CN113444733A - 一种增强玉米大斑病抗性的基因ZmBON3及其应用 - Google Patents
一种增强玉米大斑病抗性的基因ZmBON3及其应用 Download PDFInfo
- Publication number
- CN113444733A CN113444733A CN202110826649.9A CN202110826649A CN113444733A CN 113444733 A CN113444733 A CN 113444733A CN 202110826649 A CN202110826649 A CN 202110826649A CN 113444733 A CN113444733 A CN 113444733A
- Authority
- CN
- China
- Prior art keywords
- gene
- zmbon3
- corn
- resistance
- leaf blight
- 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 84
- 240000008042 Zea mays Species 0.000 title claims abstract description 66
- 235000002017 Zea mays subsp mays Nutrition 0.000 title claims abstract description 61
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 title claims abstract description 53
- 235000005822 corn Nutrition 0.000 title claims abstract description 53
- 230000002708 enhancing effect Effects 0.000 title claims abstract description 21
- 241000196324 Embryophyta Species 0.000 claims abstract description 56
- 208000035240 Disease Resistance Diseases 0.000 claims abstract description 16
- 239000002773 nucleotide Substances 0.000 claims abstract description 7
- 125000003729 nucleotide group Chemical group 0.000 claims abstract description 7
- 238000010362 genome editing Methods 0.000 claims description 23
- 239000013598 vector Substances 0.000 claims description 20
- 238000000034 method Methods 0.000 claims description 12
- 102000004169 proteins and genes Human genes 0.000 claims description 11
- 235000016383 Zea mays subsp huehuetenangensis Nutrition 0.000 claims description 10
- 235000009973 maize Nutrition 0.000 claims description 8
- 239000012620 biological material Substances 0.000 claims description 6
- 241000894006 Bacteria Species 0.000 claims description 3
- 241000219194 Arabidopsis Species 0.000 claims description 2
- 240000007594 Oryza sativa Species 0.000 claims description 2
- 235000007164 Oryza sativa Nutrition 0.000 claims description 2
- 235000021307 Triticum Nutrition 0.000 claims description 2
- 241000209140 Triticum Species 0.000 claims description 2
- 238000010367 cloning Methods 0.000 claims description 2
- 210000000056 organ Anatomy 0.000 claims description 2
- 235000009566 rice Nutrition 0.000 claims description 2
- 125000003275 alpha amino acid group Chemical group 0.000 claims 1
- 201000010099 disease Diseases 0.000 abstract description 16
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 abstract description 16
- 238000009395 breeding Methods 0.000 abstract description 4
- 230000001488 breeding effect Effects 0.000 abstract description 4
- 238000010353 genetic engineering Methods 0.000 abstract description 3
- 238000004519 manufacturing process Methods 0.000 abstract description 2
- 239000000243 solution Substances 0.000 description 19
- 102000004190 Enzymes Human genes 0.000 description 15
- 108090000790 Enzymes Proteins 0.000 description 15
- 230000008859 change Effects 0.000 description 13
- 238000011081 inoculation Methods 0.000 description 10
- 206010020649 Hyperkeratosis Diseases 0.000 description 9
- 230000001965 increasing effect Effects 0.000 description 9
- 108010054424 Copine Proteins 0.000 description 8
- 230000000694 effects Effects 0.000 description 8
- 230000006872 improvement Effects 0.000 description 8
- 238000002835 absorbance Methods 0.000 description 7
- 238000011160 research Methods 0.000 description 7
- 108091033409 CRISPR Proteins 0.000 description 6
- 108020004414 DNA Proteins 0.000 description 6
- 102000003992 Peroxidases Human genes 0.000 description 6
- 108700023158 Phenylalanine ammonia-lyases Proteins 0.000 description 6
- 238000001514 detection method Methods 0.000 description 6
- 239000000463 material Substances 0.000 description 6
- 238000012216 screening Methods 0.000 description 6
- 235000007244 Zea mays Nutrition 0.000 description 5
- 238000012163 sequencing technique Methods 0.000 description 5
- 230000009261 transgenic effect Effects 0.000 description 5
- 102000016938 Catalase Human genes 0.000 description 4
- 108010053835 Catalase Proteins 0.000 description 4
- 108010035563 Chloramphenicol O-acetyltransferase Proteins 0.000 description 4
- JEIPFZHSYJVQDO-UHFFFAOYSA-N iron(III) oxide Inorganic materials O=[Fe]O[Fe]=O JEIPFZHSYJVQDO-UHFFFAOYSA-N 0.000 description 4
- 239000000203 mixture Substances 0.000 description 4
- 230000035484 reaction time Effects 0.000 description 4
- 239000006228 supernatant Substances 0.000 description 4
- 102000001711 Copine Human genes 0.000 description 3
- 238000004364 calculation method Methods 0.000 description 3
- 230000002068 genetic effect Effects 0.000 description 3
- 108010078144 glutaminyl-glycine Proteins 0.000 description 3
- 239000001963 growth medium Substances 0.000 description 3
- 244000052769 pathogen Species 0.000 description 3
- 108040007629 peroxidase activity proteins Proteins 0.000 description 3
- 239000008055 phosphate buffer solution Substances 0.000 description 3
- 239000013612 plasmid Substances 0.000 description 3
- 241000894007 species Species 0.000 description 3
- YNJBWRMUSHSURL-UHFFFAOYSA-N trichloroacetic acid Chemical compound OC(=O)C(Cl)(Cl)Cl YNJBWRMUSHSURL-UHFFFAOYSA-N 0.000 description 3
- 241000589158 Agrobacterium Species 0.000 description 2
- MQIGTEQXYCRLGK-BQBZGAKWSA-N Ala-Gly-Pro Chemical compound C[C@H](N)C(=O)NCC(=O)N1CCC[C@H]1C(O)=O MQIGTEQXYCRLGK-BQBZGAKWSA-N 0.000 description 2
- 241000219195 Arabidopsis thaliana Species 0.000 description 2
- OQCWXQJLCDPRHV-UWVGGRQHSA-N Arg-Gly-Leu Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)NCC(=O)N[C@@H](CC(C)C)C(O)=O OQCWXQJLCDPRHV-UWVGGRQHSA-N 0.000 description 2
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- 238000010354 CRISPR gene editing Methods 0.000 description 2
- 208000003643 Callosities Diseases 0.000 description 2
- DRDSQGHKTLSNEA-GLLZPBPUSA-N Gln-Glu-Thr Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O DRDSQGHKTLSNEA-GLLZPBPUSA-N 0.000 description 2
- CCQOOWAONKGYKQ-BYPYZUCNSA-N Gly-Gly-Ala Chemical compound OC(=O)[C@H](C)NC(=O)CNC(=O)CN CCQOOWAONKGYKQ-BYPYZUCNSA-N 0.000 description 2
- 241000880493 Leptailurus serval Species 0.000 description 2
- 238000012408 PCR amplification Methods 0.000 description 2
- SRSPTFBENMJHMR-WHFBIAKZSA-N Ser-Ser-Gly Chemical compound OC[C@H](N)C(=O)N[C@@H](CO)C(=O)NCC(O)=O SRSPTFBENMJHMR-WHFBIAKZSA-N 0.000 description 2
- 108091027544 Subgenomic mRNA Proteins 0.000 description 2
- XXROXFHCMVXETG-UWVGGRQHSA-N Val-Gly-Val Chemical compound CC(C)[C@H](N)C(=O)NCC(=O)N[C@@H](C(C)C)C(O)=O XXROXFHCMVXETG-UWVGGRQHSA-N 0.000 description 2
- 235000007241 Zea diploperennis Nutrition 0.000 description 2
- 235000017556 Zea mays subsp parviglumis Nutrition 0.000 description 2
- 241000172407 Zea mays subsp. huehuetenangensis Species 0.000 description 2
- 238000000137 annealing Methods 0.000 description 2
- 244000052616 bacterial pathogen Species 0.000 description 2
- 238000005119 centrifugation Methods 0.000 description 2
- 238000002856 computational phylogenetic analysis Methods 0.000 description 2
- 238000001816 cooling Methods 0.000 description 2
- 238000012258 culturing Methods 0.000 description 2
- 238000004925 denaturation Methods 0.000 description 2
- 230000036425 denaturation Effects 0.000 description 2
- 102000054766 genetic haplotypes Human genes 0.000 description 2
- LHGVFZTZFXWLCP-UHFFFAOYSA-N guaiacol Chemical compound COC1=CC=CC=C1O LHGVFZTZFXWLCP-UHFFFAOYSA-N 0.000 description 2
- 230000009545 invasion Effects 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 239000003550 marker Substances 0.000 description 2
- 238000002156 mixing Methods 0.000 description 2
- 239000003147 molecular marker Substances 0.000 description 2
- 230000035772 mutation Effects 0.000 description 2
- 238000011392 neighbor-joining method Methods 0.000 description 2
- 239000008363 phosphate buffer Substances 0.000 description 2
- 238000012257 pre-denaturation Methods 0.000 description 2
- 108010015796 prolylisoleucine Proteins 0.000 description 2
- 230000009467 reduction Effects 0.000 description 2
- 229930000044 secondary metabolite Natural products 0.000 description 2
- 108010048397 seryl-lysyl-leucine Proteins 0.000 description 2
- 239000000725 suspension Substances 0.000 description 2
