CN114752608B - 一种高含量油菜素内酯番茄植株的培育方法 - Google Patents
一种高含量油菜素内酯番茄植株的培育方法 Download PDFInfo
- Publication number
- CN114752608B CN114752608B CN202210456566.XA CN202210456566A CN114752608B CN 114752608 B CN114752608 B CN 114752608B CN 202210456566 A CN202210456566 A CN 202210456566A CN 114752608 B CN114752608 B CN 114752608B
- Authority
- CN
- China
- Prior art keywords
- gene
- cpk27
- seq
- tomato
- content
- 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
- IXVMHGVQKLDRKH-VRESXRICSA-N Brassinolide Natural products O=C1OC[C@@H]2[C@@H]3[C@@](C)([C@H]([C@@H]([C@@H](O)[C@H](O)[C@H](C(C)C)C)C)CC3)CC[C@@H]2[C@]2(C)[C@@H]1C[C@H](O)[C@H](O)C2 IXVMHGVQKLDRKH-VRESXRICSA-N 0.000 title claims abstract description 37
- IXVMHGVQKLDRKH-KNBKMWSGSA-N brassinolide Chemical compound C1OC(=O)[C@H]2C[C@H](O)[C@H](O)C[C@]2(C)[C@H]2CC[C@]3(C)[C@@H]([C@H](C)[C@@H](O)[C@H](O)[C@@H](C)C(C)C)CC[C@H]3[C@@H]21 IXVMHGVQKLDRKH-KNBKMWSGSA-N 0.000 title claims abstract description 29
- 238000012364 cultivation method Methods 0.000 title claims abstract description 8
- 240000003768 Solanum lycopersicum Species 0.000 title description 67
- 235000007688 Lycopersicon esculentum Nutrition 0.000 claims abstract description 61
- 108090000623 proteins and genes Proteins 0.000 claims abstract description 21
- 150000001647 brassinosteroids Chemical class 0.000 claims abstract description 20
- 239000002773 nucleotide Substances 0.000 claims abstract description 9
- 125000003729 nucleotide group Chemical group 0.000 claims abstract description 9
- 102000004169 proteins and genes Human genes 0.000 claims abstract description 9
- 239000000126 substance Substances 0.000 claims abstract description 9
- 230000001105 regulatory effect Effects 0.000 claims abstract description 8
- 108700026244 Open Reading Frames Proteins 0.000 claims abstract description 6
- 241000227653 Lycopersicon Species 0.000 claims abstract 6
- 108091033409 CRISPR Proteins 0.000 claims description 26
- 238000010354 CRISPR gene editing Methods 0.000 claims description 10
- 239000012634 fragment Substances 0.000 claims description 8
- 238000000034 method Methods 0.000 claims description 7
- 241000589158 Agrobacterium Species 0.000 claims description 4
- 241000894006 Bacteria Species 0.000 claims description 4
- 238000013461 design Methods 0.000 claims description 2
- 230000002068 genetic effect Effects 0.000 claims description 2
- 230000001131 transforming effect Effects 0.000 claims description 2
- 230000003828 downregulation Effects 0.000 claims 1
- 241000196324 Embryophyta Species 0.000 abstract description 45
- 101150095452 CPK27 gene Proteins 0.000 abstract description 28
- 238000010362 genome editing Methods 0.000 abstract description 10
- 238000005516 engineering process Methods 0.000 abstract description 9
- 108020004414 DNA Proteins 0.000 description 19
- 101150031975 CYP85A1 gene Proteins 0.000 description 14
- 108091027544 Subgenomic mRNA Proteins 0.000 description 11
- 150000001646 brassinolides Chemical class 0.000 description 11
- 230000014509 gene expression Effects 0.000 description 10
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 9
- 230000015572 biosynthetic process Effects 0.000 description 9
- 239000007788 liquid Substances 0.000 description 9
- IXVMHGVQKLDRKH-YEJCTVDLSA-N (22s,23s)-epibrassinolide Chemical compound C1OC(=O)[C@H]2C[C@H](O)[C@H](O)C[C@]2(C)[C@H]2CC[C@]3(C)[C@@H]([C@H](C)[C@H](O)[C@@H](O)[C@H](C)C(C)C)CC[C@H]3[C@@H]21 IXVMHGVQKLDRKH-YEJCTVDLSA-N 0.000 description 8
- 238000012217 deletion Methods 0.000 description 8
- 230000037430 deletion Effects 0.000 description 8
- 238000012163 sequencing technique Methods 0.000 description 7
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 6
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 6
- 235000002560 Solanum lycopersicum Nutrition 0.000 description 6
- 239000000463 material Substances 0.000 description 6
- 238000003786 synthesis reaction Methods 0.000 description 6
- 230000037361 pathway Effects 0.000 description 5
- 239000006228 supernatant Substances 0.000 description 5
- 102100035102 E3 ubiquitin-protein ligase MYCBP2 Human genes 0.000 description 4
- 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 4
- 108091028043 Nucleic acid sequence Proteins 0.000 description 4
- 235000013399 edible fruits Nutrition 0.000 description 4
- 230000003647 oxidation Effects 0.000 description 4
- 238000007254 oxidation reaction Methods 0.000 description 4
- 239000013612 plasmid Substances 0.000 description 4
- 238000001179 sorption measurement Methods 0.000 description 4
- 238000007619 statistical method Methods 0.000 description 4
- 230000035882 stress Effects 0.000 description 4
- 238000012360 testing method Methods 0.000 description 4
- 239000002699 waste material Substances 0.000 description 4
- BHPQYMZQTOCNFJ-UHFFFAOYSA-N Calcium cation Chemical compound [Ca+2] BHPQYMZQTOCNFJ-UHFFFAOYSA-N 0.000 description 3
- 102000004190 Enzymes Human genes 0.000 description 3
- 108090000790 Enzymes Proteins 0.000 description 3
- 101100219321 Oryza sativa subsp. japonica CYP85A1 gene Proteins 0.000 description 3
- 150000001413 amino acids Chemical class 0.000 description 3
- 229910001424 calcium ion Inorganic materials 0.000 description 3
- 238000005119 centrifugation Methods 0.000 description 3
- 238000006243 chemical reaction Methods 0.000 description 3
- 230000001276 controlling effect Effects 0.000 description 3
- 238000011161 development Methods 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 108010054155 lysyllysine Proteins 0.000 description 3
- 239000000203 mixture Substances 0.000 description 3
- 229910052757 nitrogen Inorganic materials 0.000 description 3
- 230000002018 overexpression Effects 0.000 description 3
- 239000000047 product Substances 0.000 description 3
- 238000012795 verification Methods 0.000 description 3
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 2
- 101100165817 Arabidopsis thaliana CYP90B1 gene Proteins 0.000 description 2
- SNDBKTFJWVEVPO-WHFBIAKZSA-N Asp-Gly-Ser Chemical compound [H]N[C@@H](CC(O)=O)C(=O)NCC(=O)N[C@@H](CO)C(O)=O SNDBKTFJWVEVPO-WHFBIAKZSA-N 0.000 description 2
- RVMXMLSYBTXCAV-VEVYYDQMSA-N Asp-Pro-Thr Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N1CCC[C@H]1C(=O)N[C@@H]([C@@H](C)O)C(O)=O RVMXMLSYBTXCAV-VEVYYDQMSA-N 0.000 description 2
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 2
- 206010064571 Gene mutation Diseases 0.000 description 2
- 241000880493 Leptailurus serval Species 0.000 description 2
- 238000012408 PCR amplification Methods 0.000 description 2
- 102000001253 Protein Kinase Human genes 0.000 description 2
- HNDMFDBQXYZSRM-IHRRRGAJSA-N Ser-Val-Phe Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O HNDMFDBQXYZSRM-IHRRRGAJSA-N 0.000 description 2
- 108700005078 Synthetic Genes Proteins 0.000 description 2
- AEMPCGRFEZTWIF-IHRRRGAJSA-N Val-Leu-Lys Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCCN)C(O)=O AEMPCGRFEZTWIF-IHRRRGAJSA-N 0.000 description 2
- 238000009825 accumulation Methods 0.000 description 2
- 230000004075 alteration Effects 0.000 description 2
- 108010038633 aspartylglutamate Proteins 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- 230000000975 bioactive effect Effects 0.000 description 2
- 235000021466 carotenoid Nutrition 0.000 description 2
- 150000001747 carotenoids Chemical class 0.000 description 2
- 230000001419 dependent effect Effects 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 201000010099 disease Diseases 0.000 description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
- 235000013305 food Nutrition 0.000 description 2
- 230000013632 homeostatic process Effects 0.000 description 2
- BDAGIHXWWSANSR-UHFFFAOYSA-N methanoic acid Natural products OC=O BDAGIHXWWSANSR-UHFFFAOYSA-N 0.000 description 2
- 238000002156 mixing Methods 0.000 description 2
- 229930195732 phytohormone Natural products 0.000 description 2
- 238000002360 preparation method Methods 0.000 description 2
- 108060006633 protein kinase Proteins 0.000 description 2
- 238000003753 real-time PCR Methods 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 108010073969 valyllysine Proteins 0.000 description 2
- VYUIKSFYFRVQLF-RDIRXGJISA-N 3-Epicastasterone Natural products O=C1[C@@H]2[C@@](C)([C@@H]3[C@H]([C@H]4[C@](C)([C@H]([C@@H]([C@@H](O)[C@H](O)[C@H](C(C)C)C)C)CC4)CC3)C1)C[C@@H](O)[C@H](O)C2 VYUIKSFYFRVQLF-RDIRXGJISA-N 0.000 description 1
