N 0.000 description 1
- 229960001231 choline Drugs 0.000 description 1
- 239000001407 cinnamomum spp. Substances 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
- 238000012411 cloning technique Methods 0.000 description 1
- HUBANNPOLNYSAD-UHFFFAOYSA-N clopyralid Chemical compound OC(=O)C1=NC(Cl)=CC=C1Cl HUBANNPOLNYSAD-UHFFFAOYSA-N 0.000 description 1
- 239000003086 colorant Substances 0.000 description 1
- 235000018597 common camellia Nutrition 0.000 description 1
- 235000003488 common ragweed Nutrition 0.000 description 1
- 238000004891 communication Methods 0.000 description 1
- 238000009833 condensation Methods 0.000 description 1
- 230000005494 condensation Effects 0.000 description 1
- 230000001276 controlling effect Effects 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 238000012136 culture method Methods 0.000 description 1
- 229930186364 cyclamen Natural products 0.000 description 1
- FWFSEYBSWVRWGL-UHFFFAOYSA-N cyclohex-2-enone Chemical class O=C1CCCC=C1 FWFSEYBSWVRWGL-UHFFFAOYSA-N 0.000 description 1
- ANJTVLIZGCUXLD-DTWKUNHWSA-N cytisine Chemical compound C1NC[C@H]2CN3C(=O)C=CC=C3[C@@H]1C2 ANJTVLIZGCUXLD-DTWKUNHWSA-N 0.000 description 1
- 229940027564 cytisine Drugs 0.000 description 1
- 229930017327 cytisine Natural products 0.000 description 1
- 230000009089 cytolysis Effects 0.000 description 1
- 230000000382 dechlorinating effect Effects 0.000 description 1
- 238000012217 deletion Methods 0.000 description 1
- 230000037430 deletion Effects 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 239000000645 desinfectant Substances 0.000 description 1
- 230000004069 differentiation Effects 0.000 description 1
- 125000000118 dimethyl group Chemical group [H]C([H])([H])* 0.000 description 1
- 235000021186 dishes Nutrition 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 244000013123 dwarf bean Species 0.000 description 1
- 235000005489 dwarf bean Nutrition 0.000 description 1
- 229920001971 elastomer Polymers 0.000 description 1
- 230000000408 embryogenic effect Effects 0.000 description 1
- 239000003623 enhancer Substances 0.000 description 1
- ANJTVLIZGCUXLD-UHFFFAOYSA-N ent-cytisine Natural products C1NCC2CN3C(=O)C=CC=C3C1C2 ANJTVLIZGCUXLD-UHFFFAOYSA-N 0.000 description 1
- 230000009088 enzymatic function Effects 0.000 description 1
- 238000003028 enzyme activity measurement method Methods 0.000 description 1
- 125000004494 ethyl ester group Chemical group 0.000 description 1
- 235000008995 european elder Nutrition 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 239000013613 expression plasmid Substances 0.000 description 1
- 239000004744 fabric Substances 0.000 description 1
- 239000000835 fiber Substances 0.000 description 1
- 235000004426 flaxseed Nutrition 0.000 description 1
- 230000004907 flux Effects 0.000 description 1
- 210000002683 foot Anatomy 0.000 description 1
- 239000004459 forage Substances 0.000 description 1
- 230000005714 functional activity Effects 0.000 description 1
- ZZUFCTLCJUWOSV-UHFFFAOYSA-N furosemide Chemical compound C1=C(Cl)C(S(=O)(=O)N)=CC(C(O)=O)=C1NCC1=CC=CO1 ZZUFCTLCJUWOSV-UHFFFAOYSA-N 0.000 description 1
- 238000012215 gene cloning Methods 0.000 description 1
- 238000010353 genetic engineering Methods 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
- 239000003448 gibberellin Substances 0.000 description 1
- 235000008434 ginseng Nutrition 0.000 description 1
- 235000021384 green leafy vegetables Nutrition 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 239000011487 hemp Substances 0.000 description 1
- 238000004128 high performance liquid chromatography Methods 0.000 description 1
- 230000001900 immune effect Effects 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 238000001802 infusion Methods 0.000 description 1
- 208000014674 injury Diseases 0.000 description 1
- 238000003780 insertion Methods 0.000 description 1
- 230000037431 insertion Effects 0.000 description 1
- 238000007689 inspection Methods 0.000 description 1
- 230000002452 interceptive effect Effects 0.000 description 1
- 125000001449 isopropyl group Chemical group [H]C([H])([H])C([H])(*)C([H])([H])[H] 0.000 description 1
- JJWLVOIRVHMVIS-UHFFFAOYSA-N isopropylamine Chemical class CC(C)N JJWLVOIRVHMVIS-UHFFFAOYSA-N 0.000 description 1
- AWJUIBRHMBBTKR-UHFFFAOYSA-N isoquinoline Chemical class C1=NC=CC2=CC=CC=C21 AWJUIBRHMBBTKR-UHFFFAOYSA-N 0.000 description 1
- 235000021374 legumes Nutrition 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 231100000053 low toxicity Toxicity 0.000 description 1
- 210000001161 mammalian embryo Anatomy 0.000 description 1
- 235000005739 manihot Nutrition 0.000 description 1
- 125000000311 mannosyl group Chemical group C1([C@@H](O)[C@@H](O)[C@H](O)[C@H](O1)CO)* 0.000 description 1
- 230000001404 mediated effect Effects 0.000 description 1
- 230000037353 metabolic pathway Effects 0.000 description 1
- 150000004702 methyl esters Chemical class 0.000 description 1
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 description 1
- 229960002939 metizoline Drugs 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- DEDOPGXGGQYYMW-UHFFFAOYSA-N molinate Chemical compound CCSC(=O)N1CCCCCC1 DEDOPGXGGQYYMW-UHFFFAOYSA-N 0.000 description 1
- 230000035772 mutation Effects 0.000 description 1
- 229920003052 natural elastomer Polymers 0.000 description 1
- 229920001194 natural rubber Polymers 0.000 description 1
- 230000007935 neutral effect Effects 0.000 description 1
- 239000001711 nigella sativa Substances 0.000 description 1
- 238000010899 nucleation Methods 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 238000009401 outcrossing Methods 0.000 description 1
- 238000010979 pH adjustment Methods 0.000 description 1
- 235000011197 perejil Nutrition 0.000 description 1
- 238000005375 photometry Methods 0.000 description 1
- 230000029553 photosynthesis Effects 0.000 description 1
- 238000010672 photosynthesis Methods 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
- 239000001739 pinus spp. Substances 0.000 description 1
- 230000008654 plant damage Effects 0.000 description 1
- 239000005648 plant growth regulator Substances 0.000 description 1
- 239000003375 plant hormone Substances 0.000 description 1
- 238000004161 plant tissue culture Methods 0.000 description 1
- 230000010152 pollination Effects 0.000 description 1
- 239000000256 polyoxyethylene sorbitan monolaurate Substances 0.000 description 1
- 235000010486 polyoxyethylene sorbitan monolaurate Nutrition 0.000 description 1
- 239000011591 potassium Substances 0.000 description 1
- 229910052700 potassium Inorganic materials 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
- 239000000843 powder Substances 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- WHOKDONDRZNCBC-UHFFFAOYSA-N propan-2-yl 2-(2,4-dichlorophenoxy)acetate Chemical group CC(C)OC(=O)COC1=CC=C(Cl)C=C1Cl WHOKDONDRZNCBC-UHFFFAOYSA-N 0.000 description 1
- 230000004952 protein activity Effects 0.000 description 1
- 230000026447 protein localization Effects 0.000 description 1
- 238000001742 protein purification Methods 0.000 description 1
- 230000017854 proteolysis Effects 0.000 description 1
- 210000001938 protoplast Anatomy 0.000 description 1
- 235000014774 prunus Nutrition 0.000 description 1
- 235000015136 pumpkin Nutrition 0.000 description 1
- 229940015367 pyrethrum Drugs 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
- FFSSWMQPCJRCRV-UHFFFAOYSA-N quinclorac Chemical compound ClC1=CN=C2C(C(=O)O)=C(Cl)C=CC2=C1 FFSSWMQPCJRCRV-UHFFFAOYSA-N 0.000 description 1
- VQMSRUREDGBWKT-UHFFFAOYSA-N quinoline-4-carboxylic acid Chemical class C1=CC=C2C(C(=O)O)=CC=NC2=C1 VQMSRUREDGBWKT-UHFFFAOYSA-N 0.000 description 1
- 235000009736 ragweed Nutrition 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 238000012827 research and development Methods 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- 108091008146 restriction endonucleases Proteins 0.000 description 1
- 210000000614 rib Anatomy 0.000 description 1
- 239000012883 rooting culture medium Substances 0.000 description 1
- 239000012882 rooting medium Substances 0.000 description 1
- DCKVNWZUADLDEH-UHFFFAOYSA-N sec-butyl acetate Chemical compound CCC(C)OC(C)=O DCKVNWZUADLDEH-UHFFFAOYSA-N 0.000 description 1
- 238000002864 sequence alignment Methods 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 229910052708 sodium Inorganic materials 0.000 description 1
- MFBOGIVSZKQAPD-UHFFFAOYSA-M sodium butyrate Chemical compound [Na+].CCCC([O-])=O MFBOGIVSZKQAPD-UHFFFAOYSA-M 0.000 description 1
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 description 1
- SUKJFIGYRHOWBL-UHFFFAOYSA-N sodium hypochlorite Chemical compound [Na+].Cl[O-] SUKJFIGYRHOWBL-UHFFFAOYSA-N 0.000 description 1
- 159000000000 sodium salts Chemical class 0.000 description 1
- 125000006850 spacer group Chemical group 0.000 description 1
- 238000001228 spectrum Methods 0.000 description 1
- 235000020354 squash Nutrition 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 210000004233 talus Anatomy 0.000 description 1
- 230000008685 targeting Effects 0.000 description 1
- 101150083668 tfdD gene Proteins 0.000 description 1
- YRHRIQCWCFGUEQ-UHFFFAOYSA-N thioxanthen-9-one Chemical compound C1=CC=C2C(=O)C3=CC=CC=C3SC2=C1 YRHRIQCWCFGUEQ-UHFFFAOYSA-N 0.000 description 1
- 229940104230 thymidine Drugs 0.000 description 1
- 238000003971 tillage Methods 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- UZKQTCBAMSWPJD-UQCOIBPSSA-N trans-Zeatin Natural products OCC(/C)=C\CNC1=NC=NC2=C1N=CN2 UZKQTCBAMSWPJD-UQCOIBPSSA-N 0.000 description 1
- UZKQTCBAMSWPJD-FARCUNLSSA-N trans-zeatin Chemical compound OCC(/C)=C/CNC1=NC=NC2=C1N=CN2 UZKQTCBAMSWPJD-FARCUNLSSA-N 0.000 description 1
- GOSWTRUMMSCNCW-UHFFFAOYSA-N trans-zeatin riboside Natural products C1=NC=2C(NCC=C(CO)C)=NC=NC=2N1C1OC(CO)C(O)C1O GOSWTRUMMSCNCW-UHFFFAOYSA-N 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 238000013519 translation Methods 0.000 description 1
- 230000014621 translational initiation Effects 0.000 description 1
- 230000004102 tricarboxylic acid cycle Effects 0.000 description 1
- 150000003628 tricarboxylic acids Chemical class 0.000 description 1
- 239000012137 tryptone Substances 0.000 description 1
- 210000001113 umbilicus Anatomy 0.000 description 1
- 235000018322 upland cotton Nutrition 0.000 description 1
- 235000013311 vegetables Nutrition 0.000 description 1
- 230000003612 virological effect Effects 0.000 description 1
- 150000003722 vitamin derivatives Chemical class 0.000 description 1
- 235000020234 walnut Nutrition 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
- 238000001262 western blot Methods 0.000 description 1
- 241000228158 x Triticosecale Species 0.000 description 1
- 229940023877 zeatin Drugs 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
-
- 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
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/0004—Oxidoreductases (1.)
