CN114395566B - 甘薯ERF转录因子IbERF4在促进植物绿原酸类物质合成中的用途 - Google Patents
甘薯ERF转录因子IbERF4在促进植物绿原酸类物质合成中的用途 Download PDFInfo
- Publication number
- CN114395566B CN114395566B CN202210309189.7A CN202210309189A CN114395566B CN 114395566 B CN114395566 B CN 114395566B CN 202210309189 A CN202210309189 A CN 202210309189A CN 114395566 B CN114395566 B CN 114395566B
- Authority
- CN
- China
- Prior art keywords
- plant
- iberf4
- chlorogenic acid
- gene
- sweet potato
- 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.)
- Active
Links
- 241000196324 Embryophyta Species 0.000 title claims abstract description 104
- 235000001368 chlorogenic acid Nutrition 0.000 title claims abstract description 57
- CWVRJTMFETXNAD-FWCWNIRPSA-N 3-O-Caffeoylquinic acid Natural products O[C@H]1[C@@H](O)C[C@@](O)(C(O)=O)C[C@H]1OC(=O)\C=C\C1=CC=C(O)C(O)=C1 CWVRJTMFETXNAD-FWCWNIRPSA-N 0.000 title claims abstract description 54
- PZIRUHCJZBGLDY-UHFFFAOYSA-N Caffeoylquinic acid Natural products CC(CCC(=O)C(C)C1C(=O)CC2C3CC(O)C4CC(O)CCC4(C)C3CCC12C)C(=O)O PZIRUHCJZBGLDY-UHFFFAOYSA-N 0.000 title claims abstract description 54
- CWVRJTMFETXNAD-KLZCAUPSSA-N Neochlorogenin-saeure Natural products O[C@H]1C[C@@](O)(C[C@@H](OC(=O)C=Cc2ccc(O)c(O)c2)[C@@H]1O)C(=O)O CWVRJTMFETXNAD-KLZCAUPSSA-N 0.000 title claims abstract description 54
- CWVRJTMFETXNAD-JUHZACGLSA-N chlorogenic acid Chemical compound O[C@@H]1[C@H](O)C[C@@](O)(C(O)=O)C[C@H]1OC(=O)\C=C\C1=CC=C(O)C(O)=C1 CWVRJTMFETXNAD-JUHZACGLSA-N 0.000 title claims abstract description 54
- 229940074393 chlorogenic acid Drugs 0.000 title claims abstract description 54
- FFQSDFBBSXGVKF-KHSQJDLVSA-N chlorogenic acid Natural products O[C@@H]1C[C@](O)(C[C@@H](CC(=O)C=Cc2ccc(O)c(O)c2)[C@@H]1O)C(=O)O FFQSDFBBSXGVKF-KHSQJDLVSA-N 0.000 title claims abstract description 54
- BMRSEYFENKXDIS-KLZCAUPSSA-N cis-3-O-p-coumaroylquinic acid Natural products O[C@H]1C[C@@](O)(C[C@@H](OC(=O)C=Cc2ccc(O)cc2)[C@@H]1O)C(=O)O BMRSEYFENKXDIS-KLZCAUPSSA-N 0.000 title claims abstract description 54
- 244000017020 Ipomoea batatas Species 0.000 title claims abstract description 38
- 235000002678 Ipomoea batatas Nutrition 0.000 title claims abstract description 37
- 239000000126 substance Substances 0.000 title claims abstract description 26
- 230000015572 biosynthetic process Effects 0.000 title claims abstract description 23
- 238000003786 synthesis reaction Methods 0.000 title claims abstract description 23
- 230000001737 promoting effect Effects 0.000 title claims abstract description 13
- 108090000623 proteins and genes Proteins 0.000 claims abstract description 50
- 230000009261 transgenic effect Effects 0.000 claims abstract description 31
- 238000000034 method Methods 0.000 claims abstract description 21
- 230000014509 gene expression Effects 0.000 claims abstract description 17
- 102000004169 proteins and genes Human genes 0.000 claims abstract description 14
- 230000001105 regulatory effect Effects 0.000 claims abstract description 8
- 238000013518 transcription Methods 0.000 claims abstract description 7
- 230000035897 transcription Effects 0.000 claims abstract description 7
- 239000013604 expression vector Substances 0.000 claims abstract description 6
- 108090000364 Ligases Proteins 0.000 claims abstract description 5
- 108010011449 Long-chain-fatty-acid-CoA ligase Proteins 0.000 claims abstract description 5
- 238000003208 gene overexpression Methods 0.000 claims abstract description 4
- 230000002018 overexpression Effects 0.000 claims description 11
- UFCLZKMFXSILNL-RVXRWRFUSA-N 4,5-di-O-caffeoylquinic acid Chemical compound O([C@@H]1C[C@](O)(C[C@H]([C@@H]1OC(=O)\C=C\C=1C=C(O)C(O)=CC=1)O)C(O)=O)C(=O)\C=C\C1=CC=C(O)C(O)=C1 UFCLZKMFXSILNL-RVXRWRFUSA-N 0.000 claims description 10
- 230000037361 pathway Effects 0.000 claims description 6
- KRZBCHWVBQOTNZ-UHFFFAOYSA-N (-) 3,5-dicaffeoyl-muco-quinic acid Natural products OC1C(OC(=O)C=CC=2C=C(O)C(O)=CC=2)CC(O)(C(O)=O)CC1OC(=O)C=CC1=CC=C(O)C(O)=C1 KRZBCHWVBQOTNZ-UHFFFAOYSA-N 0.000 claims description 5
- KRZBCHWVBQOTNZ-RDJMKVHDSA-M (-)-3,5-Dicaffeoyl quinic acid Natural products O([C@@H]1CC(O)(C[C@H](C1O)OC(=O)\C=C\C=1C=C(O)C(O)=CC=1)C([O-])=O)C(=O)\C=C\C1=CC=C(O)C(O)=C1 KRZBCHWVBQOTNZ-RDJMKVHDSA-M 0.000 claims description 5
- UFCLZKMFXSILNL-AALYGJCLSA-N 3,4-Dicaffeoylquinic acid Natural products O=C(O[C@@H]1[C@H](OC(=O)/C=C/c2cc(O)c(O)cc2)C[C@](O)(C(=O)O)C[C@@H]1O)/C=C/c1cc(O)c(O)cc1 UFCLZKMFXSILNL-AALYGJCLSA-N 0.000 claims description 5
- KRZBCHWVBQOTNZ-PSEXTPKNSA-N 3,5-di-O-caffeoyl quinic acid Chemical compound O([C@@H]1C[C@](O)(C[C@H]([C@@H]1O)OC(=O)\C=C\C=1C=C(O)C(O)=CC=1)C(O)=O)C(=O)\C=C\C1=CC=C(O)C(O)=C1 KRZBCHWVBQOTNZ-PSEXTPKNSA-N 0.000 claims description 5
- -1 4-hydroxycinnamoyl Chemical group 0.000 claims description 4
- 230000000694 effects Effects 0.000 claims description 3
- 238000003976 plant breeding Methods 0.000 claims description 3
- DMZOKBALNZWDKI-MATMFAIHSA-N trans-4-coumaroyl-CoA Chemical compound O=C([C@H](O)C(C)(COP(O)(=O)OP(O)(=O)OC[C@@H]1[C@H]([C@@H](O)[C@@H](O1)N1C2=NC=NC(N)=C2N=C1)OP(O)(O)=O)C)NCCC(=O)NCCSC(=O)\C=C\C1=CC=C(O)C=C1 DMZOKBALNZWDKI-MATMFAIHSA-N 0.000 claims description 2
- 150000001875 compounds Chemical class 0.000 claims 2
- 125000003275 alpha amino acid group Chemical group 0.000 claims 1
- 239000002777 nucleoside Substances 0.000 claims 1
- 241000589158 Agrobacterium Species 0.000 abstract description 5
- 235000009508 confectionery Nutrition 0.000 abstract description 5
- 239000002299 complementary DNA Substances 0.000 abstract description 4
- 238000004458 analytical method Methods 0.000 abstract description 3
- 208000015181 infectious disease Diseases 0.000 abstract description 3
- 239000005516 coenzyme A Substances 0.000 abstract 1
- 229940093530 coenzyme a Drugs 0.000 abstract 1
- 239000013598 vector Substances 0.000 description 14
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 12
- 239000000047 product Substances 0.000 description 11
- 235000018102 proteins Nutrition 0.000 description 10
- 108020004414 DNA Proteins 0.000 description 9
- 108090000790 Enzymes Proteins 0.000 description 8