- OSWFIVFLDKOXQC-UHFFFAOYSA-N 4-(3-methoxyphenyl)aniline Chemical compound COC1=CC=CC(C=2C=CC(N)=CC=2)=C1 OSWFIVFLDKOXQC-UHFFFAOYSA-N 0.000 description 1
- UHPMCKVQTMMPCG-UHFFFAOYSA-N 5,8-dihydroxy-2-methoxy-6-methyl-7-(2-oxopropyl)naphthalene-1,4-dione Chemical compound CC1=C(CC(C)=O)C(O)=C2C(=O)C(OC)=CC(=O)C2=C1O UHPMCKVQTMMPCG-UHFFFAOYSA-N 0.000 description 1
- DKJPOZOEBONHFS-ZLUOBGJFSA-N Ala-Ala-Asp Chemical compound C[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@H](C(O)=O)CC(O)=O DKJPOZOEBONHFS-ZLUOBGJFSA-N 0.000 description 1
- VBDMWOKJZDCFJM-FXQIFTODSA-N Ala-Ala-Met Chemical compound CSCC[C@@H](C(O)=O)NC(=O)[C@H](C)NC(=O)[C@H](C)N VBDMWOKJZDCFJM-FXQIFTODSA-N 0.000 description 1
- GFBLJMHGHAXGNY-ZLUOBGJFSA-N Ala-Asn-Asp Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(O)=O)C(O)=O GFBLJMHGHAXGNY-ZLUOBGJFSA-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
- 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
- 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
- MEFILNJXAVSUTO-JXUBOQSCSA-N Ala-Leu-Thr Chemical compound C[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)O)C(O)=O MEFILNJXAVSUTO-JXUBOQSCSA-N 0.000 description 1
- BFMIRJBURUXDRG-DLOVCJGASA-N Ala-Phe-Asp Chemical compound OC(=O)C[C@@H](C(O)=O)NC(=O)[C@@H](NC(=O)[C@@H](N)C)CC1=CC=CC=C1 BFMIRJBURUXDRG-DLOVCJGASA-N 0.000 description 1
- IORKCNUBHNIMKY-CIUDSAMLSA-N Ala-Pro-Glu Chemical compound C[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CCC(O)=O)C(O)=O IORKCNUBHNIMKY-CIUDSAMLSA-N 0.000 description 1
- VJVQKGYHIZPSNS-FXQIFTODSA-N Ala-Ser-Arg Chemical compound C[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@H](C(O)=O)CCCN=C(N)N VJVQKGYHIZPSNS-FXQIFTODSA-N 0.000 description 1
- RTZCUEHYUQZIDE-WHFBIAKZSA-N Ala-Ser-Gly Chemical compound C[C@H](N)C(=O)N[C@@H](CO)C(=O)NCC(O)=O RTZCUEHYUQZIDE-WHFBIAKZSA-N 0.000 description 1
- CLOMBHBBUKAUBP-LSJOCFKGSA-N Ala-Val-His Chemical compound C[C@@H](C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC1=CN=CN1)C(=O)O)N CLOMBHBBUKAUBP-LSJOCFKGSA-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
- USFZMSVCRYTOJT-UHFFFAOYSA-N Ammonium acetate Chemical compound N.CC(O)=O USFZMSVCRYTOJT-UHFFFAOYSA-N 0.000 description 1
- 239000005695 Ammonium acetate Substances 0.000 description 1
- 241000219195 Arabidopsis thaliana Species 0.000 description 1
- 101100438273 Arabidopsis thaliana CAN1 gene Proteins 0.000 description 1
- UXJCMQFPDWCHKX-DCAQKATOSA-N Arg-Arg-Glu Chemical compound NC(N)=NCCC[C@H](N)C(=O)N[C@@H](CCCN=C(N)N)C(=O)N[C@@H](CCC(O)=O)C(O)=O UXJCMQFPDWCHKX-DCAQKATOSA-N 0.000 description 1
- OTUQSEPIIVBYEM-IHRRRGAJSA-N Arg-Asn-Tyr Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O OTUQSEPIIVBYEM-IHRRRGAJSA-N 0.000 description 1
- QAODJPUKWNNNRP-DCAQKATOSA-N Arg-Glu-Arg Chemical compound NC(N)=NCCC[C@H](N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCN=C(N)N)C(O)=O QAODJPUKWNNNRP-DCAQKATOSA-N 0.000 description 1
- GXXWTNKNFFKTJB-NAKRPEOUSA-N Arg-Ile-Ser Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CO)C(O)=O GXXWTNKNFFKTJB-NAKRPEOUSA-N 0.000 description 1
- JEOCWTUOMKEEMF-RHYQMDGZSA-N Arg-Leu-Thr Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)O)C(O)=O JEOCWTUOMKEEMF-RHYQMDGZSA-N 0.000 description 1
- MTYLORHAQXVQOW-AVGNSLFASA-N Arg-Lys-Met Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCSC)C(O)=O MTYLORHAQXVQOW-AVGNSLFASA-N 0.000 description 1
- KXOPYFNQLVUOAQ-FXQIFTODSA-N Arg-Ser-Ala Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CO)C(=O)N[C@@H](C)C(O)=O KXOPYFNQLVUOAQ-FXQIFTODSA-N 0.000 description 1
- ASQKVGRCKOFKIU-KZVJFYERSA-N Arg-Thr-Ala Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](C)C(=O)O)NC(=O)[C@H](CCCN=C(N)N)N)O ASQKVGRCKOFKIU-KZVJFYERSA-N 0.000 description 1
- CMLGVVWQQHUXOZ-GHCJXIJMSA-N Asn-Ala-Ile Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](C)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O CMLGVVWQQHUXOZ-GHCJXIJMSA-N 0.000 description 1
- MSBDSTRUMZFSEU-PEFMBERDSA-N Asn-Glu-Ile Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O MSBDSTRUMZFSEU-PEFMBERDSA-N 0.000 description 1
- SPCONPVIDFMDJI-QSFUFRPTSA-N Asn-Ile-Val Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](C(C)C)C(O)=O SPCONPVIDFMDJI-QSFUFRPTSA-N 0.000 description 1
- WIDVAWAQBRAKTI-YUMQZZPRSA-N Asn-Leu-Gly Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(C)C)C(=O)NCC(O)=O WIDVAWAQBRAKTI-YUMQZZPRSA-N 0.000 description 1
- ORJQQZIXTOYGGH-SRVKXCTJSA-N Asn-Lys-Leu Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(C)C)C(O)=O ORJQQZIXTOYGGH-SRVKXCTJSA-N 0.000 description 1
- GZXOUBTUAUAVHD-ACZMJKKPSA-N Asn-Ser-Glu Chemical compound NC(=O)C[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@H](C(O)=O)CCC(O)=O GZXOUBTUAUAVHD-ACZMJKKPSA-N 0.000 description 1
- MKJBPDLENBUHQU-CIUDSAMLSA-N Asn-Ser-Leu Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(O)=O MKJBPDLENBUHQU-CIUDSAMLSA-N 0.000 description 1
- SNYCNNPOFYBCEK-ZLUOBGJFSA-N Asn-Ser-Ser Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CO)C(O)=O SNYCNNPOFYBCEK-ZLUOBGJFSA-N 0.000 description 1
- ZLGKHJHFYSRUBH-FXQIFTODSA-N Asp-Arg-Asp Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(O)=O)C(O)=O ZLGKHJHFYSRUBH-FXQIFTODSA-N 0.000 description 1
- YNQIDCRRTWGHJD-ZLUOBGJFSA-N Asp-Asn-Ala Chemical compound OC(=O)[C@H](C)NC(=O)[C@H](CC(N)=O)NC(=O)[C@@H](N)CC(O)=O YNQIDCRRTWGHJD-ZLUOBGJFSA-N 0.000 description 1
- JUWZKMBALYLZCK-WHFBIAKZSA-N Asp-Gly-Asn Chemical compound OC(=O)C[C@H](N)C(=O)NCC(=O)N[C@@H](CC(N)=O)C(O)=O JUWZKMBALYLZCK-WHFBIAKZSA-N 0.000 description 1
- TZOZNVLBTAFJRW-UGYAYLCHSA-N Asp-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](CC(=O)O)N TZOZNVLBTAFJRW-UGYAYLCHSA-N 0.000 description 1
- XLILXFRAKOYEJX-GUBZILKMSA-N Asp-Leu-Gln Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(N)=O)C(O)=O XLILXFRAKOYEJX-GUBZILKMSA-N 0.000 description 1
- CJUKAWUWBZCTDQ-SRVKXCTJSA-N Asp-Leu-Lys Chemical compound OC(=O)C[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCCN)C(O)=O CJUKAWUWBZCTDQ-SRVKXCTJSA-N 0.000 description 1
- QNMKWNONJGKJJC-NHCYSSNCSA-N Asp-Leu-Val Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](C(C)C)C(O)=O QNMKWNONJGKJJC-NHCYSSNCSA-N 0.000 description 1
- JJQGZGOEDSSHTE-FOHZUACHSA-N Asp-Thr-Gly Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)NCC(O)=O JJQGZGOEDSSHTE-FOHZUACHSA-N 0.000 description 1
- JDDYEZGPYBBPBN-JRQIVUDYSA-N Asp-Thr-Tyr Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O JDDYEZGPYBBPBN-JRQIVUDYSA-N 0.000 description 1
- 108010027344 Basic Helix-Loop-Helix Transcription Factors Proteins 0.000 description 1
- 102000018720 Basic Helix-Loop-Helix Transcription Factors Human genes 0.000 description 1
- VYUIKSFYFRVQLF-LCILBPPUSA-N Castasterone Natural products O=C1[C@H]2[C@@](C)([C@@H]3[C@@H]([C@H]4[C@](C)([C@@H]([C@@H]([C@@H](O)[C@H](O)[C@H](C(C)C)C)C)CC4)CC3)C1)C[C@@H](O)[C@@H](O)C2 VYUIKSFYFRVQLF-LCILBPPUSA-N 0.000 description 1
- LZZYPRNAOMGNLH-UHFFFAOYSA-M Cetrimonium bromide Chemical compound [Br-].CCCCCCCCCCCCCCCC[N+](C)(C)C LZZYPRNAOMGNLH-UHFFFAOYSA-M 0.000 description 1
- 108091026890 Coding region Proteins 0.000 description 1
- ZJBWJHQDOIMVLM-WHFBIAKZSA-N Cys-Cys-Gly Chemical compound SC[C@H](N)C(=O)N[C@@H](CS)C(=O)NCC(O)=O ZJBWJHQDOIMVLM-WHFBIAKZSA-N 0.000 description 1
- NIXHTNJAGGFBAW-CIUDSAMLSA-N Cys-Lys-Ser Chemical compound C(CCN)C[C@@H](C(=O)N[C@@H](CO)C(=O)O)NC(=O)[C@H](CS)N NIXHTNJAGGFBAW-CIUDSAMLSA-N 0.000 description 1
- 108010090461 DFG peptide Proteins 0.000 description 1
- 102100030011 Endoribonuclease Human genes 0.000 description 1
- 101710199605 Endoribonuclease Proteins 0.000 description 1
- 241000588724 Escherichia coli Species 0.000 description 1
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 1
- 241000223218 Fusarium Species 0.000 description 1
- 241000223221 Fusarium oxysporum Species 0.000 description 1
- LKUWAWGNJYJODH-KBIXCLLPSA-N Gln-Ala-Ile Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](C)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O LKUWAWGNJYJODH-KBIXCLLPSA-N 0.000 description 1
- OSCLNNWLKKIQJM-WDSKDSINSA-N Gln-Ser-Gly Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CO)C(=O)NCC(O)=O OSCLNNWLKKIQJM-WDSKDSINSA-N 0.000 description 1
- UBRQJXFDVZNYJP-AVGNSLFASA-N Gln-Tyr-Ser Chemical compound C1=CC(=CC=C1C[C@@H](C(=O)N[C@@H](CO)C(=O)O)NC(=O)[C@H](CCC(=O)N)N)O UBRQJXFDVZNYJP-AVGNSLFASA-N 0.000 description 1
- LTUVYLVIZHJCOQ-KKUMJFAQSA-N Glu-Arg-Phe Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O LTUVYLVIZHJCOQ-KKUMJFAQSA-N 0.000 description 1