- C12N9/0069—Oxidoreductases (1.) acting on single donors with incorporation of molecular oxygen, i.e. oxygenases (1.13)
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H5/00—Angiosperms, i.e. flowering plants, characterised by their plant parts; Angiosperms characterised otherwise than by their botanic taxonomy
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H5/00—Angiosperms, i.e. flowering plants, characterised by their plant parts; Angiosperms characterised otherwise than by their botanic taxonomy
- A01H5/10—Seeds
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H6/00—Angiosperms, i.e. flowering plants, characterised by their botanic taxonomy
- A01H6/20—Brassicaceae, e.g. canola, broccoli or rucola
-
- 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)
-
- 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
- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
- C12N5/10—Cells modified by introduction of foreign genetic material
-
- 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
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/0004—Oxidoreductases (1.)
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
- C12Y113/00—Oxidoreductases acting on single donors with incorporation of molecular oxygen (oxygenases) (1.13)
- C12Y113/12—Oxidoreductases acting on single donors with incorporation of molecular oxygen (oxygenases) (1.13) with incorporation of one atom of oxygen (internal monooxygenases or internal mixed function oxidases)(1.13.12)
- C12Y113/12019—2-Oxuglutarate dioxygenase (ethylene-forming) (1.13.12.19)
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Genetics & Genomics (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Organic Chemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- General Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Biomedical Technology (AREA)
- Molecular Biology (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Microbiology (AREA)
- Cell Biology (AREA)
- Physics & Mathematics (AREA)
- Medicinal Chemistry (AREA)
- Biophysics (AREA)
- Plant Pathology (AREA)
- Physiology (AREA)
- Botany (AREA)
- Developmental Biology & Embryology (AREA)
- Environmental Sciences (AREA)
- Natural Medicines & Medicinal Plants (AREA)
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Enzymes And Modification Thereof (AREA)
Abstract
本发明属于生物技术领域,具体涉及一种能赋予激素类除草剂、ACCase抑制剂类除草剂抗性的蛋白、基因和用途,以及具有除草剂耐受性的植物、种子、细胞和植物部分,及其使用方法。
Description
本申请是申请日为2020年08月06日,申请号为202010784387X,发明名称为“除草剂抗性蛋白、基因和用途”的中国发明专利申请的分案申请。
技术领域
本发明属于生物技术领域,具体涉及一种除草剂抗性蛋白、基因和用途。
背景技术
杂草可以迅速耗尽土壤中的水分和各种作物所需要的养分,还与作物竞争生长空间,严重影响作物生长发育。随着农村城市化发展和农村劳动力转移,人工除草已经不经济也不现实。耕作制度的改变比如自然免耕和作物直播等轻便栽培方式的推广实施,草害问题越来越突出,严重制约农作物的高产和稳产,急需解决方案。在这种背景下除草剂的使用成为必然的选择,现在的耕地和非耕地除草越来越依赖使用除草剂。
草甘膦已经在全球广泛使用超过25年,由此导致对草甘膦和草甘膦耐性作物技术的过度依赖,长期使用单一除草剂草甘膦对野生杂草种群施加了高的选择压力,导致抗性杂草发生。据报道,已有40几种杂草表现出对草甘膦的抗性,PestManagSci 2018;74:1089–1093,包括阔叶杂草和禾本科杂草,如多种野苋菜(Amaranthus spp.)、豚草(Ambrosiaspp.)、小飞蓬(Conyza canadensis)、牛筋草(Eleusine indica)、刺撒笼(Salsolatragus)等。
2,4-二氯苯氧乙酸(2,4-D)是激素类除草剂的代表,是另一类灭生性除草剂,与草甘膦混合或单独使用可以防除草甘膦抗性杂草。2,4-D已经在耕地和非耕地用于广谱阔叶杂草控制超过65年,但仍然在全球广泛使用。另一方面,2,4-D对双子叶植物(如大豆或棉花)的选择性特别差,一般不用于敏感性双子叶作物除草;即使对禾本科作物也不十分安全。这些因素也同时限制了2,4-D在农田除草中的应用。
2,4-D有个优点,即在土壤里的半衰期较短,对后茬作物没有明显影响。根据这一线索,科学家找到了许多能够降解2,4-D化合物的土壤微生物。已经广泛研究能降解2,4-D的是真养雷氏菌(Ralstonia eutropha),其降解基因叫tfdA。TfdA编码AAD类酶,α-酮戊二酸依赖型双加氧酶(AADs,α-ketoglutarate dependent aryloxyalkanoatedioxygenases)。该类酶可降解2,4-D、二甲四氯等苯氧乙酸类除草剂,可将2,4-D脱去乙酸基团生成毒性较低的2,4-二氯苯酚(DCP)。TfdA酶蛋白有个TauD结构域负责降解2,4-D。目前已报道的2,4-D降解主要通过两条代谢途径:①在α-酮戊二酸依赖型双加氧酶基因TfdA(来自真养产碱菌JMP134)或CadABC(来自短根瘤菌HW13)的作用下,催化2,4-D脱去乙酸基团生成毒性比2,4-D低100倍的2,4-二氯苯酚(DCP),随后2,4-二氯苯酚在羟化酶TfdB的催化下被转化为3,5-二氯邻苯二酚,然后依次在TfdC、TfdD、TfdE、TfdF的作用下开环最终被降解为β-酮己二酸,进入三羧酸循环。②在球褐固氮菌(Azotobacter chroococcum)中,2,4-D在未知脱氯酶的作用下首先脱去一个氯原子生成对氯苯氧乙酸,随后氧化脱去乙酸基团生成对氯苯酚,进一步羟基化生成4-氯邻苯二酚,然后开环被降解为β-酮己二酸,最终进入三羧酸循环。还有些AAD酶,例如来源于食酸戴尔福特菌(Comamonas acidovorans)的AAD-12,可以催化其他类除草剂,如吡啶氧乙酸类的氯氟吡氧乙酸类(绿草定Triclopyr和氟草烟Fluroxypyr)以及芳氧基苯氧基丙酸酯类(AOPP)除草剂(CN101688219B;US2019/0017066Al;US10167483B2;Anne Westendorf,Dirk Benndorf,Roland H.Müller,WolfgangBabel.2002.The two enantiospecific dichlorprop/
α-ketoglutarate-dioxygenases fromDelftia acidovorans MC1–protein andsequence data ofRdpA and SdpA.Microbiol.Res.(2002)157,317–322,http://www.urbanfischer.de/journals/microbiolres;Jonathan R.Chekan,ChayanidOngpipattanakul,Terry R.Wright,Bo Zhang,J.Martin Bollinger Jr.,LaurenJ.Rajakovich,Carsten Krebs,Robert M.Cicchillo,and SatishK.Nair.2019.Molecular basis for enantioselective herbicide degradationimpartedby aryloxyalkanoate dioxygenases in transgenic plants)。
随着转基因技术的发展,AAD酶类编码基因被转入敏感植物如棉花和烟草,转基因植物获得了2,4-D抗性,这为解决草甘膦抗性杂草提供了技术方案。美国陶氏农科(DowAgroSciences)公司推出了转基因玉米、大豆、棉花等作物,分别表达AAD1、AAD12和AAD13来降解2,4-D类的除草剂(T.R.Wright et al.,Robust crop resistance to broadleafandgrass herbicides providedby aryloxyalkanoate dioxygenase transgenes.Proc.Natl.Acad.Sci.U.S.A.107,20240–20245(2010)),这使2,4-D为代表的激素类除草剂在农田除草上的应用提供了条件。
综上,除草剂的类型繁多,但目前筛选到的降解基因数目和各自的抗谱非常有限,不能满足作物除草剂抗性性状培育的需要,急需筛选更多类型的基因为转基因作物研发提供基因资源。
发明概述
为解决现有技术中存在的上述问题,本发明提供一种除草剂抗性蛋白、基因和用途。
本发明提供一种重组DNA分子,其包含下述的核酸序列:
(1)编码包含与选自以下组的至少一个氨基酸序列具有至少90%、至少95%、至少96%、至少97%、至少98%、至少99%或100%序列同一性的氨基酸序列的蛋白或其生物活性片段的核酸序列或其互补序列:SEQ ID NO:41、SEQ ID NO:51、SEQ ID NO:59、SEQ IDNO:63、SEQ ID NO:65、SEQ ID NO:67、SEQ ID NO:69、SEQ ID NO:83、SEQ ID NO:85、SEQ IDNO:87、SEQ ID NO:89、SEQ ID NO:91、SEQ ID NO:93、SEQ ID NO:95、SEQ ID NO:97、SEQ IDNO:99、SEQ ID NO:101、SEQ ID NO:103、SEQ ID NO:105、SEQ ID NO:107、SEQ ID NO:109、SEQ ID NO:111、SEQ ID NO:113、SEQ ID NO:115、SEQ ID NO:123、SEQ ID NO:143、SEQ IDNO:145、SEQ ID NO:147、SEQ ID NO:149和SEQ ID NO:151;