- JNTMAZFVYNDPLB-PEDHHIEDSA-N (2S,3S)-2-[[[(2S)-1-[(2S,3S)-2-amino-3-methyl-1-oxopentyl]-2-pyrrolidinyl]-oxomethyl]amino]-3-methylpentanoic acid Chemical compound CC[C@H](C)[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@@H]([C@@H](C)CC)C(O)=O JNTMAZFVYNDPLB-PEDHHIEDSA-N 0.000 description 1
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 1
- 108010085238 Actins Proteins 0.000 description 1
- 241000589155 Agrobacterium tumefaciens Species 0.000 description 1
- GWFSQQNGMPGBEF-GHCJXIJMSA-N Ala-Asp-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)O)NC(=O)[C@H](CC(=O)O)NC(=O)[C@H](C)N GWFSQQNGMPGBEF-GHCJXIJMSA-N 0.000 description 1
- RXTBLQVXNIECFP-FXQIFTODSA-N Ala-Gln-Gln Chemical compound C[C@H](N)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCC(N)=O)C(O)=O RXTBLQVXNIECFP-FXQIFTODSA-N 0.000 description 1
- HQJKCXHQNUCKMY-GHCJXIJMSA-N Ala-Ile-Asp Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)O)NC(=O)[C@H](C)N HQJKCXHQNUCKMY-GHCJXIJMSA-N 0.000 description 1
- KYDYGANDJHFBCW-DRZSPHRISA-N Ala-Phe-Gln Chemical compound C[C@@H](C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CCC(=O)N)C(=O)O)N KYDYGANDJHFBCW-DRZSPHRISA-N 0.000 description 1
- RMAWDDRDTRSZIR-ZLUOBGJFSA-N Ala-Ser-Asp Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(O)=O)C(O)=O RMAWDDRDTRSZIR-ZLUOBGJFSA-N 0.000 description 1
- YYAVDNKUWLAFCV-ACZMJKKPSA-N Ala-Ser-Gln Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CO)C(=O)N[C@@H](CCC(N)=O)C(O)=O YYAVDNKUWLAFCV-ACZMJKKPSA-N 0.000 description 1
- NCQMBSJGJMYKCK-ZLUOBGJFSA-N Ala-Ser-Ser Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CO)C(=O)N[C@@H](CO)C(O)=O NCQMBSJGJMYKCK-ZLUOBGJFSA-N 0.000 description 1
- VRTOMXFZHGWHIJ-KZVJFYERSA-N Ala-Thr-Arg Chemical compound [H]N[C@@H](C)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O VRTOMXFZHGWHIJ-KZVJFYERSA-N 0.000 description 1
- XQNRANMFRPCFFW-GCJQMDKQSA-N Ala-Thr-Asn Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)O)NC(=O)[C@H](C)N)O XQNRANMFRPCFFW-GCJQMDKQSA-N 0.000 description 1
- ZTKHZAXGTFXUDD-VEVYYDQMSA-N Arg-Asn-Thr Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O ZTKHZAXGTFXUDD-VEVYYDQMSA-N 0.000 description 1
- LYJXHXGPWDTLKW-HJGDQZAQSA-N Arg-Thr-Gln Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CCC(=O)N)C(=O)O)NC(=O)[C@H](CCCN=C(N)N)N)O LYJXHXGPWDTLKW-HJGDQZAQSA-N 0.000 description 1
- KXEGPPNPXOKKHK-ZLUOBGJFSA-N Asn-Asp-Ala Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](C)C(O)=O KXEGPPNPXOKKHK-ZLUOBGJFSA-N 0.000 description 1
- ACKNRKFVYUVWAC-ZPFDUUQYSA-N Asn-Ile-Lys Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCCCN)C(=O)O)NC(=O)[C@H](CC(=O)N)N ACKNRKFVYUVWAC-ZPFDUUQYSA-N 0.000 description 1
- HDHZCEDPLTVHFZ-GUBZILKMSA-N Asn-Leu-Glu Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(O)=O)C(O)=O HDHZCEDPLTVHFZ-GUBZILKMSA-N 0.000 description 1
- TZFQICWZWFNIKU-KKUMJFAQSA-N Asn-Leu-Tyr Chemical compound NC(=O)C[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@H](C(O)=O)CC1=CC=C(O)C=C1 TZFQICWZWFNIKU-KKUMJFAQSA-N 0.000 description 1
- RCFGLXMZDYNRSC-CIUDSAMLSA-N Asn-Lys-Ala Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](C)C(O)=O RCFGLXMZDYNRSC-CIUDSAMLSA-N 0.000 description 1
- VHQSGALUSWIYOD-QXEWZRGKSA-N Asn-Pro-Val Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N1CCC[C@H]1C(=O)N[C@@H](C(C)C)C(O)=O VHQSGALUSWIYOD-QXEWZRGKSA-N 0.000 description 1
- MYTHOBCLNIOFBL-SRVKXCTJSA-N Asn-Ser-Tyr Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O MYTHOBCLNIOFBL-SRVKXCTJSA-N 0.000 description 1
- GVPSCJQLUGIKAM-GUBZILKMSA-N Asp-Arg-Arg Chemical compound OC(=O)C[C@H](N)C(=O)N[C@@H](CCCN=C(N)N)C(=O)N[C@@H](CCCN=C(N)N)C(O)=O GVPSCJQLUGIKAM-GUBZILKMSA-N 0.000 description 1
- GWTLRDMPMJCNMH-WHFBIAKZSA-N Asp-Asn-Gly Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(N)=O)C(=O)NCC(O)=O GWTLRDMPMJCNMH-WHFBIAKZSA-N 0.000 description 1
- UGKZHCBLMLSANF-CIUDSAMLSA-N Asp-Asn-Leu Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(C)C)C(O)=O UGKZHCBLMLSANF-CIUDSAMLSA-N 0.000 description 1
- CRNKLABLTICXDV-GUBZILKMSA-N Asp-His-Glu Chemical compound C1=C(NC=N1)C[C@@H](C(=O)N[C@@H](CCC(=O)O)C(=O)O)NC(=O)[C@H](CC(=O)O)N CRNKLABLTICXDV-GUBZILKMSA-N 0.000 description 1
- KYQNAIMCTRZLNP-QSFUFRPTSA-N Asp-Ile-Val Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](C(C)C)C(O)=O KYQNAIMCTRZLNP-QSFUFRPTSA-N 0.000 description 1
- LTCKTLYKRMCFOC-KKUMJFAQSA-N Asp-Phe-Leu Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CC(C)C)C(O)=O LTCKTLYKRMCFOC-KKUMJFAQSA-N 0.000 description 1
- JUWISGAGWSDGDH-KKUMJFAQSA-N Asp-Phe-Lys Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)[C@@H](NC(=O)[C@@H](N)CC(O)=O)CC1=CC=CC=C1 JUWISGAGWSDGDH-KKUMJFAQSA-N 0.000 description 1
- LGGHQRZIJSYRHA-GUBZILKMSA-N Asp-Pro-Met Chemical compound CSCC[C@@H](C(=O)O)NC(=O)[C@@H]1CCCN1C(=O)[C@H](CC(=O)O)N LGGHQRZIJSYRHA-GUBZILKMSA-N 0.000 description 1
- ZQFRDAZBTSFGGW-SRVKXCTJSA-N Asp-Ser-Phe Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O ZQFRDAZBTSFGGW-SRVKXCTJSA-N 0.000 description 1
- OTKUAVXGMREHRX-CFMVVWHZSA-N Asp-Tyr-Ile Chemical compound CC[C@H](C)[C@@H](C(O)=O)NC(=O)[C@@H](NC(=O)[C@@H](N)CC(O)=O)CC1=CC=C(O)C=C1 OTKUAVXGMREHRX-CFMVVWHZSA-N 0.000 description 1
- SQIARYGNVQWOSB-BZSNNMDCSA-N Asp-Tyr-Phe Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O SQIARYGNVQWOSB-BZSNNMDCSA-N 0.000 description 1
- WAEDSQFVZJUHLI-BYULHYEWSA-N Asp-Val-Asp Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC(O)=O)C(O)=O WAEDSQFVZJUHLI-BYULHYEWSA-N 0.000 description 1
- 238000010453 CRISPR/Cas method Methods 0.000 description 1
- 241001430230 Clavibacter nebraskensis Species 0.000 description 1
- DVKQPQKQDHHFTE-ZLUOBGJFSA-N Cys-Cys-Asn Chemical compound C([C@@H](C(=O)O)NC(=O)[C@H](CS)NC(=O)[C@H](CS)N)C(=O)N DVKQPQKQDHHFTE-ZLUOBGJFSA-N 0.000 description 1
- MXZYQNJCBVJHSR-KATARQTJSA-N Cys-Lys-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](CCCCN)NC(=O)[C@H](CS)N)O MXZYQNJCBVJHSR-KATARQTJSA-N 0.000 description 1
- WTEACWBAULENKE-SRVKXCTJSA-N Cys-Phe-Asn Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)O)NC(=O)[C@H](CS)N WTEACWBAULENKE-SRVKXCTJSA-N 0.000 description 1
- CHRCKSPMGYDLIA-SRVKXCTJSA-N Cys-Phe-Ser Chemical compound [H]N[C@@H](CS)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CO)C(O)=O CHRCKSPMGYDLIA-SRVKXCTJSA-N 0.000 description 1
- 241000206602 Eukaryota Species 0.000 description 1
- RGXXLQWXBFNXTG-CIUDSAMLSA-N Gln-Arg-Ala Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](C)C(O)=O RGXXLQWXBFNXTG-CIUDSAMLSA-N 0.000 description 1
- JFOKLAPFYCTNHW-SRVKXCTJSA-N Gln-Arg-Lys Chemical compound C(CCN)C[C@@H](C(=O)O)NC(=O)[C@H](CCCN=C(N)N)NC(=O)[C@H](CCC(=O)N)N JFOKLAPFYCTNHW-SRVKXCTJSA-N 0.000 description 1
- LPYPANUXJGFMGV-FXQIFTODSA-N Gln-Gln-Ala Chemical compound C[C@@H](C(=O)O)NC(=O)[C@H](CCC(=O)N)NC(=O)[C@H](CCC(=O)N)N LPYPANUXJGFMGV-FXQIFTODSA-N 0.000 description 1
- QKCZZAZNMMVICF-DCAQKATOSA-N Gln-Leu-Glu Chemical compound NC(=O)CC[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(O)=O)C(O)=O QKCZZAZNMMVICF-DCAQKATOSA-N 0.000 description 1
- SHAUZYVSXAMYAZ-JYJNAYRXSA-N Gln-Leu-Phe Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)O)NC(=O)[C@H](CCC(=O)N)N SHAUZYVSXAMYAZ-JYJNAYRXSA-N 0.000 description 1
- QMVCEWKHIUHTSD-GUBZILKMSA-N Gln-Met-Glu Chemical compound CSCC[C@@H](C(=O)N[C@@H](CCC(=O)O)C(=O)O)NC(=O)[C@H](CCC(=O)N)N QMVCEWKHIUHTSD-GUBZILKMSA-N 0.000 description 1
- BZULIEARJFRINC-IHRRRGAJSA-N Gln-Phe-Glu Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)N[C@@H](CCC(=O)O)C(=O)O)NC(=O)[C@H](CCC(=O)N)N BZULIEARJFRINC-IHRRRGAJSA-N 0.000 description 1
- QFXNFFZTMFHPST-DZKIICNBSA-N Gln-Phe-Val Chemical compound CC(C)[C@@H](C(=O)O)NC(=O)[C@H](CC1=CC=CC=C1)NC(=O)[C@H](CCC(=O)N)N QFXNFFZTMFHPST-DZKIICNBSA-N 0.000 description 1
- UTOQQOMEJDPDMX-ACZMJKKPSA-N Gln-Ser-Asp Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(O)=O)C(O)=O UTOQQOMEJDPDMX-ACZMJKKPSA-N 0.000 description 1
- LPIKVBWNNVFHCQ-GUBZILKMSA-N Gln-Ser-Leu Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(O)=O LPIKVBWNNVFHCQ-GUBZILKMSA-N 0.000 description 1
- AUTNXSQEVVHSJK-YVNDNENWSA-N Glu-Glu-Ile Chemical compound CC[C@H](C)[C@@H](C(O)=O)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@@H](N)CCC(O)=O AUTNXSQEVVHSJK-YVNDNENWSA-N 0.000 description 1
- IQACOVZVOMVILH-FXQIFTODSA-N Glu-Glu-Ser Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CO)C(O)=O IQACOVZVOMVILH-FXQIFTODSA-N 0.000 description 1
