CN113801890B - Application of protein ZmbZIPc3 in regulation and control of salt tolerance and drought resistance of plants - Google Patents
Application of protein ZmbZIPc3 in regulation and control of salt tolerance and drought resistance of plants Download PDFInfo
- Publication number
- CN113801890B CN113801890B CN202010467043.6A CN202010467043A CN113801890B CN 113801890 B CN113801890 B CN 113801890B CN 202010467043 A CN202010467043 A CN 202010467043A CN 113801890 B CN113801890 B CN 113801890B
- Authority
- CN
- China
- Prior art keywords
- protein
- plant
- zmbzipc3
- transgenic plant
- plants
- 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
- 108090000623 proteins and genes Proteins 0.000 title claims abstract description 115
- 102000004169 proteins and genes Human genes 0.000 title claims abstract description 66
- 230000015784 hyperosmotic salinity response Effects 0.000 title claims abstract description 20
- 230000033228 biological regulation Effects 0.000 title claims description 7
- 241000196324 Embryophyta Species 0.000 claims abstract description 128
- 240000008042 Zea mays Species 0.000 claims abstract description 71
- 241000219195 Arabidopsis thaliana Species 0.000 claims abstract description 66
- 235000002017 Zea mays subsp mays Nutrition 0.000 claims abstract description 66
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 claims abstract description 54
- 235000005822 corn Nutrition 0.000 claims abstract description 54
- 239000012620 biological material Substances 0.000 claims abstract description 18
- 125000003275 alpha amino acid group Chemical group 0.000 claims abstract description 12
- 230000024346 drought recovery Effects 0.000 claims abstract description 7
- 230000001105 regulatory effect Effects 0.000 claims abstract description 6
- 210000004899 c-terminal region Anatomy 0.000 claims abstract description 3
- 108020001507 fusion proteins Proteins 0.000 claims abstract description 3
- 102000037865 fusion proteins Human genes 0.000 claims abstract description 3
- 230000009261 transgenic effect Effects 0.000 claims description 71
- 230000014509 gene expression Effects 0.000 claims description 45
- 239000013598 vector Substances 0.000 claims description 44
- 230000002018 overexpression Effects 0.000 claims description 28
- 108020004414 DNA Proteins 0.000 claims description 24
- 102000039446 nucleic acids Human genes 0.000 claims description 24
- 108020004707 nucleic acids Proteins 0.000 claims description 24
- 150000007523 nucleic acids Chemical class 0.000 claims description 24
- 238000000034 method Methods 0.000 claims description 23
- 210000004027 cell Anatomy 0.000 claims description 16
- 102000053602 DNA Human genes 0.000 claims description 13
- 210000000056 organ Anatomy 0.000 claims description 9
- 108091026890 Coding region Proteins 0.000 claims description 8
- 244000005700 microbiome Species 0.000 claims description 7
- 230000000694 effects Effects 0.000 claims description 4
- 102000010970 Connexin Human genes 0.000 claims description 2
- 108050001175 Connexin Proteins 0.000 claims description 2
- 210000001938 protoplast Anatomy 0.000 claims description 2
- 239000003513 alkali Substances 0.000 claims 1
- 230000035882 stress Effects 0.000 abstract description 18
- 150000003839 salts Chemical class 0.000 abstract description 5
- 230000001276 controlling effect Effects 0.000 abstract description 3
- 230000008723 osmotic stress Effects 0.000 abstract description 3
- 230000000087 stabilizing effect Effects 0.000 abstract description 2
- 241000219194 Arabidopsis Species 0.000 description 48
- JLIDBLDQVAYHNE-YKALOCIXSA-N (+)-Abscisic acid Chemical compound OC(=O)/C=C(/C)\C=C\[C@@]1(O)C(C)=CC(=O)CC1(C)C JLIDBLDQVAYHNE-YKALOCIXSA-N 0.000 description 44
- 230000012010 growth Effects 0.000 description 29
- 239000013604 expression vector Substances 0.000 description 24
- 230000035784 germination Effects 0.000 description 24
- 239000002609 medium Substances 0.000 description 23
- FCRACOPGPMPSHN-UHFFFAOYSA-N desoxyabscisic acid Natural products OC(=O)C=C(C)C=CC1C(C)=CC(=O)CC1(C)C FCRACOPGPMPSHN-UHFFFAOYSA-N 0.000 description 22
- 239000007787 solid Substances 0.000 description 21
- 239000001963 growth medium Substances 0.000 description 16
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 14
- 230000004083 survival effect Effects 0.000 description 13
- 235000016383 Zea mays subsp huehuetenangensis Nutrition 0.000 description 12
- 235000009973 maize Nutrition 0.000 description 12
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 10
- 238000002474 experimental method Methods 0.000 description 10
- 239000000463 material Substances 0.000 description 10
- 239000002689 soil Substances 0.000 description 9
- 241000589158 Agrobacterium Species 0.000 description 8
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 8
- 238000012163 sequencing technique Methods 0.000 description 7
- 238000012258 culturing Methods 0.000 description 6
- 230000007226 seed germination Effects 0.000 description 6
- 230000009466 transformation Effects 0.000 description 6
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 5
- 235000007244 Zea mays Nutrition 0.000 description 5
- 238000004458 analytical method Methods 0.000 description 5
- 239000002299 complementary DNA Substances 0.000 description 5
- 239000012153 distilled water Substances 0.000 description 5
- 230000008641 drought stress Effects 0.000 description 5
- 238000011160 research Methods 0.000 description 5
- 239000011780 sodium chloride Substances 0.000 description 5
- 101150090724 3 gene Proteins 0.000 description 4
- 241000589155 Agrobacterium tumefaciens Species 0.000 description 4
- 241000701489 Cauliflower mosaic virus Species 0.000 description 4
- 238000012408 PCR amplification Methods 0.000 description 4
- 239000005708 Sodium hypochlorite Substances 0.000 description 4
- 239000003623 enhancer Substances 0.000 description 4
- 230000001939 inductive effect Effects 0.000 description 4
- 239000002773 nucleotide Substances 0.000 description 4
- 125000003729 nucleotide group Chemical group 0.000 description 4
- SUKJFIGYRHOWBL-UHFFFAOYSA-N sodium hypochlorite Chemical compound [Na+].Cl[O-] SUKJFIGYRHOWBL-UHFFFAOYSA-N 0.000 description 4
- 102000004190 Enzymes Human genes 0.000 description 3
- 241000880493 Leptailurus serval Species 0.000 description 3
- 125000000539 amino acid group Chemical group 0.000 description 3
- 235000013339 cereals Nutrition 0.000 description 3
- 238000011161 development Methods 0.000 description 3
- 230000018109 developmental process Effects 0.000 description 3
- 239000012634 fragment Substances 0.000 description 3
- 238000005286 illumination Methods 0.000 description 3
- 230000001404 mediated effect Effects 0.000 description 3
- 238000003976 plant breeding Methods 0.000 description 3
- 229920001223 polyethylene glycol Polymers 0.000 description 3
- 239000011148 porous material Substances 0.000 description 3
- 230000002786 root growth Effects 0.000 description 3
- 238000013518 transcription Methods 0.000 description 3
- 230000035897 transcription Effects 0.000 description 3
- LWTDZKXXJRRKDG-KXBFYZLASA-N (-)-phaseollin Chemical compound C1OC2=CC(O)=CC=C2[C@H]2[C@@H]1C1=CC=C3OC(C)(C)C=CC3=C1O2 LWTDZKXXJRRKDG-KXBFYZLASA-N 0.000 description 2
- 102000040650 (ribonucleotides)n+m Human genes 0.000 description 2
- 101100043932 Arabidopsis thaliana SUVH4 gene Proteins 0.000 description 2
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 description 2
- 241001057636 Dracaena deremensis Species 0.000 description 2
- 206010016807 Fluid retention Diseases 0.000 description 2
- 235000007688 Lycopersicon esculentum Nutrition 0.000 description 2
- 239000005588 Oxadiazon Substances 0.000 description 2
- CHNUNORXWHYHNE-UHFFFAOYSA-N Oxadiazon Chemical compound C1=C(Cl)C(OC(C)C)=CC(N2C(OC(=N2)C(C)(C)C)=O)=C1Cl CHNUNORXWHYHNE-UHFFFAOYSA-N 0.000 description 2
- FUOGXAQMNJMBFG-WPRPVWTQSA-N Pro-Val-Gly Chemical compound OC(=O)CNC(=O)[C@H](C(C)C)NC(=O)[C@@H]1CCCN1 FUOGXAQMNJMBFG-WPRPVWTQSA-N 0.000 description 2
- 240000003768 Solanum lycopersicum Species 0.000 description 2
- 108091081024 Start codon Proteins 0.000 description 2
- UELITFHSCLAHKR-UHFFFAOYSA-N acibenzolar-S-methyl Chemical compound CSC(=O)C1=CC=CC2=C1SN=N2 UELITFHSCLAHKR-UHFFFAOYSA-N 0.000 description 2
- 108010087924 alanylproline Proteins 0.000 description 2
- 230000003321 amplification Effects 0.000 description 2
- 108010077245 asparaginyl-proline Proteins 0.000 description 2
- 239000001110 calcium chloride Substances 0.000 description 2
- 235000011148 calcium chloride Nutrition 0.000 description 2
- 229910001628 calcium chloride Inorganic materials 0.000 description 2
- 230000006866 deterioration Effects 0.000 description 2
- 235000013399 edible fruits Nutrition 0.000 description 2
- 230000006870 function Effects 0.000 description 2
- VPZXBVLAVMBEQI-UHFFFAOYSA-N glycyl-DL-alpha-alanine Natural products OC(=O)C(C)NC(=O)CN VPZXBVLAVMBEQI-UHFFFAOYSA-N 0.000 description 2
- 238000009396 hybridization Methods 0.000 description 2
- ZNJFBWYDHIGLCU-HWKXXFMVSA-N jasmonic acid Chemical compound CC\C=C/C[C@@H]1[C@@H](CC(O)=O)CCC1=O ZNJFBWYDHIGLCU-HWKXXFMVSA-N 0.000 description 2
- 238000005259 measurement Methods 0.000 description 2
- 230000007246 mechanism Effects 0.000 description 2
- 239000012528 membrane Substances 0.000 description 2
- 108020004999 messenger RNA Proteins 0.000 description 2
- 108010058731 nopaline synthase Proteins 0.000 description 2
- 238000003199 nucleic acid amplification method Methods 0.000 description 2
- 239000013612 plasmid Substances 0.000 description 2
- 108010053725 prolylvaline Proteins 0.000 description 2
- 238000003753 real-time PCR Methods 0.000 description 2
- 238000003259 recombinant expression Methods 0.000 description 2
- YGSDEFSMJLZEOE-UHFFFAOYSA-N salicylic acid Chemical compound OC(=O)C1=CC=CC=C1O YGSDEFSMJLZEOE-UHFFFAOYSA-N 0.000 description 2
- 230000005026 transcription initiation Effects 0.000 description 2
- 238000012795 verification Methods 0.000 description 2
- 238000005406 washing Methods 0.000 description 2
- BRPMXFSTKXXNHF-IUCAKERBSA-N (2s)-1-[2-[[(2s)-pyrrolidine-2-carbonyl]amino]acetyl]pyrrolidine-2-carboxylic acid Chemical compound OC(=O)[C@@H]1CCCN1C(=O)CNC(=O)[C@H]1NCCC1 BRPMXFSTKXXNHF-IUCAKERBSA-N 0.000 description 1
- KPDTZVSUQCBOAE-HTFCKZLJSA-N (2s)-2-[[(2s)-1-[(2s)-2-[[(2s)-2-[[(2s)-2-aminopropanoyl]amino]propanoyl]amino]propanoyl]pyrrolidine-2-carbonyl]amino]propanoic acid Chemical compound C[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@@H](C)C(=O)N1CCC[C@H]1C(=O)N[C@@H](C)C(O)=O KPDTZVSUQCBOAE-HTFCKZLJSA-N 0.000 description 1
- OZRFYUJEXYKQDV-UHFFFAOYSA-N 2-[[2-[[2-[(2-amino-3-carboxypropanoyl)amino]-3-carboxypropanoyl]amino]-3-carboxypropanoyl]amino]butanedioic acid Chemical compound OC(=O)CC(N)C(=O)NC(CC(O)=O)C(=O)NC(CC(O)=O)C(=O)NC(CC(O)=O)C(O)=O OZRFYUJEXYKQDV-UHFFFAOYSA-N 0.000 description 1
- SBGXWWCLHIOABR-UHFFFAOYSA-N Ala Ala Gly Ala Chemical compound CC(N)C(=O)NC(C)C(=O)NCC(=O)NC(C)C(O)=O SBGXWWCLHIOABR-UHFFFAOYSA-N 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
- CXRCVCURMBFFOL-FXQIFTODSA-N Ala-Ala-Pro Chemical compound C[C@H](N)C(=O)N[C@@H](C)C(=O)N1CCC[C@H]1C(O)=O CXRCVCURMBFFOL-FXQIFTODSA-N 0.000 description 1
- ZFXQNADNEBRERM-BJDJZHNGSA-N Ala-Ala-Pro-Pro Chemical compound C[C@H](N)C(=O)N[C@@H](C)C(=O)N1CCC[C@H]1C(=O)N1[C@H](C(O)=O)CCC1 ZFXQNADNEBRERM-BJDJZHNGSA-N 0.000 description 1
- BLGHHPHXVJWCNK-GUBZILKMSA-N Ala-Gln-Leu Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(C)C)C(O)=O BLGHHPHXVJWCNK-GUBZILKMSA-N 0.000 description 1
- YIGLXQRFQVWFEY-NRPADANISA-N Ala-Gln-Val Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](C(C)C)C(O)=O YIGLXQRFQVWFEY-NRPADANISA-N 0.000 description 1
- ZVFVBBGVOILKPO-WHFBIAKZSA-N Ala-Gly-Ala Chemical compound C[C@H](N)C(=O)NCC(=O)N[C@@H](C)C(O)=O ZVFVBBGVOILKPO-WHFBIAKZSA-N 0.000 description 1
- MPLOSMWGDNJSEV-WHFBIAKZSA-N Ala-Gly-Asp Chemical compound [H]N[C@@H](C)C(=O)NCC(=O)N[C@@H](CC(O)=O)C(O)=O MPLOSMWGDNJSEV-WHFBIAKZSA-N 0.000 description 1
- MDNAVFBZPROEHO-DCAQKATOSA-N Ala-Lys-Val Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](C(C)C)C(O)=O MDNAVFBZPROEHO-DCAQKATOSA-N 0.000 description 1
- MDNAVFBZPROEHO-UHFFFAOYSA-N Ala-Lys-Val Natural products CC(C)C(C(O)=O)NC(=O)C(NC(=O)C(C)N)CCCCN MDNAVFBZPROEHO-UHFFFAOYSA-N 0.000 description 1
- HOVPGJUNRLMIOZ-CIUDSAMLSA-N Ala-Ser-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](CO)NC(=O)[C@H](C)N HOVPGJUNRLMIOZ-CIUDSAMLSA-N 0.000 description 1
- NCQMBSJGJMYKCK-ZLUOBGJFSA-N Ala-Ser-Ser Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CO)C(=O)N[C@@H](CO)C(O)=O NCQMBSJGJMYKCK-ZLUOBGJFSA-N 0.000 description 1
- OEVCHROQUIVQFZ-YTLHQDLWSA-N Ala-Thr-Ala Chemical compound C[C@H](N)C(=O)N[C@@H]([C@H](O)C)C(=O)N[C@@H](C)C(O)=O OEVCHROQUIVQFZ-YTLHQDLWSA-N 0.000 description 1
- UISQLSIBJKEJSS-GUBZILKMSA-N Arg-Arg-Ser Chemical compound NC(N)=NCCC[C@H](N)C(=O)N[C@@H](CCCN=C(N)N)C(=O)N[C@@H](CO)C(O)=O UISQLSIBJKEJSS-GUBZILKMSA-N 0.000 description 1
- UFBURHXMKFQVLM-CIUDSAMLSA-N Arg-Glu-Ser Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CO)C(O)=O UFBURHXMKFQVLM-CIUDSAMLSA-N 0.000 description 1
- PSOPJDUQUVFSLS-GUBZILKMSA-N Arg-Met-Cys Chemical compound CSCC[C@@H](C(=O)N[C@@H](CS)C(=O)O)NC(=O)[C@H](CCCN=C(N)N)N PSOPJDUQUVFSLS-GUBZILKMSA-N 0.000 description 1
- VENMDXUVHSKEIN-GUBZILKMSA-N Arg-Ser-Arg Chemical compound NC(N)=NCCC[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@@H](CCCN=C(N)N)C(O)=O VENMDXUVHSKEIN-GUBZILKMSA-N 0.000 description 1
- ULBHWNVWSCJLCO-NHCYSSNCSA-N Arg-Val-Glu Chemical compound OC(=O)CC[C@@H](C(O)=O)NC(=O)[C@H](C(C)C)NC(=O)[C@@H](N)CCCN=C(N)N ULBHWNVWSCJLCO-NHCYSSNCSA-N 0.000 description 1
- XYOVHPDDWCEUDY-CIUDSAMLSA-N Asn-Ala-Leu Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](C)C(=O)N[C@@H](CC(C)C)C(O)=O XYOVHPDDWCEUDY-CIUDSAMLSA-N 0.000 description 1
- ORXCYAFUCSTQGY-FXQIFTODSA-N Asn-Ala-Met Chemical compound C[C@@H](C(=O)N[C@@H](CCSC)C(=O)O)NC(=O)[C@H](CC(=O)N)N ORXCYAFUCSTQGY-FXQIFTODSA-N 0.000 description 1
- PCKRJVZAQZWNKM-WHFBIAKZSA-N Asn-Asn-Gly Chemical compound NC(=O)C[C@H](N)C(=O)N[C@@H](CC(N)=O)C(=O)NCC(O)=O PCKRJVZAQZWNKM-WHFBIAKZSA-N 0.000 description 1
- DXVMJJNAOVECBA-WHFBIAKZSA-N Asn-Gly-Asn Chemical compound NC(=O)C[C@H](N)C(=O)NCC(=O)N[C@@H](CC(N)=O)C(O)=O DXVMJJNAOVECBA-WHFBIAKZSA-N 0.000 description 1
- UDSVWSUXKYXSTR-QWRGUYRKSA-N Asn-Gly-Tyr Chemical compound [H]N[C@@H](CC(N)=O)C(=O)NCC(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O UDSVWSUXKYXSTR-QWRGUYRKSA-N 0.000 description 1
- AEZCCDMZZJOGII-DCAQKATOSA-N Asn-Met-Leu Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CC(C)C)C(O)=O AEZCCDMZZJOGII-DCAQKATOSA-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
- GMUOCGCDOYYWPD-FXQIFTODSA-N Asn-Pro-Ser Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CO)C(O)=O GMUOCGCDOYYWPD-FXQIFTODSA-N 0.000 description 1
- XEDQMTWEYFBOIK-ACZMJKKPSA-N Asp-Ala-Glu Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](C)C(=O)N[C@@H](CCC(O)=O)C(O)=O XEDQMTWEYFBOIK-ACZMJKKPSA-N 0.000 description 1
- KTTCQQNRRLCIBC-GHCJXIJMSA-N Asp-Ile-Ala Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](C)C(O)=O KTTCQQNRRLCIBC-GHCJXIJMSA-N 0.000 description 1
- SPKCGKRUYKMDHP-GUDRVLHUSA-N Asp-Ile-Pro Chemical compound CC[C@H](C)[C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](CC(=O)O)N SPKCGKRUYKMDHP-GUDRVLHUSA-N 0.000 description 1
- UMHUHHJMEXNSIV-CIUDSAMLSA-N Asp-Leu-Ser Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@@H](N)CC(O)=O UMHUHHJMEXNSIV-CIUDSAMLSA-N 0.000 description 1
- WMLFFCRUSPNENW-ZLUOBGJFSA-N Asp-Ser-Ala Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](C)C(O)=O WMLFFCRUSPNENW-ZLUOBGJFSA-N 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- 101001033883 Cenchritis muricatus Protease inhibitor 2 Proteins 0.000 description 1
- 241000195493 Cryptophyta Species 0.000 description 1
- LMKYZBGVKHTLTN-NKWVEPMBSA-N D-nopaline Chemical compound NC(=N)NCCC[C@@H](C(O)=O)N[C@@H](C(O)=O)CCC(O)=O LMKYZBGVKHTLTN-NKWVEPMBSA-N 0.000 description 1
- 108010066133 D-octopine dehydrogenase Proteins 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 241000588724 Escherichia coli Species 0.000 description 1
- 241000620209 Escherichia coli DH5[alpha] Species 0.000 description 1
- 241000233866 Fungi Species 0.000 description 1
- 108700028146 Genetic Enhancer Elements Proteins 0.000 description 1
- 229930182566 Gentamicin Natural products 0.000 description 1
- CEAZRRDELHUEMR-URQXQFDESA-N Gentamicin Chemical compound O1[C@H](C(C)NC)CC[C@@H](N)[C@H]1O[C@H]1[C@H](O)[C@@H](O[C@@H]2[C@@H]([C@@H](NC)[C@@](C)(O)CO2)O)[C@H](N)C[C@@H]1N CEAZRRDELHUEMR-URQXQFDESA-N 0.000 description 1
- YJIUYQKQBBQYHZ-ACZMJKKPSA-N Gln-Ala-Ala Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](C)C(=O)N[C@@H](C)C(O)=O YJIUYQKQBBQYHZ-ACZMJKKPSA-N 0.000 description 1
- HHWQMFIGMMOVFK-WDSKDSINSA-N Gln-Ala-Gly Chemical compound OC(=O)CNC(=O)[C@H](C)NC(=O)[C@@H](N)CCC(N)=O HHWQMFIGMMOVFK-WDSKDSINSA-N 0.000 description 1
- KWUSGAIFNHQCBY-DCAQKATOSA-N Gln-Arg-Arg Chemical compound NC(=O)CC[C@H](N)C(=O)N[C@@H](CCCN=C(N)N)C(=O)N[C@@H](CCCN=C(N)N)C(O)=O KWUSGAIFNHQCBY-DCAQKATOSA-N 0.000 description 1
- WOACHWLUOFZLGJ-GUBZILKMSA-N Gln-Arg-Gln Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(N)=O)C(O)=O WOACHWLUOFZLGJ-GUBZILKMSA-N 0.000 description 1
- ULXXDWZMMSQBDC-ACZMJKKPSA-N Gln-Asp-Asp Chemical compound C(CC(=O)N)[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)N[C@@H](CC(=O)O)C(=O)O)N ULXXDWZMMSQBDC-ACZMJKKPSA-N 0.000 description 1
- QYKBTDOAMKORGL-FXQIFTODSA-N Gln-Gln-Asp Chemical compound C(CC(=O)N)[C@@H](C(=O)N[C@@H](CCC(=O)N)C(=O)N[C@@H](CC(=O)O)C(=O)O)N QYKBTDOAMKORGL-FXQIFTODSA-N 0.000 description 1
- HDUDGCZEOZEFOA-KBIXCLLPSA-N Gln-Ile-Ala Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](C)C(=O)O)NC(=O)[C@H](CCC(=O)N)N HDUDGCZEOZEFOA-KBIXCLLPSA-N 0.000 description 1
- OKARHJKJTKFQBM-ACZMJKKPSA-N Gln-Ser-Asn Chemical compound C(CC(=O)N)[C@@H](C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(=O)N)C(=O)O)N OKARHJKJTKFQBM-ACZMJKKPSA-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
- JRCUFCXYZLPSDZ-ACZMJKKPSA-N Glu-Asp-Ser Chemical compound OC(=O)CC[C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CO)C(O)=O JRCUFCXYZLPSDZ-ACZMJKKPSA-N 0.000 description 1
- WPLGNDORMXTMQS-FXQIFTODSA-N Glu-Gln-Ser Chemical compound OC(=O)CC[C@H](N)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CO)C(O)=O WPLGNDORMXTMQS-FXQIFTODSA-N 0.000 description 1
- CGOHAEBMDSEKFB-FXQIFTODSA-N Glu-Glu-Ala Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](C)C(O)=O CGOHAEBMDSEKFB-FXQIFTODSA-N 0.000 description 1
- ITBHUUMCJJQUSC-LAEOZQHASA-N Glu-Ile-Gly Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H]([C@@H](C)CC)C(=O)NCC(O)=O ITBHUUMCJJQUSC-LAEOZQHASA-N 0.000 description 1
- IRXNJYPKBVERCW-DCAQKATOSA-N Glu-Leu-Glu Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(O)=O)C(O)=O IRXNJYPKBVERCW-DCAQKATOSA-N 0.000 description 1
- QOXDAWODGSIDDI-GUBZILKMSA-N Glu-Ser-Lys Chemical compound C(CCN)C[C@@H](C(=O)O)NC(=O)[C@H](CO)NC(=O)[C@H](CCC(=O)O)N QOXDAWODGSIDDI-GUBZILKMSA-N 0.000 description 1
- QXPRJQPCFXMCIY-NKWVEPMBSA-N Gly-Ala-Pro Chemical compound C[C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)CN QXPRJQPCFXMCIY-NKWVEPMBSA-N 0.000 description 1
- JRDYDYXZKFNNRQ-XPUUQOCRSA-N Gly-Ala-Val Chemical compound CC(C)[C@@H](C(O)=O)NC(=O)[C@H](C)NC(=O)CN JRDYDYXZKFNNRQ-XPUUQOCRSA-N 0.000 description 1
- DTPOVRRYXPJJAZ-FJXKBIBVSA-N Gly-Arg-Thr Chemical compound C[C@@H](O)[C@@H](C(O)=O)NC(=O)[C@@H](NC(=O)CN)CCCN=C(N)N DTPOVRRYXPJJAZ-FJXKBIBVSA-N 0.000 description 1
- BUEFQXUHTUZXHR-LURJTMIESA-N Gly-Gly-Pro zwitterion Chemical compound NCC(=O)NCC(=O)N1CCC[C@H]1C(O)=O BUEFQXUHTUZXHR-LURJTMIESA-N 0.000 description 1
- PCPOYRCAHPJXII-UWVGGRQHSA-N Gly-Lys-Met Chemical compound [H]NCC(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCSC)C(O)=O PCPOYRCAHPJXII-UWVGGRQHSA-N 0.000 description 1
- SOEGEPHNZOISMT-BYPYZUCNSA-N Gly-Ser-Gly Chemical compound NCC(=O)N[C@@H](CO)C(=O)NCC(O)=O SOEGEPHNZOISMT-BYPYZUCNSA-N 0.000 description 1
- LCRDMSSAKLTKBU-ZDLURKLDSA-N Gly-Ser-Thr Chemical compound C[C@@H](O)[C@@H](C(O)=O)NC(=O)[C@H](CO)NC(=O)CN LCRDMSSAKLTKBU-ZDLURKLDSA-N 0.000 description 1
- DBUNZBWUWCIELX-JHEQGTHGSA-N Gly-Thr-Glu Chemical compound [H]NCC(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCC(O)=O)C(O)=O DBUNZBWUWCIELX-JHEQGTHGSA-N 0.000 description 1
- RYAOJUMWLWUGNW-QMMMGPOBSA-N Gly-Val-Gly Chemical compound NCC(=O)N[C@@H](C(C)C)C(=O)NCC(O)=O RYAOJUMWLWUGNW-QMMMGPOBSA-N 0.000 description 1
- 244000068988 Glycine max Species 0.000 description 1
- 235000010469 Glycine max Nutrition 0.000 description 1
- HVLSXIKZNLPZJJ-TXZCQADKSA-N HA peptide Chemical compound C([C@@H](C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](C(C)C)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(=O)N[C@@H](C)C(O)=O)NC(=O)[C@H]1N(CCC1)C(=O)[C@@H](N)CC=1C=CC(O)=CC=1)C1=CC=C(O)C=C1 HVLSXIKZNLPZJJ-TXZCQADKSA-N 0.000 description 1
- BIAKMWKJMQLZOJ-ZKWXMUAHSA-N His-Ala-Ala Chemical compound C[C@H](NC(=O)[C@H](C)NC(=O)[C@@H](N)Cc1cnc[nH]1)C(O)=O BIAKMWKJMQLZOJ-ZKWXMUAHSA-N 0.000 description 1
- JENKOCSDMSVWPY-SRVKXCTJSA-N His-Leu-Asn Chemical compound [H]N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(N)=O)C(O)=O JENKOCSDMSVWPY-SRVKXCTJSA-N 0.000 description 1
- DRKZDEFADVYTLU-AVGNSLFASA-N His-Val-Val Chemical compound [H]N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](C(C)C)C(O)=O DRKZDEFADVYTLU-AVGNSLFASA-N 0.000 description 1
- 102000011787 Histone Methyltransferases Human genes 0.000 description 1
- 108010036115 Histone Methyltransferases Proteins 0.000 description 1
- XHBYEMIUENPZLY-GMOBBJLQSA-N Ile-Pro-Asn Chemical compound CC[C@H](C)[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CC(N)=O)C(O)=O XHBYEMIUENPZLY-GMOBBJLQSA-N 0.000 description 1
- 108020005350 Initiator Codon Proteins 0.000 description 1
- 108010065920 Insulin Lispro Proteins 0.000 description 1
- TYYLDKGBCJGJGW-UHFFFAOYSA-N L-tryptophan-L-tyrosine Natural products C=1NC2=CC=CC=C2C=1CC(N)C(=O)NC(C(O)=O)CC1=CC=C(O)C=C1 TYYLDKGBCJGJGW-UHFFFAOYSA-N 0.000 description 1
- ZRLUISBDKUWAIZ-CIUDSAMLSA-N Leu-Ala-Asp Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@H](C(O)=O)CC(O)=O ZRLUISBDKUWAIZ-CIUDSAMLSA-N 0.000 description 1
- NTRAGDHVSGKUSF-AVGNSLFASA-N Leu-Arg-Arg Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CCCN=C(N)N)C(=O)N[C@@H](CCCN=C(N)N)C(O)=O NTRAGDHVSGKUSF-AVGNSLFASA-N 0.000 description 1
- OGCQGUIWMSBHRZ-CIUDSAMLSA-N Leu-Asn-Ser Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CO)C(O)=O OGCQGUIWMSBHRZ-CIUDSAMLSA-N 0.000 description 1
- PVMPDMIKUVNOBD-CIUDSAMLSA-N Leu-Asp-Ser Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CO)C(O)=O PVMPDMIKUVNOBD-CIUDSAMLSA-N 0.000 description 1
- FQZPTCNSNPWHLJ-AVGNSLFASA-N Leu-Gln-Lys Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCCCN)C(O)=O FQZPTCNSNPWHLJ-AVGNSLFASA-N 0.000 description 1
- IWTBYNQNAPECCS-AVGNSLFASA-N Leu-Glu-His Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@H](C(O)=O)CC1=CN=CN1 IWTBYNQNAPECCS-AVGNSLFASA-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
- GOFJOGXGMPHOGL-DCAQKATOSA-N Leu-Ser-Met Chemical compound CSCC[C@@H](C(O)=O)NC(=O)[C@H](CO)NC(=O)[C@@H](N)CC(C)C GOFJOGXGMPHOGL-DCAQKATOSA-N 0.000 description 1
- MVJRBCJCRYGCKV-GVXVVHGQSA-N Leu-Val-Gln Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCC(N)=O)C(O)=O MVJRBCJCRYGCKV-GVXVVHGQSA-N 0.000 description 1
- 102000002704 Leucyl aminopeptidase Human genes 0.000 description 1
- 108010004098 Leucyl aminopeptidase Proteins 0.000 description 1
- 108060001084 Luciferase Proteins 0.000 description 1
- FZIJIFCXUCZHOL-CIUDSAMLSA-N Lys-Ala-Ala Chemical compound OC(=O)[C@H](C)NC(=O)[C@H](C)NC(=O)[C@@H](N)CCCCN FZIJIFCXUCZHOL-CIUDSAMLSA-N 0.000 description 1
- URGPVYGVWLIRGT-DCAQKATOSA-N Lys-Met-Ala Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](C)C(O)=O URGPVYGVWLIRGT-DCAQKATOSA-N 0.000 description 1
- ODTZHNZPINULEU-KKUMJFAQSA-N Lys-Phe-Asn Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)O)NC(=O)[C@H](CCCCN)N ODTZHNZPINULEU-KKUMJFAQSA-N 0.000 description 1
- LNMKRJJLEFASGA-BZSNNMDCSA-N Lys-Phe-Leu Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CC(C)C)C(O)=O LNMKRJJLEFASGA-BZSNNMDCSA-N 0.000 description 1
- SBSIKVMCCJUCBZ-GUBZILKMSA-N Met-Asn-Arg Chemical compound CSCC[C@H](N)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@H](C(O)=O)CCCNC(N)=N SBSIKVMCCJUCBZ-GUBZILKMSA-N 0.000 description 1
- KQBJYJXPZBNEIK-DCAQKATOSA-N Met-Glu-Arg Chemical compound CSCC[C@H](N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@H](C(O)=O)CCCNC(N)=N KQBJYJXPZBNEIK-DCAQKATOSA-N 0.000 description 1
- GPAHWYRSHCKICP-GUBZILKMSA-N Met-Glu-Glu Chemical compound CSCC[C@H](N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(O)=O GPAHWYRSHCKICP-GUBZILKMSA-N 0.000 description 1
- YORIKIDJCPKBON-YUMQZZPRSA-N Met-Glu-Gly Chemical compound CSCC[C@H](N)C(=O)N[C@@H](CCC(O)=O)C(=O)NCC(O)=O YORIKIDJCPKBON-YUMQZZPRSA-N 0.000 description 1
- MIXPUVSPPOWTCR-FXQIFTODSA-N Met-Ser-Ser Chemical compound [H]N[C@@H](CCSC)C(=O)N[C@@H](CO)C(=O)N[C@@H](CO)C(O)=O MIXPUVSPPOWTCR-FXQIFTODSA-N 0.000 description 1
- WYBVBIHNJWOLCJ-UHFFFAOYSA-N N-L-arginyl-L-leucine Natural products CC(C)CC(C(O)=O)NC(=O)C(N)CCCN=C(N)N WYBVBIHNJWOLCJ-UHFFFAOYSA-N 0.000 description 1
- KZNQNBZMBZJQJO-UHFFFAOYSA-N N-glycyl-L-proline Natural products NCC(=O)N1CCCC1C(O)=O KZNQNBZMBZJQJO-UHFFFAOYSA-N 0.000 description 1