(2)SEQ ID NO:42、SEQ ID NO:52、SEQ ID NO:60、SEQ ID NO:64、SEQ ID NO:66、SEQ ID NO:68、SEQ ID NO:70、SEQ ID NO:84、SEQ ID NO:86、SEQ ID NO:88、SEQ ID NO:90、SEQ ID NO:92、SEQ ID NO:94、SEQ ID NO:96、SEQ ID NO:98、SEQ ID NO:100、SEQ IDNO:102、SEQ ID NO:104、SEQ ID NO:106、SEQ ID NO:108、SEQ ID NO:110、SEQ ID NO:112、SEQ ID NO:114、SEQ ID NO:116、SEQ ID NO:124、SEQ ID NO:144、SEQ ID NO:146、SEQ IDNO:148、SEQ ID NO:150或SEQ ID NO:152所示的核酸序列或其互补序列;
(3)在严谨条件下与(1)或(2)所示序列杂交的核酸序列;或
(4)因遗传密码的简并性而与(1)或(2)所示序列编码相同氨基酸序列的核酸序列或其互补序列。
在一个具体实施方式中,所述重组DNA分子可操作地连接至在植物细胞中有功能的异源启动子。
本发明提供一种DNA构建体,其包含可操作地连接至所述的重组DNA分子的在植物细胞中有功能的异源启动子。
在另一个具体实施方式中,所述DNA构建体存在于转基因植物的基因组中。
本发明提供一种蛋白或其生物活性片段,其由所述的重组DNA分子编码。
本发明另外提供一种蛋白或其生物活性片段,其氨基酸序列与选自以下组的至少一个氨基酸序列具有至少90%、至少95%、至少96%、至少97%、至少98%、至少99%或100%序列同一性:SEQ ID NO:41、SEQ ID NO:51、SEQ ID NO:59、SEQ ID NO:63、SEQ IDNO:65、SEQ ID NO:67、SEQ ID NO:69、SEQ ID NO:83、SEQ ID NO:85、SEQ ID NO:87、SEQ IDNO:89、SEQ ID NO:91、SEQ ID NO:93、SEQ ID NO:95、SEQ ID NO:97、SEQ ID NO:99、SEQ IDNO:101、SEQ ID NO:103、SEQ ID NO:105、SEQ ID NO:107、SEQ ID NO:109、SEQ ID NO:111、SEQ ID NO:113、SEQ ID NO:115、SEQ ID NO:123、SEQ ID NO:143、SEQ ID NO:145、SEQ IDNO:147、SEQ ID NO:149和SEQ ID NO:151。
在一个具体实施方式中,所述的蛋白或其生物活性片段,其对选自以下类型的除草剂中的至少一种具有加氧酶活性:激素类除草剂、ACCase抑制剂类除草剂。
本发明提供一种植物、种子、细胞或植物部分,其包含所述的重组DNA分子、所述的DNA构建体、或所述的蛋白或其生物活性片段。
在一个具体实施方式中,所述的植物、种子、细胞或植物部分,其包含对选自以下类型的除草剂中的至少一种的耐受性:激素类除草剂、ACCase抑制剂类除草剂。
本发明提供一种分离的多核苷酸,其中包含所述的重组DNA分子、所述的DNA构建体、或编码所述蛋白或其生物活性片段的核酸序列或其互补序列。
本发明提供一种植物基因组,其中包含所述的多核苷酸。
本发明提供一种载体,其中包含所述的多核苷酸以及与之可操作连接的同源启动子。
在一个具体实施方式中,所述启动子为诱导型启动子或植物基因组中的自身基因启动子。
本发明提供一种宿主细胞,其中包含所述的多核苷酸或所述的载体。
本发明提供一种用于产生或提高除草剂耐受性的植物或种子的方法,其包括用所述的重组DNA分子或所述的DNA构建体转化植物细胞或组织,和从所述转化的植物细胞或组织再生除草剂耐受性植物。
本发明提供通过前述方法所生产的植物或种子。
本发明提供一种用于赋予植物、种子、细胞或植物部分以除草剂耐受性的方法,所述方法包括在所述植物、种子、细胞或植物部分中表达所述的蛋白或其生物活性片段;
或者,其中包括将表达所述的蛋白或其生物活性片段的植物与另一植物杂交,以及筛选能产生或提高对除草剂耐受性的植物、种子、细胞或植物部分;
或者,其中包括对所述植物、种子、细胞或植物部分进行基因编辑,以实现在其中表达所述的蛋白或其生物活性片段。
在一个具体实施方式中,所述植物、种子、细胞或植物部分包含所述的DNA构建体。
本发明提供所述的重组DNA分子、所述的DNA构建体、所述的蛋白或其生物活性片段、所述的多核苷酸、所述的植物基因组、所述的载体或所述的宿主细胞用于产生或提高植物、种子、细胞或植物部分对除草剂耐受性的用途。
本发明提供一种用于控制植物生长区域中的杂草的方法,其包括使含有耐受除草剂的植物或种子的植物生长区域与所述除草剂接触,所述植物或种子包括前述的植物或种子、通过前述方法制备的植物或种子、或包含所述的重组DNA分子的植物或种子。
附图说明
图1.AAD-12表达菌大肠杆菌菌内降解2,4-D产生酚类物质,加入显色液后显现红色。
图2-1.筛选得到的16个能够降解2,4-D的新的AAD类似基因表达菌(编号D42、D43、D44、D45、D46、D47、D48、D49、D50、D30、D72、D73、D74、D75、D76和D58,D=QYD)菌内降解谱测试。测试化合物包括2,4-D、二甲四氯、2,4-D丁酸、麦草畏、氯氟吡氧乙酸、绿草定和属于ACCase抑制剂的精喹禾灵测试。结果显示,这些基因所编码的AAD类似酶除对2,4-D降解外,对二甲四氯也有强的降解能力;其中D42、D43、D46、D47、D48、D30、D72、D73、D74、D76和D58(D=QYD)对绿草定显示微弱降解;对其余的都不降解。pET15b空载体,作为阴性对照;AAD12是阳性对照。
图2-2.筛选得到的另外9个能够降解2,4-D的新的AAD类似基因表达菌(编号D21、D51、D52、D53、D54、D55、D56、D57、D62,D=QYD)菌内降解谱测试。测试化合物包括2,4-D、二甲四氯、2,4-D丁酸、麦草畏、氯氟吡氧乙酸、绿草定和属于ACCase抑制剂的精喹禾灵。结果显示,这些AAD类似酶除对2,4-D降解外,对二甲四氯也有更强的降解能力;但对其余的都不降解。pET15b空载体,作为阴性对照;AAD12是阳性对照。
图3.AAD类似基因在拟南芥中过表达的代表性载体。AAD类似基因的表达由pAtUBQ10(=AtUbi10启动子)拟南芥泛素启动子和甘露减合成酶MAS(mannopinesynthase)终止子调控下进行。筛选标记是潮霉素抗性基因(HygR=hpt)。
图4-1.转AAD类似基因拟南芥T1代幼苗代表性2,4-D抗性筛选试验。平板培养基中含有2,4-D化合物(0.3μmol)。Col-0:野生型拟南芥Columbia-0品种,在2,4-D的作用下其种子根受到抑制,不能生长。pQYxxxx代表用各种过表达载体蘸花转化后的T1代幼苗:没有被正常转化的(即野生型),或者虽然被正常转化但其转基因不抗2,4-D的植株,其种子根也不能生长,那些种子根能够正常长出的植株,表明是转基因,而且能够抗2,4-D。
图4-2.转AAD类似基因拟南芥T1代幼苗2,4-D丁酸钠抗性筛选试验。平板培养基中含有2,4-D丁酸钠化合物(0.7μmol)。Col-0:野生型拟南芥Columbia-0品种,在2,4-D丁酸钠的作用下其种子根受到抑制,不能生长。pQYxxxx代表用各种过表达载体蘸花转化后的T1代幼苗:没有被正常转化的(即野生型),或者虽然被正常转化但其转基因不抗2,4-D丁酸钠的植株,其种子根也不能生长,那些种子根能够正常长出的植株,表明是转基因,而且能够抗2,4-D丁酸钠。
图4-3.转AAD类似基因拟南芥T1代幼苗二甲四氯抗性筛选试验。平板培养基中含有二甲四氯化合物(0.1μmol)。Col-0:野生型拟南芥Columbia-0品种,在二甲四氯的作用下其种子根受到抑制,不能生长。pQYxxxx代表用各种过表达载体蘸花转化后的T1代幼苗:没有被正常转化的(即野生型),或者虽然被正常转化但其转基因不抗二甲四氯的植株,其种子根也不能生长,那些种子根能够正常长出的植株,表明是转基因,而且能够抗二甲四氯。
图5-1.转AAD类似基因拟南芥T2代幼苗2,4-D抗性试验。平板培养基中含有2,4-D化合物(0.3μmol)。Col-0:野生型拟南芥Columbia-0品种,其种子根不能生长。pQYxxxx代表用各种过表达载体蘸花转化后的T2代幼苗,每个载体测试两个株系:有的转基因拟南芥种子根生长正常,有的生长也不正常,这表明基因本身在2,4-D抗性上的差异,也可能是表达量上(不同转化事件)的差异。
图5-2.转AAD类似基因拟南芥T2代幼苗代表性二甲四氯抗性试验。平板培养基中含有二甲四氯化合物(0.1μmol)。Col-0:野生型拟南芥Columbia-0品种,其种子根不能生长。pQYxxxx代表用各种过表达载体蘸花转化后的T2代幼苗,每个载体测试两个株系:有的转基因拟南芥种子根生长正常,有的生长也不正常,这表明基因本身在二甲四氯抗性上的差异,也可能是表达量上(即不同转化事件)的差异。
图5-3.转AAD类似基因拟南芥T2代幼苗代表性2,4-D丁酸钠抗性试验。平板培养基中含有2,4-D丁酸钠化合物(0.7μmol)。Col-0:野生型拟南芥Columbia-0品种,其种子根不能生长。pQYxxxx代表用各种过表达载体蘸花转化后的T2代幼苗,每个载体测试两个株系:有的转基因拟南芥种子根生长正常,有的生长也不正常,这表明基因本身在2,4-D丁酸钠抗性上的差异,也可能是表达量上(即不同转化事件)的差异。
图6-1.代表性转基因拟南芥T2代苗对2,4-D(喷雾)的抗性。Col-0:野生型拟南芥Columbia-0品种,喷雾2,4-D(100克/公顷)处理10天后全死亡,而所有转基因拟南芥的所有株系均无明显伤害。
图6-2.代表性转基因拟南芥T2代苗对二甲四氯(喷雾)的抗性。Col-0:野生型拟南芥Columbia-0品种,喷雾二甲四氯(100克/公顷)处理10天后全死亡,而所有转基因拟南芥的所有株系均无明显伤害。
图6-3.代表性转基因拟南芥T2代苗对2,4-D丁酸钠(喷雾)的抗性。Col-0:野生型拟南芥Columbia-0品种,喷雾2,4-D丁酸钠(100克/公顷)处理10天后伤害严重,而所有转基因拟南芥的所有株系均无明显伤害。
图7-1.pQY2329转AAD类似基因QYD42(SEQ ID NO.83)玉米过表达载体。QYD42的表达由玉米泛素启动子和Nos终止子调控;筛选基因BlpR(草铵膦抗性基因)表达由CaMV35S启动子及其终止子调控。
图7-2.代表性转基因玉米事件T1代苗对2,4-D(喷雾)的抗性。WT(野生型):玉米B104喷雾2,4-D(4.48公斤/公顷)处理14天后茎基部肿大,而pQY2329转基因事件的茎基部无明显畸形。
图8-1.pQY2330转AAD类似基因QYD42(SEQ ID NO.83)大豆过表达载体。QYD42的表达由双CaMV35S启动子和Nos终止子调控;筛选基因BlpR(草铵膦抗性基因)表达由甘露减合成酶(mannopine synthase)MAS启动子及其终止子调控。
图8-2.代表性转基因大豆事件T1代苗对2,4-D(喷雾)的抗性。WT(野生型):大豆William82喷雾2,4-D(4.48公斤/公顷)处理10天后死亡,而pQY2330转基因事件正常,喷水对照没有差异。
发明详述
提供以下定义和方法以更好地限定本发明并指导本领域的普通技术人员实施本发明。除非另外说明,否则术语应根据相关领域的普通技术人员的常规用法来理解。
在本发明中,“激素类除草剂”是本身有除草活性的物质或者与能改变其效果的其他除草剂和/或添加剂合用的物质,其属于植物激素干扰型除草剂,在本领域中是熟知的,例如包括以下有效成分或其衍生物中的至少一种:
(1)吡啶羧酸类(Pyridine carboxylic acids):氨氯吡啶酸(毒莠定)、氯氟吡氧乙酸(氟草烟)、氯氟吡氧乙酸异辛酯、氯氨吡啶酸、二氯吡啶酸、三氯吡氧乙酸(绿草定)、氯氟吡啶酯、氟氯吡啶酯、氟氯氨草酯等;
(2)苯甲酸类(Benzoic acids):麦草畏、草灭平、草芽畏、萘草胺等;
(3)苯氧羧酸类(Phenoxycarboxylic acids):2,4-二氯苯氧乙酸(2,4-D)、2,4-二氯苯氧丁酸(2,4-D丁酸)、2,4-D异丙酸、氯甲酰草胺、二甲四氯、二甲四氯异丙酸、二甲四氯丁酸等;
(4)喹啉羧酸类(Quinoline carboxylic acids):二氯喹啉酸、氯甲喹啉酸等;
(5)其他:草除灵等。
“ACCase抑制剂类除草剂”是以乙酰辅酶A羧化酶为作用靶标的除草剂,在本领域中是熟知的,例如包括以下有效成分或其衍生物中的至少一种:
(1)芳氧苯氧丙酸类:精喹禾灵、炔草酯、氰氟草酯、禾草灵、精噁唑禾草灵、精吡氟禾草灵、高效氟吡禾草灵、噁唑酰草胺、噁草酸、喹禾糠酯等;
(2)环己烯酮类:禾草灭、烯草酮、环苯草酮、丁苯草酮、噻草酮、烯禾啶、吡喃草酮、三甲苯草酮等;
(3)苯并吡啶类:唑啉草酯等。