- RJONUNZIMUXUOI-GUBZILKMSA-N Glu-Asn-Lys Chemical compound C(CCN)C[C@@H](C(=O)O)NC(=O)[C@H](CC(=O)N)NC(=O)[C@H](CCC(=O)O)N RJONUNZIMUXUOI-GUBZILKMSA-N 0.000 description 1
- JPHYJQHPILOKHC-ACZMJKKPSA-N Glu-Asp-Asp Chemical compound OC(=O)CC[C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(O)=O)C(O)=O JPHYJQHPILOKHC-ACZMJKKPSA-N 0.000 description 1
- DSPQRJXOIXHOHK-WDSKDSINSA-N Glu-Asp-Gly Chemical compound OC(=O)CC[C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)NCC(O)=O DSPQRJXOIXHOHK-WDSKDSINSA-N 0.000 description 1
- MUSGDMDGNGXULI-DCAQKATOSA-N Glu-Glu-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@@H](N)CCC(O)=O MUSGDMDGNGXULI-DCAQKATOSA-N 0.000 description 1
- OAGVHWYIBZMWLA-YFKPBYRVSA-N Glu-Gly-Gly Chemical compound OC(=O)CC[C@H](N)C(=O)NCC(=O)NCC(O)=O OAGVHWYIBZMWLA-YFKPBYRVSA-N 0.000 description 1
- LZMQSTPFYJLVJB-GUBZILKMSA-N Glu-Leu-Cys Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CS)C(=O)O)NC(=O)[C@H](CCC(=O)O)N LZMQSTPFYJLVJB-GUBZILKMSA-N 0.000 description 1
- ATVYZJGOZLVXDK-IUCAKERBSA-N Glu-Leu-Gly Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)NCC(O)=O ATVYZJGOZLVXDK-IUCAKERBSA-N 0.000 description 1
- IVGJYOOGJLFKQE-AVGNSLFASA-N Glu-Leu-Lys Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CCCCN)C(=O)O)NC(=O)[C@H](CCC(=O)O)N IVGJYOOGJLFKQE-AVGNSLFASA-N 0.000 description 1
- WNRZUESNGGDCJX-JYJNAYRXSA-N Glu-Leu-Phe Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O WNRZUESNGGDCJX-JYJNAYRXSA-N 0.000 description 1
- KXTAGESXNQEZKB-DZKIICNBSA-N Glu-Phe-Val Chemical compound OC(=O)CC[C@H](N)C(=O)N[C@H](C(=O)N[C@@H](C(C)C)C(O)=O)CC1=CC=CC=C1 KXTAGESXNQEZKB-DZKIICNBSA-N 0.000 description 1
- HZISRJBYZAODRV-XQXXSGGOSA-N Glu-Thr-Ala Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C)C(O)=O HZISRJBYZAODRV-XQXXSGGOSA-N 0.000 description 1
- DLISPGXMKZTWQG-IFFSRLJSSA-N Glu-Thr-Val Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C(C)C)C(O)=O DLISPGXMKZTWQG-IFFSRLJSSA-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
- QSDKBRMVXSWAQE-BFHQHQDPSA-N Gly-Ala-Thr Chemical compound C[C@@H](O)[C@@H](C(O)=O)NC(=O)[C@H](C)NC(=O)CN QSDKBRMVXSWAQE-BFHQHQDPSA-N 0.000 description 1
- OCDLPQDYTJPWNG-YUMQZZPRSA-N Gly-Asn-Lys Chemical compound C(CCN)C[C@@H](C(=O)O)NC(=O)[C@H](CC(=O)N)NC(=O)CN OCDLPQDYTJPWNG-YUMQZZPRSA-N 0.000 description 1
- LHRXAHLCRMQBGJ-RYUDHWBXSA-N Gly-Glu-Phe Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)O)NC(=O)[C@H](CCC(=O)O)NC(=O)CN LHRXAHLCRMQBGJ-RYUDHWBXSA-N 0.000 description 1
- AYBKPDHHVADEDA-YUMQZZPRSA-N Gly-His-Asn Chemical compound [H]NCC(=O)N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](CC(N)=O)C(O)=O AYBKPDHHVADEDA-YUMQZZPRSA-N 0.000 description 1
- TWTPDFFBLQEBOE-IUCAKERBSA-N Gly-Leu-Gln Chemical compound [H]NCC(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(N)=O)C(O)=O TWTPDFFBLQEBOE-IUCAKERBSA-N 0.000 description 1
- PTIIBFKSLCYQBO-NHCYSSNCSA-N Gly-Lys-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)O)NC(=O)[C@H](CCCCN)NC(=O)CN PTIIBFKSLCYQBO-NHCYSSNCSA-N 0.000 description 1
- VEPBEGNDJYANCF-QWRGUYRKSA-N Gly-Lys-Lys Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)[C@@H](NC(=O)CN)CCCCN VEPBEGNDJYANCF-QWRGUYRKSA-N 0.000 description 1
- JJGBXTYGTKWGAT-YUMQZZPRSA-N Gly-Pro-Glu Chemical compound NCC(=O)N1CCC[C@H]1C(=O)N[C@@H](CCC(O)=O)C(O)=O JJGBXTYGTKWGAT-YUMQZZPRSA-N 0.000 description 1
- NVTPVQLIZCOJFK-FOHZUACHSA-N Gly-Thr-Asp Chemical compound [H]NCC(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(O)=O)C(O)=O NVTPVQLIZCOJFK-FOHZUACHSA-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
- AFMOTCMSEBITOE-YEPSODPASA-N Gly-Val-Thr Chemical compound NCC(=O)N[C@@H](C(C)C)C(=O)N[C@@H]([C@@H](C)O)C(O)=O AFMOTCMSEBITOE-YEPSODPASA-N 0.000 description 1
- 101100175482 Glycine max CG-3 gene Proteins 0.000 description 1
- BTEISVKTSQLKST-UHFFFAOYSA-N Haliclonasterol Natural products CC(C=CC(C)C(C)(C)C)C1CCC2C3=CC=C4CC(O)CCC4(C)C3CCC12C BTEISVKTSQLKST-UHFFFAOYSA-N 0.000 description 1
- NNBWMLHQXBTIIT-HVTMNAMFSA-N His-Gln-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)O)NC(=O)[C@H](CCC(=O)N)NC(=O)[C@H](CC1=CN=CN1)N NNBWMLHQXBTIIT-HVTMNAMFSA-N 0.000 description 1
- BPOHQCZZSFBSON-KKUMJFAQSA-N His-Leu-His Chemical compound CC(C)C[C@H](NC(=O)[C@@H](N)Cc1cnc[nH]1)C(=O)N[C@@H](Cc1cnc[nH]1)C(O)=O BPOHQCZZSFBSON-KKUMJFAQSA-N 0.000 description 1
- YAJQKIBLYPFAET-NAZCDGGXSA-N His-Thr-Trp Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CC1=CNC2=CC=CC=C21)C(=O)O)NC(=O)[C@H](CC3=CN=CN3)N)O YAJQKIBLYPFAET-NAZCDGGXSA-N 0.000 description 1
- 101000666730 Homo sapiens T-complex protein 1 subunit alpha Proteins 0.000 description 1
- 240000005979 Hordeum vulgare Species 0.000 description 1
- 235000007340 Hordeum vulgare Nutrition 0.000 description 1
- NULSANWBUWLTKN-NAKRPEOUSA-N Ile-Arg-Ser Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCCN=C(N)N)C(=O)N[C@@H](CO)C(=O)O)N NULSANWBUWLTKN-NAKRPEOUSA-N 0.000 description 1
- HOLOYAZCIHDQNS-YVNDNENWSA-N Ile-Gln-Glu Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCC(=O)N)C(=O)N[C@@H](CCC(=O)O)C(=O)O)N HOLOYAZCIHDQNS-YVNDNENWSA-N 0.000 description 1
- DVRDRICMWUSCBN-UKJIMTQDSA-N Ile-Gln-Val Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCC(=O)N)C(=O)N[C@@H](C(C)C)C(=O)O)N DVRDRICMWUSCBN-UKJIMTQDSA-N 0.000 description 1
- UBHUJPVCJHPSEU-GRLWGSQLSA-N Ile-Glu-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCC(=O)O)C(=O)N[C@@H]([C@@H](C)CC)C(=O)O)N UBHUJPVCJHPSEU-GRLWGSQLSA-N 0.000 description 1
- GVNNAHIRSDRIII-AJNGGQMLSA-N Ile-Lys-Lys Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCCCN)C(=O)O)N GVNNAHIRSDRIII-AJNGGQMLSA-N 0.000 description 1
- MASWXTFJVNRZPT-NAKRPEOUSA-N Ile-Met-Ala Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCSC)C(=O)N[C@@H](C)C(=O)O)N MASWXTFJVNRZPT-NAKRPEOUSA-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
- YWCJXQKATPNPOE-UKJIMTQDSA-N Ile-Val-Glu Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCC(=O)O)C(=O)O)N YWCJXQKATPNPOE-UKJIMTQDSA-N 0.000 description 1
- ZSESFIFAYQEKRD-CYDGBPFRSA-N Ile-Val-Met Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCSC)C(=O)O)N ZSESFIFAYQEKRD-CYDGBPFRSA-N 0.000 description 1
- PMGDADKJMCOXHX-UHFFFAOYSA-N L-Arginyl-L-glutamin-acetat Natural products NC(=N)NCCCC(N)C(=O)NC(CCC(N)=O)C(O)=O PMGDADKJMCOXHX-UHFFFAOYSA-N 0.000 description 1
- FADYJNXDPBKVCA-UHFFFAOYSA-N L-Phenylalanyl-L-lysin Natural products NCCCCC(C(O)=O)NC(=O)C(N)CC1=CC=CC=C1 FADYJNXDPBKVCA-UHFFFAOYSA-N 0.000 description 1
- LHSGPCFBGJHPCY-UHFFFAOYSA-N L-leucine-L-tyrosine Natural products CC(C)CC(N)C(=O)NC(C(O)=O)CC1=CC=C(O)C=C1 LHSGPCFBGJHPCY-UHFFFAOYSA-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
- WXHFZJFZWNCDNB-KKUMJFAQSA-N Leu-Asn-Tyr Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@H](C(O)=O)CC1=CC=C(O)C=C1 WXHFZJFZWNCDNB-KKUMJFAQSA-N 0.000 description 1
- MMEDVBWCMGRKKC-GARJFASQSA-N Leu-Asp-Pro Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)N1CCC[C@@H]1C(=O)O)N MMEDVBWCMGRKKC-GARJFASQSA-N 0.000 description 1
- QLQHWWCSCLZUMA-KKUMJFAQSA-N Leu-Asp-Tyr Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@H](C(O)=O)CC1=CC=C(O)C=C1 QLQHWWCSCLZUMA-KKUMJFAQSA-N 0.000 description 1
- IASQBRJGRVXNJI-YUMQZZPRSA-N Leu-Cys-Gly Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CS)C(=O)NCC(O)=O IASQBRJGRVXNJI-YUMQZZPRSA-N 0.000 description 1
- DLCXCECTCPKKCD-GUBZILKMSA-N Leu-Gln-Asn Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(N)=O)C(O)=O DLCXCECTCPKKCD-GUBZILKMSA-N 0.000 description 1
- QJUWBDPGGYVRHY-YUMQZZPRSA-N Leu-Gly-Cys Chemical compound CC(C)C[C@@H](C(=O)NCC(=O)N[C@@H](CS)C(=O)O)N QJUWBDPGGYVRHY-YUMQZZPRSA-N 0.000 description 1
- ZGUMORRUBUCXEH-AVGNSLFASA-N Leu-Lys-Gln Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCC(N)=O)C(O)=O ZGUMORRUBUCXEH-AVGNSLFASA-N 0.000 description 1
- PKKMDPNFGULLNQ-AVGNSLFASA-N Leu-Met-Arg Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CCCN=C(N)N)C(O)=O PKKMDPNFGULLNQ-AVGNSLFASA-N 0.000 description 1
- BJWKOATWNQJPSK-SRVKXCTJSA-N Leu-Met-Glu Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CCC(=O)O)C(=O)O)N BJWKOATWNQJPSK-SRVKXCTJSA-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
- 102000003960 Ligases Human genes 0.000 description 1
- 108090000364 Ligases Proteins 0.000 description 1
- UPYKUZBSLRQECL-UKMVMLAPSA-N Lycopene Natural products CC(=C/C=C/C=C(C)/C=C/C=C(C)/C=C/C1C(=C)CCCC1(C)C)C=CC=C(/C)C=CC2C(=C)CCCC2(C)C UPYKUZBSLRQECL-UKMVMLAPSA-N 0.000 description 1
