CN108018294A - A kind of pseudo-ginseng plant hormone binding-protein gene PnPhBP1 and application - Google Patents
A kind of pseudo-ginseng plant hormone binding-protein gene PnPhBP1 and application Download PDFInfo
- Publication number
- CN108018294A CN108018294A CN201810000429.9A CN201810000429A CN108018294A CN 108018294 A CN108018294 A CN 108018294A CN 201810000429 A CN201810000429 A CN 201810000429A CN 108018294 A CN108018294 A CN 108018294A
- Authority
- CN
- China
- Prior art keywords
- pnphbp1
- plant
- pseudo
- tobacco
- gene
- 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.)
- Granted
Links
- 244000131316 Panax pseudoginseng Species 0.000 title claims abstract description 24
- 235000003181 Panax pseudoginseng Nutrition 0.000 title claims abstract description 23
- 239000003375 plant hormone Substances 0.000 title claims abstract description 16
- 108091011044 hormone binding proteins Proteins 0.000 title claims abstract description 11
- 108090000623 proteins and genes Proteins 0.000 claims abstract description 40
- 102000004169 proteins and genes Human genes 0.000 claims abstract description 18
- 241000123650 Botrytis cinerea Species 0.000 claims abstract description 8
- 241000427940 Fusarium solani Species 0.000 claims abstract description 7
- 102000036124 hormone binding proteins Human genes 0.000 claims abstract description 7
- 241001529387 Colletotrichum gloeosporioides Species 0.000 claims abstract description 6
- 239000002773 nucleotide Substances 0.000 claims abstract description 4
- 125000003729 nucleotide group Chemical group 0.000 claims abstract 3
- 125000003275 alpha amino acid group Chemical group 0.000 claims abstract 2
- 241000196324 Embryophyta Species 0.000 abstract description 50
- 235000002637 Nicotiana tabacum Nutrition 0.000 abstract description 30
- 244000061176 Nicotiana tabacum Species 0.000 abstract description 28
- 230000009261 transgenic effect Effects 0.000 abstract description 10
- 241000233866 Fungi Species 0.000 abstract description 8
- 108700019146 Transgenes Proteins 0.000 abstract description 6
- 230000000843 anti-fungal effect Effects 0.000 abstract description 4
- 239000013604 expression vector Substances 0.000 abstract description 4
- 230000002401 inhibitory effect Effects 0.000 abstract description 4
- 230000002018 overexpression Effects 0.000 abstract description 4
- 239000002543 antimycotic Substances 0.000 abstract description 3
- 230000001857 anti-mycotic effect Effects 0.000 abstract description 2
- 241000208125 Nicotiana Species 0.000 abstract 2
- IXORZMNAPKEEDV-UHFFFAOYSA-N gibberellic acid GA3 Natural products OC(=O)C1C2(C3)CC(=C)C3(O)CCC2C2(C=CC3O)C1C3(C)C(=O)O2 IXORZMNAPKEEDV-UHFFFAOYSA-N 0.000 description 19
- 238000000034 method Methods 0.000 description 19
- 229930191978 Gibberellin Natural products 0.000 description 16
- 239000003448 gibberellin Substances 0.000 description 16
- 235000018102 proteins Nutrition 0.000 description 16
- 108020004414 DNA Proteins 0.000 description 13
- 238000006243 chemical reaction Methods 0.000 description 12
- 241000894006 Bacteria Species 0.000 description 11
- 239000000047 product Substances 0.000 description 11
- 239000002299 complementary DNA Substances 0.000 description 10
- 239000013612 plasmid Substances 0.000 description 10
- 201000010099 disease Diseases 0.000 description 8
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 8
- 239000012634 fragment Substances 0.000 description 8
- 238000012216 screening Methods 0.000 description 8
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 8
- 241000589158 Agrobacterium Species 0.000 description 7
- 101000900567 Pisum sativum Disease resistance response protein Pi49 Proteins 0.000 description 7
- 238000000605 extraction Methods 0.000 description 7
- 210000002966 serum Anatomy 0.000 description 7
- 241000180649 Panax notoginseng Species 0.000 description 6
- 235000003143 Panax notoginseng Nutrition 0.000 description 6
- 238000000246 agarose gel electrophoresis Methods 0.000 description 6
- 230000008569 process Effects 0.000 description 6
- 239000013598 vector Substances 0.000 description 6
- 108700026244 Open Reading Frames Proteins 0.000 description 5
- 238000009395 breeding Methods 0.000 description 5
- IXORZMNAPKEEDV-OBDJNFEBSA-N gibberellin A3 Chemical compound C([C@@]1(O)C(=C)C[C@@]2(C1)[C@H]1C(O)=O)C[C@H]2[C@]2(C=C[C@@H]3O)[C@H]1[C@]3(C)C(=O)O2 IXORZMNAPKEEDV-OBDJNFEBSA-N 0.000 description 5
- 239000003292 glue Substances 0.000 description 5
- 239000003550 marker Substances 0.000 description 5
- 238000002156 mixing Methods 0.000 description 5
- 239000007787 solid Substances 0.000 description 5
- 239000000243 solution Substances 0.000 description 5
- 239000000126 substance Substances 0.000 description 5
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 4
- 239000005980 Gibberellic acid Substances 0.000 description 4
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 4
- 101100282746 Oryza sativa subsp. japonica GID1 gene Proteins 0.000 description 4
- 101100156295 Saccharomyces cerevisiae (strain ATCC 204508 / S288c) VID30 gene Proteins 0.000 description 4
- 230000015572 biosynthetic process Effects 0.000 description 4
- 230000001488 breeding effect Effects 0.000 description 4
- 239000000872 buffer Substances 0.000 description 4
- 230000032823 cell division Effects 0.000 description 4
- 230000029087 digestion Effects 0.000 description 4
- 239000000284 extract Substances 0.000 description 4
- 238000013467 fragmentation Methods 0.000 description 4
- 238000006062 fragmentation reaction Methods 0.000 description 4
- 230000014509 gene expression Effects 0.000 description 4
- 230000012010 growth Effects 0.000 description 4
- 239000001963 growth medium Substances 0.000 description 4
- 238000005286 illumination Methods 0.000 description 4
- 239000000463 material Substances 0.000 description 4
- 239000002609 medium Substances 0.000 description 4
- 239000000203 mixture Substances 0.000 description 4
- 238000003199 nucleic acid amplification method Methods 0.000 description 4
- 238000003757 reverse transcription PCR Methods 0.000 description 4
- YGSDEFSMJLZEOE-UHFFFAOYSA-N salicylic acid Chemical compound OC(=O)C1=CC=CC=C1O YGSDEFSMJLZEOE-UHFFFAOYSA-N 0.000 description 4
- 229920001817 Agar Polymers 0.000 description 3
- 241000589155 Agrobacterium tumefaciens Species 0.000 description 3
- 102000012410 DNA Ligases Human genes 0.000 description 3
- 108010061982 DNA Ligases Proteins 0.000 description 3
- VGGSQFUCUMXWEO-UHFFFAOYSA-N Ethene Chemical compound C=C VGGSQFUCUMXWEO-UHFFFAOYSA-N 0.000 description 3
- 102100028909 Heterogeneous nuclear ribonucleoprotein K Human genes 0.000 description 3
- 108010084680 Heterogeneous-Nuclear Ribonucleoprotein K Proteins 0.000 description 3
- 208000031888 Mycoses Diseases 0.000 description 3
- 239000008272 agar Substances 0.000 description 3
- 150000001413 amino acids Chemical group 0.000 description 3
- 238000004458 analytical method Methods 0.000 description 3
- 230000008901 benefit Effects 0.000 description 3
- 102000023732 binding proteins Human genes 0.000 description 3
- 108091008324 binding proteins Proteins 0.000 description 3
- 239000003153 chemical reaction reagent Substances 0.000 description 3
- 238000012258 culturing Methods 0.000 description 3
- 230000004665 defense response Effects 0.000 description 3
- 238000005516 engineering process Methods 0.000 description 3
- 239000012530 fluid Substances 0.000 description 3
- 230000004060 metabolic process Effects 0.000 description 3
- 235000015097 nutrients Nutrition 0.000 description 3
- 239000000575 pesticide Substances 0.000 description 3
- 230000011664 signaling Effects 0.000 description 3
- 238000003786 synthesis reaction Methods 0.000 description 3
- 230000002103 transcriptional effect Effects 0.000 description 3
- 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 2
- 244000144730 Amygdalus persica Species 0.000 description 2
- 241000208340 Araliaceae Species 0.000 description 2
- 244000000626 Daucus carota Species 0.000 description 2
- 235000002767 Daucus carota Nutrition 0.000 description 2
- 102100031780 Endonuclease Human genes 0.000 description 2
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 description 2
- 101150056631 Habp2 gene Proteins 0.000 description 2