- CUXJIASLBRJOFV-LAEOZQHASA-N Glu-Gly-Ile Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)NCC(=O)N[C@@H]([C@@H](C)CC)C(O)=O CUXJIASLBRJOFV-LAEOZQHASA-N 0.000 description 1
- VMKCPNBBPGGQBJ-GUBZILKMSA-N Glu-Leu-Asn Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)O)NC(=O)[C@H](CCC(=O)O)N VMKCPNBBPGGQBJ-GUBZILKMSA-N 0.000 description 1
- YHOJJFFTSMWVGR-HJGDQZAQSA-N Glu-Met-Thr Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCSC)C(=O)N[C@@H]([C@@H](C)O)C(O)=O YHOJJFFTSMWVGR-HJGDQZAQSA-N 0.000 description 1
- VNCNWQPIQYAMAK-ACZMJKKPSA-N Glu-Ser-Ser Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CO)C(O)=O VNCNWQPIQYAMAK-ACZMJKKPSA-N 0.000 description 1
- YQPFCZVKMUVZIN-AUTRQRHGSA-N Glu-Val-Gln Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCC(N)=O)C(O)=O YQPFCZVKMUVZIN-AUTRQRHGSA-N 0.000 description 1
- FGGKGJHCVMYGCD-UKJIMTQDSA-N Glu-Val-Ile Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O FGGKGJHCVMYGCD-UKJIMTQDSA-N 0.000 description 1
- SOYWRINXUSUWEQ-DLOVCJGASA-N Glu-Val-Val Chemical compound CC(C)[C@@H](C(O)=O)NC(=O)[C@H](C(C)C)NC(=O)[C@@H](N)CCC(O)=O SOYWRINXUSUWEQ-DLOVCJGASA-N 0.000 description 1
- PUUYVMYCMIWHFE-BQBZGAKWSA-N Gly-Ala-Arg Chemical compound NCC(=O)N[C@@H](C)C(=O)N[C@H](C(O)=O)CCCN=C(N)N PUUYVMYCMIWHFE-BQBZGAKWSA-N 0.000 description 1
- KKBWDNZXYLGJEY-UHFFFAOYSA-N Gly-Arg-Pro Natural products NCC(=O)NC(CCNC(=N)N)C(=O)N1CCCC1C(=O)O KKBWDNZXYLGJEY-UHFFFAOYSA-N 0.000 description 1
- DTPOVRRYXPJJAZ-FJXKBIBVSA-N Gly-Arg-Thr Chemical compound C[C@@H](O)[C@@H](C(O)=O)NC(=O)[C@@H](NC(=O)CN)CCCN=C(N)N DTPOVRRYXPJJAZ-FJXKBIBVSA-N 0.000 description 1
- UXJHNZODTMHWRD-WHFBIAKZSA-N Gly-Asn-Ala Chemical compound [H]NCC(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](C)C(O)=O UXJHNZODTMHWRD-WHFBIAKZSA-N 0.000 description 1
- STVHDEHTKFXBJQ-LAEOZQHASA-N Gly-Glu-Ile Chemical compound [H]NCC(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O STVHDEHTKFXBJQ-LAEOZQHASA-N 0.000 description 1
- BUEFQXUHTUZXHR-LURJTMIESA-N Gly-Gly-Pro zwitterion Chemical compound NCC(=O)NCC(=O)N1CCC[C@H]1C(O)=O BUEFQXUHTUZXHR-LURJTMIESA-N 0.000 description 1
- ADZGCWWDPFDHCY-ZETCQYMHSA-N Gly-His-Gly Chemical compound OC(=O)CNC(=O)[C@@H](NC(=O)CN)CC1=CN=CN1 ADZGCWWDPFDHCY-ZETCQYMHSA-N 0.000 description 1
- PAWIVEIWWYGBAM-YUMQZZPRSA-N Gly-Leu-Ala Chemical compound NCC(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](C)C(O)=O PAWIVEIWWYGBAM-YUMQZZPRSA-N 0.000 description 1
- MHXKHKWHPNETGG-QWRGUYRKSA-N Gly-Lys-Leu Chemical compound [H]NCC(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(C)C)C(O)=O MHXKHKWHPNETGG-QWRGUYRKSA-N 0.000 description 1
- SJLKKOZFHSJJAW-YUMQZZPRSA-N Gly-Met-Glu Chemical compound CSCC[C@@H](C(=O)N[C@@H](CCC(=O)O)C(=O)O)NC(=O)CN SJLKKOZFHSJJAW-YUMQZZPRSA-N 0.000 description 1
- BMWFDYIYBAFROD-WPRPVWTQSA-N Gly-Pro-Val Chemical compound CC(C)[C@@H](C(O)=O)NC(=O)[C@@H]1CCCN1C(=O)CN BMWFDYIYBAFROD-WPRPVWTQSA-N 0.000 description 1
- GWCJMBNBFYBQCV-XPUUQOCRSA-N Gly-Val-Ala Chemical compound NCC(=O)N[C@@H](C(C)C)C(=O)N[C@@H](C)C(O)=O GWCJMBNBFYBQCV-XPUUQOCRSA-N 0.000 description 1
- RYAOJUMWLWUGNW-QMMMGPOBSA-N Gly-Val-Gly Chemical compound NCC(=O)N[C@@H](C(C)C)C(=O)NCC(O)=O RYAOJUMWLWUGNW-QMMMGPOBSA-N 0.000 description 1
- BAYQNCWLXIDLHX-ONGXEEELSA-N Gly-Val-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](C(C)C)NC(=O)CN BAYQNCWLXIDLHX-ONGXEEELSA-N 0.000 description 1
- 239000005562 Glyphosate Substances 0.000 description 1
- JBCLFWXMTIKCCB-UHFFFAOYSA-N H-Gly-Phe-OH Natural products NCC(=O)NC(C(O)=O)CC1=CC=CC=C1 JBCLFWXMTIKCCB-UHFFFAOYSA-N 0.000 description 1
- DFHVLUKTTVTCKY-PBCZWWQYSA-N His-Asn-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](CC(=O)N)NC(=O)[C@H](CC1=CN=CN1)N)O DFHVLUKTTVTCKY-PBCZWWQYSA-N 0.000 description 1
- FZKFYOXDVWDELO-KBPBESRZSA-N His-Gly-Tyr Chemical compound [H]N[C@@H](CC1=CNC=N1)C(=O)NCC(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O FZKFYOXDVWDELO-KBPBESRZSA-N 0.000 description 1
- XKIYNCLILDLGRS-QWRGUYRKSA-N His-Lys-Gly Chemical compound NCCCC[C@@H](C(=O)NCC(O)=O)NC(=O)[C@@H](N)CC1=CN=CN1 XKIYNCLILDLGRS-QWRGUYRKSA-N 0.000 description 1
- 101000631760 Homo sapiens Sodium channel protein type 1 subunit alpha Proteins 0.000 description 1
- NPROWIBAWYMPAZ-GUDRVLHUSA-N Ile-Asp-Pro Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)N1CCC[C@@H]1C(=O)O)N NPROWIBAWYMPAZ-GUDRVLHUSA-N 0.000 description 1
- LPXHYGGZJOCAFR-MNXVOIDGSA-N Ile-Glu-Leu Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCC(=O)O)C(=O)N[C@@H](CC(C)C)C(=O)O)N LPXHYGGZJOCAFR-MNXVOIDGSA-N 0.000 description 1
- GAZGFPOZOLEYAJ-YTFOTSKYSA-N Ile-Leu-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)CC)C(=O)O)N GAZGFPOZOLEYAJ-YTFOTSKYSA-N 0.000 description 1
- GLYJPWIRLBAIJH-FQUUOJAGSA-N Ile-Lys-Pro Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCCCN)C(=O)N1CCC[C@@H]1C(=O)O)N GLYJPWIRLBAIJH-FQUUOJAGSA-N 0.000 description 1
- GLYJPWIRLBAIJH-UHFFFAOYSA-N Ile-Lys-Pro Natural products CCC(C)C(N)C(=O)NC(CCCCN)C(=O)N1CCCC1C(O)=O GLYJPWIRLBAIJH-UHFFFAOYSA-N 0.000 description 1
- VZSDQFZFTCVEGF-ZEWNOJEFSA-N Ile-Phe-Tyr Chemical compound CC[C@H](C)[C@H](N)C(=O)N[C@@H](Cc1ccccc1)C(=O)N[C@@H](Cc1ccc(O)cc1)C(O)=O VZSDQFZFTCVEGF-ZEWNOJEFSA-N 0.000 description 1
- VGSPNSSCMOHRRR-BJDJZHNGSA-N Ile-Ser-Lys Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CO)C(=O)N[C@@H](CCCCN)C(=O)O)N VGSPNSSCMOHRRR-BJDJZHNGSA-N 0.000 description 1
- YCKPUHHMCFSUMD-IUKAMOBKSA-N Ile-Thr-Asp Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(=O)O)C(=O)O)N YCKPUHHMCFSUMD-IUKAMOBKSA-N 0.000 description 1
- KBDIBHQICWDGDL-PPCPHDFISA-N Ile-Thr-Leu Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(C)C)C(=O)O)N KBDIBHQICWDGDL-PPCPHDFISA-N 0.000 description 1
- IPFKIGNDTUOFAF-CYDGBPFRSA-N Ile-Val-Arg Chemical compound CC[C@H](C)[C@H](N)C(=O)N[C@@H](C(C)C)C(=O)N[C@H](C(O)=O)CCCN=C(N)N IPFKIGNDTUOFAF-CYDGBPFRSA-N 0.000 description 1
- AUIYHFRUOOKTGX-UKJIMTQDSA-N Ile-Val-Gln Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCC(=O)N)C(=O)O)N AUIYHFRUOOKTGX-UKJIMTQDSA-N 0.000 description 1
- COLNVLDHVKWLRT-QMMMGPOBSA-N L-phenylalanine Chemical compound OC(=O)[C@@H](N)CC1=CC=CC=C1 COLNVLDHVKWLRT-QMMMGPOBSA-N 0.000 description 1
- LZDNBBYBDGBADK-UHFFFAOYSA-N L-valyl-L-tryptophan Natural products C1=CC=C2C(CC(NC(=O)C(N)C(C)C)C(O)=O)=CNC2=C1 LZDNBBYBDGBADK-UHFFFAOYSA-N 0.000 description 1
- JKGHDYGZRDWHGA-SRVKXCTJSA-N Leu-Asn-Leu Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(C)C)C(O)=O JKGHDYGZRDWHGA-SRVKXCTJSA-N 0.000 description 1
- WGNOPSQMIQERPK-UHFFFAOYSA-N Leu-Asn-Pro Natural products CC(C)CC(N)C(=O)NC(CC(=O)N)C(=O)N1CCCC1C(=O)O WGNOPSQMIQERPK-UHFFFAOYSA-N 0.000 description 1
- OGCQGUIWMSBHRZ-CIUDSAMLSA-N Leu-Asn-Ser Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CO)C(O)=O OGCQGUIWMSBHRZ-CIUDSAMLSA-N 0.000 description 1
- FIJMQLGQLBLBOL-HJGDQZAQSA-N Leu-Asn-Thr Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O FIJMQLGQLBLBOL-HJGDQZAQSA-N 0.000 description 1
- BPANDPNDMJHFEV-CIUDSAMLSA-N Leu-Asp-Ala Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](C)C(O)=O BPANDPNDMJHFEV-CIUDSAMLSA-N 0.000 description 1
- WCTCIIAGNMFYAO-DCAQKATOSA-N Leu-Cys-Val Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CS)C(=O)N[C@@H](C(C)C)C(O)=O WCTCIIAGNMFYAO-DCAQKATOSA-N 0.000 description 1
- WIDZHJTYKYBLSR-DCAQKATOSA-N Leu-Glu-Glu Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(O)=O WIDZHJTYKYBLSR-DCAQKATOSA-N 0.000 description 1
- OGUUKPXUTHOIAV-SDDRHHMPSA-N Leu-Glu-Pro Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CCC(=O)O)C(=O)N1CCC[C@@H]1C(=O)O)N OGUUKPXUTHOIAV-SDDRHHMPSA-N 0.000 description 1
- CFZZDVMBRYFFNU-QWRGUYRKSA-N Leu-His-Gly Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CC1=CNC=N1)C(=O)NCC(O)=O CFZZDVMBRYFFNU-QWRGUYRKSA-N 0.000 description 1
- HMDDEJADNKQTBR-BZSNNMDCSA-N Leu-His-Tyr Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O HMDDEJADNKQTBR-BZSNNMDCSA-N 0.000 description 1
- QJXHMYMRGDOHRU-NHCYSSNCSA-N Leu-Ile-Gly Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)CC)C(=O)NCC(O)=O QJXHMYMRGDOHRU-NHCYSSNCSA-N 0.000 description 1
- FAELBUXXFQLUAX-AJNGGQMLSA-N Leu-Leu-Ile Chemical compound CC[C@H](C)[C@@H](C(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@@H](N)CC(C)C FAELBUXXFQLUAX-AJNGGQMLSA-N 0.000 description 1