在本说明书的上下文中,如果使用活性化合物的通用名称的缩写形式,则在每种情况下包括所有的常规衍生物,例如酯和盐,以及异构体,特别是光学异构体,特别是一种或多种市售形式。如果通用名称表示酯或盐,则在每种情况下还包括所有其他的常规衍生物,例如其他的酯和盐、游离酸和中性化合物,以及异构体,特别是光学异构体,特别是一种或多种市售形式。给出的化合物的化学名称表示至少一种被通用名称涵盖的化合物,通常是优选的化合物。例如,2,4-D或2,4-D丁酸衍生物包含但不限于:2,4-D或2,4-D丁酸盐如钠盐、钾盐、二甲铵盐、三乙醇铵盐、异丙胺盐、胆碱等,以及2,4-D或2,4-D丁酸酯如甲酯、乙酯、丁酯、异辛酯等;二甲四氯衍生物包含但不限于:二甲四氯钠盐、钾盐、二甲铵盐、异丙胺盐等,以及二甲四氯甲酯、乙酯、异辛酯、乙硫酯等。
在本发明中,“除草剂耐受性”和“除草剂抗性”两个术语可以互换使用,均指的是对除草剂的耐受性和对除草剂的抗性,意指植物、种子、细胞或植物部分抵抗一种或多种除草剂的毒性作用的能力。植物、种子、细胞或植物部分的除草剂耐受性可通过将植物、种子、细胞或植物部分与合适的对照进行比较来测量。例如,除草剂耐受性可通过将除草剂施加至包含编码能够赋予除草剂耐受性的蛋白质的重组DNA分子的植物(测试植物)和不包含编码能够赋予除草剂耐受性的蛋白质的重组DNA分子的植物(对照植物),且然后比较两种植物的植物损伤,其中测试植物的除草剂耐受性通过与对照植物的损伤率相比减少的损伤率指示。当与对照植物、种子、细胞或植物部分相比时,除草剂耐受性植物、种子、细胞或植物部分显示对除草剂的毒性作用的反应降低。如本文所用,“除草剂耐受性性状”是与野生型植物或对照植物相比赋予植物改善的除草剂耐受性的转基因性状。
在本发明中,“野生型”意指天然存在的,其和突变是相对来说的。
术语“蛋白”、“多肽”和“肽”在本发明中可以互换使用,指的是氨基酸残基聚合物,包括其中一个或多个氨基酸残基是天然氨基酸残基的化学类似物的聚合物。本发明的蛋白和多肽可以重组产生,也可以通过化学合成。
如本领域中所熟知的,可以从蛋白质的N和/或C末端缺失一或多个氨基酸残基而仍保留其功能活性。在本发明中,“生物活性片段”是指本发明的蛋白的一部分,其保留了蛋白的生物学活性。例如,蛋白的生物学活性片段可以是在所述蛋白质的N和/或C末端缺失了一个或多个(例如1-50个、1-25个、1-10个或1-5个,例如1、2、3、4或5个)氨基酸残基的部分,但其仍然保留了全长蛋白的生物学活性。
本发明中,“严谨条件”可以指6M尿素、0.4%SDS、0.5×SSC的条件或与其同等的杂交条件,也可以指严谨性更高的条件,例如6M尿素、0.4%SDS、0.1×SSC或与其同等的杂交条件。在各种条件中,温度可约为40℃以上,如需要严谨性更高的条件时,温度例如可约为50℃,进一步可约为65℃。
本领域技术人员十分清楚,由于遗传密码的简并性,有多种不同的核酸序列可以编码本文公开的氨基酸序列。产生编码相同蛋白质的其他核酸序列在本领域普通技术人员的能力范围内,因此本发明涵盖因遗传密码子的简并性而编码相同氨基酸序列的核酸序列。例如,为了在目标宿主生物例如植物中实现异源基因的高表达,可以对所述基因采用宿主生物偏好的密码子进行优化,以使其更好地表达。
在本发明中,“宿主生物”应理解为可以引入突变型蛋白编码核酸的任何单细胞或多细胞生物,包括例如细菌如大肠杆菌,真菌如酵母(例如酿酒酵母)、霉菌(例如曲霉菌),植物细胞和植物等。
术语“多核苷酸”和“核酸”可以互换使用,包括DNA、RNA或者其杂交体,可以是双链或单链的。术语“分离”,当提及核酸时,是指这样的核酸,所述核酸与其天然存在于其中的基因组的实质部分分开和/或与天然伴随该核酸的其他细胞组分基本上分离。例如,已经通过合成(如通过连续碱基缩合)而产生的任意核酸被认为是分离的。同样地,重组表达的核酸、克隆的核酸、通过引物延伸反应(例如PCR)产生的核酸或另外切离基因组的核酸也被认为是分离的。
术语“对照”意指为比较目的而设计的实验对照。例如,转基因植物分析中的对照植物是与实验植物(即其待测试的植物)相同类型但不含实验植物的转基因***片段、重组DNA分子或DNA构建体的植物。
术语“重组”是指为遗传工程化的结果并且因此通常不会在自然界中发现并且通过人为干预产生的非天然DNA、多肽或蛋白质。“重组DNA分子”是包含不天然存在的并且因此是人为干预的结果的DNA序列的DNA分子,例如编码工程化蛋白质的DNA分子。另一个实例是由至少两个彼此异源的DNA分子(如编码蛋白质的DNA分子和可操作地连接的异源启动子)的组合组成的DNA分子。“重组多肽”或“重组蛋白质”是包含不天然存在的氨基酸序列并且因此是人为干预的结果的多肽或蛋白质,例如工程化蛋白质。
术语“转基因”是指作为人为干预(如通过植物转化方法)的结果人工并入生物体基因组中的DNA分子。如本文所用,术语“转基因的”意指包含转基因,例如“转基因植物”是指在其基因组中包含转基因的植物,“转基因性状”是指由并入植物基因组中的转基因的存在传递或赋予的特征或表型。作为这种基因组改变的结果,所述转基因植物是与相关的野生型植物明显不同的植物,并且转基因性状是未在野生型植物中天然发现的性状。本发明的转基因植物包含通过本发明提供的重组DNA分子和工程化蛋白质。
术语“异源”是指源自不同来源且因此在自然界中通常不相关的两种或更多种物质之间的关系。例如,编码蛋白质的重组DNA分子相对于可操作地连接的启动子是异源的,如果这种组合在自然界中通常不存在。此外,当特定重组DNA分子不天然存在于所述特定细胞或生物体中时,其可相对于其所***的细胞或生物体是异源的。
术语“编码蛋白质的DNA分子”或“编码多肽的DNA分子”是指包含编码蛋白质或多肽的核苷酸序列的DNA分子。“编码蛋白质的序列”或“编码多肽的序列”意指编码蛋白质或多肽的DNA序列。“序列”意指核苷酸或氨基酸的顺序排列。编码蛋白质的序列或编码多肽的序列的边界通常由5'-末端的翻译起始密码子和3'-末端的翻译终止密码子决定。编码蛋白质的分子或编码多肽的分子可包含编码蛋白质或多肽序列的DNA序列。如本文所用,“转基因表达”、“表达转基因”、“蛋白质表达”、“多肽表达”、“表达蛋白质”和“表达多肽”意指通过将DNA分子转录成信使RNA(mRNA)并且将mRNA翻译成多肽链(其可最终折叠成蛋白质)的过程产生蛋白质或多肽。编码蛋白质的DNA分子或编码多肽的DNA分子可以可操作地连接至DNA构建体中的异源启动子,以用于在用重组DNA分子转化的细胞中表达蛋白质或多肽。如本文所用,“可操作地连接”是指以使得一个DNA分子可影响另一个DNA分子的功能的方式连接的两个DNA分子。可操作连接的DNA分子可以是单个连续分子的一部分,并且可以是或可以不是相邻的。例如,启动子与DNA构建体中的编码蛋白质的DNA分子或编码多肽的DNA分子可操作地连接,其中两个DNA分子被排列成使得所述启动子可影响转基因的表达。
术语“DNA构建体”是包含两个或更多个异源DNA序列的重组DNA分子。DNA构建体适用于转基因表达,并且可包含在载体和质粒中。DNA构建体可出于转化(即将异源DNA引入宿主细胞中)的目的用于载体中以便产生转基因植物和细胞,并且因此也可包含在转基因植物、种子、细胞或植物部分的质粒DNA或基因组DNA中。如本文所用,“载体”意指可用于植物转化目的的任何重组DNA分子。如序列表中所示的重组DNA分子可例如作为构建体的一部分***载体中,所述构建体具有可操作地连接至启动子的重组DNA分子,所述启动子在植物中起作用以驱动由所述重组DNA分子编码的工程化蛋白质的表达。用于构建DNA构建体和载体的方法是本领域中熟知的。DNA构建体或包含DNA构建体的载体的组分通常包括但不限于以下中的一种或多种:用于表达可操作地连接的DNA的合适启动子、可操作地连接的编码蛋白质非人DNA分子和3'非翻译区(3’-UTR)。适用于实践本发明的启动子包括在植物中起作用以表达可操作地连接的多核苷酸的启动子。此类启动子是多种多样的且是本领域中熟知的,并且包括诱导型的、病毒的、合成的、组成型、时间调控型的、空间调控型的和/或时空调控型的。另外的任选组分包括但不限于以下元件中的一个或多个:5'-UTR、增强子、前导序列、顺式作用元件、内含子、叶绿体转运肽(CTP)和一个或多个选择性标记转基因。
本发明的DNA构建体可包含可操作地连接至本发明提供的编码蛋白质的DNA分子的CTP分子。适用于实践本发明的CTP包括用于促进工程化蛋白分子在细胞内定位的那些。通过促进细胞内的蛋白质定位,CTP可增加工程化蛋白质的积累,保护其免受蛋白水解降解,增强除草剂耐受性水平,并且由此降低除草剂施加后的损伤水平。用于本发明的CTP分子是本领域中已知的,包括但不限于拟南芥EPSPS CTP(Klee等人,1987)、矮牵牛EPSPS CTP(della-Cioppa等人,1986)、玉米cab-m7信号序列(Becker等人,1992;PCT WO 97/41228)和豌豆谷胱甘肽还原酶信号序列(Creissen等人,1991;PCT WO 97/41228)。
本发明的重组DNA分子可通过本领域中已知的方法完全或部分地合成和修饰,特别是在期望提供适用于DNA操作的序列(如限制性酶识别位点或重组-基因克隆位点)、植物优选序列(如植物密码子使用或Kozak共有序列)或适用于DNA构建体设计的序列(如间隔区或接头序列)的情况下。如本文所用,术语“百分比序列同一性”或“%序列同一性”是指在最佳比对两个序列时(在比较窗内具有总计少于参考序列的20%的适当核苷酸或氨基酸***、缺失或空位),与测试(“主题”)序列(或其互补链)相比,参考(“查询”)序列(或其互补链)的线性多核苷酸或多肽序列中的相同核苷酸或氨基酸的百分比。用于比对比较窗口的最佳序列比对是本领域的技术人员所熟知的并且可由以下工具实施:如Smith和Waterman的局部同源性算法、Needleman和Wunsch的同源性比对算法、Pearson和Lipman的相似性搜索方法,并且由这些算法的计算机化实现方式来实施,如使用默认参数的作为Wisconsin(Accelrys Inc.,San Diego,CA)、MEGAlign(DNAStar,Inc.,1228S.Park St.,Madison,Wis.53715)和MUSCLE(3.6版)(RCEdgar,NucleicAcids Research(2004)32(5):1792-1797)的一部分可获得的GAP、BESTFIT、FASTA和TFASTA。测试序列和参考序列的比对片段的“同一性分数”是由两个比对序列共享的相同组分的数目除以参考序列片段中组分的总数,即整个参考序列或参考序列的较小限定部分。序列同一性百分比表示为同一性分数乘以100。一个或多个序列的比较可以是针对全长序列或其一部分,或针对更长的序列。
在本发明中,“植物”应理解为能够进行光合作用的任何分化的多细胞生物,特别是单子叶或双子叶植物,例如:(1)粮食作物:稻属(Oryza spp.),例如稻(Oryza sativa)、阔叶稻(Oryza latifolia)、水稻(Oryza sativa)、光稃稻(Oryza glaberrima);小麦属(Triticum spp.),例如普通小麦(Triticum aestivum)、硬粒小麦(T.Turgidumssp.durum);大麦属(Hordeum spp.),例如大麦(Hordeum vulgare)、亚利桑那大麦(Hordeum arizonicum);黑麦(Secale cereale);燕麦属(Avena spp.),例如燕麦(Avena sativa)、野燕麦(Avena fatua)、比赞燕麦(Avena byzantina)、Avena fatuavar.sativa、杂种燕麦(Avena hybrida);稗属(Echinochloa spp.),例如,珍珠粟(Pennisetum glaucum)、高粱(两色高粱(Sorghum bicolor)、高粱(Sorghum vulgare))、黑小麦、玉蜀黍或玉米、粟、稻(rice)、谷子、糜子、两色蜀黍(Sorghum bicolor)、黍子、荞麦属(Fagopyrum spp.)、黍(Panicum miliaceum)、小米(Setaria italica)、沼生菰(Zizaniapalustris)、埃塞俄比亚画眉草(Eragrostis tef)、稷(Panicum miliaceum)、龙爪稷(Eleusine coracana);(2)豆类作物:大豆属(Glycine spp.),例如大豆(Glycine max)、黄豆(Soja hispida)、Soja max)、野豌豆属(Vicia spp.)、豇豆属(Vigna spp.)、豌豆属(Pisum spp.)、芸豆(field bean)、羽扇豆属(Lupinus spp.)、蚕豆属(Vicia)、酸豆(Tamarindus indica)、兵豆(Lens culinaris)、山黧豆属(Lathyrus spp.)、扁豆属(Lablab)、蚕豆、绿豆、红豆、鹰嘴豆;(3)油料作物:花生(Arachis hypogaea)、落花生属(Arachis spp)、胡麻属(Sesamum spp.)、向日葵属(Helianthus spp.)(例如向日葵(Helianthus annuus))、油棕属(Elaeis)(例如油棕(Eiaeis guineensis)、美洲油棕(Elaeis oleifera))、大豆(soybean)、油菜(Brassicanapus)、芸苔、芝麻、芥菜(Brassicajuncea)、油菜籽油菜(oilseedrape)、油茶、油棕、油橄榄、蓖麻、欧洲油菜(Brassica napus L.)