- 206010020751 Hypersensitivity Diseases 0.000 description 2
- 240000007849 Macleaya cordata Species 0.000 description 2
- 235000002791 Panax Nutrition 0.000 description 2
- 241000208343 Panax Species 0.000 description 2
- 108010076504 Protein Sorting Signals Proteins 0.000 description 2
- 240000000528 Ricinus communis Species 0.000 description 2
- 235000004443 Ricinus communis Nutrition 0.000 description 2
- 238000012300 Sequence Analysis Methods 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 244000061458 Solanum melongena Species 0.000 description 2
- 235000002597 Solanum melongena Nutrition 0.000 description 2
- 229930006000 Sucrose Natural products 0.000 description 2
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 2
- XWYUBUYQMOUFRQ-IFFSRLJSSA-N Val-Glu-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](CCC(=O)O)NC(=O)[C@H](C(C)C)N)O XWYUBUYQMOUFRQ-IFFSRLJSSA-N 0.000 description 2
- 101001071151 Vigna radiata var. radiata Phytohormone-binding protein CSBP Proteins 0.000 description 2
- 239000000370 acceptor Substances 0.000 description 2
- OJOBTAOGJIWAGB-UHFFFAOYSA-N acetosyringone Chemical compound COC1=CC(C(C)=O)=CC(OC)=C1O OJOBTAOGJIWAGB-UHFFFAOYSA-N 0.000 description 2
- 230000004913 activation Effects 0.000 description 2
- 238000012271 agricultural production Methods 0.000 description 2
- 235000001014 amino acid Nutrition 0.000 description 2
- 229960000723 ampicillin Drugs 0.000 description 2
- AVKUERGKIZMTKX-NJBDSQKTSA-N ampicillin Chemical compound C1([C@@H](N)C(=O)N[C@H]2[C@H]3SC([C@@H](N3C2=O)C(O)=O)(C)C)=CC=CC=C1 AVKUERGKIZMTKX-NJBDSQKTSA-N 0.000 description 2
- 230000033228 biological regulation Effects 0.000 description 2
- 239000000969 carrier Substances 0.000 description 2
- 239000003795 chemical substances by application Substances 0.000 description 2
- 150000001875 compounds Chemical class 0.000 description 2
- 238000007796 conventional method Methods 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 230000018109 developmental process Effects 0.000 description 2
- 230000004069 differentiation Effects 0.000 description 2
- 239000012154 double-distilled water Substances 0.000 description 2
- 238000010195 expression analysis Methods 0.000 description 2
- 230000002538 fungal effect Effects 0.000 description 2
- 238000010353 genetic engineering Methods 0.000 description 2
- 235000011187 glycerol Nutrition 0.000 description 2
- 238000011081 inoculation Methods 0.000 description 2
- 238000003780 insertion Methods 0.000 description 2
- 229930027917 kanamycin Natural products 0.000 description 2
- SBUJHOSQTJFQJX-NOAMYHISSA-N kanamycin Chemical class O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CN)O[C@@H]1O[C@H]1[C@H](O)[C@@H](O[C@@H]2[C@@H]([C@@H](N)[C@H](O)[C@@H](CO)O2)O)[C@H](N)C[C@@H]1N SBUJHOSQTJFQJX-NOAMYHISSA-N 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 230000001404 mediated effect Effects 0.000 description 2
- 229910052757 nitrogen Inorganic materials 0.000 description 2
- 210000000056 organ Anatomy 0.000 description 2
- FJKROLUGYXJWQN-UHFFFAOYSA-N papa-hydroxy-benzoic acid Natural products OC(=O)C1=CC=C(O)C=C1 FJKROLUGYXJWQN-UHFFFAOYSA-N 0.000 description 2
- 239000013641 positive control Substances 0.000 description 2
- 238000004321 preservation Methods 0.000 description 2
- 238000004064 recycling Methods 0.000 description 2
- 230000004044 response Effects 0.000 description 2
- 238000010839 reverse transcription Methods 0.000 description 2
- 229960004889 salicylic acid Drugs 0.000 description 2
- 238000000926 separation method Methods 0.000 description 2
- 239000008223 sterile water Substances 0.000 description 2
- 239000005720 sucrose Substances 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- 238000010361 transduction Methods 0.000 description 2
- 230000026683 transduction Effects 0.000 description 2
- 230000009466 transformation Effects 0.000 description 2
- 238000005406 washing Methods 0.000 description 2
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 1
- 102000040650 (ribonucleotides)n+m Human genes 0.000 description 1
- SDZRIBWEVVRDQI-CIUDSAMLSA-N Ala-Lys-Asp Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(O)=O)C(O)=O SDZRIBWEVVRDQI-CIUDSAMLSA-N 0.000 description 1
- 235000011446 Amygdalus persica Nutrition 0.000 description 1
- 241000219194 Arabidopsis Species 0.000 description 1
- 241000219195 Arabidopsis thaliana Species 0.000 description 1
- VXLBDJWTONZHJN-YUMQZZPRSA-N Asn-His-Gly Chemical compound C1=C(NC=N1)C[C@@H](C(=O)NCC(=O)O)NC(=O)[C@H](CC(=O)N)N VXLBDJWTONZHJN-YUMQZZPRSA-N 0.000 description 1
- NCFJQJRLQJEECD-NHCYSSNCSA-N Asn-Leu-Val Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](C(C)C)C(O)=O NCFJQJRLQJEECD-NHCYSSNCSA-N 0.000 description 1
- NYGILGUOUOXGMJ-YUMQZZPRSA-N Asn-Lys-Gly Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CCCCN)C(=O)NCC(O)=O NYGILGUOUOXGMJ-YUMQZZPRSA-N 0.000 description 1
- BPTFNDRZKBFMTH-DCAQKATOSA-N Asp-Met-Lys Chemical compound CSCC[C@@H](C(=O)N[C@@H](CCCCN)C(=O)O)NC(=O)[C@H](CC(=O)O)N BPTFNDRZKBFMTH-DCAQKATOSA-N 0.000 description 1
- QOJJMJKTMKNFEF-ZKWXMUAHSA-N Asp-Val-Ser Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@H](C(C)C)NC(=O)[C@@H](N)CC(O)=O QOJJMJKTMKNFEF-ZKWXMUAHSA-N 0.000 description 1
- 229930192334 Auxin Natural products 0.000 description 1
- 241000193830 Bacillus <bacterium> Species 0.000 description 1
- 229930186147 Cephalosporin Natural products 0.000 description 1
- LZZYPRNAOMGNLH-UHFFFAOYSA-M Cetrimonium bromide Chemical compound [Br-].CCCCCCCCCCCCCCCC[N+](C)(C)C LZZYPRNAOMGNLH-UHFFFAOYSA-M 0.000 description 1
- 108091006146 Channels Proteins 0.000 description 1
- 108010082495 Dietary Plant Proteins Proteins 0.000 description 1
- 208000035240 Disease Resistance Diseases 0.000 description 1
- 108010042407 Endonucleases Proteins 0.000 description 1
- 241000305071 Enterobacterales Species 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 241000588722 Escherichia Species 0.000 description 1
- 241000588724 Escherichia coli Species 0.000 description 1
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 1
- 241000221778 Fusarium fujikuroi Species 0.000 description 1
- 101710172184 Gibberellin receptor GID1 Proteins 0.000 description 1
- HPCOBEHVEHWREJ-DCAQKATOSA-N Gln-Lys-Glu Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCC(O)=O)C(O)=O HPCOBEHVEHWREJ-DCAQKATOSA-N 0.000 description 1
- PHONAZGUEGIOEM-GLLZPBPUSA-N Glu-Glu-Thr Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O PHONAZGUEGIOEM-GLLZPBPUSA-N 0.000 description 1
- CAVMESABQIKFKT-IUCAKERBSA-N Glu-Gly-His Chemical compound C1=C(NC=N1)C[C@@H](C(=O)O)NC(=O)CNC(=O)[C@H](CCC(=O)O)N CAVMESABQIKFKT-IUCAKERBSA-N 0.000 description 1
- PJBVXVBTTFZPHJ-GUBZILKMSA-N Glu-Leu-Asp Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)O)NC(=O)[C@H](CCC(=O)O)N PJBVXVBTTFZPHJ-GUBZILKMSA-N 0.000 description 1
- IVGJYOOGJLFKQE-AVGNSLFASA-N Glu-Leu-Lys Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CCCCN)C(=O)O)NC(=O)[C@H](CCC(=O)O)N IVGJYOOGJLFKQE-AVGNSLFASA-N 0.000 description 1
- IDEODOAVGCMUQV-GUBZILKMSA-N Glu-Ser-Leu Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(O)=O IDEODOAVGCMUQV-GUBZILKMSA-N 0.000 description 1
- VIPDPMHGICREIS-GVXVVHGQSA-N Glu-Val-Leu Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC(C)C)C(O)=O VIPDPMHGICREIS-GVXVVHGQSA-N 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- VSVZIEVNUYDAFR-YUMQZZPRSA-N Gly-Ala-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](C)NC(=O)CN VSVZIEVNUYDAFR-YUMQZZPRSA-N 0.000 description 1
- FSPVILZGHUJOHS-QWRGUYRKSA-N Gly-His-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@@H](NC(=O)CN)CC1=CNC=N1 FSPVILZGHUJOHS-QWRGUYRKSA-N 0.000 description 1
- CCBIBMKQNXHNIN-ZETCQYMHSA-N Gly-Leu-Gly Chemical compound NCC(=O)N[C@@H](CC(C)C)C(=O)NCC(O)=O CCBIBMKQNXHNIN-ZETCQYMHSA-N 0.000 description 1
- 239000004471 Glycine Substances 0.000 description 1
- JBCLFWXMTIKCCB-UHFFFAOYSA-N H-Gly-Phe-OH Natural products NCC(=O)NC(C(O)=O)CC1=CC=CC=C1 JBCLFWXMTIKCCB-UHFFFAOYSA-N 0.000 description 1
- YIGCZZKZFMNSIU-RWMBFGLXSA-N His-Met-Pro Chemical compound CSCC[C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](CC2=CN=CN2)N YIGCZZKZFMNSIU-RWMBFGLXSA-N 0.000 description 1
- 206010020649 Hyperkeratosis Diseases 0.000 description 1