- UCBPDSYUVAAHCD-UWVGGRQHSA-N Leu-Pro-Gly Chemical compound CC(C)C[C@H](N)C(=O)N1CCC[C@H]1C(=O)NCC(O)=O UCBPDSYUVAAHCD-UWVGGRQHSA-N 0.000 description 1
- KWLWZYMNUZJKMZ-IHRRRGAJSA-N Leu-Pro-Leu Chemical compound CC(C)C[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CC(C)C)C(O)=O KWLWZYMNUZJKMZ-IHRRRGAJSA-N 0.000 description 1
- JIHDFWWRYHSAQB-GUBZILKMSA-N Leu-Ser-Glu Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@H](C(O)=O)CCC(O)=O JIHDFWWRYHSAQB-GUBZILKMSA-N 0.000 description 1
- VUBIPAHVHMZHCM-KKUMJFAQSA-N Leu-Tyr-Ser Chemical compound CC(C)C[C@H](N)C(=O)N[C@H](C(=O)N[C@@H](CO)C(O)=O)CC1=CC=C(O)C=C1 VUBIPAHVHMZHCM-KKUMJFAQSA-N 0.000 description 1
- YNNPKXBBRZVIRX-IHRRRGAJSA-N Lys-Arg-Leu Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(C)C)C(O)=O YNNPKXBBRZVIRX-IHRRRGAJSA-N 0.000 description 1
- KPJJOZUXFOLGMQ-CIUDSAMLSA-N Lys-Asp-Asn Chemical compound C(CCN)C[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)N[C@@H](CC(=O)N)C(=O)O)N KPJJOZUXFOLGMQ-CIUDSAMLSA-N 0.000 description 1
- SQXUUGUCGJSWCK-CIUDSAMLSA-N Lys-Asp-Cys Chemical compound C(CCN)C[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)N[C@@H](CS)C(=O)O)N SQXUUGUCGJSWCK-CIUDSAMLSA-N 0.000 description 1
- GKFNXYMAMKJSKD-NHCYSSNCSA-N Lys-Asp-Val Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](C(C)C)C(O)=O GKFNXYMAMKJSKD-NHCYSSNCSA-N 0.000 description 1
- LMMBAXJRYSXCOQ-ACRUOGEOSA-N Lys-Tyr-Phe Chemical compound NCCCC[C@H](N)C(=O)N[C@@H](Cc1ccc(O)cc1)C(=O)N[C@@H](Cc1ccccc1)C(O)=O LMMBAXJRYSXCOQ-ACRUOGEOSA-N 0.000 description 1
- DTICLBJHRYSJLH-GUBZILKMSA-N Met-Ala-Val Chemical compound CSCC[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@@H](C(C)C)C(O)=O DTICLBJHRYSJLH-GUBZILKMSA-N 0.000 description 1
- UZWMJZSOXGOVIN-LURJTMIESA-N Met-Gly-Gly Chemical compound CSCC[C@H](N)C(=O)NCC(=O)NCC(O)=O UZWMJZSOXGOVIN-LURJTMIESA-N 0.000 description 1
- HZVXPUHLTZRQEL-UWVGGRQHSA-N Met-Leu-Gly Chemical compound CSCC[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)NCC(O)=O HZVXPUHLTZRQEL-UWVGGRQHSA-N 0.000 description 1
- WPTHAGXMYDRPFD-SRVKXCTJSA-N Met-Lys-Glu Chemical compound CSCC[C@H](N)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCC(O)=O)C(O)=O WPTHAGXMYDRPFD-SRVKXCTJSA-N 0.000 description 1
- CIDICGYKRUTYLE-FXQIFTODSA-N Met-Ser-Ala Chemical compound CSCC[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@@H](C)C(O)=O CIDICGYKRUTYLE-FXQIFTODSA-N 0.000 description 1
- SOAYQFDWEIWPPR-IHRRRGAJSA-N Met-Ser-Tyr Chemical compound [H]N[C@@H](CCSC)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O SOAYQFDWEIWPPR-IHRRRGAJSA-N 0.000 description 1
- YBAFDPFAUTYYRW-UHFFFAOYSA-N N-L-alpha-glutamyl-L-leucine Natural products CC(C)CC(C(O)=O)NC(=O)C(N)CCC(O)=O YBAFDPFAUTYYRW-UHFFFAOYSA-N 0.000 description 1
- SITLTJHOQZFJGG-UHFFFAOYSA-N N-L-alpha-glutamyl-L-valine Natural products CC(C)C(C(O)=O)NC(=O)C(N)CCC(O)=O SITLTJHOQZFJGG-UHFFFAOYSA-N 0.000 description 1
- XZFYRXDAULDNFX-UHFFFAOYSA-N N-L-cysteinyl-L-phenylalanine Natural products SCC(N)C(=O)NC(C(O)=O)CC1=CC=CC=C1 XZFYRXDAULDNFX-UHFFFAOYSA-N 0.000 description 1
- 108010066427 N-valyltryptophan Proteins 0.000 description 1
- LGBVMDMZZFYSFW-HJWJTTGWSA-N Phe-Arg-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)O)NC(=O)[C@H](CCCN=C(N)N)NC(=O)[C@H](CC1=CC=CC=C1)N LGBVMDMZZFYSFW-HJWJTTGWSA-N 0.000 description 1
- HXSUFWQYLPKEHF-IHRRRGAJSA-N Phe-Asn-Arg Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)N[C@@H](CCCN=C(N)N)C(=O)O)N HXSUFWQYLPKEHF-IHRRRGAJSA-N 0.000 description 1
- DJPXNKUDJKGQEE-BZSNNMDCSA-N Phe-Asp-Phe Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O DJPXNKUDJKGQEE-BZSNNMDCSA-N 0.000 description 1
- NKLDZIPTGKBDBB-HTUGSXCWSA-N Phe-Gln-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](CCC(=O)N)NC(=O)[C@H](CC1=CC=CC=C1)N)O NKLDZIPTGKBDBB-HTUGSXCWSA-N 0.000 description 1
- XMQSOOJRRVEHRO-ULQDDVLXSA-N Phe-Leu-Arg Chemical compound NC(N)=NCCC[C@@H](C(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@@H](N)CC1=CC=CC=C1 XMQSOOJRRVEHRO-ULQDDVLXSA-N 0.000 description 1
- RSPUIENXSJYZQO-JYJNAYRXSA-N Phe-Leu-Gln Chemical compound NC(=O)CC[C@@H](C(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@@H](N)CC1=CC=CC=C1 RSPUIENXSJYZQO-JYJNAYRXSA-N 0.000 description 1
- OKQQWSNUSQURLI-JYJNAYRXSA-N Phe-Met-Val Chemical compound CC(C)[C@@H](C(=O)O)NC(=O)[C@H](CCSC)NC(=O)[C@H](CC1=CC=CC=C1)N OKQQWSNUSQURLI-JYJNAYRXSA-N 0.000 description 1
- XOHJOMKCRLHGCY-UNQGMJICSA-N Phe-Pro-Thr Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N1CCC[C@H]1C(=O)N[C@@H]([C@@H](C)O)C(O)=O XOHJOMKCRLHGCY-UNQGMJICSA-N 0.000 description 1
- IPFXYNKCXYGSSV-KKUMJFAQSA-N Phe-Ser-Lys Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)N[C@@H](CO)C(=O)N[C@@H](CCCCN)C(=O)O)N IPFXYNKCXYGSSV-KKUMJFAQSA-N 0.000 description 1
- GMWNQSGWWGKTSF-LFSVMHDDSA-N Phe-Thr-Ala Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C)C(O)=O GMWNQSGWWGKTSF-LFSVMHDDSA-N 0.000 description 1
- ORPZXBQTEHINPB-SRVKXCTJSA-N Pro-Arg-Val Chemical compound CC(C)[C@H](NC(=O)[C@H](CCCNC(N)=N)NC(=O)[C@@H]1CCCN1)C(O)=O ORPZXBQTEHINPB-SRVKXCTJSA-N 0.000 description 1
- FISHYTLIMUYTQY-GUBZILKMSA-N Pro-Gln-Gln Chemical compound NC(=O)CC[C@@H](C(O)=O)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@@H]1CCCN1 FISHYTLIMUYTQY-GUBZILKMSA-N 0.000 description 1
- DIFXZGPHVCIVSQ-CIUDSAMLSA-N Pro-Gln-Ser Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CO)C(O)=O DIFXZGPHVCIVSQ-CIUDSAMLSA-N 0.000 description 1
- BRJGUPWVFXKBQI-XUXIUFHCSA-N Pro-Leu-Ile Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O BRJGUPWVFXKBQI-XUXIUFHCSA-N 0.000 description 1
- XYSXOCIWCPFOCG-IHRRRGAJSA-N Pro-Leu-Leu Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(C)C)C(O)=O XYSXOCIWCPFOCG-IHRRRGAJSA-N 0.000 description 1
- BARPGRUZBKFJMA-SRVKXCTJSA-N Pro-Met-Arg Chemical compound CSCC[C@@H](C(=O)N[C@@H](CCCN=C(N)N)C(=O)O)NC(=O)[C@@H]1CCCN1 BARPGRUZBKFJMA-SRVKXCTJSA-N 0.000 description 1
- IIRBTQHFVNGPMQ-AVGNSLFASA-N Pro-Val-Lys Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CCCCN)C(=O)O)NC(=O)[C@@H]1CCCN1 IIRBTQHFVNGPMQ-AVGNSLFASA-N 0.000 description 1
- 101800001295 Putative ATP-dependent helicase Proteins 0.000 description 1
- 101800001006 Putative helicase Proteins 0.000 description 1
- ZUGXSSFMTXKHJS-ZLUOBGJFSA-N Ser-Ala-Ala Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](C)C(=O)N[C@@H](C)C(O)=O ZUGXSSFMTXKHJS-ZLUOBGJFSA-N 0.000 description 1
- BRKHVZNDAOMAHX-BIIVOSGPSA-N Ser-Ala-Pro Chemical compound C[C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](CO)N BRKHVZNDAOMAHX-BIIVOSGPSA-N 0.000 description 1
- VAUMZJHYZQXZBQ-WHFBIAKZSA-N Ser-Asn-Gly Chemical compound OC[C@H](N)C(=O)N[C@@H](CC(N)=O)C(=O)NCC(O)=O VAUMZJHYZQXZBQ-WHFBIAKZSA-N 0.000 description 1
- GHPQVUYZQQGEDA-BIIVOSGPSA-N Ser-Asp-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CC(=O)O)NC(=O)[C@H](CO)N)C(=O)O GHPQVUYZQQGEDA-BIIVOSGPSA-N 0.000 description 1
- OHKFXGKHSJKKAL-NRPADANISA-N Ser-Glu-Val Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](C(C)C)C(O)=O OHKFXGKHSJKKAL-NRPADANISA-N 0.000 description 1
- AEGUWTFAQQWVLC-BQBZGAKWSA-N Ser-Gly-Arg Chemical compound [H]N[C@@H](CO)C(=O)NCC(=O)N[C@@H](CCCNC(N)=N)C(O)=O AEGUWTFAQQWVLC-BQBZGAKWSA-N 0.000 description 1
- GJFYFGOEWLDQGW-GUBZILKMSA-N Ser-Leu-Gln Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CCC(=O)N)C(=O)O)NC(=O)[C@H](CO)N GJFYFGOEWLDQGW-GUBZILKMSA-N 0.000 description 1
- MUJQWSAWLLRJCE-KATARQTJSA-N Ser-Leu-Thr Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)O)C(O)=O MUJQWSAWLLRJCE-KATARQTJSA-N 0.000 description 1
- XUDRHBPSPAPDJP-SRVKXCTJSA-N Ser-Lys-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](CCCCN)NC(=O)[C@@H](N)CO XUDRHBPSPAPDJP-SRVKXCTJSA-N 0.000 description 1
- FPCGZYMRFFIYIH-CIUDSAMLSA-N Ser-Lys-Ser Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CO)C(O)=O FPCGZYMRFFIYIH-CIUDSAMLSA-N 0.000 description 1
- MQUZANJDFOQOBX-SRVKXCTJSA-N Ser-Phe-Ser Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CO)C(O)=O MQUZANJDFOQOBX-SRVKXCTJSA-N 0.000 description 1
- KQNDIKOYWZTZIX-FXQIFTODSA-N Ser-Ser-Arg Chemical compound OC[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@H](C(O)=O)CCCNC(N)=N KQNDIKOYWZTZIX-FXQIFTODSA-N 0.000 description 1
- ATEQEHCGZKBEMU-GQGQLFGLSA-N Ser-Trp-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)O)NC(=O)[C@H](CC1=CNC2=CC=CC=C21)NC(=O)[C@H](CO)N ATEQEHCGZKBEMU-GQGQLFGLSA-N 0.000 description 1
- HSWXBJCBYSWBPT-GUBZILKMSA-N Ser-Val-Val Chemical compound CC(C)[C@H](NC(=O)[C@@H](NC(=O)[C@@H](N)CO)C(C)C)C(O)=O HSWXBJCBYSWBPT-GUBZILKMSA-N 0.000 description 1
- 102100028910 Sodium channel protein type 1 subunit alpha Human genes 0.000 description 1
- TWLMXDWFVNEFFK-FJXKBIBVSA-N Thr-Arg-Gly Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)NCC(O)=O TWLMXDWFVNEFFK-FJXKBIBVSA-N 0.000 description 1
- GKMYGVQDGVYCPC-IUKAMOBKSA-N Thr-Asp-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)O)NC(=O)[C@H](CC(=O)O)NC(=O)[C@H]([C@@H](C)O)N GKMYGVQDGVYCPC-IUKAMOBKSA-N 0.000 description 1
- ZQUKYJOKQBRBCS-GLLZPBPUSA-N Thr-Gln-Gln Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CCC(=O)N)C(=O)N[C@@H](CCC(=O)N)C(=O)O)N)O ZQUKYJOKQBRBCS-GLLZPBPUSA-N 0.000 description 1
- KCRQEJSKXAIULJ-FJXKBIBVSA-N Thr-Gly-Arg Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)NCC(=O)N[C@@H](CCCNC(N)=N)C(O)=O KCRQEJSKXAIULJ-FJXKBIBVSA-N 0.000 description 1
- IMDMLDSVUSMAEJ-HJGDQZAQSA-N Thr-Leu-Asn Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(N)=O)C(O)=O IMDMLDSVUSMAEJ-HJGDQZAQSA-N 0.000 description 1
- KKPOGALELPLJTL-MEYUZBJRSA-N Thr-Lys-Tyr Chemical compound C[C@@H](O)[C@H](N)C(=O)N[C@@H](CCCCN)C(=O)N[C@H](C(O)=O)CC1=CC=C(O)C=C1 KKPOGALELPLJTL-MEYUZBJRSA-N 0.000 description 1
- DXPURPNJDFCKKO-RHYQMDGZSA-N Thr-Lys-Val Chemical compound CC(C)[C@H](NC(=O)[C@H](CCCCN)NC(=O)[C@@H](N)[C@@H](C)O)C(O)=O DXPURPNJDFCKKO-RHYQMDGZSA-N 0.000 description 1
- CGCMNOIQVAXYMA-UNQGMJICSA-N Thr-Met-Phe Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O CGCMNOIQVAXYMA-UNQGMJICSA-N 0.000 description 1
- KPNSNVTUVKSBFL-ZJDVBMNYSA-N Thr-Met-Thr Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CCSC)C(=O)N[C@@H]([C@@H](C)O)C(=O)O)N)O KPNSNVTUVKSBFL-ZJDVBMNYSA-N 0.000 description 1
- WNQJTLATMXYSEL-OEAJRASXSA-N Thr-Phe-Leu Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CC(C)C)C(O)=O WNQJTLATMXYSEL-OEAJRASXSA-N 0.000 description 1
- DOBIBIXIHJKVJF-XKBZYTNZSA-N Thr-Ser-Gln Chemical compound C[C@@H](O)[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@H](C(O)=O)CCC(N)=O DOBIBIXIHJKVJF-XKBZYTNZSA-N 0.000 description 1
- IQPWNQRRAJHOKV-KATARQTJSA-N Thr-Ser-Lys Chemical compound C[C@@H](O)[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@H](C(O)=O)CCCCN IQPWNQRRAJHOKV-KATARQTJSA-N 0.000 description 1
- NHQVWACSJZJCGJ-FLBSBUHZSA-N Thr-Thr-Ile Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O NHQVWACSJZJCGJ-FLBSBUHZSA-N 0.000 description 1
- VIWQOOBRKCGSDK-RYQLBKOJSA-N Trp-Arg-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CCCN=C(N)N)NC(=O)[C@H](CC2=CNC3=CC=CC=C32)N)C(=O)O VIWQOOBRKCGSDK-RYQLBKOJSA-N 0.000 description 1
- NOFFAYIYPAUNRM-HKUYNNGSSA-N Trp-Gly-Phe Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)O)NC(=O)CNC(=O)[C@H](CC2=CNC3=CC=CC=C32)N NOFFAYIYPAUNRM-HKUYNNGSSA-N 0.000 description 1
- DANHCMVVXDXOHN-SRVKXCTJSA-N Tyr-Asp-Asn Chemical compound NC(=O)C[C@@H](C(O)=O)NC(=O)[C@H](CC(O)=O)NC(=O)[C@@H](N)CC1=CC=C(O)C=C1 DANHCMVVXDXOHN-SRVKXCTJSA-N 0.000 description 1
- AZGZDDNKFFUDEH-QWRGUYRKSA-N Tyr-Gly-Ser Chemical compound OC[C@@H](C(O)=O)NC(=O)CNC(=O)[C@@H](N)CC1=CC=C(O)C=C1 AZGZDDNKFFUDEH-QWRGUYRKSA-N 0.000 description 1
- BBSPTGPYIPGTKH-JYJNAYRXSA-N Tyr-Met-Arg Chemical compound CSCC[C@@H](C(=O)N[C@@H](CCCN=C(N)N)C(=O)O)NC(=O)[C@H](CC1=CC=C(C=C1)O)N BBSPTGPYIPGTKH-JYJNAYRXSA-N 0.000 description 1
- SOAUMCDLIUGXJJ-SRVKXCTJSA-N Tyr-Ser-Asn Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(N)=O)C(O)=O SOAUMCDLIUGXJJ-SRVKXCTJSA-N 0.000 description 1
- AOIZTZRWMSPPAY-KAOXEZKKSA-N Tyr-Thr-Pro Chemical compound C[C@H]([C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](CC2=CC=C(C=C2)O)N)O AOIZTZRWMSPPAY-KAOXEZKKSA-N 0.000 description 1
- WQOHKVRQDLNDIL-YJRXYDGGSA-N Tyr-Thr-Ser Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CO)C(O)=O WQOHKVRQDLNDIL-YJRXYDGGSA-N 0.000 description 1
- COYSIHFOCOMGCF-WPRPVWTQSA-N Val-Arg-Gly Chemical compound CC(C)[C@H](N)C(=O)N[C@H](C(=O)NCC(O)=O)CCCN=C(N)N COYSIHFOCOMGCF-WPRPVWTQSA-N 0.000 description 1
- COYSIHFOCOMGCF-UHFFFAOYSA-N Val-Arg-Gly Natural products CC(C)C(N)C(=O)NC(C(=O)NCC(O)=O)CCCN=C(N)N COYSIHFOCOMGCF-UHFFFAOYSA-N 0.000 description 1
- BMGOFDMKDVVGJG-NHCYSSNCSA-N Val-Asp-Lys Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)N[C@@H](CCCCN)C(=O)O)N BMGOFDMKDVVGJG-NHCYSSNCSA-N 0.000 description 1
- IWZYXFRGWKEKBJ-GVXVVHGQSA-N Val-Gln-His Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CCC(=O)N)C(=O)N[C@@H](CC1=CN=CN1)C(=O)O)N IWZYXFRGWKEKBJ-GVXVVHGQSA-N 0.000 description 1
- AHHJARQXFFGOKF-NRPADANISA-N Val-Glu-Cys Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CCC(=O)O)C(=O)N[C@@H](CS)C(=O)O)N AHHJARQXFFGOKF-NRPADANISA-N 0.000 description 1
- XWYUBUYQMOUFRQ-IFFSRLJSSA-N Val-Glu-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](CCC(=O)O)NC(=O)[C@H](C(C)C)N)O XWYUBUYQMOUFRQ-IFFSRLJSSA-N 0.000 description 1
- WFENBJPLZMPVAX-XVKPBYJWSA-N Val-Gly-Glu Chemical compound CC(C)[C@H](N)C(=O)NCC(=O)N[C@H](C(O)=O)CCC(O)=O WFENBJPLZMPVAX-XVKPBYJWSA-N 0.000 description 1
- FEFZWCSXEMVSPO-LSJOCFKGSA-N Val-His-Ala Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](Cc1cnc[nH]1)C(=O)N[C@@H](C)C(O)=O FEFZWCSXEMVSPO-LSJOCFKGSA-N 0.000 description 1
- PTFPUAXGIKTVNN-ONGXEEELSA-N Val-His-Gly Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC1=CN=CN1)C(=O)NCC(=O)O)N PTFPUAXGIKTVNN-ONGXEEELSA-N 0.000 description 1
- HGJRMXOWUWVUOA-GVXVVHGQSA-N Val-Leu-Gln Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CCC(=O)N)C(=O)O)NC(=O)[C@H](C(C)C)N HGJRMXOWUWVUOA-GVXVVHGQSA-N 0.000 description 1
- KRAHMIJVUPUOTQ-DCAQKATOSA-N Val-Ser-His Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CO)C(=O)N[C@@H](CC1=CN=CN1)C(=O)O)N KRAHMIJVUPUOTQ-DCAQKATOSA-N 0.000 description 1
- VHIZXDZMTDVFGX-DCAQKATOSA-N Val-Ser-Leu Chemical compound CC(C)C[C@@H](C(=O)O)NC(=O)[C@H](CO)NC(=O)[C@H](C(C)C)N VHIZXDZMTDVFGX-DCAQKATOSA-N 0.000 description 1
- GVNLOVJNNDZUHS-RHYQMDGZSA-N Val-Thr-Lys Chemical compound [H]N[C@@H](C(C)C)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCCCN)C(O)=O GVNLOVJNNDZUHS-RHYQMDGZSA-N 0.000 description 1
- PMKQKNBISAOSRI-XHSDSOJGSA-N Val-Tyr-Pro Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC1=CC=C(C=C1)O)C(=O)N2CCC[C@@H]2C(=O)O)N PMKQKNBISAOSRI-XHSDSOJGSA-N 0.000 description 1
- BGTDGENDNWGMDQ-KJEVXHAQSA-N Val-Tyr-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](CC1=CC=C(C=C1)O)NC(=O)[C@H](C(C)C)N)O BGTDGENDNWGMDQ-KJEVXHAQSA-N 0.000 description 1
- 241000209149 Zea Species 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 230000006978 adaptation Effects 0.000 description 1
- 108010041407 alanylaspartic acid Proteins 0.000 description 1
- 108010087924 alanylproline Proteins 0.000 description 1
- 150000001413 amino acids Chemical group 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 108010013835 arginine glutamate Proteins 0.000 description 1
- 108010038850 arginyl-isoleucyl-tyrosine Proteins 0.000 description 1
- 108010018691 arginyl-threonyl-arginine Proteins 0.000 description 1
- 108010077245 asparaginyl-proline Proteins 0.000 description 1
- 108010069205 aspartyl-phenylalanine Proteins 0.000 description 1
- 108010047857 aspartylglycine Proteins 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 101150103518 bar gene Proteins 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000033228 biological regulation Effects 0.000 description 1
- 235000013339 cereals Nutrition 0.000 description 1
- 238000012512 characterization method Methods 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 238000004140 cleaning Methods 0.000 description 1
- 238000003501 co-culture Methods 0.000 description 1
- 239000012881 co-culture medium Substances 0.000 description 1
- 239000002299 complementary DNA Substances 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 230000007123 defense Effects 0.000 description 1
- 230000008260 defense mechanism Effects 0.000 description 1
- 230000001419 dependent effect Effects 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 230000004069 differentiation Effects 0.000 description 1
- 238000007865 diluting Methods 0.000 description 1
- 239000012154 double-distilled water Substances 0.000 description 1
- 230000002222 downregulating effect Effects 0.000 description 1
- 210000002257 embryonic structure Anatomy 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 238000007710 freezing Methods 0.000 description 1
- 230000008014 freezing Effects 0.000 description 1
- 230000030279 gene silencing Effects 0.000 description 1
- 108010075431 glycyl-alanyl-phenylalanine Proteins 0.000 description 1
- 108010067216 glycyl-glycyl-glycine Proteins 0.000 description 1
- XKUKSGPZAADMRA-UHFFFAOYSA-N glycyl-glycyl-glycine Natural products NCC(=O)NCC(=O)NCC(O)=O XKUKSGPZAADMRA-UHFFFAOYSA-N 0.000 description 1
- 108010033719 glycyl-histidyl-glycine Proteins 0.000 description 1
- 108010050475 glycyl-leucyl-tyrosine Proteins 0.000 description 1
- 108010010147 glycylglutamine Proteins 0.000 description 1
- 108010050848 glycylleucine Proteins 0.000 description 1
- 108010015792 glycyllysine Proteins 0.000 description 1
- 108010081551 glycylphenylalanine Proteins 0.000 description 1
- XDDAORKBJWWYJS-UHFFFAOYSA-N glyphosate Chemical compound OC(=O)CNCP(O)(O)=O XDDAORKBJWWYJS-UHFFFAOYSA-N 0.000 description 1
- 229940097068 glyphosate Drugs 0.000 description 1
- 238000000227 grinding Methods 0.000 description 1
- 229960001867 guaiacol Drugs 0.000 description 1
- 238000003306 harvesting Methods 0.000 description 1
- 238000005286 illumination Methods 0.000 description 1
- 230000001771 impaired effect Effects 0.000 description 1
- 230000006698 induction Effects 0.000 description 1
- 108010030617 leucyl-phenylalanyl-valine Proteins 0.000 description 1
- 108010025153 lysyl-alanyl-alanine Proteins 0.000 description 1
- 108010064235 lysylglycine Proteins 0.000 description 1
- 108010038320 lysylphenylalanine Proteins 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 230000001404 mediated effect Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 101150113790 nlr gene Proteins 0.000 description 1
- 230000001717 pathogenic effect Effects 0.000 description 1
- COLNVLDHVKWLRT-UHFFFAOYSA-N phenylalanine Natural products OC(=O)C(N)CC1=CC=CC=C1 COLNVLDHVKWLRT-UHFFFAOYSA-N 0.000 description 1
- 108010073025 phenylalanylphenylalanine Proteins 0.000 description 1
- 108010051242 phenylalanylserine Proteins 0.000 description 1
- 150000003904 phospholipids Chemical class 0.000 description 1
- 238000013081 phylogenetic analysis Methods 0.000 description 1
- 238000003976 plant breeding Methods 0.000 description 1
- 239000008057 potassium phosphate buffer Substances 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 108010031719 prolyl-serine Proteins 0.000 description 1
- 238000004445 quantitative analysis Methods 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 108020003175 receptors Proteins 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 238000010839 reverse transcription Methods 0.000 description 1
- 239000012883 rooting culture medium Substances 0.000 description 1
- 108010069117 seryl-lysyl-aspartic acid Proteins 0.000 description 1
- 238000002791 soaking Methods 0.000 description 1
- 238000005507 spraying Methods 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
- 108010072986 threonyl-seryl-lysine Proteins 0.000 description 1
- 230000009466 transformation Effects 0.000 description 1
- 108700026220 vif Genes Proteins 0.000 description 1
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/415—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from plants
-
- 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)
- C12N15/8201—Methods for introducing genetic material into plant cells, e.g. DNA, RNA, stable or transient incorporation, tissue culture methods adapted for transformation
- C12N15/8202—Methods for introducing genetic material into plant cells, e.g. DNA, RNA, stable or transient incorporation, tissue culture methods adapted for transformation by biological means, e.g. cell mediated or natural vector
- C12N15/8205—Agrobacterium mediated transformation
-
- 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)
- C12N15/8216—Methods for controlling, regulating or enhancing expression of transgenes in plant cells
- C12N15/8218—Antisense, co-suppression, viral induced gene silencing [VIGS], post-transcriptional induced gene silencing [PTGS]
-
- 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
- C12N15/8279—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 biotic stress resistance, pathogen resistance, disease resistance
- C12N15/8282—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 biotic stress resistance, pathogen resistance, disease resistance for fungal 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
- C12N9/14—Hydrolases (3)
- C12N9/16—Hydrolases (3) acting on ester bonds (3.1)
- C12N9/22—Ribonucleases RNAses, DNAses
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Genetics & Genomics (AREA)
- Engineering & Computer Science (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Biomedical Technology (AREA)
- Molecular Biology (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biotechnology (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Biophysics (AREA)
- Microbiology (AREA)
- Plant Pathology (AREA)
- Physics & Mathematics (AREA)
- Cell Biology (AREA)
- Medicinal Chemistry (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Gastroenterology & Hepatology (AREA)
- Botany (AREA)
- Virology (AREA)
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
Abstract
本发明提出了一种增强玉米大斑病抗性的基因ZmBON3及其应用,属于基因工程技术领域,该增强玉米大斑病抗性的基因ZmBON3的核苷酸序列如SEQ ID NO.1所示,在提高植物抗病性能中的应用。本发明提供了一种玉米大斑病抗性基因ZmBON3及其应用,该基作为一种新的植物抗病相关基因,该基因的发现丰富了新的抗病与种子基因资源,对进一步了提高玉米的抗病能力,对生产中新种质资源的培育有着重要的实际意义。
Description
技术领域
本发明涉及基因工程技术领域,具体涉及一种增强玉米大斑病抗性的基因ZmBON3及其应用。
背景技术
玉米(Zea mays L.)是世界种植面积与种植产量名列前茅的粮食作物,也是重要的饲料作物与工业原料作物。据我国国家统计局公开的数据显示,2019年我国玉米的种植面积已达41284千公顷,产量总达26077万吨,位居粮食总量首位。玉米生长过程中将受到多种不利生长的因素胁迫影响,其中,病害是影响玉米产量的重要因素。玉米大斑病是最常出现的玉米病害之一,其发病时,可造成20%左右的减产,当发病大爆发时,减产程度可上升至46%。因此,研究玉米抗病性的提升,以及相关基因的功能与抗病新种植的研创对提升玉米产量具有重要的研究价值与实际意义。
Copine基因可编码一类Ca2+依赖性的磷脂膜结合蛋白,广泛分布在真核生物中。在拟南芥的研究中发现,Copine基因AtBON1通过负调控一个NLR基因 SCN1从而行使功能,下调拟南芥对于病原菌的抵抗能力。一些研究中沉默或下调Copine基因,结果植株的表型中都具备对营养型或半营养型病原菌增强的抵抗力,这证明Copine基因是一种改良植物抗病性的潜在基因。目前尚未有研究报道 ZmBON3基因在玉米抗病反应,尤其是对玉米大斑病抗性应用的功能,因此,研究 ZmBON3基因并创制相关抗病材料新种质对进一步了解植物抗病活动机制具有深远意义,也能够为分子水平提高作物抗病性能提供更多遗传资源信息。
专利CN111153974A公开了一种玉米大斑病、南方锈病、灰斑病的抗性基因 ZmMM1,源于大刍草的抗病单倍型qLMC117和与之连锁的分子标记M2及其在筛选和培育玉米大斑病或南方锈病或灰斑病抗性品种中的应用,属于分子遗传学领域。发明公开了玉米大斑病、南方锈病、灰斑病抗性基因ZmMM1,同时,发明还公开了一个源于大刍草的抗病单倍型qLMC117(SEQ ID NO.5)以及与其紧密连锁的分子标记M2(SEQ ID NO.10)以及该标记的检测引物对(SEQ ID NO.11和 SEQ ID NO.12)和检测方法。提供了利用分子标记筛选培育玉米大斑病、南方锈病、灰斑病抗性品种的方法。
专利CN109705200A公开了一种灰斑病抗性相关蛋白ZmWAK-RLK及其编码基因和应用。发明提供的蛋白质,获自玉米自交系Y32,命名为ZmWAK-RLK蛋白,为序列表中序列1所示的蛋白质。发明还保护一种植物育种方法,包括如下步骤:增加目的植物中ZmWAK-RLK蛋白的含量和/或活性,从而提高目的植物的灰斑病抗病性。对于玉米的抗灰斑病育种具有重大的应用价值。
但上述方法操作复杂,应用效果不佳。
发明内容
本发明的目的在于提出一种增强玉米大斑病抗性的基因ZmBON3及其应用,以及提供一种新的可用于植物抗病遗传改良的基因。
本发明的技术方案是这样实现的:
本发明提供一种增强玉米大斑病抗性的基因ZmBON3,其核苷酸序列如SEQ IDNO.1所示。
本发明进一步保护上述增强玉米大斑病抗性的基因ZmBON3在提高植物抗病性能中的应用,其是以CRISPR/Cas9基因编辑方法,定点编辑受体材料中的 ZmBON3基因,使其基因功能缺失以至得到目的功能植株。
作为本发明的进一步改进,上述CRISPR/Cas9方法所使用的sgRNA序列为5 ′-GTGCACGGTGACGTGCGCGG-3′和5′-GGATGGGCGTGTAGAGGCCG-3′。
进一步的,将所述两个sgRNA作用位点通过不同表达盒串联到同一载体上,所述载体为pCXB053。
本发明进一步保护上述增强玉米对玉米大斑病抗性基因ZmBON3在玉米种质资源改良中的应用。
本发明进一步保护上述增强玉米对玉米大斑病抗性基因ZmBON3在制备抗病转基因玉米中的应用。
本发明进一步保护一种上述增强玉米大斑病抗性的基因ZmBON3的编码蛋白,其氨基酸序列如SEQ ID NO.2所示。
本发明进一步保护一种含有依据上述玉米对大斑病抗性基因ZmBON3设计的靶点序列1,其核苷酸序列如SEQ ID NO.3所示。
本发明进一步保护一种含有依据上述增强玉米大斑病抗性的基因ZmBON3 设计的靶点序列2,其核苷酸序列如SEQ ID NO.4所示。
本发明进一步保护一种含有上述增强玉米大斑病抗性的基因ZmBON3的生物材料,其特征在于,所述生物材料为表达盒、载体、工程菌或细胞。
作为本发明的进一步改进,所述载体为pCXB053载体,所述pCXB053载体的多克隆位点区域依次连接U6启动子,如权利要求5所述靶点序列1,U6启动子,如权利要求6所述靶点序列2。
作为本发明的进一步改进,所述细胞为宿主细胞,所述宿主细胞含有如权利要求7所述载体,在如权利要求1所述的增强玉米大斑病抗性的基因ZmBON3 位点处发生基因编辑。
本发明进一步保护一种一种提高植物抗病性能的方法,将上述载体导入到植物的细胞、组织和器官中,并编辑设计靶点,使其基因功能损伤。
作为本发明的进一步改进,所述植物包括玉米、水稻、小麦、拟南芥。
本发明具有如下有益效果:本发明提供了一种玉米大斑病抗性基因ZmBON3 及其应用,该基作为一种新的植物抗病相关基因,该基因的发现丰富了新的抗病与种子基因资源,对进一步了提高玉米的抗病能力,对生产中新种质资源的培育有着重要的实际意义。
附图说明
为了更清楚地说明本发明实施例或现有技术中的技术方案,下面将对实施例或现有技术描述中所需要使用的附图作简单地介绍,显而易见地,下面描述中的附图仅仅是本发明的一些实施例,对于本领域普通技术人员来讲,在不付出创造性劳动性的前提下,还可以根据这些附图获得其他的附图。
图1为ZmBON3基因与其他物种Copine基因的进化树分析图;
图2为ZmBON3基因CRISPR/Cas9载体图谱;
图3为ZmBON3基因编辑植株编辑位点突变类型示意图;
图4为ZmBON3基因编辑植株的荧光定量检测结果柱形图;
图5为玉米大斑病菌接种处理后5天内ZmBON3基因编辑植株和野生型H2O2含量的变化柱形图;
图6为玉米大斑病菌接种处理后5天内ZmBON3基因编辑植株和野生型POD 含量的变化柱形图;
图7为玉米大斑病菌接种处理后5天内ZmBON3基因编辑植株和野生型CAT 含量的变化柱形图;
图8为玉米大斑病菌接种处理后5天内ZmBON3基因编辑植株和野生型PAL 含量的变化柱形图。
具体实施方式
下面将对本发明实施例中的技术方案进行清楚、完整地描述,显然,所描述的实施例仅仅是本发明一部分实施例,而不是全部的实施例。基于本发明中的实施例,本领域普通技术人员在没有做出创造性劳动前提下所获得的所有其他实施例,都属于本发明保护的范围。
实施例所用方法如无特别说明均为本领域的技术人员所知晓的常规方法,所用的试剂等材料,如无特别说明,均为市售购买产品。
实施例1ZmBON3基因的预测
1、基于HMMER筛选玉米Copine家族基因
在Pfam上下载Copine家族的结构域序列(pfam:PF07002),利用Hmmer软件通过hmmsearch功能于玉米蛋白质序列库 (https://ftp.ncbi.nlm.nih.gov/genomes/all/GCF/902/167/145/GCF_902167 145.1_Zm-B73-REFERENCE-NAM-5.0/GCF_902167145.1_Zm-B73-REFERENCE-NAM-5.0 _protein.faa.gz)中同源比对蛋白质序列,进行数据冗余清洗后,剩下结构域完整的两条基因NP_001147038.1与NP_001167721.1。
2、基于系统进化分析ZmBON3基因
利用MAGA7.0对上述获得的基因与多物种间Copine基因进行系统进化树分析,方法选择邻间法(Neighbor-joining Method),bootstrap设定为1000,结果如图1所示,NP_001167721.1与AtBON3、OsBON3同属一个进化分支,将其命名为ZmBON3。
实施例2基因编辑载体的构建
在NCBI数据库中下载ZmBON3基因的CDS序列,将CDS序列递交至 http://crispor.tefor.net/网站,genome选择Zea mays-maize-NCBI GCF_0000050052(B73RefGen-v4),PAM选择20bp-NGG-Sp Cas9.spcas9-HF1. espcas9 1.1,选择评分高且不跨内含子的的靶点序列。根据筛选规则,获得序列 1(5’-GTGCACGGTGACGTGCGCGG-3’,如SEQID NO.3所示)和序列2(5’- GGATGGGCGTGTAGAGGCCG-3’,如SEQ ID NO.4所示),双靶点基因编辑载体 pCXB053-ZmBON3由未米生物科技(江苏)有限公司构建。
构建质粒经由质粒PCR鉴定,测序确认,载体图谱如图2所示。
注:质粒PCR测序引物:
C-BON3-F:5’-TCCCAGTCACGACGTTGTAA-3’
C-BON3-R:5’-TTCAAACAAGTGTGACAAAAA-3’
实施例3农杆菌介导的玉米愈伤遗传转化
利用冻融法将构建好的基因编辑载体pCXB053-ZmBON3转化至农杆菌EHA105 中。取长势茁壮的玉米自交系H99的未成熟幼胚,放于N6培养基上诱导愈伤组织,诱导出的愈伤组织放于黑暗中预培养三天。将愈伤组织收集到培养瓶中,以 OD=0.6-0.8的农杆菌悬浮液浸泡20min,期间每隔5min摇晃一次,结束后用滤纸吸干多余菌液,转移愈伤组织至共培养基上24℃黑暗培养3天。共培养结束后,将愈伤组织放置在草甘膦筛选培养基上,筛选培养两轮,每轮14天。将筛选培养抗性遇上转移至分化培养基上,25℃光诱导培养3周左右;将分化愈伤转移至生根培养基上至生根成苗,炼苗5天后移栽到小盆中,待适应后移栽到温室中,自交收种。
实施例4ZmBON3编辑植株的分子鉴定
1、转基因植株标记基因的PCR检测
以T1代转基因植株的叶片为材料提取植物总DNA,用Bar基因引物对:Bar-F: 5’-ATGAGCCCAGAACGACGCC-3’和Bar-R:5’-TCAAATCTCGGTGACGGGC-3’,进行PCR扩增,保留扩增结果为阳性的植株,自交收种。反应体系如表1所示。
表1 PCR扩增体系
组分 | 体积 |
Mix | 12.5μL |
Bar-F | 0.5μL |
Bar-R | 0.5μL |
DNA模板 | 1μL |
ddH<sub>2</sub>O | 50 25μL |
PCR反应程序为PCR扩增程序为95℃5min预变性,95℃30s变性,58℃35s 退火,72℃30s延伸,共30个循环。反应结束后,回收反应条带,进行测序鉴定。
2、转基因植株跨编辑位点的PCR检测
根据ZmBON3基因基因组序列靶点两侧上下100bp左右的位置设计横跨编辑位点的引物:target-F 5’-CTGTCTTTTGTCAGAGGTTAT-3’和target-R
5’--TTCTCTACATATAGCTGCCT3’。以T2代植物叶片提取的DNA为模板进行PCR 扩增体系同表1。PCR反应程序为PCR扩增程序为95℃5min预变性,95℃30s变性,58.4℃33s退火,72℃30s延伸,共30个循环。反应结束后,回收反应条带,进行测序鉴定。鉴定结果由DNAMAN软件进行比对。
测序结果显示,基因编辑载体在靶点位置引入了多个突变,结果如图3所示。
3、ZmBON3基因编辑植株荧光定量分析
提取阳性植株与野生型植株根、茎、叶部位的RNA,并反转录为cDNA作为反应模板进行荧光定量检测,内参基因为β-Actin基因,每组设三次平行重复。
注:荧光定量引物:
BON3F:5’-ACGCCCATCGAGTTGTCATT-3’
BON3R:5’-TTGGAATGGGACCAGCAGTC-3’
ActF:5’-TTGAGGTAGGATGAGACT-3’
ActR:5’-GGAGTGAAGCAGATGATT-3’
结果如图4所示,在ZmBON3基因编辑植株中,ZmBON3基因在各组织中的表达量均有显著降低,与野生型相比表达量照降低75%-80%。证明ZmBON3基因编辑植株中ZmBON3基因的功能受到损害。
4、ZmBON3基因编辑植株的抗病性分析
于人工气候室内种植检测为阳性的ZmBON3基因编辑植株与未转化的对照植株,待培养至6叶期后,喷雾法接种等体积的孢子悬浮液,接种后培养条件设置为温度25℃,光照照长度为16h,相对湿度调至85%,培养5天后采集样品,每组重复三次。
(1)H2O2含量的测定
以三氯乙酸法测定叶片中H2O2的含量。采集待测叶片2g,液氮速冻后,使用研磨机磨碎。向样品中转移2ml的三氯乙酸(浓度为0.1%),震荡混匀后于冰上静置8min,预冷离心机,待温度降至4℃后,离心10min。转移上清液至另一试管中,再加入0.25个上清液体积的磷酸钾缓冲液(10mol/L)和0.5个体积的KI 溶液(1mol/L),充分震荡以后待用。检测并记录待测液的OD390。标准曲线使用同浓度的三氯乙酸溶解20-200ng的H2O2绘制。
结果如图5所示,相比于野生型植株,ZmBON3基因编辑植株的H2O2含量更高,接菌后转基因株系和野生型植株的H2O2含量都有得到增加,但在上升幅度上, ZmBON3基因编辑植株高于野生型植株,图标中可以清晰看出,这种变化是极显著的,说明ZmBON3基因编辑植株中积累的H2O2含量较多,病菌抵抗程度增强。
(2)过氧化物酶(POD)含量变化的测定
取待叶片样本0.2克,冷冻研磨,加入质量体积比1:15加入磷酸缓冲液,震荡混匀,高速离心后转移上清液保存备用。将制备好的植物酶液0.5ml,以3ml 磷酸缓冲液稀释(100mmol/L)转移至离心管,向其中迅速添加1mlH2O2(浓度为 30%)以及1ml愈创木酚,于震荡离心机上充分混合,调零液使用制备的原始酶液。检测混合液在A470时吸光光度值的变化,酶活性计算为每分钟变化的吸光光度值。
POD酶含量的的计算公式:
式中,△OD470为反应时间内吸光值的变化;V总为酶液总体积(ml);M为样品重量;V1为测定的酶液体积(ml);t为反应时间(min)
结果如图6所示,在接菌后ZmBON3基因编辑植株中POD酶的含量迅速升高,与野生型相比变化达到极显著。说明野生型对病原体抗性弱于ZmBON3基因编辑植株。
(3)过氧化氢酶(CAT)酶含量变化的测定
取新鲜玉米叶片0.2g剪碎,剪碎的材料充分冷冻,经物理磨成粉状后,以质量体积比1:15加入磷酸缓冲液(100mmol),震荡混匀,预冷离心机,转速设置为4000rpm,离心10min后,转移上清液于新离心管,轻微离心后保存待用。
制备CAT反应预混液,将50mmol/L的磷酸缓冲液,0.1mol/L的H2O2以5:1 的体积混合,使用ddH2O作为空白管,调整分光光度计至240nm,向预混液中转移制备好的酶液,然后立即关盖度数,以30S为一个周期测定3min内吸光度的变化,以每克鲜样每分钟吸光值变化0.01为一个酶活力单位。
CAT酶含量的的计算公式:
公式中,△A为每分钟吸光度值的变化;V总为酶液总体积(mL);V1为测定用酶液体积(mL);M为样品鲜重;T为反应时间(min)
结果如图7所示,ZmBON3基因编辑植株的CAT酶含量显著高于野生型植株,这种变化在接菌后更加明显。CAT酶活的变化可以反映出植物对H2O2的敏感性,ZmBON3基因编辑植株表现出的高酶活性证明ZmBON3基因编辑植株对病原体侵入的抗性增强。
(4)苯丙氨酸解氨酶(PAL)含量变化的测定
取待测植物叶片0.2g制备粗酶液,转移0.05ml制备好的酶液向0.1ml苯丙氨酸溶液中(浓度为0.6mmol/L),使用磷酸缓冲液(0.1mol/L)定容到0.5ml,反应60min后检测OD290。数据计算公式如下:
式中:N为酶液总体积(mL);T为反应时间(min);d为比色杯直径(cm);WF为样品鲜重(g);n为反应液所用酶液的体积(mL)。
结果如图8显示,在接菌后野生型与ZmBON3基因编辑植株的PAL酶含量均有提高,但ZmBON3基因编辑植株的提升幅度大大高于野生型,且相对于野生型 PAL酶含量有显著的提高。说明ZmBON3基因编辑植株中次生代谢产物活跃,对病菌的侵害抵抗增强。
通过对ZmBON3基因编辑植株的H2O2含量,POD酶含量,CAT酶含量与苯丙氨酸解氨酶含量这一系列防御生理生化指标测定可以得出,结果发现在玉米大斑病菌的接种下,ZmBON3基因编辑植株呈现出更高的抵抗性与防御次生代谢产物活跃度,进一步验证了基因与植物的抗病调控相关,通过基因编辑丧失ZmBON3基因功能的植株对玉米大斑病的抗性有显著提高,从而为玉米品种的基因工程改良提供了新的遗传资源。
与现有技术相比,本发明提供了一种玉米大斑病抗性基因ZmBON3及其应用,该基因为一种新的植物抗病相关基因,丰富了玉米抗病防御机制的研究,对进一步了提高玉米的抗病能力,对培育抗病新品种并解决玉米大班病害问题有着重要的实际意义。
以上所述仅为本发明的较佳实施例而已,并不用以限制本发明,凡在本发明的精神和原则之内,所作的任何修改、等同替换、改进等,均应包含在本发明的保护范围之内。
序列表
<110> 吉林农业大学
<120> 一种增强玉米大斑病抗性的基因ZmBON3及其应用
<160> 4
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1770
<212> DNA
<213> 玉米(Zea mays)
<400> 1
atgggtgggt gcttctccgt gcacggtgac gtgcgcggcg gcatggaggg cgtcggcggt 60
ggcgcgagga cccagggcgg cgcggcgtcg tcgtctgcgc agcagggcgg gccgaacgat 120
gcggtcgact atttcttcca gacgcgcggg ctccgcggcc tctacacgcc catcgagttg 180
tcattttcag catcgaagtt gcggaataca gattcctttt caaagagtga tcctatgtta 240
gtggtctaca ctaacatcaa aggaaagcta gaggagattg gccgtactga agtgatttta 300
aattcactgg agccctcatg gattacaaag gttacaatgt cttaccaatt tgagattgtg 360
caaccactca tattcaggat atatgatgtt gacacaaagt accacaacac tccagtgaag 420
acattgaatt tgacacagca ggattttctg ggggtagcat gctgcaactt atcagagatt 480
gtaacaaaat ttaaccgtag cctgacattg aatcttcaga gtgattgtgg acatggtctt 540
catggaacaa tgacagtaca tgctgaagaa agtgactcct ctagaatggc agttgagatg 600
actctgcgtt gcgtgaactt ggaaaataaa gatgtgttct ccaaaagtga tcccttctta 660
agaatatcta aactggtaga gactgctggt cccattccaa tctgcaaaac ggaagtagta 720
cctgacaact tgaaccctgt ttggaggcca atcactttaa catcgcagca gtatggaagc 780
aaggataacc cactgctagt tgagtgtttt gattttaatt ccagtggtga ccatgaactg 840
ataggggcct tccagacaac tattactcag cttgaaaatc tatatacctc aaagtctgca 900
gcaatttttt acaaccacaa gggacaaaga aagatgaaag aacaattgtt tgtggacaag 960
tttcaggaaa cagtccagca tactttctta gattatatat ccagtggatt tgagctcaat 1020
ttcatggtgg ctatagattt tactgcttca aatggagacc cacgtgtacc ccagtccttg 1080
cactacattg acccctctgg cagacctaat tcgtatcaac aggcaatgct gggggttggc 1140
gaagttctgc agttttatga caatgatagg cgatttccta cgtggggttt tggagcaagg 1200
gtacatggtt atgtatcaca ttgctttaac ttgaatacag ctacaaatga ctctgaggtg 1260
gttggagttg aaggcatcat gtcagcatac acctcttctc tttacagtgt ttcccttgct 1320
gggccaacta tgttcgggcc cgtgatcaac aaagctgcag atattgcaag ccagtccctt 1380
caatattcaa acaataaata ttttgtcctt ctcattatca cggacggagt tctcactgac 1440
atccaagaaa caaaagattg cattgtaagg gcatcagacc tgccattgtc gattctcatt 1500
gtaggcgttg gaaacgctga tttcaaacaa atggagatct tagacgcgga caatggtaag 1560
cgactggaaa gctctacggg taggatcgcg acacgcgaca tcgtccagtt tgttcccatg 1620
agggaagtcc aaggtggcga gatctccgtc gtgcaatccc ttctggagga actgcccggt 1680
cagttcctgc agtacatgcg cactcgggga atcaagccac agcgggctcc ggggcttgct 1740
tcagcgcccg tttacccacc tcagcagtga 1770
<210> 2
<211> 588
<212> PRT
<213> 玉米(Zea mays)
<400> 2
Met Gly Gly Cys Phe Ser Val His Gly Asp Val Arg Gly Gly Met Glu
1 5 10 15
Gly Val Gly Gly Gly Ala Arg Thr Gln Gly Gly Ala Ala Ser Ser Ser
20 25 30
Ala Gln Gln Gly Gly Pro Asn Asp Ala Val Asp Tyr Phe Phe Gln Thr
35 40 45
Arg Gly Leu Arg Gly Leu Tyr Thr Pro Ile Glu Leu Ser Phe Ser Ala
50 55 60
Ser Lys Leu Arg Asn Thr Asp Ser Phe Ser Lys Ser Asp Pro Met Leu
65 70 75 80
Val Val Tyr Thr Asn Ile Lys Gly Lys Leu Glu Glu Ile Gly Arg Thr
85 90 95
Glu Val Ile Leu Asn Ser Leu Glu Pro Ser Trp Ile Thr Lys Val Thr
100 105 110
Met Ser Tyr Gln Phe Glu Ile Val Gln Pro Leu Ile Phe Arg Ile Tyr
115 120 125
Asp Val Asp Thr Lys Tyr His Asn Thr Pro Val Lys Thr Leu Asn Leu
130 135 140
Thr Gln Gln Asp Phe Leu Gly Val Ala Cys Cys Asn Leu Ser Glu Ile
145 150 155 160
Val Thr Lys Phe Asn Arg Ser Leu Thr Leu Asn Leu Gln Ser Asp Cys
165 170 175
Gly His Gly Leu His Gly Thr Met Thr Val His Ala Glu Glu Ser Asp
180 185 190
Ser Ser Arg Met Ala Val Glu Met Thr Leu Cys Val Asn Leu Glu Asn
195 200 205
Lys Asp Val Phe Ser Lys Ser Asp Pro Phe Leu Arg Ile Ser Lys Leu
210 215 220
Val Glu Thr Ala Gly Pro Ile Pro Ile Cys Lys Thr Glu Val Val Pro
225 230 235 240
Asp Asn Leu Asn Pro Val Trp Arg Pro Ile Thr Leu Thr Ser Gln Gln
245 250 255
Tyr Gly Ser Lys Asp Asn Pro Leu Leu Val Glu Cys Phe Asp Phe Asn
260 265 270