、卡诺拉油菜(canola);(4)纤维作物:剑麻(Agave sisalana)、棉属(棉花、海岛棉(Gossypium barbadense)、陆地棉(Gossypium hirsutum))、红麻、剑麻、蕉麻、亚麻(Linum usitatissimum)、黄麻、苎麻、***(Cannabis sativa)、火麻;(5)水果类作物:枣属(Ziziphus spp.)、香瓜属(Cucumis spp.)、鸡蛋果(Passiflora edulis)、葡萄属(Vitis spp.)、越桔属(Vaccinium spp.)、西洋梨(Pyrus communis)、李属(Prunus spp.)、番石榴属(Psidium spp.)、石榴(Punica granatum)、苹果属(Malus spp.)、西瓜(Citrullus lanatus)、柑桔属(Citrus spp.)、无花果(Ficus carica)、金桔属(Fortunella spp.)、草莓属(Fragaria spp.)、山楂属(Crataegus spp.)、柿树属(Diospyros spp.)、红仔果(Eugenia unifora)、枇杷(Eriobotryajaponica)、龙眼(Dimocarpus longan)、番木瓜(Carica papaya)、椰子属(Cocos spp.)、阳桃(Averrhoacarambola)、狲猴桃属(Actinidia spp.)、扁桃(Prunus amygdalus)、芭蕉属(Musa spp.)(香蕉)、鳄梨属(Persea spp.)(鳄梨(Persea americana))、番石榴(Psidium guajava)、曼密苹果(Mammea americana)、芒果(Mangifera indica)、橄榄(油橄榄(Oleaeuropaea))、番木瓜(Caricapapaya)、椰子(Cocos nucifera)、凹缘金虎尾(Malpighia emarginata)、人心果(Manilkara zapota)、菠萝(Ananas comosus)、番荔枝属(Annona spp.)、柑桔树(柑桔属物种(Citrus spp.))、波罗蜜属(Artocarpus spp.)、荔枝(Litchi chinensis)、茶藨子属(Ribes spp.)、悬钩子属(Rubus spp.)、梨、桃、杏、梅、杨梅、柠檬、金橘、榴莲、橙、草莓(straw berry)、蓝莓、哈密瓜、甜瓜、椰枣、胡桃树、樱桃树;(6)根茎类作物:木薯属(Manihot spp.)、甘薯(Ipomoea batatas)、芋(Colocasia esculenta)、榨菜、洋葱、荸荠、油莎草、山药;(7)蔬菜类作物:菠菜属(Spinacia spp.)、菜豆属(Phaseolus spp.)、莴苣(Lactuca sativa)、苦瓜属(Momordica spp)、欧芹(Petroselinum crispum)、辣椒属(Capsicum spp.)、茄属(Solanum spp.)(例如马铃薯(Solanum tuberosum)、红茄(Solanumintegrifolium)或蕃茄(Solanum lycopersicum))、蕃茄属(Lycopersicon spp.)(例如西红柿(Lycopersicon esculentum)、蕃茄(Lycopersicon lycopersicum)、梨形蕃茄(Lycopersicon pyriforme))、硬皮豆属(Macrotyloma spp.)、无头甘蓝(kale)、棱角丝瓜(Luffa acutangula)、小扁豆(lentil)、秋葵(okra)、洋葱(onion)、马铃薯(potato)、洋蓟(artichoke)、芦笋(asparagus)、西兰花(broccoli)、球芽甘蓝(Brussels sprouts)、卷心菜(cabbage)、胡萝卜(carrot)、花椰菜(cauliflower)、芹菜(celery)、羽衣甘蓝(collardgreens)、西葫芦(squash)、冬瓜(Benincasa hispida)、石刁柏(Asparagus officinalis)、旱芹(Apium graveolens)、苋属(Amaranthus spp.)、葱属(Allium spp.)、秋葵属(Abelmoschus spp.)、苦苣(Cichorium endivia)、南瓜属(Cucurbita spp.)、芫荽(Coriandrum sativum)、埃塞俄比亚芥(B.carinata)、萝卜(Rapbanus sativus)、芸苔属(Brassica)物种(例如例如欧洲油菜(Brassica napus)、芜菁亚种(Brassica rapa ssp.)、卡诺拉油菜(canola)、油籽油菜(oilseed rape)、芜菁油菜(turnip rape)、芥菜、甘蓝、黑芥、油菜籽油菜)、孢子甘蓝、茄科植物(茄子)、甜椒、黄瓜、丝瓜、白菜、油菜、甘蓝、葫芦、韭菜、莲、藕、生菜;(8)花卉作物:小金莲花(Tropaeolum minus)、金莲花(Tropaeolummajus)、美人蕉(Canna indica)、仙人掌属(Opuntia spp.)、万寿菊属(Tagetes spp.)、兰花、文殊兰、君子兰、朱顶红、玫瑰、月季、茉莉花、郁金香、樱花、牵牛花、金盏花、荷花、雏菊、康乃馨、矮牵牛花、郁金香、百合、梅花、水仙、迎春、报春、瑞香、山茶、白玉兰、紫玉兰、琼花、君子兰、海棠、牡丹、芍药、丁香、杜鹃、西洋杜鹃、含笑、紫荆、棣棠、锦带花、连翘、云南黄馨、金雀花、仙客来、蝴蝶兰、石斛、风信子、鸢尾、马蹄莲、金盏菊、百枝莲、四季海棠、吊钟海棠、竹节海棠、天竺葵、绿萝;(9)药用作物:红花(Carthamus tinctorius)、薄荷属(Menthaspp.)、波叶大黄(Rheum rhabarbarum)、番红花(Crocus sativus)、枸杞、玉竹、黄精、知母、麦冬、贝母、郁金、砂仁、何首乌、大黄、甘草、黄芪、人参、三七、五加、当归、川芎、北柴胡、曼佗罗、洋金花、薄荷、益母草、藿香、黄芩、夏枯草、除虫菊、银杏、金鸡纳树、天然橡胶树、苜蓿、胡椒、板蓝根、白术;(10)原料作物:橡胶、蓖麻(Ricinus communis)、油桐、桑、忽布、桦、桤木、漆树;(11)牧草作物:冰草属(Agropyron spp.)、车轴草属(Trifolium spp.)、芒(Miscanthus sinensis)、狼尾草属(Pennisetum sp.)、虉草(Phalaris arundinacea)、柳枝稷(Panicum virgatum)、草原草(prairiegrasses)、印度草(Indiangrass)、大须芒草(Big bluestem grass)、梯牧草(Phleum pratense)、草皮草(turf)、莎草科(高山嵩草、脚苔草(Carex pediformis)、低苔草)、苜蓿、梯牧草、紫花苜蓿、草木犀、紫云英、柽麻、田菁、红萍、水葫芦、紫穗槐、羽扇豆、三叶草、沙打旺、水浮莲、水花生、黑麦草;(12)糖料作物:甘蔗(甘蔗属物种(Saccharum spp.))、甜菜(Beta vulgaris);(13)饮料作物:大叶茶(Camellia sinensis)、茶(Camellia Sinensis)、茶树(tea)、咖啡(咖啡属物种(Coffeaspp.))、可可树(Theobroma cacao)、蛇麻花(啤酒花);(14)草坪植物:固沙草(Ammophilaarenaria)、早熟禾属(Poa spp.)(草地早熟禾(Poa pratensis)(蓝草))、剪股颖属物种(Agrostis spp.)(剪股颖、匍匐剪股颖(Agrostis palustris))、黑麦草属物种(Loliumspp.)(黑麦草)、羊茅属物种(Festuca spp.)(羊茅)、结缕草属物种(Zoysia spp.)(结缕草(Zoysiajaponica))、狗牙根属物种(Cynodon spp.)(百慕大草、狗牙根)、侧钝叶草(Stenotaphrum secunda tum)(圣奥古斯丁草)、雀稗属物种(Paspalum spp.)(巴哈草)、假俭草(Eremochloa ophiuroides)(百足草)、地毯草属物种(Axonopus spp.)(地毯草)、指形垂穗草(Bouteloua dactyloides)(野牛草)、垂穗草属变种物种(Bouteloua var.spp.)(格兰马草)、马唐(Digitariasanguinalis)、香附子(Cyperusrotundus)、短叶水蜈蚣(Kyllingabrevifolia)、阿穆尔莎草(Cyperusamuricus)、加拿大飞蓬(Erigeroncanadensis)、天胡荽(Hydrocotylesibthorpioides)、鸡眼草(Kummerowiastriata)、地锦(Euphorbiahumifusa)、耕地堇菜(Violaarvensis)、白颖苔草、异穗苔草、草皮草(turf);(15)树木作物:松属(Pinus spp.)、柳属(Salix sp.)、槭树属(Acer spp.)、木槿属(Hibiscus spp.)、桉属(Eucalyptus sp.)、银杏(Ginkgo biloba)、箣竹属(Bambusa sp.)、杨属(Populus spp.)、牧豆树属(Prosopis spp.)、栎属(Quercusspp.)、刺葵属(Phoenix spp.)、山毛榉属(Fagus spp.)、吉贝(Ceiba pentandra)、樟属(Cinnamomum spp.)、黄麻属(Corchorus sp.)、南方芦苇(Phragmites australis)、酸浆属(Physalis spp.)、山蚂蝗属(Desmodium spp.)、杨、常春藤、白杨、珊瑚树、银杏、栎类、臭椿、木荷、冬青、悬铃木、女贞、大叶黄扬、落叶松、黑荆树、马尾松、思茅松,云南松、南亚松、油松、红松、黑胡桃、柠檬、悬铃木、蒲桃、珙桐、木棉、爪哇木棉、洋紫荆、羊蹄甲、雨树、合欢、龙牙花、刺桐、广玉兰、苏铁、紫薇、针叶树、乔木、灌木;(16)坚果作物:巴西栗(Bertholletia excelsea)、栗属(Castanea spp.)、榛属(Corylus spp.)、山核桃属(Caryaspp.)、核桃属(Juglans spp.)、阿月浑子(Pistacia vera)、腰果(Anacardiumoccidentale)、澳洲坚果(全缘叶澳洲坚果(Macadamia integrifolia))、碧根果、夏威夷果、开心果、巴旦木以及产生坚果的植物;(17)其他:拟南芥、臂形草、蒺藜草、大狗尾草、牛筋草、Cadaba farinosa、藻类(algae)、Carex elata、观赏植物、大果假虎刺(Carissamacrocarpa)、菜蓟属(Cynara spp.)、野胡萝卜(Daucus carota)、薯蓣属(Dioscoreaspp.)、蔗茅属(Erianthus sp.)、苇状羊茅(Festuca arundinacea)、萱草(Hemerocallisfulva)、百脉根属(Lotus spp.)、Luzula sylvatica、紫苜蓿(Medicago sativa)、草木樨属(Melilotus spp.)、黑桑(Morus nigra)、烟草属(Nicotiana spp.)、木犀榄属(Oleaspp.)、鸟足豆属(Ornithopus spp.)、欧防风(Pastinaca sativa)、接骨木属(Sambucusspp.)、白芥属(Sinapis sp.)、蒲桃属(Syzygium spp.)、鸭茅状摩擦禾(Tripsacumdactyloides)、Triticosecale rimpaui、香堇(Viola odorata)等。
在本发明中,术语“植物组织”或“植物部分”包括植物细胞、原生质体、植物组织培养物、植物愈伤组织、植物块以及植物胚、花粉、胚珠、叶、茎、花、枝、幼苗、果实、核、穗、根、根尖、花药等。
在本发明中,“植物细胞”应理解为来自或发现于植物的任何细胞,其能够形成例如:未分化组织如愈伤组织,分化组织如胚胎,植物的组成部分,植物或种子。
本发明的转基因植物、子代、种子、植物细胞和植物部分还可含有一种或多种另外的转基因性状。可通过使含有包含本发明提供的重组DNA分子的转基因的植物与含有另外的转基因性状的另一种植物杂交来引入另外的转基因性状。如本文所用,“杂交”意指培育两种单独的植物以产生子代植物。因此,两种转基因植物可杂交以产生含有转基因性状的子代。如本文所用,“子代”意指亲本植物的任何传代的后代,并且转基因子代包含由本发明提供并且从至少一种亲本植物遗传的DNA构建体。或者,可通过用包含本发明提供的重组DNA分子的DNA构建体共转化所述另外的转基因性状的DNA构建体(例如,其中所有的DNA构建体呈现为用于植物转化的同一载体的部分)或通过将另外的性状***包含本发明提供的DNA构建体的转基因植物中或反之亦然(例如,通过使用关于转基因植物或植物细胞的植物转化的任何方法)来引入另外的转基因性状。此类另外的转基因性状包括但不限于增加的昆虫抗性、增加的水利用效率、增加的产量性能、增加的抗旱性、增加的种子质量、改进的营养品质、杂交种种子生产和除草剂耐受性,其中性状是相对于野生型植物或对照植物测量的。此类另外的转基因性状是本领域的技术人员已知的;例如,美国农业部(USDA)动物和植物健康检查局(APHIS)提供了此类性状的列表,并且可在它们的网站www.aphis.usda.gov上找到。