- YPQDTQJBOFOTJQ-SXTJYALSSA-N Ile-Asn-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)N[C@@H]([C@@H](C)CC)C(=O)O)N YPQDTQJBOFOTJQ-SXTJYALSSA-N 0.000 description 1
- WZDCVAWMBUNDDY-KBIXCLLPSA-N Ile-Glu-Ala Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCC(=O)O)C(=O)N[C@@H](C)C(=O)O)N WZDCVAWMBUNDDY-KBIXCLLPSA-N 0.000 description 1
- MTFVYKQRLXYAQN-LAEOZQHASA-N Ile-Glu-Gly Chemical compound [H]N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CCC(O)=O)C(=O)NCC(O)=O MTFVYKQRLXYAQN-LAEOZQHASA-N 0.000 description 1
- UWLHDGMRWXHFFY-HPCHECBXSA-N Ile-Ile-Pro Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H]([C@@H](C)CC)C(=O)N1CCC[C@@H]1C(=O)O)N UWLHDGMRWXHFFY-HPCHECBXSA-N 0.000 description 1
- RFMDODRWJZHZCR-BJDJZHNGSA-N Ile-Lys-Cys Chemical compound CC[C@H](C)[C@H](N)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CS)C(O)=O RFMDODRWJZHZCR-BJDJZHNGSA-N 0.000 description 1
- 241000880493 Leptailurus serval Species 0.000 description 1
- XIRYQRLFHWWWTC-QEJZJMRPSA-N Leu-Ala-Phe Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@H](C(O)=O)CC1=CC=CC=C1 XIRYQRLFHWWWTC-QEJZJMRPSA-N 0.000 description 1
- QDSKNVXKLPQNOJ-GVXVVHGQSA-N Leu-Gln-Val Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](C(C)C)C(O)=O QDSKNVXKLPQNOJ-GVXVVHGQSA-N 0.000 description 1
- QJXHMYMRGDOHRU-NHCYSSNCSA-N Leu-Ile-Gly Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)CC)C(=O)NCC(O)=O QJXHMYMRGDOHRU-NHCYSSNCSA-N 0.000 description 1
- TZSUCEBCSBUMDP-SRVKXCTJSA-N Leu-Leu-Ala Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](C)C(O)=O TZSUCEBCSBUMDP-SRVKXCTJSA-N 0.000 description 1
- PJWOOBTYQNNRBF-BZSNNMDCSA-N Leu-Phe-Lys Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CCCCN)C(=O)O)N PJWOOBTYQNNRBF-BZSNNMDCSA-N 0.000 description 1
- 244000045959 Lupinus luteus Species 0.000 description 1
- 235000010648 Lupinus luteus Nutrition 0.000 description 1
- HKCCVDWHHTVVPN-CIUDSAMLSA-N Lys-Asp-Ala Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](C)C(O)=O HKCCVDWHHTVVPN-CIUDSAMLSA-N 0.000 description 1
- WAIHHELKYSFIQN-XUXIUFHCSA-N Lys-Ile-Val Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](C(C)C)C(O)=O WAIHHELKYSFIQN-XUXIUFHCSA-N 0.000 description 1
- VHTOGMKQXXJOHG-RHYQMDGZSA-N Lys-Thr-Val Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C(C)C)C(O)=O VHTOGMKQXXJOHG-RHYQMDGZSA-N 0.000 description 1
- GILLQRYAWOMHED-DCAQKATOSA-N Lys-Val-Ser Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@H](C(C)C)NC(=O)[C@@H](N)CCCCN GILLQRYAWOMHED-DCAQKATOSA-N 0.000 description 1
- IHRFZLQEQVHXFA-RHYQMDGZSA-N Met-Thr-Lys Chemical compound CSCC[C@H](N)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@H](C(O)=O)CCCCN IHRFZLQEQVHXFA-RHYQMDGZSA-N 0.000 description 1
- 240000007594 Oryza sativa Species 0.000 description 1
- 235000007164 Oryza sativa Nutrition 0.000 description 1
- 101710096342 Pathogenesis-related protein Proteins 0.000 description 1
- 101710097922 Pathogenesis-related protein 10 Proteins 0.000 description 1
- KAGCQPSEVAETCA-JYJNAYRXSA-N Phe-Gln-Leu Chemical compound CC(C)C[C@@H](C(=O)O)NC(=O)[C@H](CCC(=O)N)NC(=O)[C@H](CC1=CC=CC=C1)N KAGCQPSEVAETCA-JYJNAYRXSA-N 0.000 description 1
- JEBWZLWTRPZQRX-QWRGUYRKSA-N Phe-Gly-Asp Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)NCC(=O)N[C@@H](CC(O)=O)C(O)=O JEBWZLWTRPZQRX-QWRGUYRKSA-N 0.000 description 1
- BIYWZVCPZIFGPY-QWRGUYRKSA-N Phe-Gly-Ser Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)NCC(=O)N[C@@H](CO)C(O)=O BIYWZVCPZIFGPY-QWRGUYRKSA-N 0.000 description 1
- 101710192310 Phytohormone-binding protein Proteins 0.000 description 1
- 235000006040 Prunus persica var persica Nutrition 0.000 description 1
- 108010092799 RNA-directed DNA polymerase Proteins 0.000 description 1
- 108020004511 Recombinant DNA Proteins 0.000 description 1
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical class [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 description 1
- 244000061456 Solanum tuberosum Species 0.000 description 1
- 235000002595 Solanum tuberosum Nutrition 0.000 description 1
- 102100025292 Stress-induced-phosphoprotein 1 Human genes 0.000 description 1
- 240000001705 Striga asiatica Species 0.000 description 1
- PXQUBKWZENPDGE-CIQUZCHMSA-N Thr-Ala-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)O)NC(=O)[C@H](C)NC(=O)[C@H]([C@@H](C)O)N PXQUBKWZENPDGE-CIQUZCHMSA-N 0.000 description 1
- KGKWKSSSQGGYAU-SUSMZKCASA-N Thr-Gln-Thr Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CCC(=O)N)C(=O)N[C@@H]([C@@H](C)O)C(=O)O)N)O KGKWKSSSQGGYAU-SUSMZKCASA-N 0.000 description 1
- LHEZGZQRLDBSRR-WDCWCFNPSA-N Thr-Glu-Leu Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(O)=O LHEZGZQRLDBSRR-WDCWCFNPSA-N 0.000 description 1
- KZSYAEWQMJEGRZ-RHYQMDGZSA-N Thr-Leu-Val Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](C(C)C)C(O)=O KZSYAEWQMJEGRZ-RHYQMDGZSA-N 0.000 description 1
- XSEPSRUDSPHMPX-KATARQTJSA-N Thr-Lys-Ser Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CO)C(O)=O XSEPSRUDSPHMPX-KATARQTJSA-N 0.000 description 1
- HUPLKEHTTQBXSC-YJRXYDGGSA-N Thr-Ser-Tyr Chemical compound C[C@@H](O)[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@H](C(O)=O)CC1=CC=C(O)C=C1 HUPLKEHTTQBXSC-YJRXYDGGSA-N 0.000 description 1
- SPIFGZFZMVLPHN-UNQGMJICSA-N Thr-Val-Phe Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O SPIFGZFZMVLPHN-UNQGMJICSA-N 0.000 description 1
- KHCSOLAHNLOXJR-BZSNNMDCSA-N Tyr-Leu-Leu Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(C)C)C(O)=O KHCSOLAHNLOXJR-BZSNNMDCSA-N 0.000 description 1
- 108091023045 Untranslated Region Proteins 0.000 description 1
- PMDOQZFYGWZSTK-LSJOCFKGSA-N Val-Gly-Ile Chemical compound CC[C@H](C)[C@@H](C(O)=O)NC(=O)CNC(=O)[C@@H](N)C(C)C PMDOQZFYGWZSTK-LSJOCFKGSA-N 0.000 description 1
- ZHQWPWQNVRCXAX-XQQFMLRXSA-N Val-Leu-Pro Chemical compound CC(C)C[C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](C(C)C)N ZHQWPWQNVRCXAX-XQQFMLRXSA-N 0.000 description 1
- RFKJNTRMXGCKFE-FHWLQOOXSA-N Val-Leu-Trp Chemical compound C1=CC=C2C(C[C@H](NC(=O)[C@@H](NC(=O)[C@@H](N)C(C)C)CC(C)C)C(O)=O)=CNC2=C1 RFKJNTRMXGCKFE-FHWLQOOXSA-N 0.000 description 1
- PFMSJVIPEZMKSC-DZKIICNBSA-N Val-Tyr-Glu Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC1=CC=C(C=C1)O)C(=O)N[C@@H](CCC(=O)O)C(=O)O)N PFMSJVIPEZMKSC-DZKIICNBSA-N 0.000 description 1
- 241000607479 Yersinia pestis Species 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 230000032683 aging Effects 0.000 description 1
- 108010005233 alanylglutamic acid Proteins 0.000 description 1
- 208000026935 allergic disease Diseases 0.000 description 1
- 230000003321 amplification Effects 0.000 description 1
- 230000036592 analgesia Effects 0.000 description 1
- 230000019552 anatomical structure morphogenesis Effects 0.000 description 1
- 230000003712 anti-aging effect Effects 0.000 description 1
- 230000003110 anti-inflammatory effect Effects 0.000 description 1
- 108010038633 aspartylglutamate Proteins 0.000 description 1
- 239000002363 auxin Substances 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- 230000003385 bacteriostatic effect Effects 0.000 description 1
- 238000009412 basement excavation Methods 0.000 description 1
- 230000027455 binding Effects 0.000 description 1
- 230000003115 biocidal effect Effects 0.000 description 1
- 150000001647 brassinosteroids Chemical class 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- 229960004261 cefotaxime Drugs 0.000 description 1
- GPRBEKHLDVQUJE-VINNURBNSA-N cefotaxime Chemical compound N([C@@H]1C(N2C(=C(COC(C)=O)CS[C@@H]21)C(O)=O)=O)C(=O)/C(=N/OC)C1=CSC(N)=N1 GPRBEKHLDVQUJE-VINNURBNSA-N 0.000 description 1
- 230000034303 cell budding Effects 0.000 description 1
- 230000003915 cell function Effects 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 229940124587 cephalosporin Drugs 0.000 description 1
- 150000001780 cephalosporins Chemical class 0.000 description 1
- 235000013339 cereals Nutrition 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- RCTYPNKXASFOBE-UHFFFAOYSA-M chloromercury Chemical compound [Hg]Cl RCTYPNKXASFOBE-UHFFFAOYSA-M 0.000 description 1
- 230000004087 circulation Effects 0.000 description 1
- 238000010367 cloning Methods 0.000 description 1
- 238000003501 co-culture Methods 0.000 description 1
- 238000003967 crop rotation Methods 0.000 description 1
- 238000012866 crystallographic experiment Methods 0.000 description 1
- 125000000151 cysteine group Chemical group N[C@@H](CS)C(=O)* 0.000 description 1