- 239000013612 plasmid Substances 0.000 description 8
- 206010020649 Hyperkeratosis Diseases 0.000 description 7
- 239000007788 liquid Substances 0.000 description 7
- 238000012163 sequencing technique Methods 0.000 description 6
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 5
- 102000004190 Enzymes Human genes 0.000 description 5
- 230000012010 growth Effects 0.000 description 5
- 239000002609 medium Substances 0.000 description 5
- 238000012795 verification Methods 0.000 description 5
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 4
- 108091023040 Transcription factor Proteins 0.000 description 4
- 102000040945 Transcription factor Human genes 0.000 description 4
- 239000002253 acid Substances 0.000 description 4
- 238000012258 culturing Methods 0.000 description 4
- 238000011161 development Methods 0.000 description 4
- 230000018109 developmental process Effects 0.000 description 4
- 235000013305 food Nutrition 0.000 description 4
- 239000012634 fragment Substances 0.000 description 4
- 238000011160 research Methods 0.000 description 4
- 108700039691 Genetic Promoter Regions Proteins 0.000 description 3
- 230000003321 amplification Effects 0.000 description 3
- 229960004261 cefotaxime Drugs 0.000 description 3
- 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 3
- 239000012154 double-distilled water Substances 0.000 description 3
- 235000013399 edible fruits Nutrition 0.000 description 3
- 238000001976 enzyme digestion Methods 0.000 description 3
- 239000001963 growth medium Substances 0.000 description 3
- 239000000463 material Substances 0.000 description 3
- 238000003199 nucleic acid amplification method Methods 0.000 description 3
- 239000000243 solution Substances 0.000 description 3
- 230000035882 stress Effects 0.000 description 3
- 230000009466 transformation Effects 0.000 description 3
- AAWZDTNXLSGCEK-LNVDRNJUSA-N (3r,5r)-1,3,4,5-tetrahydroxycyclohexane-1-carboxylic acid Chemical compound O[C@@H]1CC(O)(C(O)=O)C[C@@H](O)C1O AAWZDTNXLSGCEK-LNVDRNJUSA-N 0.000 description 2
- HHGYNJRJIINWAK-FXQIFTODSA-N Ala-Ala-Arg Chemical compound C[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@H](C(O)=O)CCCN=C(N)N HHGYNJRJIINWAK-FXQIFTODSA-N 0.000 description 2
- 241000894006 Bacteria Species 0.000 description 2
- AAWZDTNXLSGCEK-UHFFFAOYSA-N Cordycepinsaeure Natural products OC1CC(O)(C(O)=O)CC(O)C1O AAWZDTNXLSGCEK-UHFFFAOYSA-N 0.000 description 2
- 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 2
- 102000003960 Ligases Human genes 0.000 description 2
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 2
- 238000012408 PCR amplification Methods 0.000 description 2
- 108700023158 Phenylalanine ammonia-lyases Proteins 0.000 description 2
- AAWZDTNXLSGCEK-ZHQZDSKASA-N Quinic acid Natural products O[C@H]1CC(O)(C(O)=O)C[C@H](O)C1O AAWZDTNXLSGCEK-ZHQZDSKASA-N 0.000 description 2
- 101000702488 Rattus norvegicus High affinity cationic amino acid transporter 1 Proteins 0.000 description 2
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 2
- 229920002472 Starch Polymers 0.000 description 2
- 238000002835 absorbance Methods 0.000 description 2
- 230000008901 benefit Effects 0.000 description 2
- 239000008280 blood Substances 0.000 description 2
- 210000004369 blood Anatomy 0.000 description 2
- 238000006243 chemical reaction Methods 0.000 description 2
- 238000007796 conventional method Methods 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 239000003925 fat Substances 0.000 description 2
- 239000000706 filtrate Substances 0.000 description 2
- 230000015784 hyperosmotic salinity response Effects 0.000 description 2
- 229930027917 kanamycin Natural products 0.000 description 2
- 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 2
- 229960000318 kanamycin Drugs 0.000 description 2
- 229930182823 kanamycin A Natural products 0.000 description 2
- 229910052757 nitrogen Inorganic materials 0.000 description 2
- 108020004707 nucleic acids Proteins 0.000 description 2
- 102000039446 nucleic acids Human genes 0.000 description 2
- 150000007523 nucleic acids Chemical class 0.000 description 2
- 210000000056 organ Anatomy 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 238000000746 purification Methods 0.000 description 2
- 230000009467 reduction Effects 0.000 description 2
- 230000008929 regeneration Effects 0.000 description 2
- 238000011069 regeneration method Methods 0.000 description 2
- 108091008146 restriction endonucleases Proteins 0.000 description 2
- 238000010839 reverse transcription Methods 0.000 description 2
- 108010026333 seryl-proline Proteins 0.000 description 2
- 238000002791 soaking Methods 0.000 description 2
- 239000012086 standard solution Substances 0.000 description 2
- 235000019698 starch Nutrition 0.000 description 2
- 238000009210 therapy by ultrasound Methods 0.000 description 2
- QAIPRVGONGVQAS-DUXPYHPUSA-N trans-caffeic acid Chemical compound OC(=O)\C=C\C1=CC=C(O)C(O)=C1 QAIPRVGONGVQAS-DUXPYHPUSA-N 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- 238000005303 weighing Methods 0.000 description 2
- ACEAELOMUCBPJP-UHFFFAOYSA-N (E)-3,4,5-trihydroxycinnamic acid Natural products OC(=O)C=CC1=CC(O)=C(O)C(O)=C1 ACEAELOMUCBPJP-UHFFFAOYSA-N 0.000 description 1
- WBYWAXJHAXSJNI-VOTSOKGWSA-M .beta-Phenylacrylic acid Natural products [O-]C(=O)\C=C\C1=CC=CC=C1 WBYWAXJHAXSJNI-VOTSOKGWSA-M 0.000 description 1
- WOJJIRYPFAZEPF-YFKPBYRVSA-N 2-[[(2s)-2-[[2-[(2-azaniumylacetyl)amino]acetyl]amino]propanoyl]amino]acetate Chemical compound OC(=O)CNC(=O)[C@H](C)NC(=O)CNC(=O)CN WOJJIRYPFAZEPF-YFKPBYRVSA-N 0.000 description 1
- CWVRJTMFETXNAD-GMZLATJGSA-N 5-Caffeoyl quinic acid Natural products O[C@H]1C[C@](O)(C[C@H](OC(=O)C=Cc2ccc(O)c(O)c2)[C@@H]1O)C(=O)O CWVRJTMFETXNAD-GMZLATJGSA-N 0.000 description 1
- OPIFSICVWOWJMJ-AEOCFKNESA-N 5-bromo-4-chloro-3-indolyl beta-D-galactoside Chemical compound O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@H]1OC1=CNC2=CC=C(Br)C(Cl)=C12 OPIFSICVWOWJMJ-AEOCFKNESA-N 0.000 description 1
- 229920000936 Agarose Polymers 0.000 description 1