Ser Ser Gly Asp His Glu Leu Ile Gly Ala Phe Gln Thr Thr Ile Thr
275 280 285
Gln Leu Glu Asn Leu Tyr Thr Ser Lys Ser Ala Ala Ile Phe Tyr Asn
290 295 300
His Lys Gly Gln Arg Lys Met Lys Glu Gln Leu Phe Val Asp Lys Phe
305 310 315 320
Gln Glu Thr Val Gln His Thr Phe Leu Asp Tyr Ile Ser Ser Gly Phe
325 330 335
Glu Leu Asn Phe Met Val Ala Ile Asp Phe Thr Ala Ser Asn Gly Asp
340 345 350
Pro Arg Val Pro Gln Ser Leu His Tyr Ile Asp Pro Ser Gly Arg Pro
355 360 365
Asn Ser Tyr Gln Gln Ala Met Leu Gly Val Gly Glu Val Leu Gln Phe
370 375 380
Tyr Asp Asn Asp Arg Arg Phe Pro Thr Trp Gly Phe Gly Ala Arg Val
385 390 395 400
His Gly Tyr Val Ser His Cys Phe Asn Leu Asn Thr Ala Thr Asn Asp
405 410 415
Ser Glu Val Val Gly Val Glu Gly Ile Met Ser Ala Tyr Thr Ser Ser
420 425 430
Leu Tyr Ser Val Ser Leu Ala Gly Pro Thr Met Phe Gly Pro Val Ile
435 440 445
Asn Lys Ala Ala Asp Ile Ala Ser Gln Ser Leu Gln Tyr Ser Asn Asn
450 455 460
Lys Tyr Phe Val Leu Leu Ile Ile Thr Asp Gly Val Leu Thr Asp Ile
465 470 475 480
Gln Glu Thr Lys Asp Cys Ile Val Arg Ala Ser Asp Leu Pro Leu Ser
485 490 495
Ile Leu Ile Val Gly Val Gly Asn Ala Asp Phe Lys Gln Met Glu Ile
500 505 510
Leu Asp Ala Asp Asn Gly Lys Arg Leu Glu Ser Ser Thr Gly Arg Ile
515 520 525
Ala Thr Arg Asp Ile Val Gln Phe Val Pro Met Arg Glu Val Gln Gly
530 535 540
Gly Glu Ile Ser Val Val Gln Ser Leu Leu Glu Glu Leu Pro Gly Gln
545 550 555 560
Phe Leu Gln Tyr Met Arg Thr Arg Gly Ile Lys Pro Gln Arg Ala Pro
565 570 575
Gly Leu Ala Ser Ala Pro Val Tyr Pro Pro Gln Gln
580 585
<210> 3
<211> 20
<212> DNA
<213> 玉米(Zea mays)
<400> 3
gtgcacggtg acgtgcgcgg 20
<210> 4
<211> 20
<212> DNA
<213> 玉米(Zea mays)
<400> 4
ggatgggcgt gtagaggccg 20
Claims (10)
1.一种增强玉米大斑病抗性的基因ZmBON3,其特征在于,其核苷酸序列如SEQ ID NO.1所示。
2.如权利要求1所述增强玉米大斑病抗性的基因ZmBON3在提高植物抗病性能中的应用。
3.一种如权利要求1所述增强玉米大斑病抗性的基因ZmBON3的编码蛋白,其特征在于,其氨基酸序列如SEQ ID NO.2所示。
4.含有依据权利要求1所述玉米对大斑病抗性基因ZmBON3设计的靶点序列1,其核苷酸序列如SEQ ID NO.3所示。
5.含有依据权利要求1所述增强玉米大斑病抗性的基因ZmBON3设计的靶点序列2,其核苷酸序列如SEQ ID NO.4所示。
6.含有权利要求1所述增强玉米大斑病抗性的基因ZmBON3的生物材料,其特征在于,所述生物材料为表达盒、载体、工程菌或细胞。
7.根据权利要求6所述的生物材料,其特征在于,所述载体为pCXB053载体,所述pCXB053载体的多克隆位点区域依次连接U6启动子,如权利要求5所述靶点序列1,U6启动子,如权利要求6所述靶点序列2。
8.根据权利要求6所述的生物材料,其特征在于,所述细胞为宿主细胞,所述宿主细胞含有如权利要求7所述载体,在如权利要求1所述的增强玉米大斑病抗性的基因ZmBON3位点处发生基因编辑。
9.一种提高植物抗病性能的方法,其特征在于,将权利要求7所述载体导入到植物的细胞、组织和器官中,并编辑设计靶点,使其基因功能损伤。
10.根据权利要求9所述的方法,其特征在于,所述植物包括玉米、水稻、小麦、拟南芥。
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202110826649.9A CN113444733A (zh) | 2021-07-21 | 2021-07-21 | 一种增强玉米大斑病抗性的基因ZmBON3及其应用 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202110826649.9A CN113444733A (zh) | 2021-07-21 | 2021-07-21 | 一种增强玉米大斑病抗性的基因ZmBON3及其应用 |
Publications (1)
Publication Number | Publication Date |
---|---|
CN113444733A true CN113444733A (zh) | 2021-09-28 |
Family
ID=77817012
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202110826649.9A Pending CN113444733A (zh) | 2021-07-21 | 2021-07-21 | 一种增强玉米大斑病抗性的基因ZmBON3及其应用 |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN113444733A (zh) |
Citations (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20050251880A1 (en) * | 2004-03-29 | 2005-11-10 | Bressan Ray A | Methods and compositions for regulating plant stress tolerance |
US20060021088A1 (en) * | 2002-10-18 | 2006-01-26 | Dirk Inze | Identification of novel e2f target genes and use thereof |
CN109385432A (zh) * | 2018-09-05 | 2019-02-26 | 南京农业大学 | 小麦copine基因TaBON1和TaBON3的抗病应用 |
CN110331145A (zh) * | 2019-08-05 | 2019-10-15 | 东北林业大学 | miR156及其相关生物材料在调控植物抗病性中的应用 |
CN112626107A (zh) * | 2020-12-25 | 2021-04-09 | 中国农业大学 | 两种植物基因编辑工具 |
US20210163979A1 (en) * | 2018-04-12 | 2021-06-03 | Evogene Ltd. | Polynucleotides and polypeptides of plant and bacterial origin for protecting plants from pathogenic fungi |
-
2021
- 2021-07-21 CN CN202110826649.9A patent/CN113444733A/zh active Pending
Patent Citations (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20060021088A1 (en) * | 2002-10-18 | 2006-01-26 | Dirk Inze | Identification of novel e2f target genes and use thereof |
US20050251880A1 (en) * | 2004-03-29 | 2005-11-10 | Bressan Ray A | Methods and compositions for regulating plant stress tolerance |
US20210163979A1 (en) * | 2018-04-12 | 2021-06-03 | Evogene Ltd. | Polynucleotides and polypeptides of plant and bacterial origin for protecting plants from pathogenic fungi |
CN109385432A (zh) * | 2018-09-05 | 2019-02-26 | 南京农业大学 | 小麦copine基因TaBON1和TaBON3的抗病应用 |
CN110331145A (zh) * | 2019-08-05 | 2019-10-15 | 东北林业大学 | miR156及其相关生物材料在调控植物抗病性中的应用 |
CN112626107A (zh) * | 2020-12-25 | 2021-04-09 | 中国农业大学 | 两种植物基因编辑工具 |
Non-Patent Citations (20)
Title |
---|
SCHNABLE PS等: "Zea mays uncharacterized LOC100381409 (LOC100381409), mRNA", 《GENBANK DATABASE》 * |
SCHNABLE PS等: "Zea mays uncharacterized LOC100381409 (LOC100381409), mRNA", 《GENBANK DATABASE》, 26 June 2021 (2021-06-26), pages 001174250 * |
YANG S等: "The BON/CPN gene family represses cell death and promotes cell growth in Arabidopsis", 《PLANT J》 * |
YANG S等: "The BON/CPN gene family represses cell death and promotes cell growth in Arabidopsis", 《PLANT J》, vol. 45, no. 2, 31 December 2006 (2006-12-31), pages 166 - 179 * |
YIN X等: "Rice copine genes OsBON1 and OsBON3 function as suppressors of broad-spectrum disease resistance", 《PLANT BIOTECHNOL J》 * |
YIN X等: "Rice copine genes OsBON1 and OsBON3 function as suppressors of broad-spectrum disease resistance", 《PLANT BIOTECHNOL J》, vol. 16, no. 8, 25 February 2018 (2018-02-25) * |
ZOU B等: "Identification and analysis of copine/BONZAI proteins among evolutionarily diverse plant species", 《GENOME》 * |
ZOU B等: "Identification and analysis of copine/BONZAI proteins among evolutionarily diverse plant species", 《GENOME》, vol. 59, no. 8, 21 January 2016 (2016-01-21), pages 565 - 73 * |
丁媛: "copine基因TaBON1和TaBON3在小麦白粉病抗性调控中的功能研究", 《中国学位论文全文数据库》 * |
丁媛: "copine基因TaBON1和TaBON3在小麦白粉病抗性调控中的功能研究", 《中国学位论文全文数据库》, 19 July 2019 (2019-07-19), pages 11 * |
姚利娜等: "茶树Copine家族基因CsBON3的克隆与表达分析", 《茶叶科学》 * |
姚利娜等: "茶树Copine家族基因CsBON3的克隆与表达分析", 《茶叶科学》, vol. 37, no. 6, 31 December 2017 (2017-12-31), pages 565 - 574 * |
徐强等: "毛果杨copine/BONZAI基因家族全基因组鉴定与组织表达特异性分析", 《分子植物育种》 * |
徐强等: "毛果杨copine/BONZAI基因家族全基因组鉴定与组织表达特异性分析", 《分子植物育种》, 25 August 2020 (2020-08-25), pages 2 * |
柳兵等: "玉米自交系I18人工接种大斑病菌的mRNA表达分析", 《东北农业科学》 * |
柳兵等: "玉米自交系I18人工接种大斑病菌的mRNA表达分析", 《东北农业科学》, no. 05, 31 May 2019 (2019-05-31), pages 45 - 49 * |
蔡金兰等: "云南农业抗病育种研究近况", 《云南农业大学学报》 * |
蔡金兰等: "云南农业抗病育种研究近况", 《云南农业大学学报》, no. 02, 31 December 1993 (1993-12-31), pages 53 - 59 * |
齐拙等: "玉米BON/CPN基因家族的表达模式分析及ZmBON1基因过表达载体的构建", 《吉林农业大学学报》 * |
齐拙等: "玉米BON/CPN基因家族的表达模式分析及ZmBON1基因过表达载体的构建", 《吉林农业大学学报》, 21 July 2021 (2021-07-21), pages 1 - 13 * |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN111778265B (zh) | 玉米赤霉素氧化酶的突变基因、突变体、表达载体和应用 | |
WO2009075860A2 (en) | Transgenic plants with enhanced agronomic traits | |
CN110157718B (zh) | 一种来源于玉米的硝态氮调控基因ZmNRG2.7及其用途 | |
CN108660140B (zh) | SlSL4基因在调控番茄果实成熟中的应用 | |
CN114752579A (zh) | ZmMAPK蛋白及其编码基因在调控植物低温胁迫耐性中的应用 | |
CN111574605A (zh) | 水稻基因OsLAT5在调节敌草快的吸收积累中的应用 | |
CN112522282B (zh) | 一种调控番茄可溶性固形物含量的基因及应用 | |
CN113862265A (zh) | 一种改良水稻粒型和外观品质的方法 | |
CN112662701A (zh) | miRNA 408在调控作物镉积累中的应用 | |
CN109563519A (zh) | 使用硫氧还蛋白序列增加植物生长和产量 | |
CN109867716B (zh) | 蜡梅CpVIN3基因及其应用 | |
CN116179589B (zh) | SlPRMT5基因及其蛋白在调控番茄果实产量中的应用 | |
CN113234720B (zh) | 小麦长链非编码RNAlncR156及其在调控小麦响应干旱胁迫中的应用 | |
CN113444733A (zh) | 一种增强玉米大斑病抗性的基因ZmBON3及其应用 | |
CN114807168A (zh) | 绿豆VrMIB1基因及其应用 | |
CN104109192A (zh) | 一种小麦抗旱基因及其应用 | |
CN107090462A (zh) | 一种NF‑Y类核转录因子基因ZmNF‑YA13、其编码的蛋白及其应用 | |
CN109642239A (zh) | 使用psan序列增加植物生长和产量 | |
CN112501181A (zh) | 水稻抗逆相关基因OsTZF7及其编码蛋白与应用 | |
CN115725531A (zh) | 乙酰转移酶OsG2基因及其编码的蛋白质在调节水稻籽粒大小方面的应用 | |
CN113817750B (zh) | 一种水稻胚乳粉质相关基因OsDAAT1及其编码蛋白质和应用 | |
CN108795949A (zh) | 一种水稻叶色调控相关基因OsWSL6及其编码蛋白质和应用 | |
CN112898393B (zh) | 一种TaAOS基因及其编码的蛋白质应用 | |
CN114958870B (zh) | GmPTF1a/b基因在调控大豆结瘤中的应用 | |
CN116622761A (zh) | 玉米生长素响应蛋白iaa15的用途 |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
PB01 | Publication | ||
PB01 | Publication | ||
SE01 | Entry into force of request for substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
RJ01 | Rejection of invention patent application after publication |
Application publication date: 20210928 |
|
RJ01 | Rejection of invention patent application after publication |