含有本发明提供的转基因性状的转基因植物和子代可与本领域中通常已知的任何培育方法一起使用。在包含两种或更多种转基因性状的植物系中,转基因性状可在包含三种或更多种转基因性状的植物系中独立地分离、连接或两者的组合。还考虑与亲本植物的回交和与非转基因植物的异交,以及无性繁殖。通常用于不同性状和作物的培育方法的描述是本领域的技术人员熟知的。为了证实转基因在特定植物或种子中的存在,可进行多种测定。此类测定包括例如分子生物学测定,如DNA印迹和RNA印迹、PCR和DNA测序;生物化学测定,如例如通过免疫学方法(ELISA和蛋白质印迹)或通过酶功能检测蛋白质产物的存在;植物部分测定,如叶或根测定;以及还有通过分析整株植物的表型。
作为回交转化过程的结果实现转基因性状向植物基因型的基因渗入。其中已经基因渗入转基因性状的植物基因型可称为回交转化的基因型、系、近交植物或杂交种。类似地,缺乏所需转基因性状的植物基因型可称为未转化的基因型、系、近交植物或杂交种。
如本文所用,术语“包含”是指“包括但不限于”。
除非具体说明或暗示,如本文中所用,术语“一”、“一个/一种”和“所述”表示“至少一个”。本文提到或引用的所有专利、专利申请和出版物整体引入本文作为参考,其引用程度如同单独地个别引用一样。
具体实施方式
下面结合具体实施方式对本发明进行进一步的详细描述,给出的实施例仅为了阐明本发明,而不是为了限制本发明的范围。下述实施例中的实验方法,如无特殊说明,均为常规方法。下述实施例中所用的材料、试剂、仪器等,如无特殊说明,均可从商业途径得到。
实施例1:激素类除草剂降解侯选基因的来源、序列和合成
1.侯选基因来源
基因家族tfdA(α-酮戊二酸依赖的牛磺酸双加氧酶)来源于真产碱细菌(Ralstonia eutropha),通过含有的TauD(Taurine catabolism dioxygenase,牛磺酸分解代谢双加氧酶)结构域来降解2,4-D。迄今,基因家族数据库(Pfan Database)已有14425个基因含有TauD结构域,它们组成223个不同的组织结构,这是一个巨大的侯选基因资源,从Pfan Database和其它发表的文献中选择76个侯选基因进行测试。
2.侯选基因序列
这76个激素类除草剂降解侯选酶的氨基酸序列分别是SEQ ID NO.1、SEQ IDNO.3、SEQ ID NO.5、SEQ ID NO.7、SEQ ID NO.9、SEQ ID NO.11、SEQ ID NO.13、SEQ IDNO.15、SEQ ID NO.17、SEQ ID NO.19、SEQ ID NO.21、SEQ ID NO.23、SEQ ID NO.25、SEQ IDNO.27、SEQ ID NO.29、SEQ ID NO.31、SEQ ID NO.33、SEQ ID NO.35、SEQ ID NO.37、SEQ IDNO.39、SEQ ID NO.41、SEQ ID NO.43、SEQ ID NO.45、SEQ ID NO.47、SEQ ID NO.49、SEQ IDNO.51、SEQ ID NO.53、SEQ ID NO.55、SEQ ID NO.57、SEQ ID NO.59、SEQ ID NO.61、SEQ IDNO.63、SEQ ID NO.65、SEQ ID NO.67、SEQ ID NO.69、SEQ ID NO.71、SEQ ID NO.73、SEQ IDNO.75、SEQ ID NO.77、SEQ ID NO.79、SEQ ID NO.81、SEQ ID NO.83、SEQ ID NO.85、SEQ IDNO.87、SEQ ID NO.89、SEQ ID NO.91、SEQ ID NO.93、SEQ ID NO.95、SEQ ID NO.97、SEQ IDNO.99、SEQ ID NO.101、SEQ ID NO.103、SEQ ID NO.105、SEQ ID NO.107、SEQ ID NO.109、SEQ ID NO.111、SEQ ID NO.113、SEQ ID NO.115、SEQ ID NO.117、SEQ ID NO.119、SEQ IDNO.121、SEQ ID NO.123、SEQ ID NO.125、SEQ ID NO.127、SEQ ID NO.129、SEQ ID NO.131、SEQ ID NO.133、SEQ ID NO.135、SEQ ID NO.137、SEQ ID NO.139、SEQ ID NO.141、SEQ IDNO.143、SEQ ID NO.145、SEQ ID NO.147、SEQ ID NO.149、SEQ ID NO.151。
其编码基因序列经植物密码子编好性优化后分别是SEQ ID NO.2、SEQ ID NO.4、SEQ ID NO.6、SEQ ID NO.8、SEQ ID NO.10、SEQ ID NO.12、SEQ ID NO.14、SEQ ID NO.16、SEQ ID NO.18、SEQ ID NO.20、SEQ ID NO.22、SEQ ID NO.24、SEQ ID NO.26、SEQ IDNO.28、SEQ ID NO.30、SEQ ID NO.32、SEQ ID NO.34、SEQ ID NO.36、SEQ ID NO.38、SEQ IDNO.40、SEQ ID NO.42、SEQ ID NO.44、SEQ ID NO.46、SEQ ID NO.48、SEQ ID NO.50、SEQ IDNO.52、SEQ ID NO.54、SEQ ID NO.56、SEQ ID NO.58、SEQ ID NO.60、SEQ ID NO.62、SEQ IDNO.64、SEQ ID NO.66、SEQ ID NO.68、SEQ ID NO.70、SEQ ID NO.72、SEQ ID NO.74、SEQ IDNO.76、SEQ ID NO.78、SEQ ID NO.80、SEQ ID NO.82、SEQ ID NO.84、SEQ ID NO.86、SEQ IDNO.88、SEQ ID NO.90、SEQ ID NO.92、SEQ ID NO.94、SEQ ID NO.96、SEQ ID NO.98、SEQ IDNO.100、SEQ ID NO.102、SEQ ID NO.104、SEQ ID NO.106、SEQ ID NO.108、SEQ ID NO.110、SEQ ID NO.112、SEQ ID NO.114、SEQ ID NO.116、SEQ ID NO.118、SEQ ID NO.120、SEQ IDNO.122、SEQ ID NO.124、SEQ ID NO.126、SEQ ID NO.128、SEQ ID NO.130、SEQ ID NO.132、SEQ ID NO.134、SEQ ID NO.136、SEQ ID NO.138、SEQ ID NO.140、SEQ ID NO.142、SEQ IDNO.144、SEQ ID NO.146、SEQ ID NO.148、SEQ ID NO.150、SEQ ID NO.152。
3.侯选基因合成
计划用pET15b作为骨架载体在大肠杆菌里表达后筛选。在pET15b NdeI/BamHI位点处酶切开环后用Infusion技术克隆。因此,在上述侯选基因序列的5’端和3’端分别加上5’-GCCGCGCGGCAGCCAT-3’和5’-TGTTAGCAGCCGGATCC-3’接头序列后在南京金斯瑞人工合成。
实施例2:AAD12基因大肠杆菌过表达、纯化和显色反应体系构建
AAD12是美国陶氏益农(Dow AgroSciences LLC)发现的激素类除草剂降解基因,并转化到玉米、大豆等作物配合激素类除草剂进行商业化应用(T.R.Wright et al.,Robust crop resistance to broadleafand grass herbicides providedbyaryloxyalkanoate dioxygenase transgenes.Proc.Natl.Acad.Sci.U.S.A.107,20240–20245(2010))。AAD12能够降解2,4-D和S-手性的吡啶羧酸类除草剂,产生2,4-二氯苯酚和类似物质,这些物质显现红色。以AAD12作为模式基因建立显色反应体系,为大规模筛选激素类除草剂基因创立筛选工具。
1.载体构建
AAD12全长有293个氨基酸,分子量31.7kDa,等电点6.45,其序列来自陶氏益农发表的专利(US10167483-0004)。经植物密码子优化并分别在其5’端和3’端分别加上5’-GCCGCGCGGCAGCCAT-3’和5’-TGTTAGCAGCCGGATCC-3’接头序列后在南京金斯瑞人工合成。用Infusion克隆技术将合成的DNA在NdeI/BamHI位点克隆到pET15b骨架中形成大肠杆菌表达载体pET15b-AAD12。
2.蛋白表达和纯化
根据文献报道(Fukomori and Flausinger,Purification andcharacterization of2,4-dichlorophenoxyacetate/alpha-ketoglutarate dioxygenaseJournal ofBiological Chemistry(1993)268(32):24311-24317),将构建好的表达载体pET15b-AAD12转入大肠杆菌BL21(DE3)中,经IPTG诱导表达,收菌裂解后利用Ni-NTA柱进行纯化,SDS-PAGE检测蛋白纯化效果。
3.酶活性测试
由于AAD-12催化底物反应后,生成酚类产物,可以利用4-氨基安替比林和铁***与产物在碱性环境下显色,生成红色产物,且在510nm有吸收,直接用酶标仪读取相关参数。
4.用2,4-D等激素类除草剂为底物颜色筛选体系构建
在用2,4-D为底物测试了AAD12蛋白活性基础上,进一步测试菌内转化2,4-D的方法,构建高通量的颜色反应筛选体系,用于筛选对2,4-D等激素类除草剂有降解活性的基因。
待测质粒的转化,将AAD12表达质粒和空白对照质粒pET15b,分别转化到BL21(DE3)蛋白表达大肠杆菌菌株中。挑取单克隆,接种于有3ml LB培养基的10ml离心管后在37℃,200rpm培养6-8小时,向培养液中加入0.1mM IPTG和2mMα-酮戊二酸,再加入0.5mM 2,4-D。转到28℃继续培养过夜。第二天早上,吸取各菌液200ul离心沉淀菌体,取上清180ul到96孔酶标板中,加入pH10硼酸缓冲液20ul,然后加入2ul 8%铁***混匀后,加入2ul 2%4-氨基安替比林混匀后室温放置5min,观察颜色和检测A510。
酶活测定结果显示,2,4-D在有AAD12表达的实验组与空白载体对照组比较,实验组反应液变红。这表明实验组反应液有酚类产物生成,大肠杆菌表达的AAD12具有催化活性。利用酶标仪对A510进行测定,同样可以检测到实验组较对照组吸收值明显升高。
除对2,4-D测试外,还对另一个苯氧乙酸除草剂二甲四氯,以及2,4-D异辛酯和2,4-D丁酸进行了测试,结果表明,表达的AAD-12酶对这些底物都有降解活性,见表1。利用HPLC对各实验组与空白对照组中除草剂底物含量进行检测,发现加入酶催化反应后,各底物含量明显降低,证明各底物被降解。
表1.光度法测定AAD-12降解苯氧乙酸类除草剂活性
直接向AAD12表达菌培养液里加2,4-D,菌液也显现红色,如图1所示。这说明可以通过在培养基中添加底物的培养方法,实现菌内降解显色,以此判断候选基因产物是否降解2,4-D。这套菌内降解显色筛选体系可以提高检测通量,对多个AAD基因表达菌进行快速筛选。
实施例3:利用建成的菌内降解显色反应体系对AAD类基因进行筛选
利用与AAD12菌体内转化2,4-D为酚的颜色反应筛选方法,通过对76个AAD基因进行筛选,在这些表达AAG基因的重组大肠杆菌菌液里加2,4-D继续培养显色,检测到44个重组大肠杆菌菌液显现红色,说明这44个基因编码的AAD类似酶对2,4-D具有催化活性(表2)。
进一步测试了其中30个能够降解2,4-D的AAD基因表达菌(见图2-1,编号D42、D43、D44、D45、D46、D47、D48、D49、D50、D30、D72、D73、D74、D75、D76和D58;见图2-2,编号D21、D51、D52、D53、D54、D55、D56、D57、D62,D=QYD)菌内降解其它激素类除草剂化合物(二甲四氯、2,4-D丁酸、麦草畏、氯氟吡氧乙酸、绿草定)和属于ACCase抑制剂除草剂的精喹禾灵。另外,D26、D32、D33、D34和D35也显现红色反应。结果显示,这些酶除降解2,4-D外,还降解二甲四氯;其中D42、D43、D46、D47、D48、D30、D72、D73、D74、D76和D58(D=QYD)对绿草定显示微弱降解;对其余的都不降解。