- RLMISHABBKUNFO-WHFBIAKZSA-N Ala-Ala-Gly Chemical compound C[C@H](N)C(=O)N[C@@H](C)C(=O)NCC(O)=O RLMISHABBKUNFO-WHFBIAKZSA-N 0.000 description 1
- WKOBSJOZRJJVRZ-FXQIFTODSA-N Ala-Glu-Glu Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(O)=O WKOBSJOZRJJVRZ-FXQIFTODSA-N 0.000 description 1
- GGNHBHYDMUDXQB-KBIXCLLPSA-N Ala-Glu-Ile Chemical compound CC[C@H](C)[C@@H](C(O)=O)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](C)N GGNHBHYDMUDXQB-KBIXCLLPSA-N 0.000 description 1
- VCSABYLVNWQYQE-SRVKXCTJSA-N Ala-Lys-Lys Chemical compound NCCCC[C@H](NC(=O)[C@@H](N)C)C(=O)N[C@@H](CCCCN)C(O)=O VCSABYLVNWQYQE-SRVKXCTJSA-N 0.000 description 1
- VCSABYLVNWQYQE-UHFFFAOYSA-N Ala-Lys-Lys Natural products NCCCCC(NC(=O)C(N)C)C(=O)NC(CCCCN)C(O)=O VCSABYLVNWQYQE-UHFFFAOYSA-N 0.000 description 1
- OINVDEKBKBCPLX-JXUBOQSCSA-N Ala-Lys-Thr Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H]([C@@H](C)O)C(O)=O OINVDEKBKBCPLX-JXUBOQSCSA-N 0.000 description 1
- REWSWYIDQIELBE-FXQIFTODSA-N Ala-Val-Ser Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CO)C(O)=O REWSWYIDQIELBE-FXQIFTODSA-N 0.000 description 1
- 241000219194 Arabidopsis Species 0.000 description 1
- MFAMTAVAFBPXDC-LPEHRKFASA-N Arg-Asp-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CC(=O)O)NC(=O)[C@H](CCCN=C(N)N)N)C(=O)O MFAMTAVAFBPXDC-LPEHRKFASA-N 0.000 description 1
- YBZMTKUDWXZLIX-UWVGGRQHSA-N Arg-Leu-Gly Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(C)C)C(=O)NCC(O)=O YBZMTKUDWXZLIX-UWVGGRQHSA-N 0.000 description 1
- BSYKSCBTTQKOJG-GUBZILKMSA-N Arg-Pro-Ala Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N1CCC[C@H]1C(=O)N[C@@H](C)C(O)=O BSYKSCBTTQKOJG-GUBZILKMSA-N 0.000 description 1
- HGKHPCFTRQDHCU-IUCAKERBSA-N Arg-Pro-Gly Chemical compound NC(N)=NCCC[C@H](N)C(=O)N1CCC[C@H]1C(=O)NCC(O)=O HGKHPCFTRQDHCU-IUCAKERBSA-N 0.000 description 1
- NGYHSXDNNOFHNE-AVGNSLFASA-N Arg-Pro-Leu Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CC(C)C)C(O)=O NGYHSXDNNOFHNE-AVGNSLFASA-N 0.000 description 1
- OQPAZKMGCWPERI-GUBZILKMSA-N Arg-Ser-Val Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CO)C(=O)N[C@@H](C(C)C)C(O)=O OQPAZKMGCWPERI-GUBZILKMSA-N 0.000 description 1
- NVPHRWNWTKYIST-BPNCWPANSA-N Arg-Tyr-Ala Chemical compound NC(N)=NCCC[C@H](N)C(=O)N[C@H](C(=O)N[C@@H](C)C(O)=O)CC1=CC=C(O)C=C1 NVPHRWNWTKYIST-BPNCWPANSA-N 0.000 description 1
- UTSMXMABBPFVJP-SZMVWBNQSA-N Arg-Val-Trp Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC1=CNC2=CC=CC=C21)C(=O)O)NC(=O)[C@H](CCCN=C(N)N)N UTSMXMABBPFVJP-SZMVWBNQSA-N 0.000 description 1
- POOCJCRBHHMAOS-FXQIFTODSA-N Asn-Arg-Ser Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CO)C(O)=O POOCJCRBHHMAOS-FXQIFTODSA-N 0.000 description 1
- ACRYGQFHAQHDSF-ZLUOBGJFSA-N Asn-Asn-Asn Chemical compound NC(=O)C[C@H](N)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(N)=O)C(O)=O ACRYGQFHAQHDSF-ZLUOBGJFSA-N 0.000 description 1
- VKCOHFFSTKCXEQ-OLHMAJIHSA-N Asn-Asn-Thr Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O VKCOHFFSTKCXEQ-OLHMAJIHSA-N 0.000 description 1
- UYCPJVYQYARFGB-YDHLFZDLSA-N Asn-Phe-Val Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](C(C)C)C(O)=O UYCPJVYQYARFGB-YDHLFZDLSA-N 0.000 description 1
- 239000002028 Biomass Substances 0.000 description 1
- 108091062157 Cis-regulatory element Proteins 0.000 description 1
- 102000005870 Coenzyme A Ligases Human genes 0.000 description 1
- 241000218631 Coniferophyta Species 0.000 description 1
- 108091061403 ERF family Proteins 0.000 description 1
- OZEQPCDLCDRCGY-SOUVJXGZSA-N Gln-Phe-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CC2=CC=CC=C2)NC(=O)[C@H](CCC(=O)N)N)C(=O)O OZEQPCDLCDRCGY-SOUVJXGZSA-N 0.000 description 1
- RUFHOVYUYSNDNY-ACZMJKKPSA-N Glu-Ala-Ala Chemical compound OC(=O)[C@H](C)NC(=O)[C@H](C)NC(=O)[C@@H](N)CCC(O)=O RUFHOVYUYSNDNY-ACZMJKKPSA-N 0.000 description 1
- QQLBPVKLJBAXBS-FXQIFTODSA-N Glu-Glu-Asn Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(N)=O)C(O)=O QQLBPVKLJBAXBS-FXQIFTODSA-N 0.000 description 1
- SJPMNHCEWPTRBR-BQBZGAKWSA-N Glu-Glu-Gly Chemical compound OC(=O)CC[C@H](N)C(=O)N[C@@H](CCC(O)=O)C(=O)NCC(O)=O SJPMNHCEWPTRBR-BQBZGAKWSA-N 0.000 description 1
- PJBVXVBTTFZPHJ-GUBZILKMSA-N Glu-Leu-Asp Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)O)NC(=O)[C@H](CCC(=O)O)N PJBVXVBTTFZPHJ-GUBZILKMSA-N 0.000 description 1
- KJBGAZSLZAQDPV-KKUMJFAQSA-N Glu-Phe-Arg Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)N[C@@H](CCCN=C(N)N)C(=O)O)NC(=O)[C@H](CCC(=O)O)N KJBGAZSLZAQDPV-KKUMJFAQSA-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
- ZALGPUWUVHOGAE-GVXVVHGQSA-N Glu-Val-His Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC1=CN=CN1)C(=O)O)NC(=O)[C@H](CCC(=O)O)N ZALGPUWUVHOGAE-GVXVVHGQSA-N 0.000 description 1
- UGVQELHRNUDMAA-BYPYZUCNSA-N Gly-Ala-Gly Chemical compound [NH3+]CC(=O)N[C@@H](C)C(=O)NCC([O-])=O UGVQELHRNUDMAA-BYPYZUCNSA-N 0.000 description 1
- JBRBACJPBZNFMF-YUMQZZPRSA-N Gly-Ala-Lys Chemical compound NCC(=O)N[C@@H](C)C(=O)N[C@H](C(O)=O)CCCCN JBRBACJPBZNFMF-YUMQZZPRSA-N 0.000 description 1
- ITZOBNKQDZEOCE-NHCYSSNCSA-N Gly-Ile-Lys Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCCCN)C(=O)O)NC(=O)CN ITZOBNKQDZEOCE-NHCYSSNCSA-N 0.000 description 1
- FGPLUIQCSKGLTI-WDSKDSINSA-N Gly-Ser-Glu Chemical compound NCC(=O)N[C@@H](CO)C(=O)N[C@H](C(O)=O)CCC(O)=O FGPLUIQCSKGLTI-WDSKDSINSA-N 0.000 description 1
- FNXSYBOHALPRHV-ONGXEEELSA-N Gly-Val-Lys Chemical compound NCC(=O)N[C@@H](C(C)C)C(=O)N[C@H](C(O)=O)CCCCN FNXSYBOHALPRHV-ONGXEEELSA-N 0.000 description 1
- SQUHHTBVTRBESD-UHFFFAOYSA-N Hexa-Ac-myo-Inositol Natural products CC(=O)OC1C(OC(C)=O)C(OC(C)=O)C(OC(C)=O)C(OC(C)=O)C1OC(C)=O SQUHHTBVTRBESD-UHFFFAOYSA-N 0.000 description 1
- HTZKFIYQMHJWSQ-INTQDDNPSA-N His-Ala-Pro Chemical compound C[C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](CC2=CN=CN2)N HTZKFIYQMHJWSQ-INTQDDNPSA-N 0.000 description 1
- 206010061218 Inflammation Diseases 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
- HYMLKESRWLZDBR-WEDXCCLWSA-N Leu-Gly-Thr Chemical compound CC(C)C[C@H](N)C(=O)NCC(=O)N[C@@H]([C@@H](C)O)C(O)=O HYMLKESRWLZDBR-WEDXCCLWSA-N 0.000 description 1