- 108010079364 N-glycylalanine Proteins 0.000 description 1
- 101710202365 Napin Proteins 0.000 description 1
- 244000061176 Nicotiana tabacum Species 0.000 description 1
- 235000002637 Nicotiana tabacum Nutrition 0.000 description 1
- ZEUBEYAUIOFNBZ-UHFFFAOYSA-N O1N=NC=CC=CC=C1 Chemical compound O1N=NC=CC=CC=C1 ZEUBEYAUIOFNBZ-UHFFFAOYSA-N 0.000 description 1
- 101710089395 Oleosin Proteins 0.000 description 1
- 240000008467 Oryza sativa Japonica Group Species 0.000 description 1
- 101710163504 Phaseolin Proteins 0.000 description 1
- HHOOEUSPFGPZFP-QWRGUYRKSA-N Phe-Asn-Gly Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CC(N)=O)C(=O)NCC(O)=O HHOOEUSPFGPZFP-QWRGUYRKSA-N 0.000 description 1
- 229920002584 Polyethylene Glycol 6000 Polymers 0.000 description 1
- SKICPQLTOXGWGO-GARJFASQSA-N Pro-Gln-Pro Chemical compound C1C[C@H](NC1)C(=O)N[C@@H](CCC(=O)N)C(=O)N2CCC[C@@H]2C(=O)O SKICPQLTOXGWGO-GARJFASQSA-N 0.000 description 1
- FXGIMYRVJJEIIM-UWVGGRQHSA-N Pro-Leu-Gly Chemical compound OC(=O)CNC(=O)[C@H](CC(C)C)NC(=O)[C@@H]1CCCN1 FXGIMYRVJJEIIM-UWVGGRQHSA-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
- SVXXJYJCRNKDDE-AVGNSLFASA-N Pro-Pro-His Chemical compound C([C@@H](C(=O)O)NC(=O)[C@H]1N(CCC1)C(=O)[C@H]1NCCC1)C1=CN=CN1 SVXXJYJCRNKDDE-AVGNSLFASA-N 0.000 description 1
- PRKWBYCXBBSLSK-GUBZILKMSA-N Pro-Ser-Val Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CO)C(=O)N[C@@H](C(C)C)C(O)=O PRKWBYCXBBSLSK-GUBZILKMSA-N 0.000 description 1
- QMABBZHZMDXHKU-FKBYEOEOSA-N Pro-Tyr-Trp Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CC1=CNC2=C1C=CC=C2)C(O)=O QMABBZHZMDXHKU-FKBYEOEOSA-N 0.000 description 1
- JXVXYRZQIUPYSA-NHCYSSNCSA-N Pro-Val-Gln Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCC(N)=O)C(O)=O JXVXYRZQIUPYSA-NHCYSSNCSA-N 0.000 description 1
- 108020004511 Recombinant DNA Proteins 0.000 description 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 1
- 108010016634 Seed Storage Proteins Proteins 0.000 description 1
- MMAPOBOTRUVNKJ-ZLUOBGJFSA-N Ser-Asp-Ser Chemical compound C([C@@H](C(=O)N[C@@H](CO)C(=O)O)NC(=O)[C@H](CO)N)C(=O)O MMAPOBOTRUVNKJ-ZLUOBGJFSA-N 0.000 description 1
- BRIZMMZEYSAKJX-QEJZJMRPSA-N Ser-Glu-Trp Chemical compound C1=CC=C2C(=C1)C(=CN2)C[C@@H](C(=O)O)NC(=O)[C@H](CCC(=O)O)NC(=O)[C@H](CO)N BRIZMMZEYSAKJX-QEJZJMRPSA-N 0.000 description 1
- GJFYFGOEWLDQGW-GUBZILKMSA-N Ser-Leu-Gln Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CCC(=O)N)C(=O)O)NC(=O)[C@H](CO)N GJFYFGOEWLDQGW-GUBZILKMSA-N 0.000 description 1
- AZWNCEBQZXELEZ-FXQIFTODSA-N Ser-Pro-Ser Chemical compound OC[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CO)C(O)=O AZWNCEBQZXELEZ-FXQIFTODSA-N 0.000 description 1
- KQNDIKOYWZTZIX-FXQIFTODSA-N Ser-Ser-Arg Chemical compound OC[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@H](C(O)=O)CCCNC(N)=N KQNDIKOYWZTZIX-FXQIFTODSA-N 0.000 description 1
- SRSPTFBENMJHMR-WHFBIAKZSA-N Ser-Ser-Gly Chemical compound OC[C@H](N)C(=O)N[C@@H](CO)C(=O)NCC(O)=O SRSPTFBENMJHMR-WHFBIAKZSA-N 0.000 description 1
- BMKNXTJLHFIAAH-CIUDSAMLSA-N Ser-Ser-Leu Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(O)=O BMKNXTJLHFIAAH-CIUDSAMLSA-N 0.000 description 1
- XQJCEKXQUJQNNK-ZLUOBGJFSA-N Ser-Ser-Ser Chemical compound OC[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@@H](CO)C(O)=O XQJCEKXQUJQNNK-ZLUOBGJFSA-N 0.000 description 1
- VGQVAVQWKJLIRM-FXQIFTODSA-N Ser-Ser-Val Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CO)C(=O)N[C@@H](C(C)C)C(O)=O VGQVAVQWKJLIRM-FXQIFTODSA-N 0.000 description 1
- 244000062793 Sorghum vulgare Species 0.000 description 1
- TZKPNGDGUVREEB-FOHZUACHSA-N Thr-Asn-Gly Chemical compound C[C@@H](O)[C@H](N)C(=O)N[C@@H](CC(N)=O)C(=O)NCC(O)=O TZKPNGDGUVREEB-FOHZUACHSA-N 0.000 description 1
- ZMYCLHFLHRVOEA-HEIBUPTGSA-N Thr-Thr-Ser Chemical compound C[C@@H](O)[C@H](N)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CO)C(O)=O ZMYCLHFLHRVOEA-HEIBUPTGSA-N 0.000 description 1
- QNXZCKMXHPULME-ZNSHCXBVSA-N Thr-Val-Pro Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](C(C)C)C(=O)N1CCC[C@@H]1C(=O)O)N)O QNXZCKMXHPULME-ZNSHCXBVSA-N 0.000 description 1
- 108091023040 Transcription factor Proteins 0.000 description 1
- 102000040945 Transcription factor Human genes 0.000 description 1
- GYBVHTWOQJMYAM-HRCADAONSA-N Tyr-Met-Pro Chemical compound CSCC[C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](CC2=CC=C(C=C2)O)N GYBVHTWOQJMYAM-HRCADAONSA-N 0.000 description 1
- WTTRJMAZPDHPGS-KKXDTOCCSA-N Tyr-Phe-Ala Chemical compound C[C@H](NC(=O)[C@H](Cc1ccccc1)NC(=O)[C@@H](N)Cc1ccc(O)cc1)C(O)=O WTTRJMAZPDHPGS-KKXDTOCCSA-N 0.000 description 1
- FDKDGFGTHGJKNV-FHWLQOOXSA-N Tyr-Phe-Gln Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)N[C@@H](CCC(=O)N)C(=O)O)NC(=O)[C@H](CC2=CC=C(C=C2)O)N FDKDGFGTHGJKNV-FHWLQOOXSA-N 0.000 description 1
- CDBXVDXSLPLFMD-BPNCWPANSA-N Tyr-Pro-Ala Chemical compound OC(=O)[C@H](C)NC(=O)[C@@H]1CCCN1C(=O)[C@@H](N)CC1=CC=C(O)C=C1 CDBXVDXSLPLFMD-BPNCWPANSA-N 0.000 description 1
- 108090000848 Ubiquitin Proteins 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
- AZSHAZJLOZQYAY-FXQIFTODSA-N Val-Ala-Ser Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@@H](CO)C(O)=O AZSHAZJLOZQYAY-FXQIFTODSA-N 0.000 description 1
- GXAZTLJYINLMJL-LAEOZQHASA-N Val-Asn-Gln Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)N[C@@H](CCC(=O)N)C(=O)O)N GXAZTLJYINLMJL-LAEOZQHASA-N 0.000 description 1
- DDNIHOWRDOXXPF-NGZCFLSTSA-N Val-Asp-Pro Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)N1CCC[C@@H]1C(=O)O)N DDNIHOWRDOXXPF-NGZCFLSTSA-N 0.000 description 1
- PMXBARDFIAPBGK-DZKIICNBSA-N Val-Glu-Tyr Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@H](C(O)=O)CC1=CC=C(O)C=C1 PMXBARDFIAPBGK-DZKIICNBSA-N 0.000 description 1
- JTWIMNMUYLQNPI-WPRPVWTQSA-N Val-Gly-Arg Chemical compound CC(C)[C@H](N)C(=O)NCC(=O)N[C@H](C(O)=O)CCCNC(N)=N JTWIMNMUYLQNPI-WPRPVWTQSA-N 0.000 description 1
- GVJUTBOZZBTBIG-AVGNSLFASA-N Val-Lys-Arg Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCCN=C(N)N)C(=O)O)N GVJUTBOZZBTBIG-AVGNSLFASA-N 0.000 description 1
- WBAJDGWKRIHOAC-GVXVVHGQSA-N Val-Lys-Gln Chemical compound [H]N[C@@H](C(C)C)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCC(N)=O)C(O)=O WBAJDGWKRIHOAC-GVXVVHGQSA-N 0.000 description 1
- KISFXYYRKKNLOP-IHRRRGAJSA-N Val-Phe-Ser Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CO)C(=O)O)N KISFXYYRKKNLOP-IHRRRGAJSA-N 0.000 description 1
- LTTQCQRTSHJPPL-ZKWXMUAHSA-N Val-Ser-Asp Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(=O)O)C(=O)O)N LTTQCQRTSHJPPL-ZKWXMUAHSA-N 0.000 description 1
- YQYFYUSYEDNLSD-YEPSODPASA-N Val-Thr-Gly Chemical compound CC(C)[C@H](N)C(=O)N[C@@H]([C@@H](C)O)C(=O)NCC(O)=O YQYFYUSYEDNLSD-YEPSODPASA-N 0.000 description 1
- 230000006978 adaptation Effects 0.000 description 1
- 108010008685 alanyl-glutamyl-aspartic acid Proteins 0.000 description 1
- 108010076324 alanyl-glycyl-glycine Proteins 0.000 description 1
- 108010069020 alanyl-prolyl-glycine Proteins 0.000 description 1
- 108010041407 alanylaspartic acid Proteins 0.000 description 1
- 108010005233 alanylglutamic acid Proteins 0.000 description 1
- 108010047495 alanylglycine Proteins 0.000 description 1
- 108010013835 arginine glutamate Proteins 0.000 description 1
- 108010062796 arginyllysine Proteins 0.000 description 1
- 108010040443 aspartyl-aspartic acid Proteins 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 1
- 230000003115 biocidal effect Effects 0.000 description 1
- 238000010170 biological method Methods 0.000 description 1
- 238000009395 breeding Methods 0.000 description 1
- 230000001488 breeding effect Effects 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 108010060199 cysteinylproline Proteins 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 238000006731 degradation reaction Methods 0.000 description 1
- 239000008367 deionised water Substances 0.000 description 1
- 229910021641 deionized water Inorganic materials 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 230000029087 digestion Effects 0.000 description 1
- 238000007598 dipping method Methods 0.000 description 1
- 238000001962 electrophoresis Methods 0.000 description 1
- 210000002257 embryonic structure Anatomy 0.000 description 1
- 150000002148 esters Chemical class 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 230000004927 fusion Effects 0.000 description 1
- 238000010353 genetic engineering Methods 0.000 description 1
- 229960002518 gentamicin Drugs 0.000 description 1
- 108010049041 glutamylalanine 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
- 108010019832 glycyl-asparaginyl-glycine Proteins 0.000 description 1
- 108010051307 glycyl-glycyl-proline Proteins 0.000 description 1
- 108010010147 glycylglutamine Proteins 0.000 description 1
- 108010077515 glycylproline Proteins 0.000 description 1
- 108010087823 glycyltyrosine Proteins 0.000 description 1
- 101150054900 gus gene Proteins 0.000 description 1
- 238000003306 harvesting Methods 0.000 description 1
- 239000004009 herbicide Substances 0.000 description 1
- 108010025306 histidylleucine Proteins 0.000 description 1
- 108010085325 histidylproline Proteins 0.000 description 1
- 230000006801 homologous recombination Effects 0.000 description 1
- 238000002744 homologous recombination Methods 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 230000000977 initiatory effect Effects 0.000 description 1
- 238000003973 irrigation Methods 0.000 description 1
- 230000002262 irrigation Effects 0.000 description 1
- ZNJFBWYDHIGLCU-UHFFFAOYSA-N jasmonic acid Natural products CCC=CCC1C(CC(O)=O)CCC1=O ZNJFBWYDHIGLCU-UHFFFAOYSA-N 0.000 description 1
- 229930027917 kanamycin Natural products 0.000 description 1
- 229960000318 kanamycin Drugs 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
- 229930182823 kanamycin A Natural products 0.000 description 1
- 108010090333 leucyl-lysyl-proline Proteins 0.000 description 1
- 108010057821 leucylproline Proteins 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000003550 marker Substances 0.000 description 1
- 239000011159 matrix material Substances 0.000 description 1
- 108010005942 methionylglycine Proteins 0.000 description 1
- 238000000520 microinjection Methods 0.000 description 1
- 235000019713 millet Nutrition 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- FJKROLUGYXJWQN-UHFFFAOYSA-N papa-hydroxy-benzoic acid Natural products OC(=O)C1=CC=C(O)C=C1 FJKROLUGYXJWQN-UHFFFAOYSA-N 0.000 description 1
- 230000008506 pathogenesis Effects 0.000 description 1
- LWTDZKXXJRRKDG-UHFFFAOYSA-N phaseollin Natural products C1OC2=CC(O)=CC=C2C2C1C1=CC=C3OC(C)(C)C=CC3=C1O2 LWTDZKXXJRRKDG-UHFFFAOYSA-N 0.000 description 1
- 229940093429 polyethylene glycol 6000 Drugs 0.000 description 1
- 229920001184 polypeptide Polymers 0.000 description 1
- 238000001556 precipitation Methods 0.000 description 1
- 108090000765 processed proteins & peptides Proteins 0.000 description 1
- 102000004196 processed proteins & peptides Human genes 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 108010014614 prolyl-glycyl-proline Proteins 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 238000012113 quantitative test Methods 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 238000005215 recombination Methods 0.000 description 1
- 230000016914 response to endoplasmic reticulum stress Effects 0.000 description 1
- 238000003757 reverse transcription PCR Methods 0.000 description 1
- 229960004889 salicylic acid Drugs 0.000 description 1
- 238000005070 sampling Methods 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 230000014284 seed dormancy process Effects 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 230000035939 shock Effects 0.000 description 1
- 230000019491 signal transduction Effects 0.000 description 1
- 238000002791 soaking Methods 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 238000009331 sowing Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 230000002194 synthesizing effect Effects 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 230000002103 transcriptional effect Effects 0.000 description 1
- 238000013519 translation Methods 0.000 description 1
- 230000014621 translational initiation Effects 0.000 description 1
- 230000003827 upregulation Effects 0.000 description 1
- 239000010455 vermiculite Substances 0.000 description 1
- 229910052902 vermiculite Inorganic materials 0.000 description 1
- 235000019354 vermiculite Nutrition 0.000 description 1
- 239000013603 viral vector Substances 0.000 description 1
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
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H1/00—Processes for modifying genotypes ; Plants characterised by associated natural traits
- A01H1/02—Methods or apparatus for hybridisation; Artificial pollination ; Fertility