表2.76个AAD类似基因激素类除草剂抗性鉴定总结
/>
/>
实施例4:转AAD基因拟南芥创制和抗性测试
1.转基因拟南芥过表达载体构建
如图3所示,以pQY259为骨架载体,NcoI/BstEII为克隆位点,将这些在大肠杆菌过表达后表现菌内降解2,4-D的AAD基因,克隆到骨架载体pQY259中产生一系列拟南芥过表达载体。在拟南芥中的表达由其AtUbi10拟南芥泛素启动子和MAS(mannopine synthase)终止子调控。筛选标记是潮霉素抗性基因(HygR=hpt)。
2.拟南芥的遗传转化
农杆菌侵染液的制备:挑取活化的农杆菌(GV3101)单克隆接种于30ml YEP液体培养基【酵母粉10g/L,胰蛋白胨10g/L,NaCl 5g/L,25mg/L利福平(Rifampicin)和50mg/L卡那霉素(Karamycin)】中,28℃200rmp/min振荡培养过夜至OD600值为1.0-1.5左右后,用6000rmp/min离心10min收集菌体,弃上清。用拟南芥侵染液(蔗糖50g/L,Silwet L-77
300μL/L,无须调节pH)重新悬浮菌体至OD600=0.8左右备用。
拟南芥蘸花遗传转化:拟南芥植株转化之前注意植株是否生长良好,花序盛,无胁迫反应,第一次转化在株高20cm左右进行。若土壤干燥可适当浇一些水。转化前一天用剪刀将已长出的角果剪去。将待转化植株花序浸泡于上述农杆菌菌液中30秒到1分钟,期间轻轻搅动,浸润后的植株上应该有一层液膜。将转化完成后的植株放于黑暗环境中暗培养24小时,然后取出放于正常光照环境下生长。一周以后即可按同样的方法进行第二次转化。
待种子成熟后即可进行收获,收获后置于30℃烘箱一周左右烘干种子。
3.T1代转基因植株与抗药性植株的筛选
种子经过消毒液处理5min,用无菌水洗5遍后一部分铺于MS转基因筛选培养基(含30μg/mL潮霉素、100μg/mL羧苄青霉素)筛选转基因苗,同时另一部分铺于MS加药培养基【含250μg/mL头孢噻肟钠(Cefotaxime sodium)、0.3μM 2,4-D】筛选除草剂抗性幼苗。将铺好种子的培养皿放于4℃春化处理3天,取出后放于光照培养箱培养(温度22℃,16小时光照,8小时黑暗,光照强度100—150μmol/m2/s,湿度75%)。一周之后可观察到野生型对照幼苗的根均未伸长,生长受到抑制,而转基因幼苗根生长正常,表现出耐药性。拍照并将转基因阳性幼苗与除草剂抗性幼苗移栽至装有土壤的小花盆中培养。
结果(图4-1)显示,野生型拟南芥Col-0(Columbia-0)在含有2,4-D(0.3μmol)的平板上,不能长根。转基因拟南芥中,pQY2312(表达QYD45,SEQ ID NO.89);pQY2309(表达QYD42,SEQ ID NO.83);pQY2310(表达QYD43,SEQ ID NO.85);pQY2311(表达QYD44,SEQ IDNO.87);pQY2313(表达QYD46,SEQ ID NO.91);pQY2314(表达QYD47,SEQ ID NO.93);pQY2315(表达QYD48,SEQ ID NO.95);pQY2316(表达QYD49,SEQ ID NO.97);pQY2318(表达QYD51,SEQ ID NO.101);pQY2319(表达QYD52,SEQ ID NO.103);pQY2320(表达QYD53,SEQID NO.105);pQY2322(表达QYD55,SEQ ID NO.109);pQY2323(表达QYD56,SEQ ID NO.111);pQY2324(表达QYD57,SEQ ID NO.113),转化的拟南芥T1代苗,有少数个体长出了种子根,表现出对2,4-D的耐受性。另外,转基因T1代种子,也用分别含有2,4-D丁酸钠(0.7μmol)和二甲四氯(0.1μmol)的平板进行抗药性筛选(图4-2、4-3)。结果与2,4-D筛选相似,其中,pQY2312(表达QYD45,SEQ ID NO.89)转化的T1代苗对2,4-D抗性较弱,但对二甲四氯的抗性明显,有长根明显的植株。
将这些长出正常根的转基因拟南芥移栽到装有土壤的盆钵中继续培养,收获T2代种子进一步鉴定。
4.T2代转基因植株抗药性测试
T2代过表达AAD基因的九个载体pQY2309、pQY2310、pQY2311、pQY2312、pQY2313、pQY2314、pQY2315、pQY2316和pQY2317的转基因株系,在平板培养基上进一步测试其对2,4-D、2,4-D丁酸钠和二甲四氯除草剂的抗性。每个转化体测试两个事件。由于经过T1代抗性筛选,T2代大多数植株都表现抗性,尤其是pQY2312对2,4-D丁酸钠和二甲四氯的抗性表现明显,如图5-1、图5-2和图5-3所示。
选择代表性的四个转化体pQY2309、pQY2310、pQY2311和pQY2313的T2代苗进一步喷雾测试。来自每个载体转化所产生的T2代中随机选择4个株系,待拟南芥种子发芽、移栽和幼苗长至5-7叶时,分别叶面喷施2,4-D(100g/ha)、二甲四氯(100g/ha)和2,4-D丁酸钠(100g/ha)。非转基因的野生型Col-0作为阴性对照。喷施10天后评价抗性水平。结果发现,所有转基因拟南芥T2代幼苗均没有发生明显药害,非转基因野生型拟南芥全部死亡或几乎全部死亡,如图6-1、图6-2和图6-3所示。
实施例5:玉米过表达载体的构建与遗传转化
1.载体构建:过表达载体由pCambia3300改造而来,具有Bar抗性基因可用双丙氨膦(草铵膦)作为筛选标记。AAD类似基因QYD42,SEQ ID NO.83通过无缝克隆方式连接至pCambia3300 BamHⅠ酶切位点处,构建转基因载体pQY2329,其表达框由玉米ubiqutin-1启动子、AAD基因、Nos终止子组成,如图7-1所示。
2.玉米遗传转化:玉米转化采用农杆菌介导的幼胚转化方法(Jones T,Lowe K,Hoerster G,et al.Maize Transformation Using the Morphogenic Genes Baby Boomand Wuschel2.Methods MolBiol.2019;1864:81-93.doi:10.1007/978-1-4939-8778-8_6),将含有过表达载体pQY2329的农杆菌(EHA105)接种至YEP液体培养基(含25mg/L利福平(Rifampicin)和50mg/L卡那霉素(Karamycin),28℃220rmp/min振荡培养过夜至OD600值为1.0-1.5左右,6000rmp/min离心10min收集菌体至50ml离心管(灭菌),弃上清,用MS侵染培养基(含68.5g/L蔗糖,36g/L葡萄糖,pH 5.8)重悬至OD550=0.8。取授粉后9-12天玉米幼穗,去苞叶,底部***长柄枪型镊,放入1.6%的次氯酸钠和0.1%吐温20的消毒液中灭菌20min,无菌水洗三遍,每次5min。用手术刀削去玉米籽粒顶部2-3mm,挑出幼胚,放入50ml离心管中,加入农杆菌悬浮液,静置5min,然后除去农杆菌液体,将幼胚转入MS共培养培养基(含20g/L蔗糖,10g/L葡萄糖,100μMAS,10mg/LVC,50mg/L胸苷,8g/L琼脂,0.5g/L MESbuffer,pH 5.8)中,22℃过夜暗培养。共培养完后将幼胚转移至恢复培养基(含MS盐,0.6×N6大量盐,1.68g/LKNO3,0.6×B5微量盐,0.4×Eriksson’s维生素,1×Murashige&Skoog维生素,0.2mg/LVB1,0.3g/L水解酪蛋白,20g/L蔗糖,2g/L脯氨酸,0.6g/L葡萄糖,100mg/L头孢,150mg/L特美汀,8g/L琼脂,pH 5.8)上培养1周,之后用新鲜的恢复培养基(含2mg/L双丙氨膦)继代培养,每2周继代一次。4-6周后,将生长快速的胚性愈伤转移到分化培养基(含60g/L蔗糖,0.5mg/L玉米素,0.1mg/L赛苯隆,1mg/L 6-BA,100mg/L羧苄青霉素,8g/L琼脂,2mg/L双丙氨膦,pH 5.8)上暗培养2-3周诱导芽形成。当芽1-2cm长时,移至MS生根培养基(含40g/L蔗糖,2mg/L双丙氨膦,pH 5.8)上生芽和生根,弱光下(10-30μmol/m2/s)下培养,最后移栽温室。
实施例6:转基因玉米抗除草剂能力测试
种植过表达载体pQY2329获得的转基因玉米株系T1代苗,先用草铵膦(200mg/L)涂抹叶片确证是转基因后,与非转基因亲本B104,在苗高大约20cm时叶面喷施2,4-D(4.48kg/ha),非转基因受体亲本作为阴性对照。喷施15天后评价抗性水平,结果发现,非转基因受体亲本出现明显药害,根部结节,根系减少,植株易倒伏,而转基因玉米根系正常,未见任何生长抑制,如图7-2所示。
实施例7:大豆过表达载体的构建与遗传转化
1.载体构建:过表达载体采用pQY2330载体(见图8-1),具有Bar抗性基因,可用草铵膦抗性作为筛选标记。AAD类似基因QYD42,SEQ ID NO.83通过无缝克隆方式连接至XhoⅠ酶切位点处,构建转基因表达载体pQY2330,其表达框由双CaMV35S启动子、AAD类似基因QYD42、CaMV35S Nos终止子组成。
2.大豆遗传转化:大豆转化采用农杆菌介导的子叶节转化方法(Pareddy D,Chennareddy S,Anthony G,et al.Improved soybean transformation for efficientand high throughput transgenic production[J].Transgenic Research,2020,29(968))。将含有过表达载体pQY2330的农杆菌(EHA105)单克隆接种至5mlYEP液体培养基(含25mg/L利福平(Rifampicin)和50mg/L卡那霉素(Karamycin)),28℃220rmp/min振荡培养8小时,转移0.2-0.3ml菌液至200ml YEP液体培养基(含25mg/L利福平(Rifampicin)和50mg/L卡那霉素(Karamycin))中28℃220rmp/min过夜培养至菌达到指数增长期,4000rmp/min离心20min收集菌体至50ml离心管(灭菌),弃上清,用侵染培养基(B5盐,B5维生素,30g/L蔗糖,1.67mg/L BAP,0.25ml/L赤霉素,3.9g/L MES和200μM乙酰丁香酮,pH 5.0)悬浮菌体沉淀至OD600=0.6。大豆种子用氯气消毒,然后将种子保存在层流罩中的无盖容器中驱散多余的氯气。在培养皿中用无菌水浸泡灭菌种子,24℃黑暗处理16小时。用10号手术刀沿种子种脐纵向将大豆种子切成两半,每一半包含一半胚与子叶,胚轴远端部分被切除,大约一半至三分之一胚轴仍附着在子叶节端。将具有部分胚轴的种子外植体浸入侵染液中30分钟,然后移至含有B5盐和维生素的共培养培养基上,用滤纸覆盖,温度24℃,光周期为18小时光照6小时黑暗,光强度为80–90μmol/m2/s共培养5天。5天后,外植体用液体诱导培养基(含B5盐、B5维生素、1.11mg/L BAP、30g/L蔗糖、28mg/L硫酸亚铁、38mg/LNa2EDTA、0.6g/L MES、50mg/L卡那霉素、200mg/L头孢噻肟和100mg/L特美汀,pH 5.7)冲洗。将外植体转移至诱导培养基上(含7g/L琼脂),子叶曲面朝下,子叶节端嵌入培养基中。培养2周后,再转移到筛选培养基(含6mg/L草铵膦)培养2周,从外植体基部切取子叶,并将含有胚轴的水平“芽垫”移入继代培养基(含MS盐、30g/L蔗糖、28mg/L硫酸亚铁、0.6g/L MES、38mg/LNa2EDTA、0.1mg/LIAA、1mg/L玉米素核糖苷、0.5mg/L GA3、50mg/L天冬酰胺、100mg/L焦谷氨酸、50mg/L卡那、200mg/L头孢噻肟、50mg/L特美汀、6mg/L草铵膦、7g/L琼脂,pH 5.7)中,每2周继代一次。将芽基部切除浸入1mg/L吲哚3-丁酸中浸泡1-3分钟以促进生根。随后转移至生根培养基(含MS盐、B5维生素、20g/L蔗糖、28mg/L硫酸亚铁、0.59g/L MES、38mg/LNa2EDTA、100mg/L焦谷氨酸、50mg/L天冬酰胺、7g/L琼脂,pH 5.6)持续1-2周,然后将生根的嫩枝移栽土壤对幼苗进行驯化,最后移栽温室。在T0代苗培育过程中,用草铵膦(150mg/L)涂抹叶片进一步鉴定是否转基因。
实施例8:转基因大豆2,4-D抗性测试
得到T1代种子并经基因型鉴定和草铵膦涂抹鉴定确认转基因后,对T1苗进行喷药测试,在大豆苗高大约20cm时叶面喷施2,4-D(4.48kg/ha),非转基因受体亲本作为阴性对照。喷施10天后评价抗性水平。结果发现,非转基因受体亲本全部死亡,而转基因大豆(pQY2330)没有发生药害,未见任何生长抑制,如图8-2所示。
说明书中提及的所有出版物和专利申请均通过引用并入本文,如同每篇出版物或专利申请被单独、特别地通过引用并入本文一样。