- IZPVWNSAVUQBGP-CIUDSAMLSA-N Leu-Ser-Asp Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(O)=O)C(O)=O IZPVWNSAVUQBGP-CIUDSAMLSA-N 0.000 description 1
- ZDJQVSIPFLMNOX-RHYQMDGZSA-N Leu-Thr-Arg Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@H](C(O)=O)CCCN=C(N)N ZDJQVSIPFLMNOX-RHYQMDGZSA-N 0.000 description 1
- GQZMPWBZQALKJO-UWVGGRQHSA-N Lys-Gly-Arg Chemical compound [H]N[C@@H](CCCCN)C(=O)NCC(=O)N[C@@H](CCCNC(N)=N)C(O)=O GQZMPWBZQALKJO-UWVGGRQHSA-N 0.000 description 1
- MDDUIRLQCYVRDO-NHCYSSNCSA-N Lys-Val-Asn Chemical compound NC(=O)C[C@@H](C(O)=O)NC(=O)[C@H](C(C)C)NC(=O)[C@@H](N)CCCCN MDDUIRLQCYVRDO-NHCYSSNCSA-N 0.000 description 1
- 241000218922 Magnoliophyta Species 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
- 108010079364 N-glycylalanine Proteins 0.000 description 1
- 108020002230 Pancreatic Ribonuclease Proteins 0.000 description 1
- 102000005891 Pancreatic ribonuclease Human genes 0.000 description 1
- SWZKMTDPQXLQRD-XVSYOHENSA-N Phe-Asp-Thr Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O SWZKMTDPQXLQRD-XVSYOHENSA-N 0.000 description 1
- GDBOREPXIRKSEQ-FHWLQOOXSA-N Phe-Gln-Phe Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O GDBOREPXIRKSEQ-FHWLQOOXSA-N 0.000 description 1
- BEEVXUYVEHXWRQ-YESZJQIVSA-N Phe-His-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CC2=CN=CN2)NC(=O)[C@H](CC3=CC=CC=C3)N)C(=O)O BEEVXUYVEHXWRQ-YESZJQIVSA-N 0.000 description 1
- MYQCCQSMKNCNKY-KKUMJFAQSA-N Phe-His-Ser Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)N[C@@H](CC2=CN=CN2)C(=O)N[C@@H](CO)C(=O)O)N MYQCCQSMKNCNKY-KKUMJFAQSA-N 0.000 description 1
- FZHBZMDRDASUHN-NAKRPEOUSA-N Pro-Ala-Ile Chemical compound CC[C@H](C)[C@H](NC(=O)[C@H](C)NC(=O)[C@@H]1CCCN1)C(O)=O FZHBZMDRDASUHN-NAKRPEOUSA-N 0.000 description 1
- WPQKSRHDTMRSJM-CIUDSAMLSA-N Pro-Asp-Gln Chemical compound NC(=O)CC[C@@H](C(O)=O)NC(=O)[C@H](CC(O)=O)NC(=O)[C@@H]1CCCN1 WPQKSRHDTMRSJM-CIUDSAMLSA-N 0.000 description 1
- SEZGGSHLMROBFX-CIUDSAMLSA-N Pro-Ser-Gln Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CO)C(=O)N[C@@H](CCC(N)=O)C(O)=O SEZGGSHLMROBFX-CIUDSAMLSA-N 0.000 description 1
- FNGOXVQBBCMFKV-CIUDSAMLSA-N Pro-Ser-Glu Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CO)C(=O)N[C@@H](CCC(O)=O)C(O)=O FNGOXVQBBCMFKV-CIUDSAMLSA-N 0.000 description 1
- SXJOPONICMGFCR-DCAQKATOSA-N Pro-Ser-Lys Chemical compound C1C[C@H](NC1)C(=O)N[C@@H](CO)C(=O)N[C@@H](CCCCN)C(=O)O SXJOPONICMGFCR-DCAQKATOSA-N 0.000 description 1
- DMNANGOFEUVBRV-GJZGRUSLSA-N Pro-Trp-Gly Chemical compound N([C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)NCC(=O)O)C(=O)[C@@H]1CCCN1 DMNANGOFEUVBRV-GJZGRUSLSA-N 0.000 description 1
- 238000010802 RNA extraction kit Methods 0.000 description 1
- 238000011529 RT qPCR Methods 0.000 description 1
- SRTCFKGBYBZRHA-ACZMJKKPSA-N Ser-Ala-Glu Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](C)C(=O)N[C@@H](CCC(O)=O)C(O)=O SRTCFKGBYBZRHA-ACZMJKKPSA-N 0.000 description 1
- PYTKULIABVRXSC-BWBBJGPYSA-N Ser-Ser-Thr Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)O)C(O)=O PYTKULIABVRXSC-BWBBJGPYSA-N 0.000 description 1
- DYEGLQRVMBWQLD-IXOXFDKPSA-N Ser-Thr-Phe Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)O)NC(=O)[C@H](CO)N)O DYEGLQRVMBWQLD-IXOXFDKPSA-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
- 244000061456 Solanum tuberosum Species 0.000 description 1
- 235000002595 Solanum tuberosum Nutrition 0.000 description 1
- 229930006000 Sucrose Natural products 0.000 description 1
- 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 1
- VGYBYGQXZJDZJU-XQXXSGGOSA-N Thr-Glu-Ala Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](C)C(O)=O VGYBYGQXZJDZJU-XQXXSGGOSA-N 0.000 description 1
- ISLDRLHVPXABBC-IEGACIPQSA-N Thr-Leu-Trp Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC1=CNC2=C1C=CC=C2)C(O)=O ISLDRLHVPXABBC-IEGACIPQSA-N 0.000 description 1
- UQCNIMDPYICBTR-KYNKHSRBSA-N Thr-Thr-Gly Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)O)C(=O)NCC(O)=O UQCNIMDPYICBTR-KYNKHSRBSA-N 0.000 description 1
- 108700019146 Transgenes Proteins 0.000 description 1
- AKXBNSZMYAOGLS-STQMWFEESA-N Tyr-Arg-Gly Chemical compound NC(N)=NCCC[C@@H](C(=O)NCC(O)=O)NC(=O)[C@@H](N)CC1=CC=C(O)C=C1 AKXBNSZMYAOGLS-STQMWFEESA-N 0.000 description 1
- GAYLGYUVTDMLKC-UWJYBYFXSA-N Tyr-Asp-Ala Chemical compound OC(=O)[C@H](C)NC(=O)[C@H](CC(O)=O)NC(=O)[C@@H](N)CC1=CC=C(O)C=C1 GAYLGYUVTDMLKC-UWJYBYFXSA-N 0.000 description 1
- PAPWZOJOLKZEFR-AVGNSLFASA-N Val-Arg-Lys Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CCCN=C(N)N)C(=O)N[C@@H](CCCCN)C(=O)O)N PAPWZOJOLKZEFR-AVGNSLFASA-N 0.000 description 1
- 241000700605 Viruses Species 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 150000007513 acids Chemical class 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 238000000246 agarose gel electrophoresis Methods 0.000 description 1
- 108010087924 alanylproline Proteins 0.000 description 1
- 230000000844 anti-bacterial effect Effects 0.000 description 1
- 230000003276 anti-hypertensive effect Effects 0.000 description 1
- 230000003064 anti-oxidating effect Effects 0.000 description 1
- 230000000259 anti-tumor effect Effects 0.000 description 1
- 108010013835 arginine glutamate Proteins 0.000 description 1
- 108010052670 arginyl-glutamyl-glutamic acid Proteins 0.000 description 1
- 108010043240 arginyl-leucyl-glycine Proteins 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- 238000009412 basement excavation Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000010170 biological method Methods 0.000 description 1
- 230000033228 biological regulation Effects 0.000 description 1
- 238000010805 cDNA synthesis kit Methods 0.000 description 1
- 235000004883 caffeic acid Nutrition 0.000 description 1
- 229940074360 caffeic acid Drugs 0.000 description 1
- 150000001720 carbohydrates Chemical class 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- QAIPRVGONGVQAS-UHFFFAOYSA-N cis-caffeic acid Natural products OC(=O)C=CC1=CC=C(O)C(O)=C1 QAIPRVGONGVQAS-UHFFFAOYSA-N 0.000 description 1
- 238000003501 co-culture Methods 0.000 description 1
- 239000011248 coating agent Substances 0.000 description 1