- 239000004062 cytokinin Substances 0.000 description 1
- UQHKFADEQIVWID-UHFFFAOYSA-N cytokinin Natural products C1=NC=2C(NCC=C(CO)C)=NC=NC=2N1C1CC(O)C(CO)O1 UQHKFADEQIVWID-UHFFFAOYSA-N 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 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 1
- 238000001514 detection method Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 229930004069 diterpene Natural products 0.000 description 1
- 125000000567 diterpene group Chemical group 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 230000005611 electricity Effects 0.000 description 1
- 238000001962 electrophoresis Methods 0.000 description 1
- 238000003912 environmental pollution Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 230000006870 function Effects 0.000 description 1
- 244000053095 fungal pathogen Species 0.000 description 1
- 108010006664 gamma-glutamyl-glycyl-glycine Proteins 0.000 description 1
- 239000007789 gas Substances 0.000 description 1
- 239000000499 gel Substances 0.000 description 1
- 230000002068 genetic effect Effects 0.000 description 1
- 230000035784 germination Effects 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 108010033719 glycyl-histidyl-glycine Proteins 0.000 description 1
- 108010089804 glycyl-threonine Proteins 0.000 description 1
- 108010050848 glycylleucine Proteins 0.000 description 1
- 239000003630 growth substance Substances 0.000 description 1
- YQOKLYTXVFAUCW-UHFFFAOYSA-N guanidine;isothiocyanic acid Chemical compound N=C=S.NC(N)=N YQOKLYTXVFAUCW-UHFFFAOYSA-N 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 230000002209 hydrophobic effect Effects 0.000 description 1
- 230000009610 hypersensitivity Effects 0.000 description 1
- 230000036737 immune function Effects 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- SEOVTRFCIGRIMH-UHFFFAOYSA-N indole-3-acetic acid Chemical compound C1=CC=C2C(CC(=O)O)=CNC2=C1 SEOVTRFCIGRIMH-UHFFFAOYSA-N 0.000 description 1
- 230000006698 induction Effects 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 230000003834 intracellular effect Effects 0.000 description 1
- 108010078274 isoleucylvaline Proteins 0.000 description 1
- 239000010977 jade Substances 0.000 description 1
- -1 jasmonic Chemical compound 0.000 description 1
- 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 1
- 229960000318 kanamycin Drugs 0.000 description 1
- 229930182823 kanamycin A Natural products 0.000 description 1
- 101150044508 key gene Proteins 0.000 description 1
- 150000002596 lactones Chemical class 0.000 description 1
- 238000011031 large-scale manufacturing process Methods 0.000 description 1
- 108010076756 leucyl-alanyl-phenylalanine Proteins 0.000 description 1
- 239000003446 ligand Substances 0.000 description 1
- 230000000670 limiting effect Effects 0.000 description 1
- 238000009630 liquid culture Methods 0.000 description 1
- 108010038320 lysylphenylalanine Proteins 0.000 description 1
- 108010017391 lysylvaline Proteins 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 239000000178 monomer Substances 0.000 description 1
- 239000004570 mortar (masonry) Substances 0.000 description 1
- 230000035772 mutation Effects 0.000 description 1
- 230000009125 negative feedback regulation Effects 0.000 description 1
- 239000002777 nucleoside Substances 0.000 description 1
- 125000003835 nucleoside group Chemical group 0.000 description 1
- 238000011017 operating method Methods 0.000 description 1
- 230000036961 partial effect Effects 0.000 description 1
- 244000052769 pathogen Species 0.000 description 1
- 230000001717 pathogenic effect Effects 0.000 description 1
- 239000008188 pellet Substances 0.000 description 1
- 108010073101 phenylalanylleucine Proteins 0.000 description 1
- 230000035479 physiological effects, processes and functions Effects 0.000 description 1
- 229930195732 phytohormone Natural products 0.000 description 1
- 230000008635 plant growth Effects 0.000 description 1
- 229920001184 polypeptide Polymers 0.000 description 1
- 239000000843 powder Substances 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
- 108010053725 prolylvaline Proteins 0.000 description 1
- 238000000751 protein extraction Methods 0.000 description 1
- 239000001397 quillaja saponaria molina bark Substances 0.000 description 1
- 108020003175 receptors Proteins 0.000 description 1
- 238000011084 recovery Methods 0.000 description 1
- 230000008929 regeneration Effects 0.000 description 1
- 238000011069 regeneration method Methods 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 230000002441 reversible effect Effects 0.000 description 1
- 235000009566 rice Nutrition 0.000 description 1
- 229930182490 saponin Natural products 0.000 description 1
- 150000007949 saponins Chemical class 0.000 description 1
- 230000008117 seed development Effects 0.000 description 1
- 238000012163 sequencing technique Methods 0.000 description 1
- 230000019491 signal transduction Effects 0.000 description 1
- 235000011091 sodium acetates Nutrition 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 230000009870 specific binding Effects 0.000 description 1
- 230000003068 static effect Effects 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 230000021918 systemic acquired resistance Effects 0.000 description 1
- 108010031491 threonyl-lysyl-glutamic acid Proteins 0.000 description 1
- 238000013518 transcription Methods 0.000 description 1
- 230000035897 transcription Effects 0.000 description 1
- 108010038745 tryptophylglycine Proteins 0.000 description 1
- 108010051110 tyrosyl-lysine Proteins 0.000 description 1
- 230000034512 ubiquitination Effects 0.000 description 1
- 238000010798 ubiquitination Methods 0.000 description 1
- 238000011144 upstream manufacturing Methods 0.000 description 1
- 108010073969 valyllysine Proteins 0.000 description 1
- 238000012795 verification Methods 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
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
- C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
- C12N15/82—Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
- C12N15/8241—Phenotypically and genetically modified plants via recombinant DNA technology
- C12N15/8261—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield
- C12N15/8271—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield for stress resistance, e.g. heavy metal resistance
- C12N15/8279—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield for stress resistance, e.g. heavy metal resistance for biotic stress resistance, pathogen resistance, disease resistance
- C12N15/8282—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield for stress resistance, e.g. heavy metal resistance for biotic stress resistance, pathogen resistance, disease resistance for fungal resistance
Landscapes
- Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Molecular Biology (AREA)
- Biophysics (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Zoology (AREA)
- Biochemistry (AREA)
- Wood Science & Technology (AREA)
- General Health & Medical Sciences (AREA)
- General Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Biomedical Technology (AREA)
- Botany (AREA)
- Physics & Mathematics (AREA)
- Cell Biology (AREA)
- Plant Pathology (AREA)
- Gastroenterology & Hepatology (AREA)
- Microbiology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Medicinal Chemistry (AREA)
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
Abstract
The invention discloses a kind of pseudo-ginseng plant hormone binding-protein genePnPhBP1, its nucleotide sequence such as SEQ ID NO：Shown in 1, coding such as SEQ ID NO：The protein of amino acid sequence shown in 2, the present invention are confirmed by functional genomics relation technological researchingPnPhBP1Gene has the function of that raising plant is antimycotic, will be of the invention antimycoticPnPhBP1It is gene constructed on plant expression vector and overexpression in tobacco is transferred to, as a result transgenic tobacco plant has very strong extracorporeal antifungal activity, experimental result display overexpressionPnPhBP1Growth of the transgene tobacco to three kinds of fungies such as colletotrichum gloeosporioides Penz, Botrytis cinerea, Fusarium solani there is obvious inhibitory action.