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
- C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
- C12N15/82—Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
- C12N15/8241—Phenotypically and genetically modified plants via recombinant DNA technology
- C12N15/8261—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield
- C12N15/8271—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield for stress resistance, e.g. heavy metal resistance
- C12N15/8273—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield for stress resistance, e.g. heavy metal resistance for drought, cold, salt resistance
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Genetics & Genomics (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Molecular Biology (AREA)
- General Health & Medical Sciences (AREA)
- Biophysics (AREA)
- Biotechnology (AREA)
- Biochemistry (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- Botany (AREA)
- Biomedical Technology (AREA)
- Cell Biology (AREA)
- Physics & Mathematics (AREA)
- Plant Pathology (AREA)
- Medicinal Chemistry (AREA)
- Microbiology (AREA)
- Gastroenterology & Hepatology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Developmental Biology & Embryology (AREA)
- Environmental Sciences (AREA)
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
Abstract
The application discloses application of a protein ZmbZIPc3 in regulating and controlling salt tolerance and drought resistance of plants. The application provides application of protein and related biological materials in regulating drought resistance and/or salt tolerance of plants, wherein the protein is a protein with an amino acid sequence shown as SEQ ID NO.2 or a fusion protein obtained by connecting protein tags at N-terminal or/and C-terminal of the amino acid sequence shown as SEQ ID NO. 2. According to the application, a ZmbZIPc3 gene related to drought resistance is cloned from corn, and is transferred into Arabidopsis thaliana and corn, and the ZmbZIPc3 gene is found to improve the osmotic stress resistance and the salt stress tolerance of Arabidopsis thaliana, and improve the drought resistance and the yield of corn. Therefore, the application has important significance for improving the drought tolerance of plants, stabilizing the crop yield, identifying and cultivating new varieties of good crops.
Description
Technical Field
The present application relates to the field of biotechnology. In particular to application of a protein ZmbZIPc3 in regulating and controlling salt tolerance and drought resistance of plants.
Background
The arid and semiarid regions of the world occupy more than one third of the total land area, while the cultivated land with irrigation conditions only occupies about three percent of the total land area, thus greatly restricting the global grain production. Drought has become a major factor affecting global grain yield, and now drought areas are enlarged year by year, and natural environment deterioration leads to land desertification and grassland degradation. In the face of the increasing deterioration of plant and crop growth environments, plant biologists and agrologists are increasingly focusing on and researching plant stress resistance, and aim to analyze how plants resist various stress and mechanisms thereof so as to cultivate crop varieties with high yield, good quality and strong stress resistance.
Corn is one of three crops cultivated in the world and is used as feed, food, deep processing and industrial raw material. As an important grain crop in China, the sowing area is up to above 4200 ten thousand hectares, and the annual yield is up to 2.59 hundred million tons. Corn is a crop with large water demand, and water deficiency in critical growth period can seriously affect yield. Corn farmlands in main corn producing areas such as northwest, southwest, north China and northeast China mostly depend on dry lands with natural precipitation, and drought frequently occurs, so that the stability of corn yield is affected. The breeding of drought-resistant corn variety becomes one of the important ways to raise corn yield. The drought-resistant related mechanism of the corn is analyzed from the molecular level, and functional genes or transcription factors related to drought resistance in the corn are cloned, discovered and identified, so that the drought-resistant fine variety is cultivated efficiently by utilizing the genetic engineering means, and the method has practical significance for improving the crop yield.
Disclosure of Invention
The technical problem to be solved by the application is how to improve the stress tolerance, especially the salt tolerance and drought resistance of plants so as to further improve the growth of the plants.
In order to solve the technical problems, the application firstly provides application of a protein in regulating drought resistance and/or salt tolerance of plants, wherein the protein is derived from corn (Zea mays L.), and is named ZmbZIPc3 protein or protein ZmbZIPc3, and the protein is shown in any one of the following A1), A2) or A3):
a1 Protein with the amino acid sequence shown as SEQ ID NO. 2;
a2 Fusion protein obtained by N-terminal or/and C-terminal connexin label of the amino acid sequence shown in SEQ ID NO. 2;
a3 A protein which is obtained by substituting and/or deleting and/or adding one or more amino acid residues of the amino acid sequence shown in SEQ ID NO.2, has more than 90 percent of identity with the protein shown in A1) and has the same function.
Wherein SEQ ID NO.2 consists of 410 amino acid residues.
The protein can be synthesized artificially or obtained by synthesizing the coding gene and then biologically expressing.
In the above application, the protein tag (protein-tag) refers to a polypeptide or protein that is fusion expressed with the target protein by using DNA in vitro recombination technology, so as to facilitate the expression, detection, tracing and/or purification of the target protein. The protein tag may be a Flag tag, his tag, MBP tag, HA tag, myc tag, GST tag, and/or SUMO tag, etc.
In the above applications, identity refers to the identity of amino acid sequences. The identity of amino acid sequences can be determined using homology search sites on the internet, such as BLAST web pages of the NCBI homepage website. For example, in advanced BLAST2.1, the identity of a pair of amino acid sequences can be searched for by using blastp as a program, setting the Expect value to 10, setting all filters to OFF, using BLOSUM62 as Matrix, setting Gap existence cost, per residue gap cost and Lambda ratio to 11,1 and 0.85 (default values), respectively, and calculating, and then obtaining the value (%) of the identity.
In the above applications, the 90% identity or more may be at least 91%, 92%, 95%, 96%, 98%, 99% or 100% identity.
The application of the biological material related to the protein ZmbZIPc3 in regulating and controlling the drought resistance and/or salt tolerance of plants is also within the protection scope of the application: the biological material is any one of the following C1) -C10):
c1 A nucleic acid molecule encoding the protein zmbzip 3;
c2 An expression cassette comprising C1) said nucleic acid molecule;
c3 A recombinant vector comprising C1) said nucleic acid molecule, or a recombinant vector comprising C2) said expression cassette;
c4 A recombinant microorganism comprising C1) said nucleic acid molecule, or a recombinant microorganism comprising C2) said expression cassette, or a recombinant microorganism comprising C3) said recombinant vector;
c5 A transgenic plant cell line comprising C1) said nucleic acid molecule, or a transgenic plant cell line comprising C2) said expression cassette, or a transgenic plant cell line comprising C3) said recombinant vector;
c6 A) a transgenic plant tissue comprising C1) said nucleic acid molecule, or a transgenic plant tissue comprising C2) said expression cassette, or a transgenic plant tissue comprising C3) said recombinant vector;
c7 A transgenic plant organ comprising C1) said nucleic acid molecule, or a transgenic plant organ comprising C2) said expression cassette, or a transgenic plant organ comprising C3) said recombinant vector;
c8 A transgenic plant containing the nucleic acid molecule of C1), or a transgenic plant containing the expression cassette of C2), or a transgenic plant containing the recombinant vector of C3);
c9 A tissue culture produced by regenerable cells of the transgenic plant of C8);
c10 Protoplasts produced from the tissue culture of C9).
Wherein the nucleic acid molecule may be DNA, such as cDNA, genomic DNA, or recombinant DNA; the nucleic acid molecule may also be RNA, such as mRNA or hnRNA, etc.
In the above biological material, the nucleic acid molecule of C1) encoding the protein zmbzip 3 may specifically be any one of the following D1), D2) or D3):
d1 51-1283 DNA molecules of SEQ ID NO. 1;
d2 A DNA molecule with the coding sequence shown in the 51 st to 1283 rd positions of SEQ ID NO. 1;
d3 A DNA molecule which hybridizes under stringent conditions to the DNA molecule defined in D1) or D2) and which codes for the protein ZmbZIPc3.
Wherein, SEQ ID NO.1 is composed of 1493 nucleotides, the coding sequence is the 51 st to 1283 rd positions from the 5' end of SEQ ID NO.1, and the coding amino acid sequence is the protein shown as SEQ ID NO. 2.
The stringent conditions are hybridization and washing of the membrane 2 times at 68℃in a solution of 2 XSSC, 0.1% SDS for 5min each time, and hybridization and washing of the membrane 2 times at 68℃in a solution of 0.5 XSSC, 0.1% SDS for 15min each time.
In the above biological material, the expression cassette of C2) is an expression cassette containing a nucleic acid molecule encoding the protein ZmbZIPc3 (i.e., zmbZIPc3 gene expression cassette), which means that it is capable of being hostedA DNA molecule expressing the protein zmbzip 3 in a host cell, which may include not only a promoter for initiating transcription of the zmbzip 3 gene, but also a terminator for terminating transcription of the zmbzip 3 gene. Further, the expression cassette may also include an enhancer sequence. Promoters useful in the present application include, but are not limited to: constitutive promoters, tissue, organ and development specific promoters and inducible promoters. Examples of promoters include, but are not limited to: a constitutive promoter of cauliflower mosaic virus 35S; wound-inducible promoters from tomato, leucine aminopeptidase ("LAP", chao et al (1999) Plant Physiol 120:979-992); a chemically inducible promoter from tobacco, pathogenesis-related 1 (PR 1) (induced by salicylic acid and BTH (benzothiadiazole-7-carbothioic acid S-methyl ester); tomato protease inhibitor II promoter (PIN 2) or LAP promoter (both inducible with jasmonic acid ester); heat shock promoters (U.S. Pat. No. 5,187,267); tetracycline-inducible promoters (U.S. Pat. No. 5,057,422); seed specific promoters such as the millet seed specific promoter pF128 (CN 101063139B (China patent 2007 1 0099169.7)), seed storage protein specific promoters (e.g., phaseolin, napin, oleosin and soybean beta-glucose promoters (Beachy et al (1985) EMBO J.4:3047-3053) which may be used alone or in combination with other plant promoters all references cited herein are incorporated by reference in their entirety suitable transcription terminators include, but are not limited to, the Agrobacterium nopaline synthase terminator (NOS terminator), the cauliflower mosaic virus CaMV 35S terminator, tml terminator, the pea rbcS E9 terminator and the nopaline and octopine synthase terminators (see, e.g., odell et al (I) 985 ) Nature 313:810; rosenberg et al (1987) Gene,56:125; guerineau et al (1991) mol. Gen. Genet,262:141; proudroot (1991) Cell,64:671; sanfacon et al Genes Dev., 5:141; mogen et al (1990) Plant Cell,2:1261; munroe et al (1990) Gene,91:151; ballad et al (1989) Nucleic Acids Res.17:7891; joshi et al (1987) Nucleic Acid Res., 15:9627).
In a specific embodiment of the application, the sequence of the ZmbZIPc3 gene expression cassette is shown as SEQ ID NO.4, and the sequence sequentially comprises a promoter 35SS (36-720 th site from the 5' end of SEQ ID NO. 4), a coding sequence of the ZmbZIPc3 gene (1140-2373 th site from the 5' end of SEQ ID NO.4, corresponding to 51-1283 th sites of SEQ ID NO. 1) and a terminator PA35S (2593-2810 th site from the 5' end of SEQ ID NO. 4) on a pLeela vector.
In the above biological material, C3) the recombinant vector may contain a DNA molecule shown in SEQ ID NO.1 for encoding the protein ZmbZIPc3 (i.e., containing the ZmbZIPc3 gene).