- JEVVKJMRZMXFBT-XWDZUXABSA-N Lycophyll Natural products OC/C(=C/CC/C(=C\C=C\C(=C/C=C/C(=C\C=C\C=C(/C=C/C=C(\C=C\C=C(/CC/C=C(/CO)\C)\C)/C)\C)/C)\C)/C)/C JEVVKJMRZMXFBT-XWDZUXABSA-N 0.000 description 1
- RVOMPSJXSRPFJT-DCAQKATOSA-N Lys-Ala-Arg Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](C)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O RVOMPSJXSRPFJT-DCAQKATOSA-N 0.000 description 1
- YIBOAHAOAWACDK-QEJZJMRPSA-N Lys-Ala-Phe Chemical compound NCCCC[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@H](C(O)=O)CC1=CC=CC=C1 YIBOAHAOAWACDK-QEJZJMRPSA-N 0.000 description 1
- GQUDMNDPQTXZRV-DCAQKATOSA-N Lys-Arg-Asp Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(O)=O)C(O)=O GQUDMNDPQTXZRV-DCAQKATOSA-N 0.000 description 1
- AAORVPFVUIHEAB-YUMQZZPRSA-N Lys-Asp-Gly Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CC(O)=O)C(=O)NCC(O)=O AAORVPFVUIHEAB-YUMQZZPRSA-N 0.000 description 1
- PBIPLDMFHAICIP-DCAQKATOSA-N Lys-Glu-Glu Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(O)=O PBIPLDMFHAICIP-DCAQKATOSA-N 0.000 description 1
- GNLJXWBNLAIPEP-MELADBBJSA-N Lys-His-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CC2=CN=CN2)NC(=O)[C@H](CCCCN)N)C(=O)O GNLJXWBNLAIPEP-MELADBBJSA-N 0.000 description 1
- VMTYLUGCXIEDMV-QWRGUYRKSA-N Lys-Leu-Gly Chemical compound OC(=O)CNC(=O)[C@H](CC(C)C)NC(=O)[C@@H](N)CCCCN VMTYLUGCXIEDMV-QWRGUYRKSA-N 0.000 description 1
- LJADEBULDNKJNK-IHRRRGAJSA-N Lys-Leu-Val Chemical compound CC(C)C[C@H](NC(=O)[C@@H](N)CCCCN)C(=O)N[C@@H](C(C)C)C(O)=O LJADEBULDNKJNK-IHRRRGAJSA-N 0.000 description 1
- ZJWIXBZTAAJERF-IHRRRGAJSA-N Lys-Lys-Arg Chemical compound NCCCC[C@H](N)C(=O)N[C@@H](CCCCN)C(=O)N[C@H](C(O)=O)CCCN=C(N)N ZJWIXBZTAAJERF-IHRRRGAJSA-N 0.000 description 1
- RIJCHEVHFWMDKD-SRVKXCTJSA-N Lys-Lys-Asn Chemical compound NCCCC[C@H](N)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(N)=O)C(O)=O RIJCHEVHFWMDKD-SRVKXCTJSA-N 0.000 description 1
- HVAUKHLDSDDROB-KKUMJFAQSA-N Lys-Lys-Leu Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(C)C)C(O)=O HVAUKHLDSDDROB-KKUMJFAQSA-N 0.000 description 1
- KJIXWRWPOCKYLD-IHRRRGAJSA-N Lys-Lys-Met Chemical compound CSCC[C@@H](C(=O)O)NC(=O)[C@H](CCCCN)NC(=O)[C@H](CCCCN)N KJIXWRWPOCKYLD-IHRRRGAJSA-N 0.000 description 1
- OBZHNHBAAVEWKI-DCAQKATOSA-N Lys-Pro-Asn Chemical compound NCCCC[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CC(N)=O)C(O)=O OBZHNHBAAVEWKI-DCAQKATOSA-N 0.000 description 1
- PDIDTSZKKFEDMB-UWVGGRQHSA-N Lys-Pro-Gly Chemical compound [H]N[C@@H](CCCCN)C(=O)N1CCC[C@H]1C(=O)NCC(O)=O PDIDTSZKKFEDMB-UWVGGRQHSA-N 0.000 description 1
- SUZVLFWOCKHWET-CQDKDKBSSA-N Lys-Tyr-Ala Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](C)C(O)=O SUZVLFWOCKHWET-CQDKDKBSSA-N 0.000 description 1
- RMKJOQSYLQQRFN-KKUMJFAQSA-N Lys-Tyr-Asp Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CC(O)=O)C(O)=O RMKJOQSYLQQRFN-KKUMJFAQSA-N 0.000 description 1
- GILLQRYAWOMHED-DCAQKATOSA-N Lys-Val-Ser Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@H](C(C)C)NC(=O)[C@@H](N)CCCCN GILLQRYAWOMHED-DCAQKATOSA-N 0.000 description 1
- MCNGIXXCMJAURZ-VEVYYDQMSA-N Met-Asp-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](CC(=O)O)NC(=O)[C@H](CCSC)N)O MCNGIXXCMJAURZ-VEVYYDQMSA-N 0.000 description 1
- OFNCSQNBSWGGNV-DCAQKATOSA-N Met-Cys-His Chemical compound CSCC[C@H](N)C(=O)N[C@@H](CS)C(=O)N[C@H](C(O)=O)CC1=CNC=N1 OFNCSQNBSWGGNV-DCAQKATOSA-N 0.000 description 1
- WWWGMQHQSAUXBU-BQBZGAKWSA-N Met-Gly-Asn Chemical compound CSCC[C@H](N)C(=O)NCC(=O)N[C@H](C(O)=O)CC(N)=O WWWGMQHQSAUXBU-BQBZGAKWSA-N 0.000 description 1
- JZNGSNMTXAHMSV-AVGNSLFASA-N Met-His-Arg Chemical compound CSCC[C@@H](C(=O)N[C@@H](CC1=CN=CN1)C(=O)N[C@@H](CCCN=C(N)N)C(=O)O)N JZNGSNMTXAHMSV-AVGNSLFASA-N 0.000 description 1
- OBCRZLRPJFNLAN-DCAQKATOSA-N Met-His-Asp Chemical compound [H]N[C@@H](CCSC)C(=O)N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](CC(O)=O)C(O)=O OBCRZLRPJFNLAN-DCAQKATOSA-N 0.000 description 1
- WXJXYMFUTRXRGO-UWVGGRQHSA-N Met-His-Gly Chemical compound CSCC[C@H](N)C(=O)N[C@H](C(=O)NCC(O)=O)CC1=CNC=N1 WXJXYMFUTRXRGO-UWVGGRQHSA-N 0.000 description 1
- UNPGTBHYKJOCCZ-DCAQKATOSA-N Met-Lys-Ala Chemical compound CSCC[C@H](N)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](C)C(O)=O UNPGTBHYKJOCCZ-DCAQKATOSA-N 0.000 description 1
- YYEIFXZOBZVDPH-DCAQKATOSA-N Met-Lys-Asp Chemical compound CSCC[C@H](N)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(O)=O)C(O)=O YYEIFXZOBZVDPH-DCAQKATOSA-N 0.000 description 1
- UFOWQBYMUILSRK-IHRRRGAJSA-N Met-Lys-His Chemical compound CSCC[C@H](N)C(=O)N[C@@H](CCCCN)C(=O)N[C@H](C(O)=O)CC1=CNC=N1 UFOWQBYMUILSRK-IHRRRGAJSA-N 0.000 description 1
- LCPUWQLULVXROY-RHYQMDGZSA-N Met-Lys-Thr Chemical compound CSCC[C@H](N)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H]([C@@H](C)O)C(O)=O LCPUWQLULVXROY-RHYQMDGZSA-N 0.000 description 1
- 108010002311 N-glycylglutamic acid Proteins 0.000 description 1
- 108010066427 N-valyltryptophan Proteins 0.000 description 1
- 108091005461 Nucleic proteins Chemical group 0.000 description 1
- APJPXSFJBMMOLW-KBPBESRZSA-N Phe-Gly-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)CNC(=O)[C@@H](N)CC1=CC=CC=C1 APJPXSFJBMMOLW-KBPBESRZSA-N 0.000 description 1
- MSHZERMPZKCODG-ACRUOGEOSA-N Phe-Leu-Phe Chemical compound C([C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC=1C=CC=CC=1)C(O)=O)C1=CC=CC=C1 MSHZERMPZKCODG-ACRUOGEOSA-N 0.000 description 1
- YCCUXNNKXDGMAM-KKUMJFAQSA-N Phe-Leu-Ser Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(O)=O YCCUXNNKXDGMAM-KKUMJFAQSA-N 0.000 description 1
- AHXPYZRZRMQOAU-QXEWZRGKSA-N Pro-Asn-Val Chemical compound CC(C)[C@H](NC(=O)[C@H](CC(N)=O)NC(=O)[C@@H]1CCCN1)C(O)=O AHXPYZRZRMQOAU-QXEWZRGKSA-N 0.000 description 1
- XUSDDSLCRPUKLP-QXEWZRGKSA-N Pro-Asp-Val Chemical compound CC(C)[C@@H](C(O)=O)NC(=O)[C@H](CC(O)=O)NC(=O)[C@@H]1CCCN1 XUSDDSLCRPUKLP-QXEWZRGKSA-N 0.000 description 1
- KIPIKSXPPLABPN-CIUDSAMLSA-N Pro-Glu-Asn Chemical compound NC(=O)C[C@@H](C(O)=O)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@@H]1CCCN1 KIPIKSXPPLABPN-CIUDSAMLSA-N 0.000 description 1
- MRYUJHGPZQNOAD-IHRRRGAJSA-N Pro-Leu-Lys Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CCCCN)C(=O)O)NC(=O)[C@@H]1CCCN1 MRYUJHGPZQNOAD-IHRRRGAJSA-N 0.000 description 1
- ZUZINZIJHJFJRN-UBHSHLNASA-N Pro-Phe-Ala Chemical compound C([C@@H](C(=O)N[C@@H](C)C(O)=O)NC(=O)[C@H]1NCCC1)C1=CC=CC=C1 ZUZINZIJHJFJRN-UBHSHLNASA-N 0.000 description 1
- SNSYSBUTTJBPDG-OKZBNKHCSA-N Pro-Trp-Pro Chemical compound C1C[C@H](NC1)C(=O)N[C@@H](CC2=CNC3=CC=CC=C32)C(=O)N4CCC[C@@H]4C(=O)O SNSYSBUTTJBPDG-OKZBNKHCSA-N 0.000 description 1
- QDDJNKWPTJHROJ-UFYCRDLUSA-N Pro-Tyr-Tyr Chemical compound C([C@@H](C(=O)O)NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@H]1NCCC1)C1=CC=C(O)C=C1 QDDJNKWPTJHROJ-UFYCRDLUSA-N 0.000 description 1
- 102000012515 Protein kinase domains Human genes 0.000 description 1
- 108050002122 Protein kinase domains Proteins 0.000 description 1
- 238000010802 RNA extraction kit Methods 0.000 description 1
- 238000011529 RT qPCR Methods 0.000 description 1
- BTKUIVBNGBFTTP-WHFBIAKZSA-N Ser-Ala-Gly Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](C)C(=O)NCC(O)=O BTKUIVBNGBFTTP-WHFBIAKZSA-N 0.000 description 1
- XNCUYZKGQOCOQH-YUMQZZPRSA-N Ser-Leu-Gly Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(=O)NCC(O)=O XNCUYZKGQOCOQH-YUMQZZPRSA-N 0.000 description 1
- UBRMZSHOOIVJPW-SRVKXCTJSA-N Ser-Leu-Lys Chemical compound OC[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCCN)C(O)=O UBRMZSHOOIVJPW-SRVKXCTJSA-N 0.000 description 1
- RCOUFINCYASMDN-GUBZILKMSA-N Ser-Val-Met Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCSC)C(O)=O RCOUFINCYASMDN-GUBZILKMSA-N 0.000 description 1
- 101710113029 Serine/threonine-protein kinase Proteins 0.000 description 1
- 241000208292 Solanaceae Species 0.000 description 1
- 229930182558 Sterol Natural products 0.000 description 1
- 102100038410 T-complex protein 1 subunit alpha Human genes 0.000 description 1
- ZUUDNCOCILSYAM-KKHAAJSZSA-N Thr-Asp-Val Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](C(C)C)C(O)=O ZUUDNCOCILSYAM-KKHAAJSZSA-N 0.000 description 1
- GKWNLDNXMMLRMC-GLLZPBPUSA-N Thr-Glu-Gln Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CCC(=O)O)C(=O)N[C@@H](CCC(=O)N)C(=O)O)N)O GKWNLDNXMMLRMC-GLLZPBPUSA-N 0.000 description 1
- MECLEFZMPPOEAC-VOAKCMCISA-N Thr-Leu-Lys Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCCN)C(=O)O)N)O MECLEFZMPPOEAC-VOAKCMCISA-N 0.000 description 1
- MXDOAJQRJBMGMO-FJXKBIBVSA-N Thr-Pro-Gly Chemical compound C[C@@H](O)[C@H](N)C(=O)N1CCC[C@H]1C(=O)NCC(O)=O MXDOAJQRJBMGMO-FJXKBIBVSA-N 0.000 description 1
- MQVGIFJSFFVGFW-XEGUGMAKSA-N Trp-Ala-Glu Chemical compound [H]N[C@@H](CC1=CNC2=C1C=CC=C2)C(=O)N[C@@H](C)C(=O)N[C@@H](CCC(O)=O)C(O)=O MQVGIFJSFFVGFW-XEGUGMAKSA-N 0.000 description 1
- MVHHTXAUJCIOMZ-WDSOQIARSA-N Trp-Arg-Lys Chemical compound C1=CC=C2C(=C1)C(=CN2)C[C@@H](C(=O)N[C@@H](CCCN=C(N)N)C(=O)N[C@@H](CCCCN)C(=O)O)N MVHHTXAUJCIOMZ-WDSOQIARSA-N 0.000 description 1
- WVRUKYLYMFGKAN-IHRRRGAJSA-N Tyr-Glu-Glu Chemical compound OC(=O)CC[C@@H](C(O)=O)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@@H](N)CC1=CC=C(O)C=C1 WVRUKYLYMFGKAN-IHRRRGAJSA-N 0.000 description 1
- HVPPEXXUDXAPOM-MGHWNKPDSA-N Tyr-Ile-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H]([C@@H](C)CC)NC(=O)[C@@H](N)CC1=CC=C(O)C=C1 HVPPEXXUDXAPOM-MGHWNKPDSA-N 0.000 description 1
- AVIQBBOOTZENLH-KKUMJFAQSA-N Tyr-Leu-Cys Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CS)C(=O)O)NC(=O)[C@H](CC1=CC=C(C=C1)O)N AVIQBBOOTZENLH-KKUMJFAQSA-N 0.000 description 1
- ASQFIHTXXMFENG-XPUUQOCRSA-N Val-Ala-Gly Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](C)C(=O)NCC(O)=O ASQFIHTXXMFENG-XPUUQOCRSA-N 0.000 description 1
- QHDXUYOYTPWCSK-RCOVLWMOSA-N Val-Asp-Gly Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)NCC(=O)O)N QHDXUYOYTPWCSK-RCOVLWMOSA-N 0.000 description 1
- HHSILIQTHXABKM-YDHLFZDLSA-N Val-Asp-Phe Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](Cc1ccccc1)C(O)=O HHSILIQTHXABKM-YDHLFZDLSA-N 0.000 description 1
- OVLIFGQSBSNGHY-KKHAAJSZSA-N Val-Asp-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](CC(=O)O)NC(=O)[C@H](C(C)C)N)O OVLIFGQSBSNGHY-KKHAAJSZSA-N 0.000 description 1
- SCBITHMBEJNRHC-LSJOCFKGSA-N Val-Asp-Val Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)N[C@@H](C(C)C)C(=O)O)N SCBITHMBEJNRHC-LSJOCFKGSA-N 0.000 description 1
- VCAWFLIWYNMHQP-UKJIMTQDSA-N Val-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(C)C)N VCAWFLIWYNMHQP-UKJIMTQDSA-N 0.000 description 1
- LAYSXAOGWHKNED-XPUUQOCRSA-N Val-Gly-Ser Chemical compound CC(C)[C@H](N)C(=O)NCC(=O)N[C@@H](CO)C(O)=O LAYSXAOGWHKNED-XPUUQOCRSA-N 0.000 description 1
- KVRLNEILGGVBJX-IHRRRGAJSA-N Val-His-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@@H](NC(=O)[C@@H](N)C(C)C)CC1=CN=CN1 KVRLNEILGGVBJX-IHRRRGAJSA-N 0.000 description 1
- FTKXYXACXYOHND-XUXIUFHCSA-N Val-Ile-Leu Chemical compound CC(C)[C@H](N)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC(C)C)C(O)=O FTKXYXACXYOHND-XUXIUFHCSA-N 0.000 description 1
- LYERIXUFCYVFFX-GVXVVHGQSA-N Val-Leu-Glu Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CCC(=O)O)C(=O)O)NC(=O)[C@H](C(C)C)N LYERIXUFCYVFFX-GVXVVHGQSA-N 0.000 description 1
- DOFAQXCYFQKSHT-SRVKXCTJSA-N Val-Pro-Pro Chemical compound CC(C)[C@H](N)C(=O)N1CCC[C@H]1C(=O)N1[C@H](C(O)=O)CCC1 DOFAQXCYFQKSHT-SRVKXCTJSA-N 0.000 description 1
- VVIZITNVZUAEMI-DLOVCJGASA-N Val-Val-Gln Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](C(C)C)C(=O)N[C@H](C(O)=O)CCC(N)=O VVIZITNVZUAEMI-DLOVCJGASA-N 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 239000003463 adsorbent Substances 0.000 description 1
- 108010086434 alanyl-seryl-glycine Proteins 0.000 description 1
- 108010005233 alanylglutamic acid Proteins 0.000 description 1
- 108010044940 alanylglutamine Proteins 0.000 description 1
- 108010087924 alanylproline Proteins 0.000 description 1
- KOSRFJWDECSPRO-UHFFFAOYSA-N alpha-L-glutamyl-L-glutamic acid Natural products OC(=O)CCC(N)C(=O)NC(CCC(O)=O)C(O)=O KOSRFJWDECSPRO-UHFFFAOYSA-N 0.000 description 1
- 235000019257 ammonium acetate Nutrition 0.000 description 1
- 229940043376 ammonium acetate Drugs 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 238000000137 annealing Methods 0.000 description 1
- 239000008346 aqueous phase Substances 0.000 description 1
- 108010008355 arginyl-glutamine Proteins 0.000 description 1
- 108010001271 arginyl-glutamyl-arginine Proteins 0.000 description 1
- 108010077245 asparaginyl-proline Proteins 0.000 description 1
- 108010040443 aspartyl-aspartic acid Proteins 0.000 description 1
- 108010093581 aspartyl-proline Proteins 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- 230000033228 biological regulation Effects 0.000 description 1
- 230000001851 biosynthetic effect Effects 0.000 description 1
- 230000006696 biosynthetic metabolic pathway Effects 0.000 description 1
- 239000011575 calcium Substances 0.000 description 1
- VYUIKSFYFRVQLF-YLNAYWRASA-N castasterone Chemical compound C([C@@H]1C(=O)C2)[C@H](O)[C@H](O)C[C@]1(C)[C@@H]1[C@@H]2[C@@H]2CC[C@H]([C@H](C)[C@@H](O)[C@H](O)[C@@H](C)C(C)C)[C@@]2(C)CC1 VYUIKSFYFRVQLF-YLNAYWRASA-N 0.000 description 1
- 210000004027 cell Anatomy 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 210000000349 chromosome Anatomy 0.000 description 1
- 238000003776 cleavage reaction Methods 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
- 238000012258 culturing Methods 0.000 description 1
- 238000004925 denaturation Methods 0.000 description 1
- 230000036425 denaturation Effects 0.000 description 1
- 239000012154 double-distilled water Substances 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 238000001506 fluorescence spectroscopy Methods 0.000 description 1
- 235000019253 formic acid Nutrition 0.000 description 1
- 108010006664 gamma-glutamyl-glycyl-glycine Proteins 0.000 description 1
- 238000012224 gene deletion Methods 0.000 description 1
- 238000003209 gene knockout Methods 0.000 description 1
- 238000012239 gene modification Methods 0.000 description 1
- 230000030279 gene silencing Effects 0.000 description 1
- 238000012226 gene silencing method Methods 0.000 description 1
- 108010078144 glutaminyl-glycine Proteins 0.000 description 1
- 108010055341 glutamyl-glutamic acid Proteins 0.000 description 1
- XBGGUPMXALFZOT-UHFFFAOYSA-N glycyl-L-tyrosine hemihydrate Natural products NCC(=O)NC(C(O)=O)CC1=CC=C(O)C=C1 XBGGUPMXALFZOT-UHFFFAOYSA-N 0.000 description 1
- 108010072405 glycyl-aspartyl-glycine Proteins 0.000 description 1
- 108010079413 glycyl-prolyl-glutamic acid Proteins 0.000 description 1
- 108010089804 glycyl-threonine Proteins 0.000 description 1
- 108010087823 glycyltyrosine Proteins 0.000 description 1
- 108010037850 glycylvaline Proteins 0.000 description 1
- 238000000227 grinding Methods 0.000 description 1
- 238000004128 high performance liquid chromatography Methods 0.000 description 1
- 108010025306 histidylleucine Proteins 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 229930027917 kanamycin Natural products 0.000 description 1
- 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 1
- 229960000318 kanamycin Drugs 0.000 description 1
- 229930182823 kanamycin A Natural products 0.000 description 1
- 108010034529 leucyl-lysine Proteins 0.000 description 1
- 108010090333 leucyl-lysyl-proline Proteins 0.000 description 1
- 108010012058 leucyltyrosine Proteins 0.000 description 1
- OAIJSZIZWZSQBC-GYZMGTAESA-N lycopene Chemical compound CC(C)=CCC\C(C)=C\C=C\C(\C)=C\C=C\C(\C)=C\C=C\C=C(/C)\C=C\C=C(/C)\C=C\C=C(/C)CCC=C(C)C OAIJSZIZWZSQBC-GYZMGTAESA-N 0.000 description 1
- 229960004999 lycopene Drugs 0.000 description 1
- 235000012661 lycopene Nutrition 0.000 description 1
- 239000001751 lycopene Substances 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 108010056582 methionylglutamic acid Proteins 0.000 description 1
- 108010085203 methionylmethionine Proteins 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 230000035772 mutation Effects 0.000 description 1
- 238000002414 normal-phase solid-phase extraction Methods 0.000 description 1
- 150000007523 nucleic acids Chemical group 0.000 description 1
- 230000036542 oxidative stress Effects 0.000 description 1
- 239000008188 pellet Substances 0.000 description 1
- 108010073025 phenylalanylphenylalanine Proteins 0.000 description 1
- 108010051242 phenylalanylserine Proteins 0.000 description 1
- 230000003711 photoprotective effect Effects 0.000 description 1
- 230000000243 photosynthetic effect Effects 0.000 description 1
- 230000037039 plant physiology Effects 0.000 description 1
- 239000002244 precipitate Substances 0.000 description 1
- 239000002243 precursor Substances 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 238000000197 pyrolysis Methods 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 238000006722 reduction reaction Methods 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- 230000003938 response to stress Effects 0.000 description 1
- 230000007017 scission Effects 0.000 description 1
- 108010026333 seryl-proline Proteins 0.000 description 1
- 230000019491 signal transduction Effects 0.000 description 1
- 238000009331 sowing Methods 0.000 description 1
- 238000005507 spraying Methods 0.000 description 1
- 150000003432 sterols Chemical class 0.000 description 1
- 235000003702 sterols Nutrition 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 230000008685 targeting Effects 0.000 description 1
- 238000010998 test method Methods 0.000 description 1
- 108010061238 threonyl-glycine Proteins 0.000 description 1
- ZCIHMQAPACOQHT-ZGMPDRQDSA-N trans-isorenieratene Natural products CC(=C/C=C/C=C(C)/C=C/C=C(C)/C=C/c1c(C)ccc(C)c1C)C=CC=C(/C)C=Cc2c(C)ccc(C)c2C ZCIHMQAPACOQHT-ZGMPDRQDSA-N 0.000 description 1
- 230000009261 transgenic effect Effects 0.000 description 1
- 108010080629 tryptophan-leucine Proteins 0.000 description 1
- 108010079202 tyrosyl-alanyl-cysteine Proteins 0.000 description 1
- 238000001195 ultra high performance liquid chromatography Methods 0.000 description 1
- 108010015385 valyl-prolyl-proline Proteins 0.000 description 1
- 235000013311 vegetables Nutrition 0.000 description 1
- 238000003260 vortexing Methods 0.000 description 1
- 239000003643 water by type Substances 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/10—Transferases (2.)
- C12N9/12—Transferases (2.) transferring phosphorus containing groups, e.g. kinases (2.7)
-
- 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
- C12N15/8242—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
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
- C12Y207/00—Transferases transferring phosphorus-containing groups (2.7)
- C12Y207/11—Protein-serine/threonine kinases (2.7.11)
- C12Y207/11017—Ca2+/Calmodulin-dependent protein kinase (2.7.11.17)
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Genetics & Genomics (AREA)
- Engineering & Computer Science (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Molecular Biology (AREA)
- Biotechnology (AREA)
- General Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Microbiology (AREA)
- Physics & Mathematics (AREA)
- Plant Pathology (AREA)
- Biophysics (AREA)
- Cell Biology (AREA)
- Virology (AREA)
- Medicinal Chemistry (AREA)
- Nutrition Science (AREA)
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
本发明公开了一种高含量油菜素内酯番茄植株的培育方法,属于生物技术领域。本发明提供了CPK27基因作为负调控因子在提高植物油菜素内酯含量中的应用,所述CPK27基因的蛋白编码区的核苷酸序列如SEQID NO.1所示或与SEQ ID NO.1所示序列具有至少70%同源性且编码的蛋白在功能上等价。本发明首次公开CPK27基因在提高植物油菜素内酯类物质中的新用途,通过基因编辑技术缺失CPK27基因功能,可显著提高植物油菜素内酯类物质含量。本发明为培育高含量油菜素内酯类物质和具备广谱逆境抗性的番茄品种提供了重要的基因资源。
Description
技术领域
本发明涉及生物技术领域,具体涉及CPK27基因作为负调控因子在提高植物尤其是番茄内源油菜素内酯含量中的应用。
背景技术
番茄(Solanum lycopersicum),茄科番茄属植物,作为食用蔬果被全球性广泛种植。油菜素内酯类物质(Brassinosteroids,BRs)是一类天然存在于植物体内的甾醇类激素,其中油菜素甾酮(Castasterone,CS)和芸薹甾内酯(Brassinolide,BL)是最具生物活性的两类物质。研究表明油菜素内酯类物质能促进番茄生长发育和品质形成。
研究表明,外源喷施BL可显著提高番茄等作物的光合效率(Li等,DWARFoverexpression induces alteration in phytohormone homeostasis,development,architecture and carotenoid accumulation in tomato.Plant BiotechnologyJournal 2016,14,1021-1033)。外源喷施BL还能提高番茄果实中的番茄红素含量,并延缓采后番茄果实的低温损伤(Aghdam等,Enhancement of chilling stress tolerance oftomato fruit by postharvest brassinolidetreatment.Food and BioprocessTechnology 2014,7,909-914)。
值得注意的是,油菜素内酯类及其信号途径同样在番茄应答逆境胁迫过程中起着关键的作用。番茄过表达油菜素甾酮合成基因CYP85A1能有效降低因低温产生的氧化胁迫程度,增强低温抗性(Fang等,Brassinosteroids Act as a Positive Regulator ofPhotoprotection in Response to Chilling Stress.Plant Physiology 2019,180,2061-2076.)。外源喷施BL还增强了植物对尖孢镰刀菌的抗性(Ali等,Brassinosteroidenhances resistance to Fusarium diseases of barley.Phytopathology 103,1260–1267)。
由以上可看出,油菜素内酯类物质在促进番茄生长发育、品质形成和抗病抗逆中均发挥了积极的作用。因此,通过基因编辑技术提高番茄油菜素内酯类物质可创制高抗优质的番茄种质,改良现有番茄品种,达到番茄生产提质增效的目的。
油菜素内酯类物质的生物合成途径复杂多样,由早期和晚期C-22氧化途径、早期和晚期C-6氧化途径、早期C-22氧化途径与晚期C-6氧化途径之间的捷径组成。油菜甾醇(Campestrol)是芸薹甾内酯生物合成的前体。在这一系列的反应过程中,编码关键酶的DWF4和CYP85A1等基因发挥了关键的作用。通过影响合成酶的基因表达可进一步实现对植株内源油菜素内酯类物质含量的调控。有研究表明,过表达番茄CYP85A1基因可显著增加番茄内源油菜素内酯类物质含量(Li等,DWARF overexpression induces alteration inphytohormone homeostasis,development,architecture and carotenoid accumulationin tomato.Plant Biotechnology Journal 2016,14,1021-1033)。拟南芥中的bHLH型转录因子TCP1可结合在DWF4的启动子上促进其基因表达进而提高油菜素内酯类物质含量(Guo等,TCP1modulates brassinosteroid biosynthesis by regulating the expression ofthe key biosynthetic gene DWARF4 in Arabidopsis thaliana.Plant Cell2010,22:1161–1173)。
植物体内油菜素内酯类物质的合成受到多种因素的影响,调控合成的其他元件急需深入挖掘。CPK是植物一种钙离子依赖型蛋白激酶,其在调控油菜素内酯合成过程中的作用鲜有报道,因此,基于番茄CPK27基因进而创制高含量油菜素内酯类物质的番茄植株具有理论和实际应用价值。
发明内容
本发明的目的是提供一种可调控植物内源油菜素内酯类物质合成的基因,通过基因改造达到提高植物内源油菜素内酯类物质合成的目的,为培育高含量油菜素内酯类物质作物种质提供依据。
为实现上述目的,本发明采用如下技术方案:
本发明提供了CPK27基因作为负调控因子在提高植物油菜素内酯含量中的应用,所述CPK27基因的蛋白编码区的核苷酸序列如SEQ ID NO.1所示或与SEQ ID NO.1所示序列具有至少70%同源性且编码的蛋白在功能上等价。
核苷酸序列如SEQ ID NO.1所示的CPK27基因位于番茄的第11染色体上,编码一种钙离子依赖型蛋白激酶,属于丝氨酸/苏氨酸蛋白激酶,由N段可变结构域、蛋白激酶域、自我抑制结构域和Ca2+结合域组成,当与钙离子结合时,该蛋白被激活。
所述应用包括:利用生物学技术使得所述CPK27基因功能缺失,进而提高植物内源油菜素内酯类物质的含量。
具体的,利用基因突变、基因敲除、基因干扰或基因沉默技术导致所述CPK27基因的表达降低或缺失,从而获得高含量油菜素内酯的突变体植株。
进一步的,所述油菜素内酯包括油菜素甾酮(Castasterone,CS)和芸薹甾内酯(Brassinolide,BL)。
进一步的,所述CPK27基因编码的蛋白质负调控油菜素甾酮合成基因CYP85A1(基因登录号为:Solyc02g089160)的表达。
本发明通过基因编辑技术制备得到定点编辑CPK27基因的番茄cpk27缺失突变体植株。研究发现,番茄CPK27基因突变后,CYP85A1基因表达量显著提高,番茄油菜素甾酮和芸薹甾内酯含量显著提高;因此,敲除番茄CPK27基因有利于提高番茄内源油菜素内酯类物质含量,可用于创制高含量油菜素内酯类物质的番茄种质。
进一步的,所述植物为番茄。
番茄CPK27基因编码的蛋白质由533个氨基酸组成,其氨基酸序列如SEQ ID NO.2所示。
本发明还提供了一种提高番茄植株内源油菜素内酯含量的培育方法,包括以下步骤:
(1)在番茄CPK27基因的蛋白编码区选取含有PAM结构的靶标片段,以靶标片段PAM结构前20个碱基为依据,进行引物设计,构建CRISPR/Cas9载体;所述番茄CPK27基因的核苷酸序列如SEQ ID NO.3所示;
(2)构建含步骤(1)所述CRISPR/Cas9载体的农杆菌基因工程菌;
(3)将步骤(2)所述基因工程菌转化番茄子叶,获得不含外源Cas9蛋白且靶标序列发生变异的稳定遗传的纯合突变体株系。
进一步的,所述靶标片段PAM结构前20个碱基如SEQ ID NO.4或SEQ ID NO.5所示。
进一步的,构建CRISPR/Cas9载体的引物对的核苷酸序列如SEQ ID NO.6和SEQ IDNO.7所示,或者如SEQ ID NO.8和SEQ ID NO.9所示。
本发明具有的有益效果:
(1)本发明首次公开CPK27基因在提高植物油菜素内酯类物质中的新用途,通过基因编辑技术缺失CPK27基因功能,可显著提高植物油菜素内酯类物质含量。本发明为培育高含量油菜素内酯类物质和具备广谱逆境抗性的番茄品种提供了重要的基因资源。
(2)本发明利用基因编辑技术提供了一种高含量油菜素内酯类物质番茄种质的培育方法,与转基因技术相比,不会引入外源基因,并得到了高含量油菜素内酯类物质番茄纯合植株。
附图说明
图1为油菜素内酯类物质合成途径中最后两个步骤反应式,其中6-deoxocastasterone(6-deoxoCS)在CYP85A1基因编码的酶作用下,转化为有生物活性的油菜素甾酮(Castasterone,CS),并进一步合成芸薹甾内酯(Brassinolide,BL)。
图2为番茄cpk27缺失突变体T2代(纯合、不含外源Cas9片段)的核酸序列和蛋白序列与野生型番茄的比较,其中,番茄cpk27有两种突变类型,在sgRNA的位置分别***了碱基A和T,导致编码的蛋白质提前终止于第140位氨基酸位置;对照代表野生型番茄,cpk27#2、cpk27#6分别代表突变体植株的两种株系。
图3为野生型番茄和cpk27缺失突变体正常环境下的生长表型,其中(A)为植株照片,从左到右依次为对照、cpk27#2、cpk27#6,可以看出,突变体植株的株高显著高于野生型番茄;(B)为植株高度比较图,小写字母a,b代表不同植株间在5%水平上的差异显著,所用统计方法为Turkey’s test。
图4为野生型番茄和cpk27缺失突变体中CS的含量,小写字母a,b代表不同植株间在5%水平上的差异显著,所用统计方法为Turkey’s test。
图5为野生型番茄和cpk27缺失突变体中BL的含量,小写字母a,b代表不同植株间在5%水平上的差异显著,所用统计方法为Turkey’s test。
图6为CYP85A1基因在野生型番茄和cpk27缺失突变体中的表达量,可以看出,野生型番茄的CYP85A1基因表达量远远低于cpk27缺失突变体。小写字母a,b代表不同植株间在5%水平上的差异显著,所用统计方法为Turkey’s test。
具体实施方式
下面结合具体实施例对本发明做进一步说明。以下实施例仅用于说明本发明,不用来限制本发明的适用范围。在不背离本发明精神和本质的情况下,对本发明方法、步骤或条件所做的修改或替换,均属于本发明的范围。
下述实施例中所使用的试验方法如无特殊说明,均为常规方法;所使用的材料、试剂等,如无特殊说明,为可从商业途径得到的试剂和材料。
下述实施例中采用的番茄材料为常规野生型番茄品种Condine Red。
实施例1:含2个特异sgRNA的CRISPR/Cas9载体的构建
在SGN网站https://solgenomics.net/上找到CPK27基因的DNA全长序列如SEQ IDNO.3所示,将其序列输入http://crispr.hzau.edu.cn/cgi-bin/CRISPR2/CRISPR网站,查找PAM序列,即5‘-NGG-3’的序列(N代表A、T、C、G任一种核苷酸),将NGG之前20bp的序列定义为sgRNA,寻找特异性靶向基因蛋白编码区的sgRNA。该特异性靶向CPK27基因的蛋白编码区的2个sgRNA的DNA序列分别如SEQ ID NO.4和SEQ ID NO.5所示。
设计CRISPR引物,如下所示:
sgRNA1-F:5’-GCGGTCTCTATTGAACAAAGCACCAGTGGTCTAGTG-3’(SEQ ID NO.6);
sgRNA1-R:
5’-GCGGTCTCTCCCTCACAAAGTTCCATCACTGCACCAGCCGGGAAT CG-3’(SEQ ID NO.7);
sgRNA2-F:
5’-GCGGTCTCTTACCGTTTTAGAGCTAGAAATAGCAAGTTAA-3’(SEQ ID NO.8);
sgRNA2-R:
5’-GCGGTCTCTAAACATCAAACAACTCACCCCCCTTGCACCAGCCGG GAATCG-3’(SEQ IDNO.9);
上述引物分别以pBAtC-tRNA质粒为模板进行PCR扩增,产物经普通DNA纯化试剂盒纯化后,用BsaI酶将扩增片段与pHEE401质粒连接,产物42℃转化大肠杆菌涂板,抗性为卡那霉素。
挑单克隆菌落,用pHEE401载体通用前引物M13-F:5’-TGTAAAACGACGGCCAGT-3’(SEQ ID NO.10)和通用后引物M13-R:5’-GGTATTGGTTTATCTCATCGGAACTGCA-3’(SEQ IDNO.11),进行PCR验证。
将条带大小正确的菌液送至测序公司测序,测序结果显示载体包含2个sgRNA序列,提质粒后电击入农杆菌GV3101感受态,28℃培养两天后挑斑进行PCR验证,获得可用于构建其CRISPR/Cas9基因编辑材料的农杆菌菌株。
实施例2:CPK27基因突变体材料制备与鉴定
在播种培养基播种已消毒的种子,7天后切子叶。农杆菌侵染法将实施例1制备的最终质粒转化入子叶中,利用植物细胞全能性,获得T0代基因编辑番茄。T0代基因编辑番茄苗检测:利用CTAB法提取T0代植株的基因组DNA并以其为模板,在包含sgRNA的DNA序列前后约200bp处设计如下引物,进行PCR扩增测序验证:
验证前引物:5’-TGGTGCAACTCAAGCATCTG-3’(SEQ ID NO.12);
验证后引物:5’-GGATGGTGAAATCCTGTGTG-3’(SEQ ID NO.13);
所得PCR产物送测序公司测序。测序结果与该段基因原序列利用Snapgene软件进行比对,选取sgRNA序列发生碱基缺失或减少、且测序显示单峰的植株,进行自交繁种,获得T0代的种子。
上述T0代种子种植于生长室中,获得T1代植株。利用上述同样方法检测T1代植株的sgRNA序列碱基编辑情况。同时,利用Cas9基因引物对T1代植株的DNA进行PCR扩增,检测是否含有Cas9序列。选取sgRNA发生变异,但不含Cas9的T1代植株,确定为基因编辑植株的两个株系,分别命名为cpk27#2和cpk27#6,其基因编辑位点如图2所示。
cpk27#2比对照植株多一个碱基A,cpk27#6比对照植株多一个碱基T。对上述两个株系进一步自交繁种,获得T1代种子。上述两个株系T1代种子播种后,获得不含外源基因Cas9,且sgRNA发生变异的稳定遗传的T2代植株。如图3所示,突变体植株的株高显著高于野生型番茄。