Description
Technical field
The present invention relates to molecular biology and genetic engineering relation technological researching field, a kind of particularly pseudo-ginseng plant swashs
Plain binding-protein genePnPhBP1And application.
Background technology
Plant disease is very a stubborn problem, especially fungal disease in agricultural production, and it is always sick to account for plant
Harmful 80%, seriously affects the yield and quality of crops.Traditional pest control method achieves certain effect, first, relying on
Traditional breeding way cultivates resistant variety, second, the use of chemical pesticide, third, taking the cropping systems such as crop rotation.However, these
Method all there are it is more or less the drawbacks of, such as cycle that resistant variety is cultivated is long, chemical pesticide residual is high and easily causes environment
Pollution, cropping system adjustment are then time-consuming and laborious, so the method for Traditional control plant disease cannot be solved the problems, such as thoroughly.With
The foundation and development of recombinant DNA technology, first-stage success has been obtained using technique for gene engineering to cultivate disease-resistant plants new varieties, and
It is expected to fundamentally solve the problems, such as fungal disease.
For plant by the defence process that disease-resistant gene mediates there are a series of Physiology and biochemistries and molecular biology reaction, these are anti-
Should be since the hypersensitivity (Hypersensitive Response, HR) pathogen infection point, and extend to distant organs
System resistant or acquired resistance (Systemic Acquired Resistance, SAR), are limited by the tune of signaling transduction network
Control.Plant hormone（phytohormones）The signal system of wide participation defense responses, such as salicylic acid, jasmonic, ethene
Deng being amplified by the expression and signal that regulate and control key gene, finally induce the expression and metabolism of a series of Defense response genes
Change and produce resistance.Plant hormone is the chemical messenger for coordinating many cell functions, including（But it is not limited to）Ten main
Species：Auximone, the basic element of cell division, gibberellin, abscisic acid, Brassinosteroids, ethene, jasmonate, polypeptide swash
Element, salicylic acid and witchweed lactone (Santner A, Calderon-Villalobos L I, Estelle M. Plant
hormones are versatile chemical regulators of plant growth. Nat Chem Biol.
2009, 5: 301-307).In the signaling transduction network of plant hormone mediation, various plants hormonebinding protein
(phytohormone-binding proteins, PhBP) also plays an important role.
Gibberellin（Such as gibberellic acid）It is diterpene Fourth Ring or five rings growth regulator, induction seed development, sprouting, organ extend
With (Yamaguchi S. Gibberellin metabolism and its regulation. Annu Rev Plant of blooming
Biol. 2008, 59: 225-251).Gibberellin is first red mould（Gibberella fujikuroi）In be found, it is a kind of
The fungal pathogens of rice, cause the racking of stem and ultimately result in plant and wither and dead.It is red mould that plant produces endogenous
Element, the level of its intracellular is adjusted by negative feedback loop, in addition, the concentration of auxin and ethene also adjusts the water of cell inner gibberellin
It is flat（Fleet C M, Sun T P. A DELLAcate balance: the role of gibberellin in plant
morphogenesis. Curr Opin Plant Biol. 2005, 8: 77-85; Yamaguchi S. Gibberellin
metabolism and its regulation. Annu Rev Plant Biol. 2008, 59: 225-251）.Gibberellin
Acceptor is referred to as insensitive 1 albumen of short stem of gibberellin（GID1）,gid1The afunction mutation of gene causes plant to downgrade (Peng
J, Richards D E, Hartley N M, Murphy G P, Devos K M, Flintham J E, Beales J,
Fish L J, Worland A J, Pelica F, Sudhakar D, Christou P, Snape J W, Gale M D,
Harberd N P. ‘Green revolution’ genes encode mutant gibberellins response
modulators. Nature. 1999, 400: 256-261).Murase et al. is to arabidopsis（Arabidopsis thaliana）The compound of GID1 and gibberellin studied in structure (Murase K, Hirano Y, Sun T P,
Hakoshima T. Gibberellin-induced DELLA recognition by the gibberellin
receptor GID1. Nature. 2008, 456: 459-463).GID1 acceptors can be combined with conservative Asp-Glu-
The DELLA albumen of Leu-Leu-Ala N- end sequences, they are the negative-feedback regu- lation agent of gibberellin reaction
（Schwechheimer C. Understanding gibberellic acid signaling--are we there yet.
Curr Opin Plant Biol. 2008, 11: 9-15; Schwechheimer C, Willige B C. Shedding
light on gibberellic acid signaling. Curr Opin Plant Biol. 2009, 12: 57-62）.
The gibberellin combined with GID1 starts the formation of GID1-DELLA compounds, and DELLA albumen cannot be re-used as gibberellin dependence
The transcription inhibitory factor of gene, but by ubiquitination and be degraded.
Pathogenesis-related proteins 10（pathogenesis related protein 10）Very little（Highest only 19 kDa）, lead to
Be often monomer, subacidity kytoplasm plant specific proteins (Fernandes H, Michalska K, Sikorski M,
Jaskolski M. Structural and functional aspects of PR-10 proteins. FEBS J.
2013, 280: 1169-1199).The structural conservation of PR10 albumen is established well, has typical PR10 foldings
It is folded.The most prominent features that PR10 is folded are to form big hydrophobic cavity between spiral α 3 and beta sheet, it is clear that are PR-10 ligands
Binding site.Crystallographic Study confirms possibility (the Fernandes H, Bujacz that PR10 albumen is combined with the basic element of cell division
A, Bujacz G, Jelen F, Jasinski M, Kachlicki P, Otlewski J, Sikorski M M,
Jaskolski M. Cytokinin-induced structural adaptability of a Lupinus luteus
PR-10 protein. FEBS J. 2009, 276: 1596-1609).The very low PR10 of some sequence similarities is accredited as
Basic element of cell division binding proteins specific (Cytokinin-Specific Binding Proteins, CSBP) subfamily, this
Outside, two CSBP MtCSBP and VrCSBP are proved also to be combined with gibberellin（Ruszkowski M, Sliwiak J,
Ciesielska A, Barciszewski J, Sikorski M, Jaskolski M. Specific binding of
gibberellic acid by cytokinin-specific binding proteins: a new aspect of
plant hormone-binding proteins with the PR-10 fold. Acta Crystallographica.
2014, 70(Pt 7): 2032-2041）.The studies above show PR10 be not only with basic element of cell division specific bond so that
Ruszkowski et al. suggests with this term of PhBP substitution CSBP.
Pseudo-ginseng [Panax notoginseng(Burk) F.H. Chen] it is Araliaceae (Araliaceae) Panax
(Panax) herbaceous plant, also known as pseudo-ginseng and invaluable, it is the important traditional rare traditional Chinese medicine in China, most early in Southwestern China portion
The ethnic group in area（Strong, seedling, precious jade, the Yi nationality, distributed over Yunnan, Sichuan and Guizhou）Middle application, then with the exchange between various nationalities and army, the propagation of merchants,
Just gradually it is passed to central plain area.Effective active composition in pseudo-ginseng is mainly saponin(e, has Repercusion analgesia, anti-inflammatory, anti-aging
Effect, can adjust immune function of human body, dispelling fatigue, slows down aging, arasaponin can also be effectively improved memory, be to go through
The long medicine-food two-purpose natural resources of history.Although going up year by year to the demand of pseudo-ginseng, the yield of pseudo-ginseng is without significantly
Rise, and the Panax notoginseng Growth cycle is long, multiple disease is one of the main reasons.The resistant heredity breeding research weak foundation of pseudo-ginseng,
Have become the limiting factor of pseudo-ginseng industry sound development.The molecular level mechanism of disease resistance of pseudo-ginseng is furtherd investigate, is especially disclosed disease-resistant
The signal transduction and the disease-resistant functional gene of excavation of defense response, it will help pseudo-ginseng resistant heredity breeding work carries steadily
Rise.
The content of the invention
The object of the present invention is to provide the full-length gene that a kind of clone from pseudo-ginseng obtains coded plant hormonebinding proteinPnPhBP1, plant hormone binding-protein genePnPhBP1Nucleotide sequence such as SEQ ID NO：Shown in 1, the gene cDNA total length
Sequence is 623bp, 5 ' non-translational regions of open reading frame, 56bp comprising a 465bp, the 3 ' non-translational regions of 102bp, coding
Such as SEQ ID NO：The protein of amino acid sequence shown in 2.
In the present inventionPnPhBP1The code area of gene is sequence table SEQ ID NO：Nucleosides in 1 shown in 57-521
Acid sequence.