- 238000000576 coating method 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
- 238000001514 detection method Methods 0.000 description 1
- 235000014113 dietary fatty acids Nutrition 0.000 description 1
- 230000004069 differentiation Effects 0.000 description 1
- 230000024346 drought recovery Effects 0.000 description 1
- 230000008641 drought stress Effects 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 238000004043 dyeing Methods 0.000 description 1
- 238000001962 electrophoresis Methods 0.000 description 1
- 210000002257 embryonic structure Anatomy 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 239000003623 enhancer Substances 0.000 description 1
- 235000019197 fats Nutrition 0.000 description 1
- 229930195729 fatty acid Natural products 0.000 description 1
- 239000000194 fatty acid Substances 0.000 description 1
- 150000004665 fatty acids Chemical class 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 235000013373 food additive Nutrition 0.000 description 1
- 239000002778 food additive Substances 0.000 description 1
- 230000008014 freezing Effects 0.000 description 1
- 238000007710 freezing Methods 0.000 description 1
- 108010049041 glutamylalanine Proteins 0.000 description 1
- VPZXBVLAVMBEQI-UHFFFAOYSA-N glycyl-DL-alpha-alanine Natural products OC(=O)C(C)NC(=O)CN VPZXBVLAVMBEQI-UHFFFAOYSA-N 0.000 description 1
- 108010027668 glycyl-alanyl-valine Proteins 0.000 description 1
- 108010078326 glycyl-glycyl-valine Proteins 0.000 description 1
- 108010037850 glycylvaline Proteins 0.000 description 1
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 description 1
- 239000010931 gold Substances 0.000 description 1
- 229910052737 gold Inorganic materials 0.000 description 1
- 239000008187 granular material Substances 0.000 description 1
- 238000003306 harvesting Methods 0.000 description 1
- 229910052500 inorganic mineral Inorganic materials 0.000 description 1
- 229960000367 inositol Drugs 0.000 description 1
- CDAISMWEOUEBRE-GPIVLXJGSA-N inositol Chemical compound O[C@H]1[C@H](O)[C@@H](O)[C@H](O)[C@H](O)[C@@H]1O CDAISMWEOUEBRE-GPIVLXJGSA-N 0.000 description 1
- 210000003734 kidney Anatomy 0.000 description 1
- 108010057821 leucylproline Proteins 0.000 description 1
- 239000003446 ligand Substances 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- 108010009298 lysylglutamic acid Proteins 0.000 description 1
- 108010064235 lysylglycine Proteins 0.000 description 1
- 229910052943 magnesium sulfate Inorganic materials 0.000 description 1
- 230000000442 meristematic effect Effects 0.000 description 1
- 230000002503 metabolic effect Effects 0.000 description 1
- 238000000520 microinjection Methods 0.000 description 1
- 239000011707 mineral Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 239000002773 nucleotide Substances 0.000 description 1
- 125000003729 nucleotide group Chemical group 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 239000003921 oil Substances 0.000 description 1
- 235000019198 oils Nutrition 0.000 description 1
- 230000000144 pharmacologic 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
- 229930015704 phenylpropanoid Natural products 0.000 description 1
- 150000002995 phenylpropanoid derivatives Chemical class 0.000 description 1
- 235000013824 polyphenols Nutrition 0.000 description 1
- 229920001282 polysaccharide Polymers 0.000 description 1
- 239000005017 polysaccharide Substances 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 238000012257 pre-denaturation Methods 0.000 description 1
- 239000003755 preservative agent Substances 0.000 description 1
- 230000002335 preservative effect Effects 0.000 description 1
- 108010090894 prolylleucine Proteins 0.000 description 1
- 238000004445 quantitative analysis Methods 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 238000003753 real-time PCR Methods 0.000 description 1
- 238000003259 recombinant expression Methods 0.000 description 1
- 230000006798 recombination Effects 0.000 description 1
- 238000005215 recombination Methods 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- 239000012882 rooting medium Substances 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- CDAISMWEOUEBRE-UHFFFAOYSA-N scyllo-inosotol Natural products OC1C(O)C(O)C(O)C(O)C1O CDAISMWEOUEBRE-UHFFFAOYSA-N 0.000 description 1
- 230000024053 secondary metabolic process Effects 0.000 description 1
- 229930000044 secondary metabolite Natural products 0.000 description 1
- 230000009758 senescence Effects 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 239000008107 starch Substances 0.000 description 1
- 239000008223 sterile water Substances 0.000 description 1
- 239000005720 sucrose Substances 0.000 description 1
- 230000004083 survival effect Effects 0.000 description 1
- 238000004114 suspension culture Methods 0.000 description 1
- 238000010998 test method Methods 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- NGSWKAQJJWESNS-ZZXKWVIFSA-N trans-4-coumaric acid Chemical compound OC(=O)\C=C\C1=CC=C(O)C=C1 NGSWKAQJJWESNS-ZZXKWVIFSA-N 0.000 description 1
- CWVRJTMFETXNAD-NXLLHMKUSA-N trans-5-O-caffeoyl-D-quinic acid Chemical compound O[C@H]1[C@H](O)C[C@](O)(C(O)=O)C[C@H]1OC(=O)\C=C\C1=CC=C(O)C(O)=C1 CWVRJTMFETXNAD-NXLLHMKUSA-N 0.000 description 1
- WBYWAXJHAXSJNI-VOTSOKGWSA-N trans-cinnamic acid Chemical compound OC(=O)\C=C\C1=CC=CC=C1 WBYWAXJHAXSJNI-VOTSOKGWSA-N 0.000 description 1
- 238000000870 ultraviolet spectroscopy Methods 0.000 description 1
- 108010073969 valyllysine Proteins 0.000 description 1
- 239000011782 vitamin Substances 0.000 description 1
- 229940088594 vitamin Drugs 0.000 description 1
- 229930003231 vitamin Natural products 0.000 description 1
- 235000013343 vitamin Nutrition 0.000 description 1
- 238000005406 washing Methods 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)
- C12N15/8241—Phenotypically and genetically modified plants via recombinant DNA technology
- C12N15/8242—Phenotypically and genetically modified plants via recombinant DNA technology with non-agronomic quality (output) traits, e.g. for industrial processing; Value added, non-agronomic traits