以下实施例均以上述两个纯合株系T2代植株作为材料进行实验。
实施例3:番茄CPK27基因对内源油菜素内酯类物质含量的影响
制备MCX@BBII:将1.0g商用MCX固相萃取吸附剂放入装有7mLBBII溶液的15mL管中。剧烈搅拌3min后,以10,000g离心3min,弃上清,用7mL 60%乙腈洗涤。最后,物料在60℃的真空中干燥保存。
番茄苗种植在生长室中,35天后取幼嫩的叶片0.1g,用液氮充分研磨后加入1mL乙腈(HPLC级)和1ng/μL的标样,放置在摇床上4℃震荡过夜。4℃,12,000g离心10min后,上清转移至含有0.3g MCX@BBII和3mL双蒸水的离心管中,充分涡旋。4℃,10,000g离心10min后,用5mL含有0.5%甲酸的90%丙酮重悬浮沉淀,充分涡旋。4℃,10,000g离心10min后,弃上清,加入1.2mL丙酮和0.3g醋酸铵,涡旋1min后10,000g离心3分钟。吸上清液于新的1.5mL管中,氮气吹干后用100μL 45%乙腈溶解,上机检测。油菜素内酯类物质含量分析采用ACQUITY超高效液相色谱联用Waters Xevo TQ-XS三重四重质谱仪。结果如图4和图5所示。
结果表明:cpk27缺失突变体中有生物活性的CS和BL含量显著高于野生型。
实施例4:番茄CPK27基因对内源油菜素内酯类物质合成基因CYP85A1的影响
取叶片样品0.2g,用总RNA提取试剂盒(Tiangen,DP419)按照Trizol裂解法提取RNA,具体步骤为:
(1)取0.1g叶片在液氮中磨碎,加1mL裂解液,后用均浆仪混匀;
(2)将均浆样品在15-30℃放置5min;
(3)4℃,12,000rpm离心5min,去上清;
(4)加入200μL氯仿,剧烈震荡15s,室温放置3min;
(5)4℃,12,000rpm离心10min,将上部水相转移到新的1.5mL管中,进行下一步操作;
(6)缓慢加入0.5倍体积的无水乙醇,混合均匀。将得到的溶液和沉淀一起转入吸附柱CR3,4℃,12,000rpm离心30s,弃废液;
(7)向吸附柱CR3中加入500μL去蛋白液RD,4℃,12,000rpm离心30s,弃废液;
(8)向吸附柱CR3中加入600μL漂洗液RW,室温静止2min,4℃,12000rpm离心30s,弃废液;
(9)重复操作步骤(8);
(10)将吸附柱放入2mL收集管中,4℃,12000rpm离心2min,去废液;使用HiScriptII QRT SuperMix(Vazyme Biotech,Nanjing,China)将RNA反转录成cDNA,进行qRT-PCR实验,实时荧光定量PCR(qRT-PCR)采用480II Real-TimePCR detection system(Roche,Swiss),并使用AceQ qPCR SYBR Green Master Mix kits(Vazyme Biotech,Nanjing,China)。PCR反应条件为:95℃3min;95℃变性15s,58℃退火15s,72℃延伸30s,40个循环。在每个循环的延伸末期采集荧光数据。
根据番茄的CYP85A1基因序列设计特异性引物:
CYP85A1基因前引物:5’-ATGAAGCGAAAGGACTGGTC(SEQ ID NO.14);
CYP85A1基因后引物:5’-TGCACCCCTCATGTACTTGT(SEQ ID NO.15);
结果如图6所示,cpk27基因缺失突变体的CYP85A1基因表达量提高了100倍左右,表明CPK27负调控CYP85A1的表达进而抑制CS和BL的含量。
序列表
<110> 浙江大学
浙江大学山东(临沂)现代农业研究院
<120> 一种高含量油菜素内酯番茄植株的培育方法
<160> 15
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1602
<212> DNA
<213> 番茄(Solanum lycopersicum)
<400> 1
atggggaatt gctgtgggac acctggtaat tcttctgaga ataagaagaa gaagaacaaa 60
ccaaaccctt ttgctcttga ttatggtgca actcaagcat ctggaggtga tggaaacaag 120
cttgttgtgt tgaaagatcc aacaggacac aatattcaag aaaaatatga tcttggttgt 180
gagctaggaa gaggagaatt tggggttaca tatttatgta ctgatgttga tacaggggac 240
aaatatgctt gcaaatcgat atcaaagaag aaacttagga ctgctgtaga tatagatgat 300
gttaggagag aagttgagat catgaagcat ttgcctaaac atcctaatat tgtgaccttg 360
aaggacactt atgaggatga taatgcagtg catattgtga tggaactttg tgaggggggt 420
gagttgtttg ataggattgt tgccagggga cattatacag agagagcagc tgcggttatt 480
atgaagacta tagtagaagt tgttcagatg tgtcatatgc atggagtaat gcatcgtgat 540
ctcaaacctg agaactttct gtttggtaac aagaaagaaa ctgctccttt gaaggctatt 600
gactttggat tatcagtgtt ctttaaaccc ggggaacgct ttaatgagat tgtgggaagt 660
ccttattaca tggctccaga ggtcctaaaa cgcaattatg gcccggaggt tgatgtctgg 720
agtgctggag ttatccttta tattcttctt tgtggtgttc cacctttctg ggcagagact 780
gaacaaggag tagcccaagc aattattcgg tctgtggttg atttcaagag agatccatgg 840
cctaaggttt ctgataatgc aaaggatctt gtaaagaaga tgcttgaccc ggatccaact 900
cgacgactca cagctcagca ggttcttgag cacacctggt tacaaaatat aaagaaagca 960
ccaaatgtct cattgggtga gactgtgaaa gcaaggctta aacaattttc agtaatgaac 1020
aagctcaaga aaagagctct aacgattatg gccgagtttt tatcagcgga agaagtggct 1080
ggaatgaagg acgcatttga tatgatggat acaggaaaga aaggcaagat taaccttgga 1140
gaacttaaaa atggtctgca aaagcttggc catcagatcc ctgatgttga tcttcagatt 1200
cttatggaag ctgcggatgt tgatggagat ggaagcttaa attatgggga gtttgttgct 1260
gtatctgttc atctcagaaa aatggcaaat gatgagcacc tgcacaaagc attttcagtt 1320
tttgacagag atcagagtgg ttacatagaa atcgaggagc tgcgcagtgc cttgagtgat 1380
gaggatggtg gcaacagtga ggaagtcatc aatgccatta tgcatgatgt tgacactgac 1440
aaggatggcc gcattagtta cgaggaattt gctgcaatga tgaaggctgg cacagattgg 1500
agaaaagcat cgagacagta ttctcgtgaa agattcaaca gtctcagctt aaaattgatg 1560
agagatggct cgatacaagt cggaaaggaa gaaggtagat ga 1602
<210> 2
<211> 533
<212> PRT
<213> 番茄(Solanum lycopersicum)
<400> 2
Met Gly Asn Cys Cys Gly Thr Pro Gly Asn Ser Ser Glu Asn Lys Lys
1 5 10 15
Lys Lys Asn Lys Pro Asn Pro Phe Ala Leu Asp Tyr Gly Ala Thr Gln
20 25 30
Ala Ser Gly Gly Asp Gly Asn Lys Leu Val Val Leu Lys Asp Pro Thr
35 40 45
Gly His Asn Ile Gln Glu Lys Tyr Asp Leu Gly Cys Glu Leu Gly Arg
50 55 60
Gly Glu Phe Gly Val Thr Tyr Leu Cys Thr Asp Val Asp Thr Gly Asp
65 70 75 80
Lys Tyr Ala Cys Lys Ser Ile Ser Lys Lys Lys Leu Arg Thr Ala Val
85 90 95
Asp Ile Asp Asp Val Arg Arg Glu Val Glu Ile Met Lys His Leu Pro
100 105 110
Lys His Pro Asn Ile Val Thr Leu Lys Asp Thr Tyr Glu Asp Asp Asn
115 120 125
Ala Val His Ile Val Met Glu Leu Cys Glu Gly Gly Glu Leu Phe Asp
130 135 140
Arg Ile Val Ala Arg Gly His Tyr Thr Glu Arg Ala Ala Ala Val Ile
145 150 155 160
Met Lys Thr Ile Val Glu Val Val Gln Met Cys His Met His Gly Val
165 170 175
Met His Arg Asp Leu Lys Pro Glu Asn Phe Leu Phe Gly Asn Lys Lys
180 185 190
Glu Thr Ala Pro Leu Lys Ala Ile Asp Phe Gly Leu Ser Val Phe Phe
195 200 205
Lys Pro Gly Glu Arg Phe Asn Glu Ile Val Gly Ser Pro Tyr Tyr Met
210 215 220
Ala Pro Glu Val Leu Lys Arg Asn Tyr Gly Pro Glu Val Asp Val Trp
225 230 235 240
Ser Ala Gly Val Ile Leu Tyr Ile Leu Leu Cys Gly Val Pro Pro Phe
245 250 255
Trp Ala Glu Thr Glu Gln Gly Val Ala Gln Ala Ile Ile Arg Ser Val
260 265 270
Val Asp Phe Lys Arg Asp Pro Trp Pro Lys Val Ser Asp Asn Ala Lys
275 280 285
Asp Leu Val Lys Lys Met Leu Asp Pro Asp Pro Thr Arg Arg Leu Thr
290 295 300
Ala Gln Gln Val Leu Glu His Thr Trp Leu Gln Asn Ile Lys Lys Ala
305 310 315 320
Pro Asn Val Ser Leu Gly Glu Thr Val Lys Ala Arg Leu Lys Gln Phe
325 330 335
Ser Val Met Asn Lys Leu Lys Lys Arg Ala Leu Thr Ile Met Ala Glu
340 345 350
Phe Leu Ser Ala Glu Glu Val Ala Gly Met Lys Asp Ala Phe Asp Met
355 360 365
Met Asp Thr Gly Lys Lys Gly Lys Ile Asn Leu Gly Glu Leu Lys Asn
370 375 380
Gly Leu Gln Lys Leu Gly His Gln Ile Pro Asp Val Asp Leu Gln Ile
385 390 395 400
Leu Met Glu Ala Ala Asp Val Asp Gly Asp Gly Ser Leu Asn Tyr Gly
405 410 415
Glu Phe Val Ala Val Ser Val His Leu Arg Lys Met Ala Asn Asp Glu
420 425 430
His Leu His Lys Ala Phe Ser Val Phe Asp Arg Asp Gln Ser Gly Tyr
435 440 445
Ile Glu Ile Glu Glu Leu Arg Ser Ala Leu Ser Asp Glu Asp Gly Gly
450 455 460
Asn Ser Glu Glu Val Ile Asn Ala Ile Met His Asp Val Asp Thr Asp
465 470 475 480
Lys Asp Gly Arg Ile Ser Tyr Glu Glu Phe Ala Ala Met Met Lys Ala
485 490 495
Gly Thr Asp Trp Arg Lys Ala Ser Arg Gln Tyr Ser Arg Glu Arg Phe
500 505 510
Asn Ser Leu Ser Leu Lys Leu Met Arg Asp Gly Ser Ile Gln Val Gly
515 520 525
Lys Glu Glu Gly Arg
530
<210> 3
<211> 4914
<212> DNA
<213> 番茄(Solanum lycopersicum)
<400> 3
atggggaatt gctgtgggac acctggtaat tcttctgaga ataagaagaa gaagaacaaa 60
ccaaaccctt ttgctcttga ttatggtgca actcaagcat ctggaggtga tggaaacaag 120
cttgttgtgt tgaaagatcc aacaggacac aatattcaag aaaaatatga tcttggttgt 180
gagctaggaa gaggagaatt tggggttaca tatttatgta ctgatgttga tacaggggac 240
aaatatgctt gcaaatcgat atcaaagaag aaacttagga ctgctgtaga tatagatgat 300
gttaggagag aagttgagat catgaagcat ttgcctaaac atcctaatat tgtgaccttg 360
aaggacactt atgaggatga taatgcagtg catattgtga tggaactttg tgaggggggt 420
gagttgtttg ataggattgt tgccagggga cattatacag agagagcagc tgcggttatt 480
atgaagacta tagtagaagt tgttcaggta cagttttact aattaatgct tgtctttgct 540
tttcatctat aattgttact tgtttgttta tttcatgtaa tttaggggtt gctaatttag 600
gattatgtta atgaagtttg aattgacaat gccaactgaa agtgtcaatt tagactagaa 660
aggaatgttg agattgaagt ttcatgtttg taagcagaat gaatgttctt gtcgaagaat 720
ctcttttaag aatatctttc ccacttcttc agtaaatttt cctactttct tatgtcgata 780
actgccactc cttttctgag tttgcctgca ctctcttggt agataggaca ctccctttct 840
tagctcagga tctactttgc ttgtgtaaat gagtgcttaa tgtttttatt tgattcattt 900
tacacgttga gatcctatag catttaatta gactagattt tatcttccaa tatcactaaa 960
cctagatgtc aaatatagta aaaagttatg ttgctcgaac tcttcaaaga tatcgacatg 1020
tgtgtgtcag atcctccaaa agtagtgcat ttttggagga tccgacgggg gtggggcaac 1080
aattttggag agttcgagca acttaggtaa aaagtgtatt catgctctaa atgttgttag 1140
gttttctatc tttcttatct tgtcttgtga attttgcttt cttggcaact gtttcattgt 1200
cctatatttt tctcatgcaa atacctttaa aattatatac tttcgtgcca accatttaaa 1260
tgaaacttaa ggaagctaat cacacaggat ttcaccatcc aaatgtcgaa aatgaatcgc 1320
aatttgtgat agttttagac agagagctta tttttataga tgcaattaga ataatagaaa 1380
tgtaaaataa ctatcagtaa ccagtaagat gccggtcttt atggtctatt ttcagagggg 1440
gagatctatt gtagcttgct ttcagcatca gttagtatag cataggactg ttgctaacag 1500
ggttcaattt taaactgtat ttggtgaaca taatgacatg agagctcaac ttacataact 1560
acagattaga gagagacatt cattttatgg aatggattta agaagttcat gtttctacat 1620
ttagttgtta taagaacagg gtggtaactg gtaaccgacg caaacatgat ggctaattat 1680
cccaagttta tacaatttaa tctatgtggt tgctcataca tctttggtca atgttcactt 1740
tacagttgca ttatgtttcc tcaagaaatg gctgttcttc tttcaatttt caacttacgg 1800
agtttcaatt gacattgcag atgtgtcata tgcatggagt aatgcatcgt gatctcaaac 1860