The plant expression vector can be used for constructing a recombinant vector containing the ZmbZIPc3 gene expression cassette. The plant expression vector may be a Gateway system vector or a binary agrobacterium vector, etc., such as pweela, pGWB411, pGWB412, pGWB405, pBin438, pCAMBIA1300, pCAMBIA1302, pCAMBIA2300, pCAMBIA2301, pCAMBIA1301, pBI121, pCAMBIA1391-Xa, or pCAMBIA1391-Xb. When ZmbZIPc3 is used to construct recombinant vectors, any one of enhanced, constitutive, tissue-specific or inducible promoters, such as cauliflower mosaic virus (CAMV) 35S promoter, ubiquitin gene Ubiqutin promoter (pUbi) and the like, can be added before the transcription initiation nucleotide thereof, and can be used alone or in combination with other plant promoters; in addition, when the gene of the present application is used to construct a plant expression vector, enhancers, including translational enhancers or transcriptional enhancers, may be used, and these enhancers may be ATG initiation codon or adjacent region initiation codon, etc., but must be identical to the reading frame of the coding sequence to ensure proper translation of the entire sequence. The sources of the translational control signals and initiation codons are broad, and can be either natural or synthetic. The translation initiation region may be derived from a transcription initiation region or a structural gene.
In order to facilitate the identification and selection of transgenic plant cells or plants, the plant expression vectors used may be processed, for example, by adding genes encoding enzymes or luminescent compounds which produce a color change (GUS gene, luciferase gene, etc.), antibiotic markers with resistance (gentamicin markers, kanamycin markers, etc.), or anti-chemical marker genes (e.g., anti-herbicide genes), etc., which may be expressed in plants.
In a specific embodiment of the application, the recombinant vector is a pLeela 35SS: zmbZIPc3 overexpression vector or a pU130 35SS: zmbZIPc3 overexpression vector. ZmbZIPc3 over-expression vector is a vector which is obtained by replacing the sequence between attR1 and attR2 sites of the pLeela vector with a DNA molecule shown in SEQ ID NO.3 and keeping other sequences unchanged; the pU130 35SS is obtained by inserting a ZmbZIPc3 gene expression cassette shown in SEQ ID No.4 between HindIII recognition sites of the pU130 vector and keeping other sequences of the pU130 vector unchanged.
Of the above biological materials, C4) the recombinant microorganism may be specifically yeast, bacteria, algae and fungi.
In the above biological material, the transgenic plant organ of C7) may be the root, stem, leaf, flower, fruit and seed of the transgenic plant.
In the above biological material, C9) the tissue culture may be derived from roots, stems, leaves, flowers, fruits, seeds, pollen, embryos and anthers.
In the above biological materials, none of the transgenic plant cell lines, transgenic plant tissues and transgenic plant organs include propagation material.
The application of the protein ZmbZIPc3 or the biological material thereof in plant breeding is also within the protection scope of the application.
Among the above applications, the plant breeding may be specifically performed by crossing a plant containing the protein zmbzip 3 or a biological material related to the protein zmbzip 3 (e.g., the protein zmbzip 3 encoding gene zmbzip 3) with other plants for plant breeding.
In the application, the drought resistance of the regulation plant is improved, and the salt tolerance of the regulation plant is improved.
The application further provides a method for cultivating transgenic plants with high drought resistance and/or salt tolerance.
The method for cultivating transgenic plants with high drought resistance and/or salt tolerance comprises the steps of increasing the expression quantity of a gene of a protein ZmbZIPc3 in a target plant and/or the content of the protein ZmbZIPc3 and/or the activity of the protein ZmbZIPc3 to obtain transgenic plants; the transgenic plants have drought and/or salt tolerance higher than the plants of interest.
In the above method, the method for increasing the expression level of the gene of the protein ZmbZIPc3 in the target plant and/or the content of the protein ZmbZIPc3 and/or the activity of the protein ZmbZIPc3 is to express or overexpress the protein ZmbZIPc3 in the target plant.
In the method, the method of expression or over-expression is to introduce the coding gene of the protein ZmbZIPc3 into a target plant.
In the above method, the gene encoding the protein zmbzip 3 may be introduced into a target plant by a plant expression vector carrying the zmbzip 3 gene of the application. The plant expression vector carrying ZmbZIPc3 of the application may be transformed into plant cells or tissues by using conventional biological methods such as Agrobacterium-mediated, ti-plasmid, ri-plasmid, plant viral vector, direct DNA transformation, microinjection, electric conduction, etc., and the transformed plant cells or tissues may be cultivated into plants.
In the method, the nucleotide sequence of the coding gene of the protein ZmbZIPc3 is a DNA molecule shown in SEQ ID NO. 1.
In a specific embodiment of the application, the plant expression vector carrying the gene ZmbZIPc3 of the application can be a pLeela 35SS: zmbZIPc3 over-expression vector or a pU130 35SS: zmbZIPc3 over-expression vector. ZmbZIPc3 over-expression vector is a vector which is obtained by replacing the sequence between attR1 and attR2 sites of the pLeela vector with a DNA molecule shown in SEQ ID NO.3 and keeping other sequences unchanged; the pU130 35SS is obtained by inserting a ZmbZIPc3 gene expression cassette shown in SEQ ID No.4 between HindIII recognition sites of the pU130 vector and keeping other sequences of the pU130 vector unchanged.
In the application, the improvement of drought resistance or high drought resistance of plants is particularly characterized by sensitivity to ABA (abscisic acid), high plant survival rate, low plant wilting rate, fast root growth, small aperture opening and the like under drought conditions.
In the present application, the plant is any one of the following E1) to E6):
e1 Dicotyledonous plants;
e2 Monocotyledonous plants;
e3 Cruciferous plants;
e4 Arabidopsis thaliana (Arabidopsis thaliana);
e5 A gramineous plant;
e6 Corn (Zea mays).
The above protein zmbzip 3 and biological materials related to the above protein zmbzip 3 are also within the scope of the application.
The application clones and obtains ZmbZIPc3 gene related to drought resistance from corn. The ZmbZIPc3 gene is transferred into the Arabidopsis thaliana, and the ZmbZIPc3 gene can improve the osmotic stress resistance, improve the salt stress tolerance of the Arabidopsis thaliana, reduce the pore opening of the Arabidopsis thaliana, slow down the water loss rate and facilitate drought resistance and water retention of plants; overexpression of the zmbzip 3 gene in maize has been found to: the ZmbZIPc3 gene affects the growth and development of corn in germination and seedling stage, so as to raise the drought tolerance and yield of corn. Therefore, the application has important significance for improving the drought tolerance of plants, stabilizing the crop yield, identifying and cultivating new varieties of good crops.
Drawings
FIG. 1 shows the expression level identification (A) of ZmZIPc 3-transferred Arabidopsis thaliana and the expression level identification (B) of ZmZIPc 3-transferred corn.
FIG. 2 shows the comparison of growth (a and b) and root length (c) of Arabidopsis thaliana transformed with ZmZIPc 3 under osmotic stress.
FIG. 3 shows the growth of Arabidopsis thaliana transformed with ZmZIPc 3 under salt stress.
FIG. 4 shows germination of Arabidopsis thaliana seeds transformed to ZmZIPc 3 under ABA treatment conditions; wherein A is the germination condition of ZmZIPc 3 Arabidopsis seeds transferred in a 1/2MS culture medium without ABA, B is the germination condition of ZmZIPc 3 Arabidopsis seeds transferred in a 1/2MS culture medium added with 0.25 mu M ABA, C is the germination condition of ZmZIPc 3 Arabidopsis seeds transferred in a 1/2MS culture medium added with 0.5 mu M ABA, and D is the germination condition of ZmZIPc 3 Arabidopsis seeds transferred in a 1/2MS culture medium added with 1 mu M ABA.
FIG. 5 shows the seedling growth of ZmZIPc 3-transformed Arabidopsis under ABA treatment conditions; wherein A is the growth condition of ZmZIPc 3 Arabidopsis seedlings transferred in a 1/2MS culture medium without ABA, B is the growth condition of ZmZIPc 3 Arabidopsis seedlings transferred in a 1/2MS culture medium added with 0.1 mu M ABA, and C is the growth condition of ZmZIPc 3 Arabidopsis seedlings transferred in a 1/2MS culture medium added with 1 mu M ABA.
FIG. 6 shows the growth of Arabidopsis thaliana transformed with ZmZIPc 3 under drought stress.
FIG. 7 is a comparison result of pore opening degrees of Arabidopsis thaliana leaves transformed with ZmbZIPc3 under drought stress.
FIG. 8 shows seed germination (A) and drought resistance index (B) under drought stress during germination of ZmZIPc 3 overexpressing corn.
FIG. 9 shows root observations under drought stress in maize germination over-expressing ZmZIPc 3.
FIG. 10 is an identification of growth under drought stress in maize seedlings over-expressing ZmZIPc 3; a is that seedlings grow to a three-leaf one-heart period; b is the growth condition after 72 hours of first rehydration treatment; c is that the second drought treatment enters a wilting state; d is the growth condition of the second rehydration treatment for 72 h.
In FIGS. 1-7, co1 represents wild type Arabidopsis Col-0,2-1 represents ZmZIPc 3-transferred Arabidopsis strain OE2-1, 5-6 represents ZmZIPc 3-transferred Arabidopsis strain OE 5-6 and 10-4 represents ZmZIPc 3-transferred Arabidopsis strain OE 10-4.
In FIGS. 1, 8-10, WT represents wild-type maize C01, C3-1 represents ZmZIPc 3-transferred Arabidopsis line OE C3-1, C3-3 represents ZmZIPc 3-transferred Arabidopsis line OE C3-3, C3-5 represents ZmZIPc 3-transferred Arabidopsis line OE C3-5 and C3-7 represents ZmZIPc 3-transferred Arabidopsis line OE C3-7.
Detailed Description
The following detailed description of the application is provided in connection with the accompanying drawings that are presented to illustrate the application and not to limit the scope thereof. The examples provided below are intended as guidelines for further modifications by one of ordinary skill in the art and are not to be construed as limiting the application in any way.
The experimental methods in the following examples are conventional methods unless otherwise specified. Materials, reagents and the like used in the examples described below are commercially available unless otherwise specified.
The quantitative tests in the following examples were all performed in triplicate, and the results were averaged.
The pENTR/D-TOPO vector in the following examples is a product of Invitrogen corporation.
The plant expression vector pLeela in the examples described below is described in non-patent document "A novel role for histone methyltransferase KYP/SUVH4 in the control of Arabidopsis primary seed dormancy, new Phytologist (2012) 193:605-616", which is publicly available from plant research at the national academy of sciences, to repeat the experiments of the application, and is not useful for other purposes. .
Maize inbred line B73 in the following examples is described in non-patent document "Zea mays l.; zmbZIP60 mRNA is spliced in maize in response to ER stress, BMC Research Notes (2012) 5: the public was available from the national academy of sciences plant institute to repeat the experiments of the application and was not available for other uses.
pU130 vectors in the examples described below are described in the literature "Development of Novel Glyphosate-Tolerant Japonica Rice Lines: A Step Toward Commercial Release. Cui Y, et al front. Plant Sci. (2016) 7:1218", and are available to the public from national academy of sciences plant research, and this biomaterial is used only for repeated experiments in connection with the application and is not used for other purposes; the public is also available from the national seed group company life science and technology center.
Agrobacterium tumefaciens GV3101 (Agrobacterium tumefaciens strain GV 3101) in the following examples: as described in the document "Binary Agrobacterium vectors for plant transformation, M Bevanin, nucleic Acids Research (1984) 12 (22): 8711-8721", the public is available from the national academy of sciences plant study, and the biomaterial is used only for repeated experiments related to the present application and is not used for other purposes.
Arabidopsis Col-0 in the following examples is described in the literature "Arabidopsis, a use of the strain Meyerowitz EM, cell (1989): 263-270", and is available to the public from plant research institute of national academy of sciences, and this biomaterial is used only for repeated experiments related to the present application and is not used for other purposes.
Example 1 obtaining ZmbZIPc3 Gene
1. cDNA acquisition
Total RNA of the trefoil-heart maize inbred line B73 was extracted and reverse transcribed into cDNA.
2. PCR amplification
And (3) taking the cDNA obtained in the step (1) as a template, and carrying out PCR amplification by adopting a primer F and a primer R to obtain a PCR product. The primer sequences were as follows:
F:5'–CCTCGATCCGGAATGGAGCG-3'
R:5'-CAGTGAGCAGCCAGCACACG-3'
3. sequencing of PCR products
The gene shown in SEQ ID NO.1 was designated as ZmbZIPc3 gene. Sequencing the PCR product obtained in the step 2. Sequencing results showed that: the PCR product is 1382bp DNA fragment, the nucleotide sequence of the DNA fragment is shown as 38 th to 1419 th positions of SEQ ID NO.1, and the DNA fragment comprises the coding sequence of ZmbZIPc3 gene shown as 51 st to 1283 rd positions of SEQ ID NO. 1. The protein coded by the ZmbZIPc3 gene is named as ZmbZIPc3 protein, the ZmbZIPc3 protein consists of 410 amino acid residues, and the amino acid sequence of the ZmbZIPc3 protein is shown as SEQ ID NO. 2.
Example 2 obtaining ZmbZIc 3-transformed Arabidopsis thaliana and phenotypic analysis thereof
1. Obtaining ZmZIPc 3-transformed Arabidopsis thaliana
1. Obtaining recombinant expression vectors
(1) PCR amplification was performed using the cDNA obtained in step 1 of example 1 as a template and the primers F and R of example 1 to obtain a PCR product. The PCR product contains the coding sequence of ZmbZIPc3 gene shown in 51-1283 th position of the 5' end of SEQ ID NO. 1.
(2) And (3) connecting the PCR product obtained in the step (1) into an intermediate vector pENTR/D-TOPO by using a Gateway method, and then recombining the PCR product into a pLeela vector by using a gateway@enzyme Mix of Invitrogen company by using a homologous recombination method to construct the pLeela 35SS: zmZIPc 3 over-expression vector. Specific methods refer to the detailed steps in the gateway@enzyme Mix specification.
(3) The pLeela 35SS is transformed into escherichia coli DH5 alpha by using ZmbZIc3 over-expression vector, cultured overnight at 37 ℃, and monoclonal is selected for colony PCR verification and sequencing.
Sequencing results showed that: zmbZIPc3 is obtained by replacing the sequence between attR1 and attR2 sites of the pLeela vector with the DNA molecule shown in SEQ ID NO.3, and keeping other sequences unchanged. The over-expression vector is a vector obtained by recombining the DNA molecule shown in SEQ ID NO.3 into a pLeela vector. Wherein, the DNA molecule shown in the 51 st to 1283 rd positions of SEQ ID NO.3 is CDS sequence of ZmbZIPc3 gene (corresponding to the 51 st to 1283 rd positions of SEQ ID NO. 1), namely the ZmbZIPc3 gene is successfully connected into the pLeela vector. The pLeela 35SS is that the ZmbZIPc3 over-expression vector expresses the ZmbZIPc3 protein.
2. Acquisition of recombinant Agrobacterium
Transferring the pLeela 35SS:: zmbZIPc3 over-expression vector into agrobacterium tumefaciens GV3101 by using an electric shock method to obtain recombinant agrobacterium tumefaciens GV3101/pLeela 35SS:: zmbZIPc3, and performing amplified culture after colony PCR identification.