The global cDNA fragment of an antimycotic related gene for present invention separation clone pseudo-ginseng, utilizes Agrobacterium tumefaciems
(Agrobacterium tumefaciens) target gene is transferred in recipient plant and overexpression by mediation, by further
Whether the experimental verification gene has fungal resistance, and nosomycosis is resisted for the later-stage utilization improvement of genes tobacco and other plant
Harmful ability lays the foundation, this unnamed gene is by inventorPnPhBP1。
The present invention relates to separation to includePnPhBP1DNA fragmentation and identify its function, wherein the DNA fragmentation such as sequence
Table SEQ ID NO：Shown in 1, sequence analysis is carried out to the gene, is foundPnPhBP1Full-length cDNA is 623 bp, includes one
The open reading frame (ORF) of 465 bp, the 5 ' non-translational regions (untranslated region, UTR) of 56bp, the 3 ' of 102 bp
UTR, wherein ORF encode a protein with 154 amino acid.PnPhBP1The protein of coding has PR10 albumen
Conserved motifs glycine-rich loop, BLASTp retrieval results show PnPhBP1 and carrot (Daucus carota), peach
(Prunus persica), macleaya cordata (Macleaya cordata) and castor-oil plant (Ricinus communis) plant hormone knot
The similitude of hop protein is respectively 76%, 66%, 62% and 61%, shows that it belongs to the plant hormone associated proteins in pseudo-ginseng.Super table
Up to sequence table SEQ ID NO：Protein shown in 1 can strengthen tobacco to Botrytis cinerea (Botrytis cinerea), eggplant it is rotten
Sickle-like bacteria (Fusarium solani), colletotrichum gloeosporioides Penz (Colletotrichum gloeosporioides) resistance.
The present invention is by pseudo-ginseng plant hormone binding-protein genePnPhBP1Apply and improving tobacco to Botrytis cinerea, eggplant corruption
In sickle-like bacteria, colletotrichum gloeosporioides Penz resistance, concrete operations are as follows：
（1）Using amplificationPnPhBP1Special primer, total serum IgE is extracted from Roots of Panax Notoginseng, it is anti-by Transcription-Polymerase Chain formula
(reverse transcription-polymerase chain reaction, RT-PCR) is answered to amplifyPnPhBP1Coding
Area, is subsequently attached on pMD-18T carriers, and the clone with target gene is obtained through sequencing；
（2）Use restriction enzymePstI andEcoRI digestions pMD-18T-PnPhBP1Carrier, purpose base is obtained by glue reclaim
Because of fragment, with same endonuclease digestion plant expression vector pCAMBIA2300s, glue reclaim obtain needed for carrier large fragment, then
It will be obtainedPnPhBP1Genetic fragment is connected with pCAMBIA2300s fragments, builds plant overexpression vector, afterwards by constructed by
Recombinant vector expressed by Agrobacterium tumefaciens mediated be transferred in tobacco；
（3）Transformant is screened with the resistance marker having on recombinant vector T-DNA, and detects to obtain by PCR and RT-PCR
Real transfer-gen plant, the inhibitory activity that analysis genetically modified plants albumen grows fungi, finally filters out to fungus resistant
The transfer-gen plant being remarkably reinforced.
The present invention provides a kind of new method to improve the resistance of plant against fungal disease, is trained by genetic engineering means
Traditional breeding method can be overcome the shortcomings of by educating disease-resistant plants, and not only breeding cycle shortens, but also easy to operate, is readily available highly resistance material
Material.The present invention is from pseudo-ginsengPnPhBP1Gene can strengthen the resistance of plant against fungal, can be with by the channel genes tobacco
Produce new varieties and new material with fungus resistant.Resistance plant kind and material are cultivated with bright using technique for gene engineering
Aobvious advantage and the importance do not replaced.It can be not only that the offers such as large-scale production crop, flowers are convenient, a large amount of to reduce
The use of chemical pesticide, can also be that agricultural production is cost-effective, reduce environmental pollution, therefore the present invention has a vast market
Application prospect.
Brief description of the drawings
Fig. 1 is the present inventionPnPhBP1The PCR testing result schematic diagrames of transgene tobacco genomic DNA, in figure：Marker
For DL2000 DNA Marker (the precious biology in Dalian)；Positive control is plasmid pMD-18T-PnPhBP1Production is tied for the PCR of template
Thing；WT is the product that non-transgenic tobacco (wild type) STb gene is template PCR；
Fig. 2 is of the invention positivePnPhBP1In transgene tobaccoPnPhBP1The expression analysis result figure of transcriptional level；In figure：
Marker is DL2000 DNA Marker (the precious biology in Dalian)；WT is that non-transgenic tobacco total serum IgE reverse transcription cDNA is template
PCR product；Positive control is plasmid pMD-18T-PnPhBP1For the PCR product of template；
Fig. 3 is the present inventionPnPhBP1Transgene tobacco In Vitro Bacteriostatic design sketch；A, b, c are Botrytis cinerea, glue respectively in figure
Spore anthrax-bacilus, Fusarium solani；WT is the total protein of wild-type tobacco；Buffer is blank control, i.e., (is used for without protein control
Extract the buffer solution of albumen).
Embodiment
The present invention is described in further detail below by drawings and examples, but the scope of the present invention is not limited to
The content, method is conventional method unless otherwise specified in embodiment, and the reagent used is routine unless otherwise specified
Commercial reagent or the reagent prepared according to a conventional method.
Embodiment 1：PnPhBP1Full-length gene is cloned and sequence analysis
Pseudo-ginseng is inoculated with Fusarium solani, total serum IgE is extracted with the root after 24 h of inoculation, is ground the Roots of Panax Notoginseng after inoculation with liquid nitrogen
Into powder, then it is transferred in centrifuge tube, total serum IgE is extracted using guanidine isothiocyanate method, using reverse transcriptase M-MLV (promega)
It is by the first chains of templated synthesis cDNA, reaction system and operating process of total serum IgE：5 μ g total serum IgEs are taken, sequentially add 50 ng
oligo（dT）、2 μL dNTP（2.5mM each）, DEPC water to reaction volume be 14.5 μ L；After mixing, 70 DEG C of heat denatureds
After 5 min 4 μ L 5 × First-stand buffer, 0.5 μ L are then sequentially added in 5 min of cooled on ice rapidly
RNasin（200U）、1 μL M-MLV（200U）, mix and centrifuge in short-term, 42 DEG C of 1.5 h of warm bath, 70 DEG C of heating 10 after taking-up
Min, terminates reaction.The synthesis of the first chains of cDNA is placed on -20 DEG C and saves backup.
Using the first chain cDNA of synthesis as template, amplifying target genesPnPhBP1, upstream and downstream primer sequence used is respectivelyAnd。
Using AdvantageTM2 PCR Enzyme（Clontech）Amplify target gene；PCR reaction conditions：95℃ 1 min；94
DEG C 30 s, 61 DEG C of 30 s, 72 DEG C of 40 s, 30 circulations；72℃ 5 min；Reaction system（10 μL）For 1 μ L cDNA, 1
μL 10×Advantage 2 PCR Buffer、0.5 μL 50×dNTP Mix （10mM each）, 0.2 μ L forward primers
（10 μM）, 0.2 μ L reverse primers（10 μM）、0.2 μL Advantage 2 PCR Polymerase Mix、6.9 μL
PCR-Grade water；After PCR, take 5 μ L to be used for agarose gel electrophoresis, with detect the specificity of amplified production and
Size.
Resulting PCR product only has a DNA band, therefore directly carries out TA clones to PCR product, and the kit used is
pMD18-T vector kit（The precious biology in Dalian）, reaction system and operating process are：1.5 μ L PCR products are taken, are sequentially added
1 μL pMD18-T vector（50 ng/μL）With 2.5 μ L 2 × Ligation solution I, mixing is placed on 16 DEG C of mistakes
Night reacts.Connection product is transferred in bacillus coli DH 5 alpha using heat-shock transformed method.Using containing ampicillin
（Ampicillin, Amp）LB solid medium screening positive clones, select several single bacterium colonies, shake after bacterium with amplificationPnPhBP1Special primer identify multiple cloning sites insertionPnPhBP1Clone, the clone identified is sequenced, most
Obtain eventuallyPnPhBP1Full-length cDNA is 623bp, passes through NCBI ORF finder（http://
www.ncbi.nlm.nih.gov/gorf/gorf.html）Analysis finds that it includes the opening code-reading frame of a 465bp（See sequence
List）,PnPhBP1One protein PnPhBP1 containing 154 amino acid of coding, its molecular weight is about 16.99 KDa, waits electricity
Point about 5.08, contains 1 cysteine residues.Analyzed by bioinformatics software SignalP 4.1PnPhBP1Coding
Protein sequence, detects whether it has N-terminal signal peptide.The result is shown inPnPhBP1In do not detect the presence of signal peptide, table
Bright PnPhBP1 is a kind of non-secreted protein.