- C12N15/8243—Phenotypically and genetically modified plants via recombinant DNA technology with non-agronomic quality (output) traits, e.g. for industrial processing; Value added, non-agronomic traits involving biosynthetic or metabolic pathways, i.e. metabolic engineering, e.g. nicotine, caffeine
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Genetics & Genomics (AREA)
- Engineering & Computer Science (AREA)
- Chemical & Material Sciences (AREA)
- Biomedical Technology (AREA)
- Organic Chemistry (AREA)
- Biotechnology (AREA)
- Molecular Biology (AREA)
- Wood Science & Technology (AREA)
- General Engineering & Computer Science (AREA)
- Zoology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Health & Medical Sciences (AREA)
- Biophysics (AREA)
- Biochemistry (AREA)
- Microbiology (AREA)
- Plant Pathology (AREA)
- Physics & Mathematics (AREA)
- Cell Biology (AREA)
- Botany (AREA)
- Gastroenterology & Hepatology (AREA)
- Medicinal Chemistry (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Nutrition Science (AREA)
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
Abstract
本发明公开了甘薯ERF转录因子IbERF4在促进植物绿原酸类物质合成中的用途,从甘薯中分离出编码ERF转录因子基因的完整cDNA,连接到植物表达载体上,利用农杆菌侵染法转化植物,获得的转基因植株,对转基因植株进行了抗逆性分析,结果表明IbERF4蛋白通过与绿原酸合成途径中4‑羟基桂皮酰辅酶A连接酶基因启动子结合激活其表达,IbERF4基因过表达转基因植株中4‑羟基桂皮酰辅酶A连接酶基因的表达水平上调,能够促进植物绿原酸类物质的合成,对于提高植物体内绿原酸类物质含量具有十分重要的意义。
Description
技术领域
本发明属于生物技术领域,具体涉及甘薯ERF转录因子IbERF4在促进植物绿原酸类物质合成中的一个新用途。
背景技术
转录因子是能与真核基因启动子区域中的顺式作用元件发生特异性结合,从而调控目的基因表达的蛋白质分子。AP2/ERF是植物中广泛存在的一类转录因子超家族,该家族蛋白含有AP2/ERF结构域,与植物的生长发育及逆境胁迫响应密切相关。IbERF4是一个从甘薯中分离出来的AP2/ERF家族转录因子,该基因受盐、干旱胁迫诱导表达。研究表明,在拟南芥中过量表达IbERF4明显削弱了转基因植株的耐盐性和耐旱性,在甘薯中过量表达IbERF4则会削弱转基因甘薯植株的耐盐性。本发明的发明人在公开号为CN104862320A的专利申请中已经报道了其负责调控植物耐逆与促进植物衰老的功能,然而,有关IbERF4在甘薯生长发育过程中的作用仍不清楚,随着研究的深入,本发明将继续探索其在甘薯生长发育过程中的作用。
绿原酸(Chlorogenic acid, CGA)是植物苯丙素类次生代谢产物之一,狭义上是指由咖啡酸(Caffe acid)与奎宁酸(Quinic acid)组成的咖啡单宁酸(5-O-Caffeoylquinic acid)。实际上,绿原酸在植物中通常是以几种异构体共存的形式存在。因此,广义上绿原酸代表由奎尼酸与反式肉桂酸(trans-Cinnamic acids)缩合而成的酯类化合物家族。目前,已报道的绿原酸合成途径在不同植物间存在一定差异,但基本包括苯丙氨酸解氨酶(L-phenylalaninammonia-lyase,PAL)、肉桂酰-4-羟化酶(Cinnamate-4-hydroxylase,C4H)、4-羟基桂皮酰辅酶A连接酶(4-coumaroyl-CoA-ligase,4CL)。近年已发现绿原酸具抗氧化、抗高血压、抗菌、抗肿瘤、抗辐射、降血糖、降血脂、抗炎、补肾、保肝等多种药理作用。除了药用外,绿原酸还可用于食品工业作食品和果品的保鲜剂。
以甘薯为代表的薯类作物,单位面积的生物能产量高于其它栽培作物,且具有耐瘠耐旱、适应性广、块根(茎)淀粉率高等特点,是世界上重要的粮食、饲料和工业原料作物。相比常见主粮作物,甘薯不仅含有丰富的蛋白质、糖类、维生素和矿物质,同时也因含有大量功能性成分而具有重要的药用价值,如绿原酸等。甘薯一方面可以作为日常生活快速便捷摄取绿原酸的来源,另一方面也因其巨大的生物量可用来开发高附加值的产品。因此,分析、挖掘和利用绿原酸合成调控相关基因将有助于提高植物绿原酸类物质的合成及其营养价值,促进植物绿原酸次生代谢工程的应用。
在前期的研究过程中,发明人已经找到了一些甘薯绿原酸合成途径关键酶基因,如公开号为CN111690672A的“甘薯绿原酸合成途径关键酶基因IbPAL2及应用”以及公开号为CN111690670A的“甘薯绿原酸合成途径关键酶基因IbHCT1及应用”,研究发现,对于控制同一性状的基因可能不止一个,继续寻找与绿原酸合成相关的基因,对甘薯生长发育研究具有重要的意义。
发明内容
本发明的目的在于提供一种甘薯ERF转录因子IbERF4的一个新用途,该转录因子能够促进植物绿原酸类物质合成。
还提供一种提高植物绿原酸含量的方法,通过转基因技术将IbERF4基因克隆到植物体内,获得转基因植株,该转基因植株绿原酸类物质含量提高。
为了实现上述目的,本发明的技术方案概述如下:
甘薯ERF转录因子IbERF4在促进植物绿原酸类物质合成中的用途,所述ERF转录因子IbERF4基因的核苷酸序列如SEQ ID NO.1 所示,其编码蛋白序列如SEQ ID NO.2所示。
其中,IbERF4蛋白通过与绿原酸合成途径中4-羟基桂皮酰辅酶A连接酶(4-coumarate:coenzyme A ligase, 4CL)基因启动子结合激活其表达;IbERF4基因过表达转基因植株中4-羟基桂皮酰辅酶A连接酶基因的表达水平上调。
所述绿原酸类物质包括绿原酸、异绿原酸A、异绿原酸B或异绿原酸C。
含有ERF转录因子IbERF4的过表达载体pCAMBIA1305-2×35s-IbERF4同样具有促进植物绿原酸类物质合成的作用。
为了提高植物的优良性状,本发明还公开了一种提高植物体内绿原酸含量的方法,将所述的IbERF4基因导入目的植物,得到转基因植物,所述转基因植物中的绿原酸类物质含量高于目的植物。
具体地,IbERF4基因具体可通过所述过表达载体导入所述目的植物。所述方法中,所述过表达载体可通过使用Ti质粒、Ri质粒、植物病毒载体、直接DNA转化、显微注射、电导、农杆菌介导等常规生物学方法转化植物细胞或组织,并将转化的植物组织培育成植株。
另外,也公开了一种植物育种方法,所述方法为如下(1)和/或(2):
(1)通过增加目的植物中IbERF4蛋白的活性,获得绿原酸类物质含量高于目的植物的植株;
(2)通过促进目的植物中IbERF4基因的表达,获得绿原酸类物质含量高于目的植物的植株。
其中,促进目的植物中IbERF4基因的表达的实现方式包括将IbERF4基因导入目的植物或引入强启动子和/或增强子。
本发明中,对于适用于本发明的植物没有特别的限制,只要其适合进行基因的转化操作即可,如各种农作物、花卉植物、或林业植物等。所述的植物比如可以是(不限于):双子叶植物、单子叶植物或裸子植物。
作为一种实施方式,所述的“植物”包括但不限于:甘薯,凡是具有该基因或者与之同源的基因均适用。尤其适用于需要提高绿原酸类物质含量的植物,在实际的应用过程中,对于需要提高绿原酸类物质含量的植物,均可以通过转基因的方式培育转入该基因的株系。
本发明中所说的“植物”包括整株植物,其亲本和子代植株以及植物的不同部位,包括种子、果实、芽、茎、叶、根(包括块茎)、花、组织和器官,在这些不同的部分均有我们目的基因或者核酸。这里所提及的“植物”也包括植物细胞、悬浮培养物、愈伤组织、胚、分生组织区、配子体、孢子体、花粉和小孢子,同样,其中每种前述对象包含目的基因/核酸。
本发明包括任何植物细胞,或任何由其中的方法获得或可获得的植物,以及所有的植物部分及其繁殖体。本专利也包含由任何前述方法所获得的转染细胞、组织、器官或完整植物。唯一的要求是子代表现出相同的的基因型或表型特征,使用本专利中的方法获得的子代特性相同。
本发明还扩展到如上所述的植物的可收获的部分,但不限于种子、叶、果实、花、茎、根、根茎、块茎和球茎。同时进一步涉及植株收获后的其他衍生物,如干燥颗粒或粉末、油、脂肪和脂肪酸、淀粉或蛋白质。本发明还涉及由相关植物获得的食品或食品添加剂。
本发明的优点:
本发明找到了一个甘薯ERF转录因子IbERF4的新用途,从甘薯中分离出编码ERF转录因子基因的完整cDNA,连接到植物表达载体上,利用农杆菌侵染法转化植物,获得的转基因植株,对转基因植株进行了抗逆性分析,结果表明IbERF4蛋白通过与绿原酸合成途径中4-羟基桂皮酰辅酶A连接酶基因启动子结合激活其表达,IbERF4基因过表达转基因植株中4-羟基桂皮酰辅酶A连接酶基因的表达水平上调,能够促进植物绿原酸类物质的合成,对于提高植物体内绿原酸类物质含量具有十分重要的意义。
对于一些需要提高植物体内绿原酸类物质含量的植物,可以通过导入该基因的方式培育一些新品种,对于植物育种具有重大的应用价值。
附图说明
图1是IbERF4过表达转基因代表家系IbERF4表达水平;
图2是IbERF4过表达转基因代表家系绿原酸类物质含量;
图3是IbERF4过表达转基因代表家系4CL表达水平;
图4是IbERF4与4CL启动子结合。
具体实施方式
下面结合具体实施例来进一步描述本发明,本发明的优点和特点将会随着描述而更为清楚。但下述实施例中所涉及的具体实验方法如无特殊说明,均为常规方法或按照制造厂商说明书建议的条件实施。
若未特别指明,实施例中所用技术手段为本领域技术人员所熟知的常规手段。下述实施例中的试验方法,如无特别说明,均为常规方法。如无特殊说明,所采用的试剂及材料,均可以从市场中购买获得。
除非另行定义,文中所使用的所有专业与科学用语与本领域熟练人员所熟悉的意义相同。此外,任何与所记载内容相似或均等的方法及材料皆可应用于本发明中。文中所述的较佳实施方法与材料仅作示范之用。
实施例1 IbERF4过表达转基因甘薯的获得及功能
1. IbERF4过表达转基因甘薯的获得
利用液氮冻融法将构建的过表达载体pCAMBIA1305-2×35s-IbERF4(江苏省农业科学院)转入农杆菌EHA105。将阳性克隆接种到50 mLYEP (含100 μg/mL Rif、100 μg/mLKan)液体培养基中,28℃、180 rpm继续培养至OD600至0.6~0.8。4000 rpm离心10 min,弃培养基,收集菌体。用MS1D液体培养基(4.4 g/l MS+0.4 mg/l VB1+30 g/l sucrose+肌醇0.1g/l +1 mg/l 2,4D)+0.1% As 将菌体稀释至OD600 0.3~0.5,制备成甘薯转化侵染液。将甘薯愈伤浸入其中20 min,期间遮光置于摇床缓慢摇晃。超声波超声10 sec后,倒掉侵染液,用消毒滤纸吸干愈伤表面水分,将干燥的已侵染愈伤转入MS1D(含0.1% As)培养基上共培2~3d(黑暗)。用无菌水洗净愈伤表面农杆菌,将愈伤转移至筛选培养基上(MS1D+10mg/L潮霉素+400 mg/L cefotaxime),暗培4-6周后转移至再生培养基。将潮霉素筛选后的愈伤转移至再生培养基MSCH (4.4 g/L MS+10 mg/L 潮霉素+200mg/L cefotaxime),待分化成苗后将苗转移至生根培养基SBMC (4.4 g/L MS+200 mg/L cefotaxime+0.3 mg/LVB1)。
将分化成苗的转基因苗移栽至盆钵,成活后(约7~14d)提取叶片DNA进行PCR鉴定,1%琼脂糖凝胶电泳检测PCR产物。对经PCR检测为阳性的转基因植株进一步取样,提取RNA进行Real-time PCR定量分析,以验证IbERF4过表达效应(图1)。
IbERF4基因的功能验证
以转基因受体徐薯29号(由中国农业科学院甘薯研究所徐州市农业科学院选育)及其转化过表达空载体植株(EV)为对照,取实施例1获得的IbERF4过表达转基因植株和对照植株相同部位叶片进行绿原酸含量测定,每植株设3次重复。具体操作步骤如下:准确称取烘干后的甘薯叶0.2g于100mL具塞锥形瓶中,加50%甲醇20mL浸泡24h,超声0.5h后过滤、洗涤。重复操作2次,合并滤液,用50%甲醇定容于50mL容量瓶中,稀释10倍后待测。
绿原酸标准曲线绘制:准确称取绿原酸和异绿原酸A、异绿原酸B、异绿原酸C标准品(购自北京索莱宝科技有限公司)各5.0mg,用50%甲醇溶液在超声条件下使其完全溶解,并定容至25mL,得到浓度各为0.2mg/mL的混合标准品溶液。分别取0.5、1.0、1.5、2.0和2.5mL标准品溶液置于25mL棕色容量瓶中,用50%甲醇溶液定容。在吸收光波长为326 nm的高效液相色谱仪中测定吸光度,以浓度为x坐标,吸光度为y坐标绘制。