ctgagaactt tctgtttggt aacaagaaag aaactgctcc tttgaaggct attgactttg 1920
gattatcagt gttctttaaa cccggtaatg caataagtaa tgatagtgtg ttgtaacttt 1980
ttttcatgct aaaaaagatt aatgttggaa tcatctttta atatgtttaa caggggaacg 2040
ctttaatgag attgtgggaa gtccttatta catggctcca gaggtcctaa aacgcaatta 2100
tggcccggag gttgatgtct ggagtgctgg agttatcctt tatattcttc tttgtggtgt 2160
tccacctttc tgggcaggtc cgtttctttt atttgccttc tttaggtaac aagtaatacc 2220
tcggttcacc gacaaataac aaagtaaatc aaccttggtg catactattc atttcttaga 2280
acttggtaat ttattaatga gtgtcaaaat gaaatggaat ttgaaacata ttgatgcgtt 2340
gttaacatcg tcgatgacta ggtgagggtg gctgcaagag tgaatcccac ctcttcctgg 2400
accaattcaa aatcaaagtt cctttttttc cttttggcct gtgatagtca gtactttgtg 2460
cttccattaa tttactcaca gctttctcag taattgccta aaattctact gaatatgtgc 2520
agagactgaa caaggagtag cccaagcaat tattcggtct gtggttgatt tcaagagaga 2580
tccatggcct aaggtttctg ataatgcaaa ggatcttgta aagaagatgc ttgacccgga 2640
tccaactcga cgactcacag ctcagcaggt tcttggtaat gtttgttaat ccttgagatg 2700
ctatactttc atttgcttgc tgtttgttct ctcttagttt cataagttca attttgtgac 2760
ttttttgggg ctttcgtgca tatacatgaa tgcagagcac acctggttac aaaatataaa 2820
gaaagcacca aatgtctcat tgggtgagac tgtgaaagca aggcttaaac aattttcagt 2880
aatgaacaag ctcaagaaaa gagctctaac ggtaaaatat tcccttttca tttgcagtta 2940
catatcatta gcgttctgca ccttctcaga aggatatctt catatgccta attcttaaag 3000
cccaaaataa ggacaaaaga ttttctgcct tctcacaaaa gtttcgggtg ttatattctt 3060
tcttcattgc ttgcacagat tatggccgag tttttatcag cggaagaagt ggctggaatg 3120
aaggacgcat ttgatatgat ggatacagga aagaaaggca agattaacct tggagaactt 3180
aaaaatggtc tgcaaaagct tggccatcag atccctgatg ttgatcttca gattcttatg 3240
gaagctgtaa gcattctcat gctcttcatt tcacttgatt ggattttctt tcaactttac 3300
gaagactaaa tatatatatt tataagtcaa gcgtctaata ttattataat tgttcccttt 3360
tttatcttct gcttcacttt tataggcgga tgttgatgga gatggaagct taaattatgg 3420
ggagtttgtt gctgtatctg ttcatctcag aaaaatggca aatgatgagc acctgcacaa 3480
agcattttca gtttttgaca gagatcagag tggttacata gaaatcgagg agctgcgcag 3540
tgccttgagt gatgaggatg gtggcaacag tgaggaagtc atcaatgcca ttatgcatga 3600
tgttgacact gacaaggtta gaattttttt tgtgtccatt catgtatatc tactggattt 3660
catactagtg catatacttc ttttataaat gtctctttaa ttgcttgtgg aaccataaat 3720
acccccctta tcttgttccg cagctccaac tacacacgaa actatgttga agacgtatta 3780
cccccttgta cttgtaaaat gtgtatcttc cgtcatcctg agaggcatat atgatgtaac 3840
caaatgtgaa catttaagta tatgcatgtg ctgcctcatg tcatttaccc ctttttccct 3900
tttcacttct cttccatctc cgcaacatgt taatgtcctc tagcaaatct gaaaaaagat 3960
ggagtgagct tctcaacgga aaaaatcgga ttcaccctag ttttagtttt ctggcaacat 4020
aaacagctca agtcgataca aaaaaacaaa tgttcctatc aaaaaactca aattagtaga 4080
aaggaaaaga agggtcatag gtaagggaag aacaaaaagg aatagaaaga cccttcgggg 4140
tggcccagtt atttgggctt ggggcttggg gcttgggact tccatgttgg aggtctcaag 4200
ttcaaaaccc cttgccaatg aaagcaacgg gttttccttc tacgttgact gtcgagctcg 4260
tcgcaccggg cttacctagt gcggtttacc tctcctttgc gtgctatagg aataggggtt 4320
ttaccctgtg tgcactcaaa gggtagcgac tgcggatttc ccaaaagaaa aaggaagaga 4380
aaggtaaaga aaacatttta aaaaagagta ggatggttat aatagaagaa atgctacaat 4440
attttttcac ctcaggcgct ttaagagagt aaatactcta cgttacatgg cgtcccaatg 4500
ttgtcgtagt tacattttgg acaagaacag ggagtaatag gcctcccgca aagttaaaat 4560
gtagtgggga atgggggaat aattgaggaa ggtatttata tattttctcc tttctaaatc 4620
atattttctt ttccattcac tgctcttctg ttcaacctta tcattcattt ggactaaatg 4680
ttgaattgat tttgagcttt tagtttgtta ccacagcttc aaagttcatc agttacattg 4740
tctataacca tgcaggatgg ccgcattagt tacgaggaat ttgctgcaat gatgaaggct 4800
ggcacagatt ggagaaaagc atcgagacag tattctcgtg aaagattcaa cagtctcagc 4860
ttaaaattga tgagagatgg ctcgatacaa gtcggaaagg aagaaggtag atga 4914
<210> 4
<211> 20
<212> DNA
<213> 番茄(Solanum lycopersicum)
<400> 4
gtgatggaac tttgtgaggg 20
<210> 5
<211> 20
<212> DNA
<213> 番茄(Solanum lycopersicum)
<400> 5
aggggggtga gttgtttgat 20
<210> 6
<211> 36
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 6
gcggtctcta ttgaacaaag caccagtggt ctagtg 36
<210> 7
<211> 47
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 7
gcggtctctc cctcacaaag ttccatcact gcaccagccg ggaatcg 47
<210> 8
<211> 40
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 8
gcggtctctt accgttttag agctagaaat agcaagttaa 40
<210> 9
<211> 51
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 9
gcggtctcta aacatcaaac aactcacccc ccttgcacca gccgggaatc g 51
<210> 10
<211> 18
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 10
tgtaaaacga cggccagt 18
<210> 11
<211> 28
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 11
ggtattggtt tatctcatcg gaactgca 28
<210> 12
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 12
tggtgcaact caagcatctg 20
<210> 13
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 13
ggatggtgaa atcctgtgtg 20
<210> 14
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 14
atgaagcgaa aggactggtc 20
<210> 15
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 15
tgcacccctc atgtacttgt 20
Claims (2)
1.CPK27基因作为负调控因子在提高番茄油菜素内酯含量中的应用,其特征在于,所述应用包括:利用生物学技术使得所述CPK27基因功能缺失,进而提高番茄内源油菜素内酯类物质的含量;所述CPK27基因的蛋白编码区的核苷酸序列如SEQ ID NO.1所示;所述油菜素内酯包括油菜素甾酮和芸薹甾内酯;所述CPK27基因编码的蛋白质负调控油菜素甾酮合成基因CYP85A1的表达。
2.一种提高番茄内源油菜素内酯含量的培育方法,其特征在于,包括以下步骤:
(1)在番茄CPK27基因的蛋白编码区选取含有PAM结构的靶标片段,以靶标片段PAM结构前20个碱基为依据,进行引物设计,构建CRISPR/Cas9载体;所述番茄CPK27基因的核苷酸序列如SEQ ID NO.3所示;所述靶标片段PAM结构前20个碱基如SEQ ID NO.4或SEQ ID NO.5所示;构建CRISPR/Cas9载体的引物对的核苷酸序列如SEQ ID NO.6和SEQ ID NO.7所示,或者如SEQ ID NO.8和SEQ ID NO.9所示;
(2)构建含步骤(1)所述CRISPR/Cas9载体的农杆菌基因工程菌;
(3)将步骤(2)所述基因工程菌转化番茄子叶,获得不含外源Cas9蛋白且靶标序列发生变异的稳定遗传的纯合突变体株系。
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202210456566.XA CN114752608B (zh) | 2022-04-24 | 2022-04-24 | 一种高含量油菜素内酯番茄植株的培育方法 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202210456566.XA CN114752608B (zh) | 2022-04-24 | 2022-04-24 | 一种高含量油菜素内酯番茄植株的培育方法 |
Publications (2)
Publication Number | Publication Date |
---|---|
CN114752608A CN114752608A (zh) | 2022-07-15 |
CN114752608B true CN114752608B (zh) | 2023-12-15 |
Family
ID=82334061
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202210456566.XA Active CN114752608B (zh) | 2022-04-24 | 2022-04-24 | 一种高含量油菜素内酯番茄植株的培育方法 |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN114752608B (zh) |
Families Citing this family (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN118207227A (zh) * | 2022-12-15 | 2024-06-18 | 中国农业科学院农业基因组研究所 | 多核苷酸、蛋白质、生物材料及其在提升植物果实品质中的应用 |
Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN103796996A (zh) * | 2011-09-16 | 2014-05-14 | 先正达参股股份有限公司 | 植物生长调节化合物 |
-
2022
- 2022-04-24 CN CN202210456566.XA patent/CN114752608B/zh active Active
Patent Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN103796996A (zh) * | 2011-09-16 | 2014-05-14 | 先正达参股股份有限公司 | 植物生长调节化合物 |
Non-Patent Citations (3)
Title |
---|
Involvement of calcium-dependent protein kinase in rice(Oryzasativa L.) laminain clination caused by brassinolide;Yang,G.X.,andKomatsu,S;Plant Cell Physiol.;第41卷;摘要 * |
PREDICTED: Solanum lycopersicum calcium-dependent protein kinase 7 (LOC101247097), mRNA;Genbank;Genbank;LOCUS、FEATURES、ORIGIN部分 * |
SlCBL1基因调控番茄果实成熟发育的分子机理;王媛花;贾思振;颜志明;冯英娜;;西北植物学报(第11期);全文 * |
Also Published As
Publication number | Publication date |
---|---|
CN114752608A (zh) | 2022-07-15 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN107205354A (zh) | 单倍体诱导物 | |
CN108822194B (zh) | 一个植物淀粉合成相关蛋白OsFLO10及其编码基因与应用 | |
CN110684796B (zh) | CRISPR-Cas9特异性敲除大豆脂肪氧化酶基因的方法及其应用 | |
CN108660140B (zh) | SlSL4基因在调控番茄果实成熟中的应用 | |
CN110088284A (zh) | 高水平蒂巴因罂粟及其生产的方法 | |
CN111690625A (zh) | 具有除草剂抗性的乙酰乳酸合酶突变蛋白及其应用 | |
CN111574605B (zh) | 水稻基因OsLAT5在调节敌草快的吸收积累中的应用 | |
CN114846022A (zh) | 通过阻抑基因的下调增强作物的疾病抗性 | |
CN114752608B (zh) | 一种高含量油菜素内酯番茄植株的培育方法 | |
CN108395472A (zh) | 一种控制稻类粒长和粒重的基因及其应用 | |
CN112280786B (zh) | 一种养分高效利用耐除草剂玉米连hh2823转化事件及其特异性鉴定方法和应用 | |
CN113646326A (zh) | 用于抗植物病害的基因 | |
CN113621625A (zh) | 芝麻SiERF103基因在增强植物抗性中的应用 | |
CN117511975A (zh) | LecRK27基因在提高果实均一性和产量中的应用 | |
CN109971763A (zh) | 花期调控基因cmp1和相关的载体及其应用 | |
CN109776664A (zh) | 一种控制稻类小粒和半矮秆的基因及其应用 | |
CN111560055B (zh) | 水稻基因OsLAT3在调节敌草快的吸收累积中的应用 | |
CN114540373A (zh) | 一种降低水稻籽粒中镉含量的基因及其应用 | |
CN110343159B (zh) | 一种绿豆开花基因VrELF3的表达载体的应用 | |
CN110407922B (zh) | 水稻耐冷基因qSCT11及其应用 | |
CN101142318B (zh) | 细胞数目多核苷酸和多肽以及使用其的方法 | |
CN108795949B (zh) | 一种水稻叶色调控相关基因OsWSL6及其编码蛋白质和应用 | |
CN106349353B (zh) | 一种调控植物淀粉合成相关蛋白OsFSE及其编码基因与应用 | |
CN102203243A (zh) | 转基因植物 | |
NL2028064B1 (en) | Gene for controlling small grain and semi-dwarf of oryza sativa and application thereof |
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 |