Embodiment 2：Plant overexpression vector is built
Using a small amount of extraction agent boxes of SanPrep pillar Plasmid DNA（Give birth to work in Shanghai）Extraction insertionPnPhBP1Escherichia coli matter
Grain pMD-18T-PnPhBP1And the plasmid of plant expression vector pCAMBIA2300s, take 1 μ L to be used for agarose gel electrophoresis
With detection extraction plasmid integrality and concentration level；Use restriction enzymePstI（TaKaRa）WithEcoRI（TaKaRa）
Respectively to plasmid pMD-18T-PnPhBP1Double digestion is carried out with pCAMBIA2300s（100 μ L systems）, reaction system and operation
Process is：Take 20 μ L pMD-18T-PnPhBP1With pCAMBIA2300s plasmids, sequentially add 10 μ L 10 × K buffer, 4
μLPstI、6 μLEcoRI、60 μL ddH2O, centrifuges in short-term after mixing, is placed in 37 DEG C of reaction overnights；By all digestion products points
Electrophoresis is carried out in Ago-Gel, it is then rightPnPhBP1Fragment and pCAMBIA2300s carriers large fragment carry out glue and return respectively
Receive；Take 1 μ L recovery products to detect the size and concentration of recycling fragment by agarose gel electrophoresis, it is standby to be placed in -20 DEG C of preservations
With.
Utilize T4 DNA Ligase（TaKaRa）, by recyclingPnPhBP1DNA fragmentation and pCAMBIA2300s carrier-pellets
Section connects, reaction system（20 μL）It is with operating process：Take 10 μ LPnPhBP1DNA fragmentation sequentially adds 2 μ L
PCAMBIA2300s carrier DNAs, 2 μ L 10 × T4 DNA Ligase Buffer, 1 μ L T4 DNA Ligase, 5 μ L
ddH2O, centrifuges in short-term after mixing, then 16 DEG C of water-bath reaction overnights.Then connection product is transferred to greatly using heat-shock transformed method
In enterobacteria DH5 α, with containing 50 mg/L kanamycins（Kanamycin, Km）Solid medium screening positive clone.Select
Single bacterium colony shakes bacterium, is expanded by template of bacterium solutionPnPhBP1Special primer carry out PCR, pick outPnPhBP1With
The clone that pCAMBIA2300s is successfully connected, the bacterial strain detected are placed in -80 DEG C and save backup if the positive, addition glycerine.
Extract and purify the pCAMBIA2300s- in above-mentioned Escherichia coliPnPhBP1Plasmid, then will with frozen-thawed method
The plant expression vector pCAMBIA2300s- of above-mentioned structurePnPhBP1It is transferred in agrobacterium tumefaciens lba4404 competent cell.
Operating procedure is：Take 2 μ g pCAMBIA2300s-PnPhBP1Plasmid is added in the centrifuge tube containing 200 μ L competent cells,
5 min of ice bath after gently mixing, then continues in liquid nitrogen and freezes 1 min, be then immediately placed in 37 DEG C of water-baths 5 min, Zhi Houli
That is 2 min of ice bath, adds 800 μ L LB Liquid Cultures and is based on 28 DEG C of 4 h of shaken cultivation.By the Agrobacterium after activation be applied to containing
On the LB solid mediums of 50 mg/L Km, 28 DEG C of static gas wave refrigerators.Picking individual colonies shake bacterium, then with amplificationPnPhBP1It is special
Property primer carry out PCR, detect pCAMBIA2300s-PnPhBP1Whether it is transferred in Agrobacterium, for positive colony, adds glycerine
- 80 DEG C are placed on to save backup.
Embodiment 3：Agriculture bacillus mediated Genetic Transformation in Higher Plants and genetically modified plants screening
The transgene receptor of this experiment is tobacco, by tobacco seed with 75% alcohol soak 30s, with after sterile water washing with 0.1
The HgCl of %2Soak 8 min, then again with sterile water washing several times, be seeded on 1/2 MS culture mediums, 28 DEG C of light cultures 6
D, illumination box is gone to after germination（25 DEG C, 16h/d illumination）, monthly use 1/2MS culture mediums subculture once later.
That preservation is taken out from -80 DEG C of refrigerators contains pCAMBIA2300s-PnPhBP1The Agrobacterium LBA4404 bacterium of plasmid
Kind, it is inoculated in the LB fluid nutrient mediums that 5 mL contain 50 mg/L Km and 20 mg/L rifampins, 28 DEG C of cultures are muddy to culture medium
It is turbid.Draw on bacterium solution to the LB solid mediums containing 50 mg/L Km of 1 mL muddinesses, 28 DEG C of 48 h of culture；Then LB is consolidated
Agrobacterium on body culture medium scrapes to be inoculated in the MGL fluid nutrient mediums for the acetosyringone for being attached with 20 mg/L in right amount, and 28
DEG C shaken cultivation 2-3 h are to activate Agrobacterium.
Tobacco aseptic seedling leaf is taken to be cut into 1 cm2The leaf dish of left and right, is completely soaked in the above-mentioned MGL containing activation Agrobacterium
In fluid nutrient medium, immerged time is 15 min, and the bacterium solution of blade surface is blotted with aseptic filter paper, leaf dish is placed in co-cultivation base
Upper carry out incubated at room temperature, the co-cultivation base of Transformation of tobacco is MS+0.02 mg/L 6-BA+2.1 mg/L NAA+30 g/L
Sucrose+6 g/L agar, co-cultures 2 days under 22 DEG C of no light conditions.
Leaf dish after co-cultivation is gone into seedling differentiation in the MS screening and culturing mediums added with antibiotic, while screening transgenic
Plant.Tobacco screening and culturing medium for MS+0.5 mg/L 6-BA+0.1 mg/L NAA+30 g/L sucrose+6 g/L agar+
50 mg/L Km+200 mg/L cephalosporins（Cefotaxime sodium salt, Cef）；Blake bottle is turned during screening and culturing
Move to illumination box culture（25 DEG C, 16h/d illumination, 8h/d dark）, used after tobacco length budding containing 50 mg/L Km and
The MS culture medium squamous subcultures of 200 mg/L Cef, because tobacco callus differentiation rate is higher, therefore need to regeneration plant into advance one
Step screening, tobacco regrowth, which is moved on the MS culture mediums containing 50 mg/L Km, makes it take root, and finally selects and takes root preferably
Regrowth is further to be detected.
Using the genomic DNA of CTAB methods extraction transgenic tobacco plant blade, 1 μ L are taken to lead to the genomic DNA of extraction
Cross agarose gel electrophoresis and detect its integrality and concentration, expanded by template of the genomic DNA of transfer-gen plantPnPhBP1
Special primer carry out PCR, after PCR, take 8 μ L products to be used for agarose gel electrophoresis to detect positive transgenic plant,
The amplification of Partial Tobacco transfer-gen plant as shown in Figure 1,PnPhBP1Transgene tobacco screens 53 plants of positive transgenics altogether
Plant.
Embodiment 4：In transgene tobaccoPnPhBP1Expression analysis and transfer-gen plant antifungal activity analysis
Take positive transgenic single plant and non-transgenic tobacco（Wild type）Tender leaf extraction total serum IgE, reverse transcription generation cDNA the
One chain, and expanded as templatePnPhBP1Special primer carry out PCR, according in each transgenosis single plant of PCR interpretations of resultPnPhBP1The method of the expression of transcriptional level, Total RNAs extraction and RT-PCR are in the same manner as in Example 1, after PCR terminates, take
5 μ L are used for agarose gel electrophoresis, and the testing result of part single plant as shown in Fig. 2, detect in 36 transgenosis single plants altogetherPnPhBP1In transcriptional level great expression, the numbering of these single plants is 1~36.
Several fungies that laboratory preserves are inoculated in PDA solid mediums（200 g/L potatos, 15 g/L agar, 20
G/L glucose）On, 28 DEG C of light cultures, add albumen when colony growth to diameter is about 2 ~ 3cm, analyze transfer-gen plant body
Outer antifungal activity.The albumen that other living contaminantses are extracted in order to prevent, whole vegetable protein extraction process are sterile behaviour
Make, take 1 g transgene tobacco single plants first（Numbering is respectively 7,12,28,35）And wild-type leaves are put into mortar, 1 is added
ML protein extracts（1M NaCl, 0.1M sodium acetates, 1% PVP, pH6）, it is fully ground；It is transferred in 1.5 mL centrifuge tubes, mixes
Stand overnight for 4 DEG C afterwards, 4 DEG C of 30 min of centrifugation（12,000 g/min）, supernatant is taken in 1.5 new mL centrifuge tubes, and is taken appropriate
Total protein concentration is measured with UV detector.The total protein concentration of transgenosis and WT lines is adjusted to 0.5 μ g/ μ
L, then takes 20 μ L drops on the aseptic filter paper of each fungi culture medium respectively, except addition is different on the tablet of each fungi
The total protein of transgenic tobacco plant, while the total protein of parallel addition wild-type tobacco and blank control（Extract used in albumen
Solution）, the situation of each processing fungi growth is after a few days observed in 28 DEG C of cultures, and is evaluated accordinglyPnPhBP1Transgene tobacco
Extracorporeal antifungal activity, the results are shown in Figure 3,PnPhBP1Transgene tobacco albumen is to Botrytis cinerea, Fusarium solani, glue
The growth of spore anthrax-bacilus has very strong inhibitory action.