结果表明(图2),IbERF4过表达转基因植株中绿原酸类物质含量明显高于对照(X29和EV),包括绿原酸、异绿原酸A(ICGA-A)、异绿原酸B(ICGA-B)和异绿原酸C(ICGA-C)。
实施例2 IbERF4过表达转基因植株中4CL表达水平检测
为了更进一步从分子水平上说明IbERF4基因在促进植物绿原酸类物质合成中的用途,继续进行了如下实验:
以转基因受体徐薯29号(由中国农业科学院甘薯研究所徐州市农业科学院选育)为对照,取实施例1获得的IbERF4过表达转基因植株和对照植株相同部位叶片,液氮速冻后置于-80 °C冰箱备用。取一部分样品用于转录组测序分析,结果表明苯丙氨酸代谢途径中4CL基因在IbERF4过表达转基因植株中的表达水平明显上调。另一部分样品用于对转录组测序结果进行验证,具体为:使用RNAprep Pure多糖多酚植物总RNA 提取试剂盒(北京天根)提取总RNA后,用NanoDrop 2000紫外-可见分光光度计(Thermo Fisher USA)检测RNA浓度及质量,1%琼脂糖电泳检测RNA完整性。取1 μg RNA作为模板,使用PrimeScript™ 1stStrand cDNA Synthesis Kit (Takara)反转录合成第一链cDNA,-20 °C保存。将反转录合成的cDNA稀释5倍后作为模板,利用SYBR premix Ex TaqTM (TaKaRa公司)试剂盒,在ABIStepOnePlus上扩增进行qRT-PCR反应。扩增程序为: 95 °C 60 s 预变性;95 °C 15 s,60°C 15 s,72 °C 45 s,40个循环;95 °C 15 s,60 °C 1 min。IbTublin基因(F:CAACTACCAGCCACCAACTGT,R:CAAGATCCTCACGAGCTTCAC)的表达量作为内参,计算出4CL基因(F:GGCGACTCATTTGATGGCTT,R:AATTGCCATCTCCTGACCCA)的相对表达量。样本和内参分别设3次重复。
结果表明(图3),IbERF4过表达转基因家系中4CL表达水平明显上调。
实施例3 IbERF4蛋白与4CL启动子区DRE元件互作
1.甘薯IbERF4基因与载体pB42AD(南京农业大学)的连接
以实施例1得到的pCAMBIA1305-2×35s-IbERF4质粒为模板,采用引物
ERF4AD-F:TGCCTCTCCCGAATTCATGGCGGTGAAGGGCAGA
ERF4AD-R:CGAGTCGGCCGAATTCAGCTTCCGTGGGTGGAGC
在IbERF4前后引入含酶切位点EcoR I的重组序列,利用TAKARA公司KOD Plus Neo聚合酶进行扩增,具体步骤如下:
Plasmid DNA 1 μL
10 pmol ERF4AD-F 1.5 μL
10 pmol ERF4AD-R 1.5 μL
2mM dNTPs 5 μL
25 mM MgSO4 3 μL
KOD Plus Neo (1.0 U/μl) 1 μL
10×PCR Buffer for KOD Plus Neo 5 μL
ddH2O Up to 50 μL
反应条件如下:94℃,2min;98℃,10sec;58℃,30sec;68℃,lmin;68℃,10min;34个循环。使用OMEGA DNA纯化试剂盒将PCR扩增产物纯化。使用限制性内切酶EcoR I将载体pB42AD酶切后,将纯化后的PCR产物与质粒的酶切产物相连接(50 °C 15 min),连接体系如下:
pB42AD酶切后空载体片段 2 μL
IbERF4加酶切位点PCR产物片段 1 μL
5×In-fusion HD Enzyme Premix 1 μL
ddH2O up to 5 μL
连接产物转化E-Coli.Trans1-T1(北京全式金生物),涂布于含100mg/ml浓度卡纳霉素抗性的LB平板上。37℃培养,12h后挑取单菌落进行菌落PCR验证,将菌落PCR验证阳性的菌,摇菌提取质粒,酶切鉴定得到目的条带,最后送生工生物工程(上海)测序公司测序,结果表明载体IbERF4-pB42AD构建正确。
2.甘薯4CL基因启动子区(p4CL)与载体pLacZi(南京农业大学)的连接
利用EasyPure® Plant Genomic DNA Kit (含RNase A)(全式金生物技术有限公司)提取甘薯叶片中DNA,具体操作步骤参考试剂盒说明书。以提取获得的甘薯叶片DNA为模板,采用引物
p4CLlaczi-F:ATCTGTCGACCTCGAGCCGTTATACACCGTCCCTGT
p4CLlaczi-R:GAGCACATGCCTCGAGACGAACGTTTCGGACACATAA
在4CL前后引入含酶切位点Xho I的重组序列,利用TAKARA公司KOD Plus Neo聚合酶进行扩增,具体步骤同1.1。使用OMEGA DNA纯化试剂盒将PCR扩增产物纯化。使用限制性内切酶Xho I将载体pLacZi酶切后,将纯化后的PCR产物与质粒的酶切产物相连接(50 °C10 min),连接体系如下:
pLacZi酶切后空载体片段 2 μL
p4CL加酶切位点PCR产物片段 1 μL
5×In-fusion HD Enzyme Premix 1 μL
ddH2O up to 5 μL
连接产物转化E-Coli.Trans1-T1(北京全式金生物),涂布于含100mg/ml浓度卡纳霉素抗性的LB平板上。37℃培养,12h后挑取单菌落进行菌落PCR验证,将菌落PCR验证阳性的菌,摇菌提取质粒,酶切鉴定得到目的条带,最后送生工生物工程(上海)测序公司测序,结果表明载体p4CL-pLacZi构建正确。
3.载体共转化
利用EGY48感受态细胞(上海唯地生物技术有限公司)进行载体共转化,具体操作步骤参考说明书。将转化后的菌液涂于SD-Trp-Ura培养基上,30℃培养3 d后,将每个板的酵母单克隆划线于SD-Trp-Ura/Gal/Raf+X-gal上,30℃培养3 d后,观察划线酵母状态。
结果表明(图4),IbERF4能够直接与4CL启动子结合,从而直接调节其表达水平。
以上所述之实施例,只是本发明的较佳实施例而已,仅仅用以解释本发明,并非限制本发明实施范围,对于本技术领域的技术人员来说,当然可根据本说明书中所公开的技术内容,通过置换或改变的方式轻易做出其它的实施方式,故凡在本发明的原理上所作的变化和改进等,均应包括于本发明申请专利范围内。
序列表
<110> 江苏省农业科学院
<120> 甘薯ERF转录因子IbERF4在促进植物绿原酸类物质合成中的用途
<130> 2022
<160> 2
<170> SIPOSequenceListing 1.0
<210> 1
<211> 684
<212> DNA
<213> Ipomoea batatas
<400> 1
atggcggtga agggcagaga ggagggcggc gttaagggcg gagccgggaa agtgaacggc 60
attaaggagg tgcactacag aggcgtgagg aagaggccgt ggggccgcta cgccgccgaa 120
atccgcgatc cagccaagaa gagccgcgtg tggctcggga cttttgacac ggcggaggag 180
gcggcgcgtg cgtatgatgc cgccgcccgg gagtttcgtg gggctaaggc gaagacgaac 240
ttccactcgc cgtcggagaa tcgcagcccc agtcagagca gcacggtgga gtcctccggg 300
agcgagacga ccggccacgc gccgcagttc ccgctcgagc tggatctcac gcgccgcctt 360
ggttccgccg aggccgccgg cgttagatcc gtcaataata ataacaatac attccagttc 420
tttcatcctc agccggccgt cgccgttctc cccaacggac agccggttct gcttttcgag 480
acgctgtggc ggccgggcgc cgtgagccgc ccactcccgg atcagttcga ggcggcgccg 540
gcgattccat ccaaacgccc cgctctcagc gactcaagca ccttctccgt cgtcgaagag 600
aacaacttcg tcggcgccgg cgccggcgtt gcagagaaac gccttaacct cgatcttaac 660
cttgctccac ccacggaagc ttga 684
<210> 2
<211> 227
<212> PRT
<213> Ipomoea batatas
<400> 2
Met Ala Val Lys Gly Arg Glu Glu Gly Gly Val Lys Gly Gly Ala Gly
1 5 10 15
Lys Val Asn Gly Ile Lys Glu Val His Tyr Arg Gly Val Arg Lys Arg
20 25 30
Pro Trp Gly Arg Tyr Ala Ala Glu Ile Arg Asp Pro Ala Lys Lys Ser
35 40 45
Arg Val Trp Leu Gly Thr Phe Asp Thr Ala Glu Glu Ala Ala Arg Ala
50 55 60
Tyr Asp Ala Ala Ala Arg Glu Phe Arg Gly Ala Lys Ala Lys Thr Asn
65 70 75 80
Phe His Ser Pro Ser Glu Asn Arg Ser Pro Ser Gln Ser Ser Thr Val
85 90 95
Glu Ser Ser Gly Ser Glu Thr Thr Gly His Ala Pro Gln Phe Pro Leu
100 105 110
Glu Leu Asp Leu Thr Arg Arg Leu Gly Ser Ala Glu Ala Ala Gly Val
115 120 125
Arg Ser Val Asn Asn Asn Asn Asn Thr Phe Gln Phe Phe His Pro Gln
130 135 140
Pro Ala Val Ala Val Leu Pro Asn Gly Gln Pro Val Leu Leu Phe Glu
145 150 155 160
Thr Leu Trp Arg Pro Gly Ala Val Ser Arg Pro Leu Pro Asp Gln Phe
165 170 175
Glu Ala Ala Pro Ala Ile Pro Ser Lys Arg Pro Ala Leu Ser Asp Ser
180 185 190
Ser Thr Phe Ser Val Val Glu Glu Asn Asn Phe Val Gly Ala Gly Ala
195 200 205
Gly Val Ala Glu Lys Arg Leu Asn Leu Asp Leu Asn Leu Ala Pro Pro
210 215 220
Thr Glu Ala
225
Claims (7)
1.甘薯ERF转录因子IbERF4在促进甘薯绿原酸类物质合成中的用途,其特征在于,甘薯ERF转录因子IbERF4基因的核苷酸序列如SEQ ID NO.1 所示,其氨基酸序列如SEQ ID NO.2所示。
2.根据权利要求1所述的用途,其特征在于,IbERF4基因过表达转基因植株中4-羟基桂皮酰辅酶A连接酶基因的表达水平上调。
3.根据权利要求1所述的用途,其特征在于,IbERF4蛋白通过与绿原酸合成途径中4-羟基桂皮酰辅酶A连接酶基因启动子结合激活其表达。
4.根据权利要求1所述的用途,其特征在于,所述绿原酸类物质包括绿原酸、异绿原酸A、异绿原酸B或异绿原酸C。
5.根据权利要求1所述的用途,含有权利要求1中所述IbERF4基因的过表达载体为pCAMBIA1305-2×35s-IbERF4。
6.一种提高植物体内绿原酸含量的方法,其特征在于,将权利要求1中所述的IbERF4基因导入目的植物,得到转基因植物,所述转基因植物中的绿原酸类物质含量高于目的植物,所述目的植物为甘薯。
7.一种植物育种方法,其特征在于,所述方法为如下(1)和/或(2):
(1)通过增加目的植物中IbERF4蛋白的活性,获得绿原酸类物质含量高于目的植物的植株;
(2)通过促进目的植物中IbERF4基因的表达,获得绿原酸类物质含量高于目的植物的植株;所述目的植物为甘薯。
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202210309189.7A CN114395566B (zh) | 2022-03-28 | 2022-03-28 | 甘薯ERF转录因子IbERF4在促进植物绿原酸类物质合成中的用途 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202210309189.7A CN114395566B (zh) | 2022-03-28 | 2022-03-28 | 甘薯ERF转录因子IbERF4在促进植物绿原酸类物质合成中的用途 |
Publications (2)
Publication Number | Publication Date |
---|---|
CN114395566A CN114395566A (zh) | 2022-04-26 |
CN114395566B true CN114395566B (zh) | 2022-05-24 |
Family
ID=81234556
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202210309189.7A Active CN114395566B (zh) | 2022-03-28 | 2022-03-28 | 甘薯ERF转录因子IbERF4在促进植物绿原酸类物质合成中的用途 |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN114395566B (zh) |