尽管为清楚理解起见,前述发明已通过举例说明和实施例的方式较为详细地进行了描述,但显而易见的是,可以在所附权利要求书的范围内实施某些改变和修改,这样的改变和修改均在本发明的范围之内。
Claims (10)
1.一种重组DNA分子,其包含下述的核酸序列:
(1)编码包含与选自以下组的至少一个氨基酸序列具有至少90%、至少95%、至少96%、至少97%、至少98%、至少99%或100%序列同一性的氨基酸序列的蛋白或其生物活性片段的核酸序列或其互补序列:SEQ ID NO:85、SEQ ID NO:41、SEQ ID NO:51、SEQ IDNO:59、SEQ ID NO:63、SEQ ID NO:65、SEQ ID NO:67、SEQ ID NO:69、SEQ ID NO:87、SEQIDNO:89、SEQ ID NO:91、SEQ ID NO:93、SEQ ID NO:95、SEQ ID NO:97、SEQ ID NO:99、SEQID NO:101、SEQ ID NO:103、SEQ ID NO:105、SEQ ID NO:107、SEQ ID NO:109、SEQID NO:111、SEQ ID NO:113、SEQ ID NO:115、SEQ ID NO:123、SEQ ID NO:143、SEQ IDNO:145、SEQID NO:147、SEQ ID NO:149和SEQ ID NO:151;
(2)SEQ ID NO:86、SEQ ID NO:42、SEQ ID NO:52、SEQ ID NO:60、SEQ ID NO:64、SEQID NO:66、SEQ ID NO:68、SEQ ID NO:70、SEQ ID NO:88、SEQ ID NO:90、SEQ IDNO:92、SEQID NO:94、SEQ ID NO:96、SEQ ID NO:98、SEQ ID NO:100、SEQ ID NO:102、SEQ ID NO:104、SEQ ID NO:106、SEQ ID NO:108、SEQ ID NO:110、SEQ ID NO:112、SEQID NO:114、SEQ IDNO:116、SEQ ID NO:124、SEQ ID NO:144、SEQ ID NO:146、SEQ IDNO:148、SEQ ID NO:150或SEQ ID NO:152所示的核酸序列或其互补序列;
(3)在严谨条件下与(1)或(2)所示序列杂交的核酸序列;或
(4)因遗传密码的简并性而与(1)或(2)所示序列编码相同氨基酸序列的核酸序列或其互补序列;
优选地,所述重组DNA分子可操作地连接至在植物细胞中有功能的异源启动子。
2.一种DNA构建体,其包含可操作地连接至如权利要求1所述的重组DNA分子的在植物细胞中有功能的异源启动子;优选地,其存在于转基因植物的基因组中。
3.一种分离的多核苷酸,其中包含如权利要求1所述的重组DNA分子、如权利要求2所述的DNA构建体。
4.一种植物基因组,其中包含如权利要求3所述的多核苷酸。
5.一种载体,其中包含如权利要求3所述的多核苷酸以及与之可操作连接的同源启动子;优选地,所述启动子为诱导型启动子或植物基因组中的自身基因启动子。
6.一种用于产生或提高除草剂耐受性的植物或种子的方法,其包括用如权利要求1所述的重组DNA分子或如权利要求2所述的DNA构建体转化植物细胞或组织,和从所述转化的植物细胞或组织再生除草剂耐受性植物。
7.一种用于赋予植物、种子、细胞或植物部分以除草剂耐受性的方法,所述方法包括在所述植物、种子、细胞或植物部分中表达蛋白或其生物活性片段;
或者,其中包括将表达蛋白或其生物活性片段的植物与另一植物杂交,以及筛选能产生或提高对除草剂耐受性的植物、种子、细胞或植物部分;
或者,其中包括对所述植物、种子、细胞或植物部分进行基因编辑,以实现在其中表达蛋白或其生物活性片段;
所述蛋白由权利要求1所述重组DNA分子编码。
8.如权利要求1所述的重组DNA分子、如权利要求2所述的DNA构建体、如权利要求3所述的多核苷酸、如权利要求4所述的植物基因组或如权利要求5所述的载体用于产生或提高植物、种子、细胞或植物部分对除草剂耐受性的用途。
9.一种用于控制植物生长区域中的杂草的方法,其包括使含有耐受除草剂的植物或种子的植物生长区域与所述除草剂接触,所述植物或种子包括通过权利要求6或7的方法制备的植物或种子、或包含如权利要求1所述的重组DNA分子的植物或种子。
10.根据权利要求6、7或9所述的方法、根据权利要求8所述的用途,其特征在于,所述除草剂选自以下类型中的至少一种:激素类除草剂、ACCase抑制剂类除草剂;优选地,所述除草剂选自以下类型中的至少一种:吡啶羧酸类、苯甲酸类、苯氧羧酸类和芳氧苯氧丙酸类;优选地,所述除草剂选自以下化合物中的至少一种:氯氟吡氧乙酸、绿草定、麦草畏、2,4-D、2,4-D丁酸、2,4-D丁酸钠、2,4-D异辛酯、二甲四氯和精喹禾灵。
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202410244759.8A CN118147174A (zh) | 2020-08-06 | 2020-08-06 | 除草剂抗性蛋白、基因和用途 |
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202410244759.8A CN118147174A (zh) | 2020-08-06 | 2020-08-06 | 除草剂抗性蛋白、基因和用途 |
CN202010784387.XA CN114085849B (zh) | 2020-08-06 | 2020-08-06 | 除草剂抗性蛋白、基因和用途 |
Related Parent Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202010784387.XA Division CN114085849B (zh) | 2020-08-06 | 2020-08-06 | 除草剂抗性蛋白、基因和用途 |
Publications (1)
Publication Number | Publication Date |
---|---|
CN118147174A true CN118147174A (zh) | 2024-06-07 |
Family
ID=80117012
Family Applications (2)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202010784387.XA Active CN114085849B (zh) | 2020-08-06 | 2020-08-06 | 除草剂抗性蛋白、基因和用途 |
CN202410244759.8A Pending CN118147174A (zh) | 2020-08-06 | 2020-08-06 | 除草剂抗性蛋白、基因和用途 |
Family Applications Before (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202010784387.XA Active CN114085849B (zh) | 2020-08-06 | 2020-08-06 | 除草剂抗性蛋白、基因和用途 |
Country Status (4)
Country | Link |
---|---|
US (1) | US20240043859A1 (zh) |
CN (2) | CN114085849B (zh) |
BR (1) | BR112023002148A2 (zh) |
WO (1) | WO2022028440A1 (zh) |
Family Cites Families (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP5285906B2 (ja) * | 2004-04-30 | 2013-09-11 | ダウ アグロサイエンシズ リミテッド ライアビリティー カンパニー | 除草剤抵抗性遺伝子 |
US8278505B2 (en) * | 2007-05-09 | 2012-10-02 | Dow Agrosciences, Llc. | Herbicide resistance genes for resistance to aryloxyalkanoate herbicides |
-
2020
- 2020-08-06 CN CN202010784387.XA patent/CN114085849B/zh active Active
- 2020-08-06 CN CN202410244759.8A patent/CN118147174A/zh active Pending
-
2021
- 2021-08-04 WO PCT/CN2021/110411 patent/WO2022028440A1/zh active Application Filing
- 2021-08-04 BR BR112023002148A patent/BR112023002148A2/pt unknown
- 2021-08-04 US US18/019,498 patent/US20240043859A1/en active Pending
Also Published As
Publication number | Publication date |
---|---|
US20240043859A1 (en) | 2024-02-08 |
WO2022028440A1 (zh) | 2022-02-10 |
CN114085849A (zh) | 2022-02-25 |
BR112023002148A2 (pt) | 2023-03-14 |
CN114085849B (zh) | 2024-03-29 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN113403289B (zh) | 突变型对羟苯基丙酮酸双氧化酶、其编码核酸以及应用 | |
CN101460626B (zh) | 选择转化细胞的方法 | |
CN103013938B (zh) | 除草剂抗性蛋白质、其编码基因及用途 | |
CN104611307B (zh) | 除草剂抗性蛋白质、其编码基因及用途 | |
IL233858A (en) | Glyphosate-resistant plants and related methods | |
CN103060279B (zh) | 除草剂抗性蛋白质、其编码基因及用途 | |
CN105746255B (zh) | 除草剂耐受性蛋白质的用途 | |
CN113249344B (zh) | 抗除草剂突变蛋白、核酸及其应用 | |
CN105724139B (zh) | 除草剂耐受性蛋白质的用途 | |
CN105766992B (zh) | 除草剂耐受性蛋白质的用途 | |
CN108330116A (zh) | 除草剂耐受性蛋白质、其编码基因及用途 | |
CN102399794A (zh) | 一种棉花epsp合成酶突变体基因及其应用 | |
CN113249345B (zh) | 抗除草剂多肽、核酸及其应用 | |
CN104611308B (zh) | 除草剂抗性蛋白质、其编码基因及用途 | |
CA2818581C (en) | Herbicide resistance gene and use thereof | |
CN101812462B (zh) | 水稻GT转录因子家族基因OsGTγ-1在控制水稻耐盐性中的应用 | |
CN103013939B (zh) | 除草剂抗性蛋白质、其编码基因及用途 | |
CN105724140B (zh) | 除草剂耐受性蛋白质的用途 | |
CN104313033A (zh) | 百脉根抗逆相关转录因子及其编码基因和应用 | |
CA2570830A1 (en) | Microbial glyphosate resistant 5-enolpyruvylshikimate-3-phosphate synthases | |
CN104611306B (zh) | 除草剂抗性蛋白质、其编码基因及用途 | |
CN118147174A (zh) | 除草剂抗性蛋白、基因和用途 | |
WO2022205974A1 (zh) | 具有三酮类除草剂抗性的hir突变体及其在植物育种中的应用 | |
CN103740666A (zh) | 除草剂抗性蛋白质、其编码基因及用途 | |
CN111139228B (zh) | 一种抗草甘膦植物epsps酶双突变体及其克隆、表达与应用 |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
PB01 | Publication | ||
SE01 | Entry into force of request for substantive examination | ||
SE01 | Entry into force of request for substantive examination |