3. Obtaining transgenic Arabidopsis thaliana
The Arabidopsis thaliana Col-0 was infected with the recombinant Agrobacterium GV3101/pLeela 35SS:: zmZIPc 3 obtained in step 2 by the Arabidopsis thaliana flower dipping method (specific method is referred to non-patent literature "Clough SJ, bent AF (1998). Flora dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thiana. Plant J16,735-743"), to obtain T 0 Transgenic arabidopsis seed. Then the obtained T 0 The transgenic arabidopsis seeds are placed on a 1/2MS solid culture medium containing 10 mug/mL oxadiazon for screening positive seedlings, and the positive seedlings with resistance are transferred to a culture room for culture (the culture condition is that the temperature is 22 ℃ and the light is 16 h/the dark is 8 h) to obtain T 1 Transgenic arabidopsis seed. And then T is taken 1 Culturing the transgenic arabidopsis seeds on 1/2MS culture medium solid containing 10 mug/mL oxadiazon, and selecting T with a survival rate of 3/4 (i.e. a separation ratio of 3:1) 1 Culturing the survival seedling in a culture room, and harvesting T 2 Transgenic arabidopsis seed. Finally T is set 2 The transgenic Arabidopsis seeds are placed in a medium containing 10 mug/mL oxadiazonIn 1/2MS solid medium, the total surviving T is added 2 Transferring the survival seedling to a culture room for culturing to obtain T 3 The generation transgenic arabidopsis seeds (namely homozygous lines) are cultured to obtain three T 3 The homozygous lines of the transgenic Arabidopsis thaliana are respectively named as T 3 The transgenic Arabidopsis homozygous lines OE2-1, OE 5-6 and OE10-4 were used for further experiments.
4. Identification of transgenic Arabidopsis thaliana
Selecting T obtained in step 3 3 Leaf total RNA is extracted from homozygous strains OE2-1, OE 5-6 and OE10-4 of the generation transgenic Arabidopsis thaliana and wild Arabidopsis thaliana Col-0 respectively, and RT-PCR amplification is carried out by using primers RT-ZmZIPc 3-F and RT-ZmZIPc 3-R as well as primers 18S-F and 18S-R (18S genes are internal reference genes) as templates, and electrophoresis analysis is carried out, so that the expression quantity of ZmZIPc 3 genes in the transgenic Arabidopsis thaliana is analyzed.
The primer sequences were as follows:
RT-ZmbZIPc3-F:5'-AACTGGAGGCACAGGTAGCACA-3';
RT-ZmbZIPc3-R:5'-TATCAGAGGCGGAGTCAGAAGG-3';
18S-F:5’-CTTAGTTGGTGGAGCGATTTG-3’;
18S-R:5’-CCTGTTATTGCCTCAAACTTCC-3。
the results are shown in FIG. 1A, from which it can be seen that: the wild type Arabidopsis thaliana Col-0 (Col) has no target band, the transgenic Arabidopsis thaliana OE2-1, the transgenic Arabidopsis thaliana OE 5-6 and the transgenic Arabidopsis thaliana OE10-4 have target bands, the brightness of the transgenic Arabidopsis thaliana OE is OE2-1 > OE10-4 > OE 5-6, the positive transformation of the transgenic Arabidopsis thaliana OE2-1, the transgenic Arabidopsis thaliana OE 5-6 and the transgenic Arabidopsis thaliana OE10-4 by the positive transformation of the transgenic Arabidopsis thaliana OE2-1, the transgenic Arabidopsis thaliana OE 5-6 and the transgenic Arabidopsis thaliana OE10-4 is OE10-4, and the expression level of the ZmbZIPc3 genes is OE2-1 > OE 5-6 >.
2. Phenotypic analysis of ZmZIPc 3-transformed Arabidopsis thaliana
The plant materials were as follows: wild type Arabidopsis thaliana Col-0 (Col), T 3 Generation positive ZmZIPc 3 Arabidopsis homozygous strains OE2-1, OE 5-6 and OE 10-4.
1. The plant material seeds are treated with 75% ethanol for 1min, then sterilized with 10% sodium hypochlorite for 15min, then washed with sterilized distilled water for 5 times, and then treated at a low temperature of 4 ℃ for 3 days. Low and lowAfter the temperature treatment, the seeds are respectively sown on 1/2MS solid culture medium, 1/2MS solid culture medium which is permeated by 25 percent PEG overnight, 60mM CaCl is added 2 1/2MS solid medium of (C), 150mM NaCl was added 2 On a 1/2MS solid culture medium, culturing at 22+ -3deg.C under 50% humidity under 16h light/8 h dark condition, observing growth phenotype about 2 weeks, and counting root growth.
Growth phenotype and root length of 1/2MS solid Medium and 25% PEG-infiltrated overnight 1/2MS solid Medium in 1/2MS solid Medium, wild type Arabidopsis thaliana Col-0 (Col) with T is shown in FIG. 2 3 The growth phenotype and root length of the generation positive ZmZIPc 3-transformed Arabidopsis homozygous strains OE2-1, OE 5-6 and OE10-4 are equivalent; whereas T was compared to wild-type Arabidopsis thaliana Col-0 (Col) in 1/2MS solid medium infiltrated with 25% PEG overnight 3 The growth phenotype and root length of the generation positive ZmZIPc 3-transformed Arabidopsis thaliana homozygous strains OE2-1, OE 5-6 and OE10-4 have obvious advantages.
1/2MS solid Medium and 60mM CaCl added thereto 2 The growth phenotype and root length of the 1/2MS solid medium is shown in FIG. 3, and wild type Arabidopsis thaliana Col-0 (Col) and T in the 1/2MS solid medium 3 The growth phenotype and root length of the generation positive ZmZIPc 3-transformed Arabidopsis homozygous lines OE2-1, OE 5-6 and OE10-4 are comparable (shown in FIG. 3A); after 60mM CaCl was added 2 In the 1/2MS solid medium of (2), strain OE2-1 grew slightly weaker than wild-type under stress of 60mM CaCl2 compared to wild-type Arabidopsis thaliana Col-0 (Col), strain OE 5-6 and OE10-4 grew slightly stronger than wild-type (shown in FIG. 3B), and thus it could not be concluded whether ZmbZIPc3 overexpressing transgenic Arabidopsis thaliana was sensitive to CaCl2 stress.
1/2MS solid Medium and 150mM NaCl added 2 The growth phenotype and root length of the 1/2MS solid medium is shown in FIG. 3, and wild type Arabidopsis thaliana Col-0 (Col) and T in the 1/2MS solid medium 3 The growth phenotype and root length of the generation positive ZmZIPc 3-transformed Arabidopsis homozygous lines OE2-1, OE 5-6 and OE10-4 are comparable (shown in FIG. 3A); at the time of adding 150mM NaCl 2 T under stress of 150mM NaCl in 1/2MS solid medium 3 Generation positive ZmZIPc 3 arabidopsis thaliana homozygous strain OE2-1, OE5-6 and OE10-4 grew stronger than wild-type Arabidopsis thaliana Col-0 (Col) (shown as C in FIG. 3), and therefore ZmZIPc 3 over-expressed transgenic Arabidopsis thaliana was presumed to be resistant to NaCl stress.
The results show that the ZmbZIPc 3-transferred Arabidopsis thaliana strain has a more obvious PEG-resistant phenotype and salt stress-resistant phenotype.
2. The plant material seeds are treated with 75% ethanol for 1min, then sterilized with 10% sodium hypochlorite for 15min, then washed with sterilized distilled water for 5 times, and then treated at a low temperature of 4 ℃ for 3 days. After low temperature treatment, seeds were sown on 1/2MS (CK) and 1/2MS medium supplemented with 0.25. Mu.M, 0.5. Mu.M, and 1. Mu.M ABA, respectively, and cultured for 10-12d, and germination rates were counted daily using a stereoscopic microscope (brand: OPTEC). As a result, as shown in FIG. 4, T was performed on 1/2MS medium without ABA (shown as A in FIG. 4) 3 The germination rate of the generation positive ZmZIPc 3-transferred arabidopsis thaliana homozygous strain OE2-1 is slightly lower than that of the wild arabidopsis thaliana Col-0 (Col), but the germination rate can reach 100% after 4-5 d; t on 1/2MS medium supplemented with 0.25. Mu.M ABA (shown in FIG. 4B), 2 3 The germination rate of the generation positive ZmZIPc 3-transferred arabidopsis thaliana homozygous strain OE2-1 is 60% -70% in 3-5d, which is obviously lower than that of the wild arabidopsis thaliana Col-0 (80% -90%); t on 1/2MS medium (shown as C in FIG. 4) supplemented with 0.5. Mu.M ABA 3 The germination rate of the generation positive ZmZIPc 3-transferred arabidopsis thaliana homozygous lines OE2-1 and OE 5-6 is only about 45% at the time of 5d, and is obviously lower than that of the wild type arabidopsis thaliana Col-0 (80%); wild-type Arabidopsis Col-0 and T on 1/2MS medium supplemented with 1. Mu.M ABA (shown as D in FIG. 4) 3 The germination rate of the generation positive ZmZIPc 3-transferred Arabidopsis thaliana strain is inhibited, and the germination rate of the ZmZIPc 3-transferred Arabidopsis thaliana strain OE2-1 in 3-7d is obviously lower than that of the wild type strain. The above results indicate T 3 The germination rate of the generation positive ZmZIPc 3-transformed Arabidopsis thaliana homozygous lines OE2-1, OE 5-6 and OE10-4 is sensitive to ABA compared with that of wild Arabidopsis thaliana Col-0.
Will T 3 Seeds of the generation positive ZmbZIPc3 arabidopsis lines OE2-1, OE 5-6 and OE10-4 and wild arabidopsis Col-0 are respectively sown on a 1/2MS culture medium and a 1/2MS culture medium containing 0.1 mu M and 1 mu M ABA after 3d of suction expansion and low-temperature placement, and the seedlings are observed for growth after two weeks of culture, and are photographed. As a result, the result is shown in FIG. 5, at 1/2MS medium and 1/2MS medium containing 0.1. Mu.M ABA, T 3 The generation positive ZmZIPc 3-transformed Arabidopsis lines OE2-1, OE 5-6 and OE10-4 have no obvious difference from the growth conditions of the wild Arabidopsis Col-0 (shown in A and B in FIG. 5); on 1/2MS medium containing 1. Mu.M ABA, T is compared with wild type Arabidopsis Col-0 3 The generation positive ZmZIPc 3-transformed Arabidopsis strain OE2-1 was significantly inhibited (shown as C in FIG. 5). Shows that the seedling stage growth of the ZmZIPc 3-transferred arabidopsis thaliana homozygous strain is sensitive to ABA compared with wild type arabidopsis thaliana Col-0 (Col).
The results show that the ZmbZIPc3 gene plays a positive regulation role in the ABA signal transduction pathway.
3. The plant material seeds are treated with 75% ethanol for 1min, then sterilized with 10% sodium hypochlorite for 15min, then washed with sterilized distilled water for 5 times, and then treated at a low temperature of 4 ℃ for 3 days. The seeds are respectively sown on 1/2MS solid culture medium after low temperature treatment, are placed on a condition of 22+/-3 ℃ and 50% humidity and 16h illumination/8 h darkness for culture, arabidopsis thaliana with consistent size is selected and transferred into soil (nutrient soil: vermiculite=1:2 is filled into eight-hole trays, each hole is filled with 80g of soil, 6L of water is used for soaking soil for 4h, and then the seeds are poured into redundant water, 16 seedlings are transplanted in each hole, each 4 holes are a group, and wild arabidopsis thaliana Col-0 and T are respectively used as a group 3 The generation positive ZmZIPc 3 arabidopsis homozygous strains OE2-1, OE 5-6 and OE10-4 are respectively provided with an drought group and a control group, and are placed in a constant temperature culture room with 20+/-3 ℃ and 16h illumination/8 h darkness. The result of normal water application of the control group is shown in FIG. 6, and the control group is transformed into ZmZIPc 3 Arabidopsis homozygous strains OE2-1, OE 5-6 and OE10-4, which are consistent with the growth vigor of wild Arabidopsis Col-0. And (3) automatically transferring the drought group into soil, stopping watering, and starting to enter drought when the soil grows normally for 16d, observing and photographing the growth state, and observing and recording when the soil is drought for 7d and the soil is drought for 14d respectively. As shown in FIG. 6, when drought group plants drought to 14d, wild type Arabidopsis thaliana Col-0 leaves have all wilted, and the wilting rate of the over-expressed ZmbZIPc3 transgenic strain is OE2-1<OE 10-4<After rehydration for 3d, the wild arabidopsis thaliana Col-0 plant has a rehydration survival rate of 13%, an OE2-1 survival rate of 82%, an OE 5-6 survival rate of 38% and an OE10-4 survival rate of 69%. The rehydration survival rate is sequentially OE2-1>OE 10-4>OE 2-1>Col, the trend is consistent with the expression level of ZmbZIPc3 genes, which shows that the drought resistance of the transgenic Arabidopsis strain over-expressing ZmbZIPc3 is obviously better than that of wild Arabidopsis Col-0.
4. The plant material seeds are treated with 75% ethanol for 1min, then sterilized with 10% sodium hypochlorite for 15min, and then washed with sterilized distilled water for 5 times, and then treated at a low temperature of 4 ℃ for 3 days. After low temperature treatment, the seeds are respectively sown on 1/2MS solid culture medium and are cultured under the conditions of 22+/-3 ℃, 50% humidity, 16h illumination and 8h darkness. Selecting wild type Arabidopsis Col-0 (WT), T 3 The generation of positive ZmZIPc 3 arabidopsis homozygous strains OE2-1, OE 5-6 and OE10-4 are arabidopsis with equivalent growth vigor, each plant material is divided into three groups, the isolated leaves are subjected to drought treatment for different time, including drought 0, 0.25, 0.5, 1, 2 and 4 hours, sampling is fixed by FAA and decolored, water-filled tablets are manufactured, and the air hole condition is observed under a positive microscope (Nikco 80 i). The results of the drought 0, 1, 4h stomatal conditions are shown in FIG. 7, T 3 The leaf pore opening of the generation positive ZmbZIPc 3-transformed arabidopsis homozygous lines OE2-1, OE 5-6 and OE10-4 is small, and the water loss rate of the plant is low, so that the water retention and drought resistance functions of the plant are facilitated due to the over-expression of ZmbZIPc3.
In conclusion, zmbZIPc3 is related to stress resistance such as drought resistance, salt stress and the like.
Example 3 obtaining ZmZIPc 3-transformed maize and phenotypic analysis
1. Obtaining ZmZIPc 3-transformed corn
1. Obtaining recombinant expression vectors
(1) The pLeela 35SS in example 2 is used as a template, and a ZmbZIPc3 overexpression vector is used for PCR amplification by using a primer F2 and a primer R2 to obtain the ZmbZIPc3 gene expression cassette. The primer sequences were as follows:
F:5'-CCCAAGCTTGGGTTCCCGCCTTCGGTTTGG-3';
R:5'-CCCAAGCTTGGGCCTTGCTCGTCGGTGATGTA-3'。
the sequence of ZmbZIPc3 gene expression cassette is shown as SEQ ID NO.4, and comprises a promoter 35SS (36-720 positions from 5 'end of SEQ ID NO. 4), a coding sequence of ZmbZIPc3 gene (1141-2373 positions from 5' end of SEQ ID NO.4, corresponding 51-1283 positions from 5 'end of SEQ ID NO. 1) and a terminator PA35S (2593-2810 positions from 5' end of SEQ ID NO. 4) on a pLeela carrier in sequence.
(2) The ZmbZIPc3 gene expression cassette is inserted into the pU130 vector by a single digestion method of HindIII (TAKARA) to construct pU130 35SS, wherein the ZmbZIPc3 over-expression vector is obtained.
(3) pU130 35SS, zmbZIPc3 over-expression vector was transformed into E.coli DH 5. Alpha. And cultured overnight at 37℃and the monoclonal was picked for colony PCR verification and sequencing.
Sequencing results showed that: pU130 35SS, zmbZIPc3 over-expression vector is obtained by inserting ZmbZIPc3 gene expression cassette shown in SEQ ID No.4 between HindIII recognition sites of pU130 vector and keeping other sequences of pU130 vector unchanged. pU130 35SS that ZmbZIPc3 protein is expressed by the ZmbZIPc3 over-expression vector.
2. Obtaining transgenic maize
The pU130 35SS with correct sequence is that ZmbZIPc3 over-expression vector entrusts China seed group limited company life science and technology center to carry out corn transformation acceptor corn plant by adopting agrobacterium-mediated method, and the acceptor corn plant is corn C01 (belonging to PB group) (purchased from China seed group limited company life science and technology center) to obtain T0 generation ZmbZIPc3 corn; the corn seeds obtained by the test crossing of the T0 generation ZmbZIPc 3-transformed corn and the recipient corn thereof are T1 generation ZmbZIPc 3-transformed corn, and the genotype of the T1 generation ZmbZIPc 3-transformed corn is Aa: aa=1: 1. through continuous selfing, a homozygous strain with the genotype of T3 generation AA is obtained, and ZmbZIPc 3-transformed corn (namely T3 generation homozygous ZmbZIPc 3-transformed corn) seeds identified as the genotype of T3 generation AA are subjected to the following ZmbZIPc3 gene expression level measurement.
3. Measurement of ZmbZIPc3 Gene expression of transgenic corn
Total RNAs of T3 generation homozygous ZmbZIPc 3-transformed corn and wild corn C01 leaf of three leaves are respectively extracted, and half-quantitative PCR (real-time PCR) amplification is carried out by using a primer RT-ZmbZIPc3-F and a primer RT-ZmbZIPc3-R as well as a primer 18S-F and a primer 18S-R (18S gene is an internal reference gene) by taking the total RNAs as templates, so that the expression amount of the ZmbZIPc3 gene in the ZmbZIPc 3-transformed corn is analyzed (the primer sequence is consistent with that in example 2).
As a result, as shown in FIG. 1B, the expression level of ZmbZIPc3 gene in wild type corn C01 (WT) is used as a reference, the expression level of ZmbZIPc3 gene in ZmbZIPc3 transgenic corn is larger than that of wild type corn C01 (WT), wherein the expression level OEc 3-3> OEc3-1 > OEc3-5 > OEc3-7 > OEc3-2 of the positive T3-generation ZmbZIPc3 transgenic corn strain is higher than that of ZmbZIPc3 gene in ZmbZIPc3 transgenic corn, and the over-expression ZmbZIPc3 homozygous strains C3-1, C3-3, C3-5 and C3-7 are finally selected for subsequent experiments.
2. Phenotypic analysis of ZmbZIPc 3-transformed maize
The plant materials were as follows: wild type maize C01 plants (WT) and over-expressed ZmbZIPc3 homozygous lines C3-1, C3-3, C3-5, C3-7.
1. Wild corn C01 (WT), and the homozygous lines C3-1, C3-3, C3-5 and C3-7 of the over-expression ZmZIPc 3 are selected for germination drought experiments. Reference is made to the local standard of Hebei province in the technical Specification for drought resistance identification of maize. Stress cultures (i.e., treatment groups) were performed separately: each of the replicates was 20, 4, 20ul of polyethylene glycol-6000 solution (distilled water as solvent) was added, and the double-layer filter paper was used as a bud bed. Control culture (i.e., control group): one repetition of 20, 4 repetitions, 20 μl deionized water was added and the double layer filter paper was used as the bud bed. A round culture dish with the diameter of 10cm is used for culturing in a constant temperature incubator (25 ℃ C. Constant temperature care/dark culture for 16h/8 h), and the germination condition and drought resistance index of seeds are observed at 2, 4, 6 and 8 d.
The seed germination drought resistance index is calculated according to the following formula (1):
GDRI=PIs/PIc×100%………………………………………………………(1)
PI=1.00×nd2+0.75×nd4+0.50×nd6+0.25×nd8……………………………… (2)
PI c =1.00×nd2 ck+ 0.75×nd4 ck +0.50×nd6 ck +0.25×nd8 ck ……………(3)
PI s =1.00×nd2 s +0.75×nd4 s +0.50×nd6 s +0.25×nd8 s …………………………(4)
wherein:
PI-seed germination index, nd2, nd4, nd6, nd8 are seed germination rates at 2d, 4d, 6d, and 8d, respectively;
PI C -a control seed germination index (ck in formula (3) represents control conditions);
PI S -seed germination index under stress conditions (s in formula (4) represents stress conditions);
GDRI-seed germination drought resistance index.
The seed germination condition and the seed germination drought resistance index result are shown as A and B in FIG. 8, and the drought resistance indexes of the over-expression ZmZIPc 3 homozygous strains OEc 3-1, OEc 3-5 and OEc 3-7 are obviously higher than that of wild corn C01 (WT); root growth of different corn lines was observed on day 9 of germination, and as shown in FIG. 9, the main roots of the over-expressed ZmZIPc 3 homozygous lines OEc 3-1, OEc 3-5 and OEc 3-7 were significantly longer than that of wild type corn C01 (WT) and the lateral roots were also greater than that of wild type corn C01 (WT) under stress culture conditions.
The above shows that the drought resistance of the strain ZmbZIPc3 over-expressed in germination period is better than that of wild corn C01.
2. Selecting wild corn C01 (WT), over-expression ZmbZIPc3 homozygous strain OEc 3-1, OEc 3-5 and OEc 3-7, and performing seedling drought culture. Soil planting and culturing are adopted, a control group and a treatment group are respectively arranged in each plant line in a basin with the specification of 16cm multiplied by 16cm, the seedling base number of each plant line is 20, and three times of repetition are arranged. Wherein the control group is normally watered in the whole growth period, the seedlings of the treatment group grow to about the three-leaf one-heart period and start drought treatment (A in figure 10), namely, the watering is stopped, the wilting condition of the seedlings is started to be observed and recorded, when the seedlings all show the wilting condition, the first rehydration treatment is carried out, and the survival condition of the seedlings is observed after 72 hours of rehydration (B in figure 10). And after the first rehydration is finished, the second drought treatment is started, and the second rehydration treatment is also carried out after the seedlings are all in a wilting state (C in fig. 10), and the survival rate of the seedlings is observed for 72 hours, wherein the result is that, as shown in D in fig. 10, after two drought rehydration, the survival rate of each strain is OE C3-5 (60%) > OE C3-7 (40%) > WT (10%), the repeated drought rehydration survival rate of the over-expression ZmbZIPc3 homozygous strain is higher than that of wild corn C01 (WT), the trend is consistent with the expression amount trend of each strain, and the drought resistance of the over-expression ZmbZIPc3 strain in the seedling stage is better than that of the wild corn C01 (WT).
The experiment shows that the over-expression of ZmbZIPc3 can improve drought tolerance of corn in germination period and seedling period.
The present application is described in detail above. It will be apparent to those skilled in the art that the present application can be practiced in a wide range of equivalent parameters, concentrations, and conditions without departing from the spirit and scope of the application and without undue experimentation. While the application has been described with respect to specific embodiments, it will be appreciated that the application may be further modified. In general, this application is intended to cover any variations, uses, or adaptations of the application following, in general, the principles of the application and including such departures from the present disclosure as come within known or customary practice within the art to which the application pertains. The application of some of the basic features may be done in accordance with the scope of the claims that follow.
SEQUENCE LISTING
<110> institute of plant Material at national academy of sciences
Application of <120> protein ZmbZIPc3 in regulation of drought resistance and/or salt tolerance of plants
<130> GNCFY200882
<160> 4
<170> PatentIn version 3.5
<210> 1
<211> 1493
<212> DNA
<213> corn (Zea mays)
<400> 1
ccctctccga ggattctgcc cagcagcgcg gcgatccgcc tcgatccgga atggagcgcg 60
tcttctccat ggaggagatc cccaacccct actgggcccc gccgcaccct caaccggcgg 120
ccggcggcgc tgttgctgca ccaggtggag taggaggagc gggggacgcg gcgggcgcga 180
tgaaccggtg cccatctgag tggtacttcc agaagttcct cgaggaggcc gtgctcgaca 240
gtccgggtcc cgtcgccggc gtgggtagaa gcagcggaca agctggagtt gaggcggcgg 300
agagcaagcc gctgggcgcc gcggcgccgg cgtccgtctc gagctcggtc gttgaccccg 360
tcgagtacaa cgcgatgctc aagcagaagc tggagaagga cctcgctgcc atcgctatgt 420
ggagggcttc tggtgcagca cctccagatc tttctgcgac tgctgcttcc ttgccaagtg 480
tcggtgctcc gcatgcagct cctcttaaac ccgtcggagg tactgaaagt ctagttcaaa 540
acatgctagc tggtgctcca gtaggggggt caggtccaca tgtagtacaa attgctgata 600
tccctgttaa gcaaaccact agctcttcct cacgagagca gtcagatgat gatgacatgg 660
aaggcgatgc tgagacaaat ggaaatggaa accctgttca acaaagacaa cagagaagga 720
agcaatccaa ccgggaatca gccaggcgtt cgagaagcag aaaggcggct cacttgaatg 780
aactggaggc acaggtagca caattaagag tcgaaaactc ttcgctgcta aggcggcttg 840
ctgatgttaa ccagaagttc aatgaagctg ctgttgacaa tagggtgcta aaggcagacg 900
tcgaaacctt aagagcaaag gtgaagatgg cagaggactc ggtgaagcgg gtaacaggca 960
tgaacgcatt gtaccctgcc gtgtcggata tgtcttccct cagcatgcca ttcaatggct 1020
ccccttctga ctccgcctct gatagcaccg tcccggtaca agatgacctg aacagttact 1080
tcgccaaccc aagcgaaatc ggatgcaaca acggctacat gccagatata gcctcctcgg 1140
ttcaacagga cgacaatttt gtcaacgggt atcaggctgc tggcaagatg ggcagaacag 1200
actcgctgca gcgggtggca agcctggagc acctccagaa aagaatgtgc ggaggcccgg 1260
cttcatccgg ctcgacctcc taggtttcat atgaaaaagt ccctggtgcc taacggcaag 1320
aatgctaaat taacaatttc aacatgcttt gtactggaca ttgataatcc tgcgatgcgc 1380
attagtgcgt tatggacttg tgggtcaatt gctatttggc gtgtgctggc tgctcactgt 1440
agtgagcaga acaagttcca gtttctatcg aagagcaccc tctccttttc gct 1493
<210> 2
<211> 410
<212> PRT
<213> corn (Zea mays)
<400> 2
Met Glu Arg Val Phe Ser Met Glu Glu Ile Pro Asn Pro Tyr Trp Ala
1 5 10 15
Pro Pro His Pro Gln Pro Ala Ala Gly Gly Ala Val Ala Ala Pro Gly
20 25 30
Gly Val Gly Gly Ala Gly Asp Ala Ala Gly Ala Met Asn Arg Cys Pro
35 40 45
Ser Glu Trp Tyr Phe Gln Lys Phe Leu Glu Glu Ala Val Leu Asp Ser
50 55 60
Pro Gly Pro Val Ala Gly Val Gly Arg Ser Ser Gly Gln Ala Gly Val
65 70 75 80
Glu Ala Ala Glu Ser Lys Pro Leu Gly Ala Ala Ala Pro Ala Ser Val
85 90 95
Ser Ser Ser Val Val Asp Pro Val Glu Tyr Asn Ala Met Leu Lys Gln
100 105 110
Lys Leu Glu Lys Asp Leu Ala Ala Ile Ala Met Trp Arg Ala Ser Gly
115 120 125
Ala Ala Pro Pro Asp Leu Ser Ala Thr Ala Ala Ser Leu Pro Ser Val
130 135 140
Gly Ala Pro His Ala Ala Pro Leu Lys Pro Val Gly Gly Thr Glu Ser
145 150 155 160
Leu Val Gln Asn Met Leu Ala Gly Ala Pro Val Gly Gly Ser Gly Pro
165 170 175
His Val Val Gln Ile Ala Asp Ile Pro Val Lys Gln Thr Thr Ser Ser
180 185 190
Ser Ser Arg Glu Gln Ser Asp Asp Asp Asp Met Glu Gly Asp Ala Glu
195 200 205
Thr Asn Gly Asn Gly Asn Pro Val Gln Gln Arg Gln Gln Arg Arg Lys
210 215 220
Gln Ser Asn Arg Glu Ser Ala Arg Arg Ser Arg Ser Arg Lys Ala Ala
225 230 235 240
His Leu Asn Glu Leu Glu Ala Gln Val Ala Gln Leu Arg Val Glu Asn
245 250 255
Ser Ser Leu Leu Arg Arg Leu Ala Asp Val Asn Gln Lys Phe Asn Glu
260 265 270
Ala Ala Val Asp Asn Arg Val Leu Lys Ala Asp Val Glu Thr Leu Arg
275 280 285
Ala Lys Val Lys Met Ala Glu Asp Ser Val Lys Arg Val Thr Gly Met
290 295 300
Asn Ala Leu Tyr Pro Ala Val Ser Asp Met Ser Ser Leu Ser Met Pro
305 310 315 320
Phe Asn Gly Ser Pro Ser Asp Ser Ala Ser Asp Ser Thr Val Pro Val
325 330 335
Gln Asp Asp Leu Asn Ser Tyr Phe Ala Asn Pro Ser Glu Ile Gly Cys
340 345 350
Asn Asn Gly Tyr Met Pro Asp Ile Ala Ser Ser Val Gln Gln Asp Asp
355 360 365
Asn Phe Val Asn Gly Tyr Gln Ala Ala Gly Lys Met Gly Arg Thr Asp
370 375 380
Ser Leu Gln Arg Val Ala Ser Leu Glu His Leu Gln Lys Arg Met Cys
385 390 395 400
Gly Gly Pro Ala Ser Ser Gly Ser Thr Ser
405 410
<210> 3
<211> 1387
<212> DNA
<213> Artificial sequence (Artificial Sequence)
<400> 3
ggaatggagc gcgtcttctc catggaggag atccccaacc cctactgggc cccgccgcac 60
cctcaaccgg cggccggcgg cgctgttgct gcaccaggtg gagtaggagg agcgggggac 120
gcggcgggcg cgatgaaccg gtgcccatct gagtggtact tccagaagtt cctcgaggag 180
gccgtgctcg acagtccggg tcccgtcgcc ggcgtgggta gaagcagcgg acaagctgga 240
gttgaggcgg cggagagcaa gccgctgggc gccgcggcgc cggcgtccgt ctcgagctcg 300
gtcgttgacc ccgtcgagta caacgcgatg ctcaagcaga agctggagaa ggacctcgct 360
gccatcgcta tgtggagggc ttctggtgca gcacctccag atctttctgc gactgctgct 420
tccttgccaa gtgtcggtgc tccgcatgca gctcctctta aacccgtcgg aggtactgaa 480
agtctagttc aaaacatgct agctggtgct ccagtagggg ggtcaggtcc acatgtagta 540
caaattgctg atatccctgt taagcaaacc actagctctt cctcacgaga gcagtcagat 600
gatgatgaca tggaaggcga tgctgagaca aatggaaatg gaaaccctgt tcaacaaaga 660
caacagagaa ggaagcaatc caaccgggaa tcagccaggc gttcgagaag cagaaaggcg 720
gctcacttga atgaactgga ggcacaggta gcacaattaa gagtcgaaaa ctcttcgctg 780
ctaaggcggc ttgctgatgt taaccagaag ttcaatgaag ctgctgttga caatagggtg 840
ctaaaggcag acgtcgaaac cttaagagca aaggtgaaga tggcagagga ctcggtgaag 900
cgggtaacag gcatgaacgc attgtaccct gccgtgtcgg atatgtcttc cctcagcatg 960
ccattcaatg gctccccttc tgactccgcc tctgatagca ccgtcccggt acaagatgac 1020
ctgaacagtt acttcgccaa cccaagcgaa atcggatgca acaacggcta catgccagat 1080
atagcctcct cggttcaaca ggacgacaat tttgtcaacg ggtatcaggc tgctggcaag 1140
atgggcagaa cagactcgct gcagcgggtg gcaagcctgg agcacctcca gaaaagaatg 1200
tgcggaggcc cggcttcatc cggctcgacc tcctaggttt catatgaaaa agtccctggt 1260
gcctaacggc aagaatgcta aattaacaat ttcaacatgc tttgtactgg acattgataa 1320
tcctgcgatg cgcattagtg cgttatggac ttgtgggtca attgctattt ggcgtgtgct 1380
ggctgct 1387
<210> 4
<211> 3018
<212> DNA
<213> Artificial sequence (Artificial Sequence)
<400> 4
aagcttgggt tcccgccttc ggtttgggcg cgcccggttc agaagaccag agggctattg 60
agacttttca acaaagggta atatcgggaa acctcctcgg attccattgc ccagctatct 120
gtcacttcat cgaaaggaca gtagaaaagg aagatggctt ctacaaatgc catcattgcg 180
ataaaggaaa ggctatcgtt caagatgcct ctaccgacag tggtcccaaa gatggacccc 240
catccacgag gaacatcgtg gaaaaagaag acgttccaac cacgtcttca aagcaagtgg 300
attgatgtga tacatggtgg agcacgacac tctcgtctac tccaagaata tcaaagatac 360
agtctcagaa gaccagaggg ctattgagac ttttcaacaa agggtaatat cgggaaacct 420
cctcggattc cattgcccag ctatctgtca cttcatcgaa aggacagtag aaaaggaaga 480
tggcttctac aaatgccatc attgcgataa aggaaaggct atcgttcaag atgcctctac 540
cgacagtggt cccaaagatg gacccccacc cacgaggaac atcgtggaaa aagaagacgt 600
tccaaccacg tcttcaaagc aagtggattg atgtgatatc tccactgacg taagggatga 660
cgcacaatcc cactatcctt cgcaagaccc ttcctctata taaggaagtt catttcattt 720
ggagaggacc tcgagaaaga ggatccacct gaggcctccg ttccatgggc tagaagcttc 780
tcctcctctg ctaacgtaag cctctctgtt ttttttctct gtttcttttg aaatgaatcc 840
aattagtgat gataatctgt gtttgatgta tcattgattt aacatcttga caatgaatcg 900
tgatcggaag tgataaagtt atgggtcaac ggtttcaaag agagagaaag acttttagag 960
tcaactctcg actctttctt aattatgtta ttgctatttg tctcttttct tgaagtctga 1020
acaattcttg ggattgtttt gcaggttcta gcttctccaa ccacaggaat tcatcgcccg 1080
ggactgggtt atcacaagtt tgtacaaaaa agcaggctcc gcggccgccc ccttcaccgg 1140
aatggagcgc gtcttctcca tggaggagat ccccaacccc tactgggccc cgccgcaccc 1200
tcaaccggcg gccggcggcg ctgttgctgc accaggtgga gtaggaggag cgggggacgc 1260
ggcgggcgcg atgaaccggt gcccatctga gtggtacttc cagaagttcc tcgaggaggc 1320
cgtgctcgac agtccgggtc ccgtcgccgg cgtgggtaga agcagcggac aagctggagt 1380
tgaggcggcg gagagcaagc cgctgggcgc cgcggcgccg gcgtccgtct cgagctcggt 1440
cgttgacccc gtcgagtaca acgcgatgct caagcagaag ctggagaagg acctcgctgc 1500
catcgctatg tggagggctt ctggtgcagc acctccagat ctttctgcga ctgctgcttc 1560
cttgccaagt gtcggtgctc cgcatgcagc tcctcttaaa cccgtcggag gtactgaaag 1620
tctagttcaa aacatgctag ctggtgctcc agtagggggg tcaggtccac atgtagtaca 1680
aattgctgat atccctgtta agcaaaccac tagctcttcc tcacgagagc agtcagatga 1740
tgatgacatg gaaggcgatg ctgagacaaa tggaaatgga aaccctgttc aacaaagaca 1800
acagagaagg aagcaatcca accgggaatc agccaggcgt tcgagaagca gaaaggcggc 1860
tcacttgaat gaactggagg cacaggtagc acaattaaga gtcgaaaact cttcgctgct 1920
aaggcggctt gctgatgtta accagaagtt caatgaagct gctgttgaca atagggtgct 1980
aaaggcagac gtcgaaacct taagagcaaa ggtgaagatg gcagaggact cggtgaagcg 2040
ggtaacaggc atgaacgcat tgtaccctgc cgtgtcggat atgtcttccc tcagcatgcc 2100
attcaatggc tccccttctg actccgcctc tgatagcacc gtcccggtac aagatgacct 2160
gaacagttac ttcgccaacc caagcgaaat cggatgcaac aacggctaca tgccagatat 2220
agcctcctcg gttcaacagg acgacaattt tgtcaacggg tatcaggctg ctggcaagat 2280
gggcagaaca gactcgctgc agcgggtggc aagcctggag cacctccaga aaagaatgtg 2340
cggaggcccg gcttcatccg gctcgacctc ctaggtttca tatgaaaaag tccctggtgc 2400
ctaacggcaa gaatgctaaa ttaacaattt caacatgctt tgtactggac attgataatc 2460
ctgcgatgcg cattagtgcg ttatggactt gtgggtcaat tgctatttgg cgtgtgctgg 2520
ctgctaaggg tgggcgcgcc gacccagctt tcttgtacaa agtggtgata acttccgatc 2580
gattctagac tagttctaga gtccgcaaaa atcaccagtc tctctctaca aatctatctc 2640
tctctatttt tctccagaat aatgtgtgag tagttcccag ataagggaat tagggttctt 2700
atagggtttc gctcatgtgt tgagcatata agaaaccctt agtatgtatt tgtatttgta 2760
aaatacttct atcaataaaa tttctaattc ctaaaaccaa aatccagtga ccgggcggcc 2820
gccaccgcgg tggaggggga tcagattgtc gtttcccgcc ttcagtttaa actatcagtg 2880
tttgacagga tatattggcg ggtaaaccta agagaaaaga gcgtttatta gaataatcgg 2940
atatttaaaa gggcgtgaaa aggtttatcc gttcgtccat ttgtatgtgt acatcaccga 3000
cgagcaaggc ccaagctt 3018
Claims (6)
1. The application of the protein shown in the following A1) or A2) in regulating drought resistance and/or salt tolerance of plants:
a1 Protein with the amino acid sequence shown as SEQ ID NO. 2;
a2 Fusion protein obtained by N-terminal or/and C-terminal connexin label of the amino acid sequence shown in SEQ ID NO. 2;
the regulation of drought resistance and/or salt tolerance of plants is to improve the drought resistance and/or salt tolerance of plants;
the plant is corn or Arabidopsis thaliana.
2. Use of a biological material related to the protein of claim 1 for modulating drought and/or salt tolerance in a plant; the biological material is any one of the following C1) -C10):
c1 A nucleic acid molecule encoding the protein of claim 1;
c2 An expression cassette comprising C1) said nucleic acid molecule;
c3 A recombinant vector comprising C1) said nucleic acid molecule, or a recombinant vector comprising C2) said expression cassette;
c4 A recombinant microorganism comprising C1) said nucleic acid molecule, or a recombinant microorganism comprising C2) said expression cassette, or a recombinant microorganism comprising C3) said recombinant vector;
c5 A transgenic plant cell line comprising C1) said nucleic acid molecule, or a transgenic plant cell line comprising C2) said expression cassette, or a transgenic plant cell line comprising C3) said recombinant vector;
c6 A) a transgenic plant tissue comprising C1) said nucleic acid molecule, or a transgenic plant tissue comprising C2) said expression cassette, or a transgenic plant tissue comprising C3) said recombinant vector;
c7 A transgenic plant organ comprising C1) said nucleic acid molecule, or a transgenic plant organ comprising C2) said expression cassette, or a transgenic plant organ comprising C3) said recombinant vector;
c8 A transgenic plant containing the nucleic acid molecule of C1), or a transgenic plant containing the expression cassette of C2), or a transgenic plant containing the recombinant vector of C3);
c9 A tissue culture produced by regenerable cells of the transgenic plant of C8);
c10 Protoplasts produced from the tissue culture of C9);
the regulation of drought resistance and/or salt tolerance of plants is to improve the drought resistance and/or salt tolerance of plants;
the plant is corn or Arabidopsis thaliana.
3. The use according to claim 2, characterized in that: c1 A nucleic acid molecule encoding the protein according to claim 1 as shown in D1) or D2) below:
d1 A DNA molecule shown in SEQ ID NO. 1;
d2 A DNA molecule with the coding sequence shown as SEQ ID NO. 1.
4. A method of growing transgenic plants with high drought and/or salt tolerance, characterized by: the method comprises increasing the expression level of the gene of the protein of claim 1 and/or the content of the protein and/or the activity of the protein in a plant of interest to obtain a transgenic plant; the transgenic plant has drought and/or alkali tolerance higher than the plant of interest; the plant is corn or Arabidopsis thaliana.
5. The method according to claim 4, wherein: the method for increasing the expression level of the gene of the protein of claim 1 and/or the content of the protein and/or the activity of the protein in a plant of interest is to express or overexpress the protein of claim 1 in a plant of interest.
6. The method according to claim 5, wherein: the method of expression or overexpression is to introduce a gene encoding the protein of claim 1 into a plant of interest.
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202010467043.6A CN113801890B (en) | 2020-05-28 | 2020-05-28 | Application of protein ZmbZIPc3 in regulation and control of salt tolerance and drought resistance of plants |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202010467043.6A CN113801890B (en) | 2020-05-28 | 2020-05-28 | Application of protein ZmbZIPc3 in regulation and control of salt tolerance and drought resistance of plants |
Publications (2)
Publication Number | Publication Date |
---|---|
CN113801890A CN113801890A (en) | 2021-12-17 |
CN113801890B true CN113801890B (en) | 2023-10-31 |
Family
ID=78943664
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202010467043.6A Active CN113801890B (en) | 2020-05-28 | 2020-05-28 | Application of protein ZmbZIPc3 in regulation and control of salt tolerance and drought resistance of plants |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN113801890B (en) |
Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN110684088A (en) * | 2018-07-04 | 2020-01-14 | 中国科学院植物研究所 | Protein ZmbZIPa3 and application of coding gene thereof in regulating and controlling plant growth and development and stress tolerance |
-
2020
- 2020-05-28 CN CN202010467043.6A patent/CN113801890B/en active Active
Patent Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN110684088A (en) * | 2018-07-04 | 2020-01-14 | 中国科学院植物研究所 | Protein ZmbZIPa3 and application of coding gene thereof in regulating and controlling plant growth and development and stress tolerance |
Non-Patent Citations (3)
Title |
---|
bZIP转录因子与植物抗逆性研究进展;曹红利等;《南方农业学报》;20120815;全文 * |
Yilmaz,A.等.Zea mays subsp. mays clone UT3221 bZIP-type transcription factor mRNA, partial cds.《Genbank》.2012, * |
Zea mays subsp. mays clone UT3221 bZIP-type transcription factor mRNA, partial cds;Yilmaz,A.等;《Genbank》;20121016;全文 * |
Also Published As
Publication number | Publication date |
---|---|
CN113801890A (en) | 2021-12-17 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
US10538779B2 (en) | Constitutive photomorphogenesis 1 (COP1) nucleic acid sequence from Zea mays and its use thereof | |
JP4365021B2 (en) | Plant stress tolerance and aging delay | |
US9809827B2 (en) | Transgenic maize | |
US11299744B2 (en) | Transgenic plants expressing type 2C protein phosphatase abscisic acid (PP2CABA) proteins and uses thereof | |
US20160068860A1 (en) | Transgenic plants | |
CN113563442B (en) | Drought-resistant related protein IbSPB1, and coding gene and application thereof | |
CN114560919B (en) | Plant drought tolerance related transcription factor VcMYB and coding gene and application thereof | |
CN113980106B (en) | Small peptide for regulating and controlling sizes of plant seeds and organs, and coding gene and application thereof | |
CN117264964A (en) | Application of wheat TaGSKB protein and encoding gene thereof in regulation and control of plant stress tolerance | |
US10017779B2 (en) | Gene implicated in abiotic stress tolerance and growth accelerating and use thereof | |
CN111153979A (en) | Drought-resistant related protein IbBT4, and coding gene and application thereof | |
WO2013056677A1 (en) | USE OF OsPP18 GENE IN CONTROLLING RICE DROUGHT RESISTANCE | |
CN111434678B (en) | Plant dehydration response element encoding protein and application of encoding gene thereof in low nitrogen stress resistance | |
CN114591409B (en) | Application of TaDTG6 protein in improving drought resistance of plants | |
CN113929758B (en) | Potassium ion transporter protein HbRSAR1 and application thereof in regulating potassium transport of plants | |
CN111116721A (en) | Plant stress resistance-related transcription factor PwNAC30, and coding gene and application thereof | |
CN113801890B (en) | Application of protein ZmbZIPc3 in regulation and control of salt tolerance and drought resistance of plants | |
CN103361323B (en) | Application of rice ssg genes in improvement of plant salt tolerance | |
CN113773374B (en) | Transcription factor ZmbZIPa6 and coding gene and application thereof | |
CN113832160B (en) | ZmbZIPf3 gene, protein coded by same and application thereof | |
CN114645032B (en) | Application of 4 RAF proteins and encoding genes thereof in drought resistance of plants | |
CN112979775B (en) | Method for cultivating pre-sprouting resistant transgenic wheat and related biological material thereof | |
CN114644691B (en) | EIP1 protein, coding gene thereof and drought resisting application | |
CN118546985A (en) | Plant drought-tolerance related protein TaHIN-6A and coding gene and application thereof | |
CN118562876A (en) | BnaHsfA2 gene and application of overexpression vector thereof in improving freezing resistance of rape |
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 |