Families Citing this family (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN115807003A (zh) * | 2022-08-24 | 2023-03-17 | 中国烟草总公司郑州烟草研究院 | NtERF4基因及其在总黄酮与绿原酸合成调控中应用 |
Citations (13)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN101693739A (zh) * | 2009-10-15 | 2010-04-14 | 中国科学院植物研究所 | 绿原酸合成相关蛋白及其编码基因与应用 |
WO2011014741A1 (en) * | 2009-07-31 | 2011-02-03 | Artemis Health, Inc. | Methods and compositions for cell stabilization |
CN102271496A (zh) * | 2008-12-01 | 2011-12-07 | 维亚拉克什亚生物科学(新西兰)有限公司 | 提高植物对环境应激的耐受性的方法和组合物 |
WO2013178705A1 (fr) * | 2012-05-29 | 2013-12-05 | Plant Advanced Technologies Pat | Enzymes de bioconversion de l'acide chrologenique en au moins un acide di-, tri- ou tetra-cafeoylquinique |
CN104024415A (zh) * | 2011-11-14 | 2014-09-03 | 巴斯夫植物科学有限公司 | 具有增强的产量相关性状的植物及其制备方法 |
CN104862320A (zh) * | 2015-04-28 | 2015-08-26 | 江苏省农业科学院 | 一种编码甘薯ERF转录因子的IbERF4基因及应用 |
CN105671058A (zh) * | 2016-03-24 | 2016-06-15 | 江苏省农业科学院 | 编码甘薯erf转录因子的基因及应用 |
CN106222182A (zh) * | 2016-08-11 | 2016-12-14 | 江苏省农业科学院 | 编码甘薯ERF转录因子的IbERF5基因及应用 |
CN109207499A (zh) * | 2018-09-28 | 2019-01-15 | 江苏省农业科学院 | 甘薯绿原酸合成途径中关键酶基因IbPAL1及应用 |
CN111690670A (zh) * | 2020-07-23 | 2020-09-22 | 江苏省农业科学院 | 甘薯绿原酸合成途径关键酶基因IbHCT1及应用 |
CN111690672A (zh) * | 2020-07-23 | 2020-09-22 | 江苏省农业科学院 | 甘薯绿原酸合成途径关键酶基因IbPAL2及应用 |
CN112105257A (zh) * | 2018-03-05 | 2020-12-18 | 奥驰亚客户服务有限公司 | 用于生产具有改变的生物碱含量和期望的叶质量的烟草植物和制品的组合物和方法 |
WO2021180959A1 (fr) * | 2020-03-13 | 2021-09-16 | Universite De Lorraine | Enzyme pour la conversion d'acide chlorogénique en acide isochlorogénique |
Family Cites Families (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP3701042A4 (en) * | 2017-10-23 | 2021-08-11 | The Broad Institute, Inc. | SYSTEMS, PROCEDURES AND COMPOSITIONS FOR TARGETED NUCLEIC ACID EDITING |
-
2022
- 2022-03-28 CN CN202210309189.7A patent/CN114395566B/zh active Active
Patent Citations (13)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN102271496A (zh) * | 2008-12-01 | 2011-12-07 | 维亚拉克什亚生物科学(新西兰)有限公司 | 提高植物对环境应激的耐受性的方法和组合物 |
WO2011014741A1 (en) * | 2009-07-31 | 2011-02-03 | Artemis Health, Inc. | Methods and compositions for cell stabilization |
CN101693739A (zh) * | 2009-10-15 | 2010-04-14 | 中国科学院植物研究所 | 绿原酸合成相关蛋白及其编码基因与应用 |
CN104024415A (zh) * | 2011-11-14 | 2014-09-03 | 巴斯夫植物科学有限公司 | 具有增强的产量相关性状的植物及其制备方法 |
WO2013178705A1 (fr) * | 2012-05-29 | 2013-12-05 | Plant Advanced Technologies Pat | Enzymes de bioconversion de l'acide chrologenique en au moins un acide di-, tri- ou tetra-cafeoylquinique |
CN104862320A (zh) * | 2015-04-28 | 2015-08-26 | 江苏省农业科学院 | 一种编码甘薯ERF转录因子的IbERF4基因及应用 |
CN105671058A (zh) * | 2016-03-24 | 2016-06-15 | 江苏省农业科学院 | 编码甘薯erf转录因子的基因及应用 |
CN106222182A (zh) * | 2016-08-11 | 2016-12-14 | 江苏省农业科学院 | 编码甘薯ERF转录因子的IbERF5基因及应用 |
CN112105257A (zh) * | 2018-03-05 | 2020-12-18 | 奥驰亚客户服务有限公司 | 用于生产具有改变的生物碱含量和期望的叶质量的烟草植物和制品的组合物和方法 |
CN109207499A (zh) * | 2018-09-28 | 2019-01-15 | 江苏省农业科学院 | 甘薯绿原酸合成途径中关键酶基因IbPAL1及应用 |
WO2021180959A1 (fr) * | 2020-03-13 | 2021-09-16 | Universite De Lorraine | Enzyme pour la conversion d'acide chlorogénique en acide isochlorogénique |
CN111690670A (zh) * | 2020-07-23 | 2020-09-22 | 江苏省农业科学院 | 甘薯绿原酸合成途径关键酶基因IbHCT1及应用 |
CN111690672A (zh) * | 2020-07-23 | 2020-09-22 | 江苏省农业科学院 | 甘薯绿原酸合成途径关键酶基因IbPAL2及应用 |
Non-Patent Citations (9)
Title |
---|
Bian,X..Ipomoea batatas cultivar Sushu 16 ethylene response factor 4 (ERF4) mRNA, complete cds.《Genbank database》.2017, * |
Liu Yi等.Integrated transcriptome, small RNA and degradome sequencing approaches proffer insights into chlorogenic acid biosynthesis in leafy sweet potato.《PloS one》.2021,第16卷(第1期), * |
Shutao He等.Genome-wide identification, phylogeny and expression analysis of AP2/ERF transcription factors family in sweet potato.《BMC Genomics》.2021,第22卷 * |
Xu Jing等.Comparative transcriptome and weighted correlation network analyses reveal candidate genes involved in chlorogenic acid biosynthesis in sweet potato.《Scientific Reports》.2022,第12卷 * |
Yang Yu等.A novel ethylene‑responsive factor IbERF4 from sweetpotato negatively regulates abiotic stress.《Plant Biotechnology Reports》.2020,第14卷 * |
丁安然等.苹果MdERF113基因克隆及响应轮纹病菌侵染的表达分析.《青岛农业大学学报(自然科学版)》.2018,(第03期), * |
张斌斌等.基于果肉单体酚和总酚含量评价桃果实抗氧化能力.《园艺学报》.2018,(第05期), * |
边小峰等.甘薯抗逆相关基因IbERF3的克隆与表达分析.《华北农学报》.2014,(第06期), * |
阮先乐等.甘薯AP2基因家族的生物信息学分析.《分子植物育种》.2017,(第06期), * |
Also Published As
Publication number | Publication date |
---|---|
CN114395566A (zh) | 2022-04-26 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN108864267B (zh) | 甘薯类胡萝卜素合成和耐盐抗旱相关蛋白IbARF5及其编码基因与应用 | |
CN109081865B (zh) | 毛竹PeVQ28蛋白及其编码基因与应用 | |
CN111909941B (zh) | 一种百合转录因子基因LrWRKY-L1及应用 | |
CN113563442A (zh) | 抗旱相关蛋白IbSPB1及其编码基因与应用 | |
CN113265408A (zh) | 一种三七DOF转录因子基因PnDof1及其应用 | |
CN114350684B (zh) | 一种苹果MdERF-073基因、蛋白及应用 | |
CN114395566B (zh) | 甘薯ERF转录因子IbERF4在促进植物绿原酸类物质合成中的用途 | |
CN114540357A (zh) | 玉米长链非编码RNA lncRNA25659及其用途 | |
CN113845578A (zh) | 调控植物原花青素合成的myb类转录因子及其编码基因和应用 | |
CN111690665A (zh) | 一种分离自砂梨的具有抗黑斑病功能的基因PpHSP21及其应用 | |
CN116590310A (zh) | 梨转录因子PbbHLH164在促进果实成熟中的应用 | |
CN114214332B (zh) | 一种天目地黄花青素相关基因RcMYB1及其应用 | |
CN111961124B (zh) | 一种植物早熟蛋白及其编码基因与应用 | |
CN111996197B (zh) | 一种杜梨耐盐基因、蛋白及重组载体和应用 | |
WO2007061146A1 (en) | A method for producing chinese cabbage transformant using tissues of flower stalk and a transformant with promoted soft rot resistance obtained from the method | |
CN111154772A (zh) | 梨糖转运基因PbSWEET4及其应用 | |
CN116640775B (zh) | 能够增强MBW复合体调控花青苷合成能力的龙眼DlMYB15基因 | |
CN116606865B (zh) | 一种红皮龙眼DlMYBrp基因及其应用 | |
CN114908106B (zh) | 一种玫瑰耐盐基因RrLBD40及其应用 | |
CN108148849A (zh) | 一种苹果MdPHR1基因及其制备方法和应用 | |
CN118064497B (zh) | PfbZIP52基因的过表达载体及构建方法和在烟草中的应用 | |
KR100996667B1 (ko) | 벼의 캘러스 또는 분화중인 캘러스에서 특이적으로 발현하는 유전자의 프로모터 및 전사인자 | |
CN114561404B (zh) | 苹果MdSHN1基因及在提高植物耐涝性中的应用 | |
CN111793641B (zh) | 甜樱桃PavSS或PavSPS基因在调控果实着色或果实成熟软化中的用途 | |
KR20100006228A (ko) | 식물 스트레스 저항성을 증가시키는 AtUBPH1 및AtUBPH2 유전자의 돌연변이체 및 상기 유전자가도입된 성장을 촉진시키는 형질전환 식물체 |
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 | ||
GR01 | Patent grant | ||
GR01 | Patent grant |