Sequence table
<110>Kunming University of Science and Technology
<120>A kind of pseudo-ginseng plant hormone binding-protein gene PnPhBP1 and application
<160> 4
<170> SIPOSequenceListing 1.0
<210> 1
<211> 623
<212> DNA
<213>Pseudo-ginseng (Panax notoginseng)
<400> 1
agtaaatgta cctgaacttg tactactaat tatctttagg gattgcatca agaggtatga 60
caaaagaact aaaaacccaa acaaaggtta gtgttgggat tgaagtcttg tggggggctc 120
tagctaagga tataaacatt gtgcttccaa gaattattcc aaatttggtt aaagatgcag 180
aagtgcttga aggacatggc ggccttggta ctgtttttct cttcaagttt ggctctgatg 240
tatcaacatt tggggatcag aaggaaaaga ttgtcgaact tgatgagtcc ctgcatctaa 300
ttgggcttca agtaatagaa ggcggtcatc tgaatcatgg ctttacttca tacaaaacgg 360
tttttcaact aacagcaata acagagttgg agacactagt tgatatgaag gtggtgtatg 420
agattgaagc agaagaaact catatgccag tggagactac aaagtccgca cttgctttta 480
taaaatgtgt agaaacatat ctgttaaaca aaggatccta gacgaatctt gtctaaattc 540
attcctgaat ttaggttctc acagtccttt gctgtgatca aattcatttt tcaaaaaaaa 600
aaaaaaaaaa aaaaaaaaaa aaa 623
<210> 2
<211> 154
<212> PRT
<213>Pseudo-ginseng (Panax notoginseng)
<400> 2
Met Thr Lys Glu Leu Lys Thr Gln Thr Lys Val Ser Val Gly Ile Glu
1 5 10 15
Val Leu Trp Gly Ala Leu Ala Lys Asp Ile Asn Ile Val Leu Pro Arg
20 25 30
Ile Ile Pro Asn Leu Val Lys Asp Ala Glu Val Leu Glu Gly His Gly
35 40 45
Gly Leu Gly Thr Val Phe Leu Phe Lys Phe Gly Ser Asp Val Ser Thr
50 55 60
Phe Gly Asp Gln Lys Glu Lys Ile Val Glu Leu Asp Glu Ser Leu His
65 70 75 80
Leu Ile Gly Leu Gln Val Ile Glu Gly Gly His Leu Asn His Gly Phe
85 90 95
Thr Ser Tyr Lys Thr Val Phe Gln Leu Thr Ala Ile Thr Glu Leu Glu
100 105 110
Thr Leu Val Asp Met Lys Val Val Tyr Glu Ile Glu Ala Glu Glu Thr
115 120 125
His Met Pro Val Glu Thr Thr Lys Ser Ala Leu Ala Phe Ile Lys Cys
130 135 140
Val Glu Thr Tyr Leu Leu Asn Lys Gly Ser
145 150
<210> 3
<211> 25
<212> DNA
<213>Artificial sequence (Artificial)
<400> 3
tatctttagg gattgcatca agagg 25
<210> 4
<211> 25
<212> DNA
<213>Artificial sequence (Artificial)
<400> 4
acagcaaagg actgtgagaa cctaa 25
Claims (3)
- A kind of 1. pseudo-ginseng plant hormone binding-protein genePnPhBP1, it is characterised in that：Nucleotide sequence such as SEQ ID NO：1 It is shown, coding such as SEQ ID NO：The protein of amino acid sequence shown in 2.
- 2. pseudo-ginseng plant hormone binding-protein gene according to claim 1PnPhBP1, it is characterised in that：PnPhBP1Volume Code area is SEQ ID NO：Nucleotide sequence in 1 shown in 57-521.
- 3. the pseudo-ginseng plant hormone binding-protein gene described in claim 1 or 2PnPhBP1Tobacco is being improved to Botrytis cinerea (Botrytis cinerea), Fusarium solani (Fusarium solani), colletotrichum gloeosporioides Penz (Colletotrichum gloeosporioides) application in resistance.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810000429.9A CN108018294B (en) | 2018-01-02 | 2018-01-02 | Panax notoginseng plant hormone binding protein genePnPhBP1And applications |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810000429.9A CN108018294B (en) | 2018-01-02 | 2018-01-02 | Panax notoginseng plant hormone binding protein genePnPhBP1And applications |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN108018294A true CN108018294A (en) | 2018-05-11 |
| CN108018294B CN108018294B (en) | 2020-11-17 |
Family
ID=62072585
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201810000429.9A Active CN108018294B (en) | 2018-01-02 | 2018-01-02 | Panax notoginseng plant hormone binding protein genePnPhBP1And applications |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN108018294B (en) |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| AU2003226907A1 (en) * | 2002-03-27 | 2003-10-08 | Leukotech A/S | Method for the preparation of recombinant mammalian heparin-binding protein (hbp) |
| CN101775070A (en) * | 2010-01-14 | 2010-07-14 | 中国农业科学院生物技术研究所 | Plant stress tolerance correlative protein, encoding gene and application thereof |
| CN102234322A (en) * | 2010-04-27 | 2011-11-09 | 中国农业科学院作物科学研究所 | Protein GmNF-YA1 related with stress tolerance of plants, and coding gene and application thereof |
| CN106244599A (en) * | 2016-09-21 | 2016-12-21 | 昆明理工大学 | A kind of Radix Notoginseng pathogenesis-related proteins 1 family gene PnPR1 2 and application |
-
2018
- 2018-01-02 CN CN201810000429.9A patent/CN108018294B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| AU2003226907A1 (en) * | 2002-03-27 | 2003-10-08 | Leukotech A/S | Method for the preparation of recombinant mammalian heparin-binding protein (hbp) |
| CN101775070A (en) * | 2010-01-14 | 2010-07-14 | 中国农业科学院生物技术研究所 | Plant stress tolerance correlative protein, encoding gene and application thereof |
| CN102234322A (en) * | 2010-04-27 | 2011-11-09 | 中国农业科学院作物科学研究所 | Protein GmNF-YA1 related with stress tolerance of plants, and coding gene and application thereof |
| CN106244599A (en) * | 2016-09-21 | 2016-12-21 | 昆明理工大学 | A kind of Radix Notoginseng pathogenesis-related proteins 1 family gene PnPR1 2 and application |
Non-Patent Citations (1)
| Title |
|---|
| 杨丹等: "三七病程相关蛋白PR10-2 基因的克隆、表达及功能初步分析", 《中国中药杂志》 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN108018294B (en) | 2020-11-17 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN105441460B (en) | A kind of Ming River lily WRKY transcription factor genes LrWRKY1 and application | |
| CN110818783B (en) | Lilium regale WRKY transcription factor gene LrWRKY2 and application thereof | |
| CN110747202B (en) | Lilium regale WRKY transcription factor gene LrWRKY11 and application thereof | |
| CN105861517B (en) | A kind of Radix Notoginseng antibacterial peptide gene PnSN1 and its application | |
| CN110818782A (en) | Lilium regale WRKY transcription factor gene LrWRKY3 and application thereof | |
| CN107746846B (en) | IbABF4 gene for coding sweet potato bZIP transcription factor and application thereof | |
| CN106244599B (en) | A kind of 1 family gene PnPR1-2 of Radix Notoginseng pathogenesis-related proteins and application | |
| CN108251432A (en) | Radix Notoginseng class PR gene PnPRlike and application | |
| CN103194457B (en) | Lilium regale germin-like protein gene LrGLP2 and application thereof | |
| CN103194456B (en) | Lilium regale antifungal gene Lr14-3-3 and application thereof | |
| CN107267526B (en) | Radix Notoginseng myb transcription factor gene PnMYB2 and its application | |
| CN113150094A (en) | EjAP2L gene related to loquat flower development and encoding protein and application thereof | |
| CN104878019A (en) | Yangbi walnut germin-like protein gene JsGLP1 and application thereof | |
| CN106892973A (en) | Plant adversity resistance related protein GhMYB4 and encoding gene and application | |
| CN103320448B (en) | Lilium regle bZIP transcription factor LrbZIP1 and application | |
| CN107365794B (en) | Application of panax notoginseng chitinase gene PnCHI1 | |
| CN107904238B (en) | Thick boisiana is with high salt, drought-inducible promoter IpLEA-PRO and its application | |
| CN110607307A (en) | Sedum lineare salt-tolerant gene SlNAC and application thereof | |
| CN107267525B (en) | Application of panax notoginseng polygalacturonase inhibitor protein gene PnPGIP | |
| CN113481210B (en) | Application of cotton GhDof1.7 gene in promotion of salt tolerance of plants | |
| CN104152465B (en) | Lilium regale cytochrome b5 gene LrCyt-b5 and application thereof | |
| CN108018294A (en) | A kind of pseudo-ginseng plant hormone binding-protein gene PnPhBP1 and application | |
| CN108707610B (en) | Notoginseng defensein antibacterial peptide genePnDEFL1And applications | |
| CN107287212B (en) | Halogeton sativus salt-tolerant gene HgS3 and application thereof | |
| CN113604477B (en) | Lilium regale defensin antibacterial peptide gene LrDEF1 and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |