CN107354166A - A kind of glucanase gene PnGlu1 of pseudo-ginseng β 1,3 and its application - Google Patents
A kind of glucanase gene PnGlu1 of pseudo-ginseng β 1,3 and its application Download PDFInfo
- Publication number
- CN107354166A CN107354166A CN201710540033.9A CN201710540033A CN107354166A CN 107354166 A CN107354166 A CN 107354166A CN 201710540033 A CN201710540033 A CN 201710540033A CN 107354166 A CN107354166 A CN 107354166A
- Authority
- CN
- China
- Prior art keywords
- pnglu1
- plant
- ginseng
- pseudo
- 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
- 108090000623 proteins and genes Proteins 0.000 title claims abstract description 38
- 244000131316 Panax pseudoginseng Species 0.000 title claims abstract description 22
- 235000003181 Panax pseudoginseng Nutrition 0.000 title claims abstract description 22
- 241000894006 Bacteria Species 0.000 claims abstract description 23
- 241000427940 Fusarium solani Species 0.000 claims abstract description 9
- 241001529387 Colletotrichum gloeosporioides Species 0.000 claims abstract description 7
- 235000009754 Vitis X bourquina Nutrition 0.000 claims abstract description 6
- 235000012333 Vitis X labruscana Nutrition 0.000 claims abstract description 6
- 240000006365 Vitis vinifera Species 0.000 claims abstract description 6
- 235000014787 Vitis vinifera Nutrition 0.000 claims abstract description 6
- 239000002773 nucleotide Substances 0.000 claims abstract description 4
- 125000003729 nucleotide group Chemical group 0.000 claims abstract description 3
- 101000763602 Manilkara zapota Thaumatin-like protein 1 Proteins 0.000 claims description 17
- 101000763586 Manilkara zapota Thaumatin-like protein 1a Proteins 0.000 claims description 17
- 101000966653 Musa acuminata Glucan endo-1,3-beta-glucosidase Proteins 0.000 claims description 17
- 102000004169 proteins and genes Human genes 0.000 claims description 16
- 241000555706 Botryosphaeria dothidea Species 0.000 claims description 2
- 241000233732 Fusarium verticillioides Species 0.000 claims description 2
- 125000003275 alpha amino acid group Chemical group 0.000 claims 1
- 241000196324 Embryophyta Species 0.000 abstract description 55
- 244000061176 Nicotiana tabacum Species 0.000 abstract description 32
- 235000002637 Nicotiana tabacum Nutrition 0.000 abstract description 32
- 241000233866 Fungi Species 0.000 abstract description 13
- 108010001682 Dextranase Proteins 0.000 abstract description 12
- 230000009261 transgenic effect Effects 0.000 abstract description 9
- 108700019146 Transgenes Proteins 0.000 abstract description 6
- 230000000843 anti-fungal effect Effects 0.000 abstract description 5
- 230000002018 overexpression Effects 0.000 abstract description 5
- 239000013604 expression vector Substances 0.000 abstract description 4
- 230000002401 inhibitory effect Effects 0.000 abstract description 4
- 101710096342 Pathogenesis-related protein Proteins 0.000 abstract description 3
- 230000001857 anti-mycotic effect Effects 0.000 abstract description 3
- 239000002543 antimycotic Substances 0.000 abstract description 3
- 238000000034 method Methods 0.000 description 18
- 108020004414 DNA Proteins 0.000 description 15
- 201000010099 disease Diseases 0.000 description 14
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 14
- 235000018102 proteins Nutrition 0.000 description 14
- 238000006243 chemical reaction Methods 0.000 description 11
- 239000013612 plasmid Substances 0.000 description 10
- 239000002299 complementary DNA Substances 0.000 description 9
- 238000000605 extraction Methods 0.000 description 9
- 239000000047 product Substances 0.000 description 9
- 210000002421 cell wall Anatomy 0.000 description 8
- 238000012216 screening Methods 0.000 description 8
- 241000589158 Agrobacterium Species 0.000 description 7
- 150000001413 amino acids Chemical group 0.000 description 7
- 239000012634 fragment Substances 0.000 description 7
- 230000008569 process Effects 0.000 description 7
- 210000002966 serum Anatomy 0.000 description 7
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 7
- 235000003143 Panax notoginseng Nutrition 0.000 description 6
- 241000180649 Panax notoginseng Species 0.000 description 6
- 238000002156 mixing Methods 0.000 description 6
- 239000013598 vector Substances 0.000 description 6
- 229920002498 Beta-glucan Polymers 0.000 description 5
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 5
- 238000000246 agarose gel electrophoresis Methods 0.000 description 5
- 238000004458 analytical method Methods 0.000 description 5
- 230000002538 fungal effect Effects 0.000 description 5
- 239000003292 glue Substances 0.000 description 5
- 239000007788 liquid Substances 0.000 description 5
- 239000003550 marker Substances 0.000 description 5
- 239000000463 material Substances 0.000 description 5
- 244000052769 pathogen Species 0.000 description 5
- 230000001717 pathogenic effect Effects 0.000 description 5
- 230000004044 response Effects 0.000 description 5
- 239000007787 solid Substances 0.000 description 5
- 239000000243 solution Substances 0.000 description 5
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 4
- 208000035240 Disease Resistance Diseases 0.000 description 4
- 229920001503 Glucan Polymers 0.000 description 4
- 244000068988 Glycine max Species 0.000 description 4
- 235000010469 Glycine max Nutrition 0.000 description 4
- 240000004713 Pisum sativum Species 0.000 description 4
- 241000607479 Yersinia pestis Species 0.000 description 4
- 230000015572 biosynthetic process Effects 0.000 description 4
- 239000000872 buffer Substances 0.000 description 4
- 210000004027 cell Anatomy 0.000 description 4
- 230000004665 defense response Effects 0.000 description 4
- 230000029087 digestion Effects 0.000 description 4
- 244000000004 fungal plant pathogen Species 0.000 description 4
- 239000001963 growth medium Substances 0.000 description 4
- 230000007062 hydrolysis Effects 0.000 description 4
- 238000006460 hydrolysis reaction Methods 0.000 description 4
- 238000005286 illumination Methods 0.000 description 4
- 239000002609 medium Substances 0.000 description 4
- 229920001542 oligosaccharide Polymers 0.000 description 4
- 150000002482 oligosaccharides Chemical class 0.000 description 4
- 108090000765 processed proteins & peptides Proteins 0.000 description 4
- 238000003757 reverse transcription PCR Methods 0.000 description 4
- 238000003786 synthesis reaction Methods 0.000 description 4
- 229920001817 Agar Polymers 0.000 description 3
- 241000589155 Agrobacterium tumefaciens Species 0.000 description 3
- UHOVQNZJYSORNB-UHFFFAOYSA-N Benzene Chemical compound C1=CC=CC=C1 UHOVQNZJYSORNB-UHFFFAOYSA-N 0.000 description 3
- 102000012410 DNA Ligases Human genes 0.000 description 3
- 108010061982 DNA Ligases Proteins 0.000 description 3
- 102000004190 Enzymes Human genes 0.000 description 3
- 108090000790 Enzymes Proteins 0.000 description 3
- 208000031888 Mycoses Diseases 0.000 description 3
- 235000010582 Pisum sativum Nutrition 0.000 description 3
- 239000008272 agar Substances 0.000 description 3
- 235000001014 amino acid Nutrition 0.000 description 3
- 210000000170 cell membrane Anatomy 0.000 description 3
- 239000003153 chemical reaction reagent Substances 0.000 description 3
- 238000012258 culturing Methods 0.000 description 3
- 230000006378 damage Effects 0.000 description 3
- 238000005516 engineering process Methods 0.000 description 3
- 239000012530 fluid Substances 0.000 description 3
- 238000013467 fragmentation Methods 0.000 description 3
- 238000006062 fragmentation reaction Methods 0.000 description 3
- 238000010353 genetic engineering Methods 0.000 description 3
- 230000001404 mediated effect Effects 0.000 description 3
- 239000000203 mixture Substances 0.000 description 3
- 238000003199 nucleic acid amplification method Methods 0.000 description 3
- 235000015097 nutrients Nutrition 0.000 description 3
- 102000004196 processed proteins & peptides Human genes 0.000 description 3
- 238000010839 reverse transcription Methods 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- 238000012360 testing method Methods 0.000 description 3
- 230000002103 transcriptional effect Effects 0.000 description 3
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 2
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 2
- 101100150621 Arabidopsis thaliana GEBP gene Proteins 0.000 description 2
- 229920002101 Chitin Polymers 0.000 description 2
- 108010022172 Chitinases Proteins 0.000 description 2
- 102000012286 Chitinases Human genes 0.000 description 2
- 101000866354 Cryptopygus antarcticus Glucan endo-1,3-beta-glucosidase Proteins 0.000 description 2
- 241000588724 Escherichia coli Species 0.000 description 2
- WHUUTDBJXJRKMK-UHFFFAOYSA-N Glutamic acid Natural products OC(=O)C(N)CCC(O)=O WHUUTDBJXJRKMK-UHFFFAOYSA-N 0.000 description 2
- 108090000604 Hydrolases Proteins 0.000 description 2
- 102000004157 Hydrolases Human genes 0.000 description 2
- 102100034343 Integrase Human genes 0.000 description 2
- 240000007049 Juglans regia Species 0.000 description 2
- 235000009496 Juglans regia Nutrition 0.000 description 2
- SITLTJHOQZFJGG-UHFFFAOYSA-N N-L-alpha-glutamyl-L-valine Natural products CC(C)C(C(O)=O)NC(=O)C(N)CCC(O)=O SITLTJHOQZFJGG-UHFFFAOYSA-N 0.000 description 2
- 241000233616 Phytophthora capsici Species 0.000 description 2
- 108010029485 Protein Isoforms Proteins 0.000 description 2
- 102000001708 Protein Isoforms Human genes 0.000 description 2
- 108010092799 RNA-directed DNA polymerase Proteins 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 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
- 244000299461 Theobroma cacao Species 0.000 description 2
- 235000009470 Theobroma cacao Nutrition 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
- 108010087924 alanylproline Proteins 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
- 230000000844 anti-bacterial effect Effects 0.000 description 2
- 230000003385 bacteriostatic effect Effects 0.000 description 2
- 230000008901 benefit Effects 0.000 description 2
- 238000009395 breeding Methods 0.000 description 2
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 description 2
- 239000000969 carrier Substances 0.000 description 2
- 239000003795 chemical substances by application Substances 0.000 description 2
- 238000007796 conventional method Methods 0.000 description 2
- 238000003967 crop rotation Methods 0.000 description 2
- 238000001514 detection 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
- 230000000694 effects Effects 0.000 description 2
- 239000005712 elicitor Substances 0.000 description 2
- 238000010195 expression analysis Methods 0.000 description 2
- 244000053095 fungal pathogen Species 0.000 description 2
- 230000014509 gene expression Effects 0.000 description 2
- 235000011187 glycerol Nutrition 0.000 description 2
- 230000003301 hydrolyzing effect Effects 0.000 description 2
- 108091006086 inhibitor proteins Proteins 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
- 238000004519 manufacturing process Methods 0.000 description 2
- 229910052757 nitrogen Inorganic materials 0.000 description 2
- 108010004568 plant pathogenesis-related proteins Proteins 0.000 description 2
- 229920001184 polypeptide Polymers 0.000 description 2
- 239000013641 positive control Substances 0.000 description 2
- 238000004321 preservation Methods 0.000 description 2
- 238000011084 recovery Methods 0.000 description 2
- 230000009467 reduction Effects 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
- 230000009466 transformation Effects 0.000 description 2
- 238000005406 washing Methods 0.000 description 2
- 102000040650 (ribonucleotides)n+m Human genes 0.000 description 1
- 108020005345 3' Untranslated Regions Proteins 0.000 description 1
- 108020003589 5' Untranslated Regions Proteins 0.000 description 1
- YIQAOPNCIJVKDN-XKNYDFJKSA-N Ala-Asn-Asp-Pro Chemical compound C[C@H](N)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(O)=O)C(=O)N1CCC[C@H]1C(O)=O YIQAOPNCIJVKDN-XKNYDFJKSA-N 0.000 description 1
- ZIBWKCRKNFYTPT-ZKWXMUAHSA-N Ala-Asn-Val Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](C(C)C)C(O)=O ZIBWKCRKNFYTPT-ZKWXMUAHSA-N 0.000 description 1
- VGPWRRFOPXVGOH-BYPYZUCNSA-N Ala-Gly-Gly Chemical compound C[C@H](N)C(=O)NCC(=O)NCC(O)=O VGPWRRFOPXVGOH-BYPYZUCNSA-N 0.000 description 1
- PVQLRJRPUTXFFX-CIUDSAMLSA-N Ala-Met-Gln Chemical compound CSCC[C@H](NC(=O)[C@H](C)N)C(=O)N[C@@H](CCC(N)=O)C(O)=O PVQLRJRPUTXFFX-CIUDSAMLSA-N 0.000 description 1
- GMGWOTQMUKYZIE-UBHSHLNASA-N Ala-Pro-Phe Chemical compound C[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@H](C(O)=O)CC1=CC=CC=C1 GMGWOTQMUKYZIE-UBHSHLNASA-N 0.000 description 1
- MSWSRLGNLKHDEI-ACZMJKKPSA-N Ala-Ser-Glu Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CO)C(=O)N[C@@H](CCC(O)=O)C(O)=O MSWSRLGNLKHDEI-ACZMJKKPSA-N 0.000 description 1
- JJHBEVZAZXZREW-LFSVMHDDSA-N Ala-Thr-Phe Chemical compound C[C@@H](O)[C@H](NC(=O)[C@H](C)N)C(=O)N[C@@H](Cc1ccccc1)C(O)=O JJHBEVZAZXZREW-LFSVMHDDSA-N 0.000 description 1
- YXXPVUOMPSZURS-ZLIFDBKOSA-N Ala-Trp-Leu Chemical compound C1=CC=C2C(C[C@@H](C(=O)N[C@@H](CC(C)C)C(O)=O)NC(=O)[C@H](C)N)=CNC2=C1 YXXPVUOMPSZURS-ZLIFDBKOSA-N 0.000 description 1
- VHAQSYHSDKERBS-XPUUQOCRSA-N Ala-Val-Gly Chemical compound C[C@H](N)C(=O)N[C@@H](C(C)C)C(=O)NCC(O)=O VHAQSYHSDKERBS-XPUUQOCRSA-N 0.000 description 1
- 241001124076 Aphididae Species 0.000 description 1
- KGSJCPBERYUXCN-BPNCWPANSA-N Arg-Ala-Tyr Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](C)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O KGSJCPBERYUXCN-BPNCWPANSA-N 0.000 description 1
- NABSCJGZKWSNHX-RCWTZXSCSA-N Arg-Arg-Thr Chemical compound NC(N)=NCCC[C@@H](C(=O)N[C@@H]([C@H](O)C)C(O)=O)NC(=O)[C@@H](N)CCCN=C(N)N NABSCJGZKWSNHX-RCWTZXSCSA-N 0.000 description 1
- JSHVMZANPXCDTL-GMOBBJLQSA-N Arg-Asp-Ile Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O JSHVMZANPXCDTL-GMOBBJLQSA-N 0.000 description 1
- GXXWTNKNFFKTJB-NAKRPEOUSA-N Arg-Ile-Ser Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CO)C(O)=O GXXWTNKNFFKTJB-NAKRPEOUSA-N 0.000 description 1
- ZDBWKBCKYJGKGP-DCAQKATOSA-N Arg-Leu-Ala Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](C)C(O)=O ZDBWKBCKYJGKGP-DCAQKATOSA-N 0.000 description 1
- OTZMRMHZCMZOJZ-SRVKXCTJSA-N Arg-Leu-Glu Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(O)=O)C(O)=O OTZMRMHZCMZOJZ-SRVKXCTJSA-N 0.000 description 1
- HGKHPCFTRQDHCU-IUCAKERBSA-N Arg-Pro-Gly Chemical compound NC(N)=NCCC[C@H](N)C(=O)N1CCC[C@H]1C(=O)NCC(O)=O HGKHPCFTRQDHCU-IUCAKERBSA-N 0.000 description 1
- PFOYSEIHFVKHNF-FXQIFTODSA-N Asn-Ala-Arg Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](C)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O PFOYSEIHFVKHNF-FXQIFTODSA-N 0.000 description 1
- ZZXMOQIUIJJOKZ-ZLUOBGJFSA-N Asn-Asn-Ala Chemical compound OC(=O)[C@H](C)NC(=O)[C@H](CC(N)=O)NC(=O)[C@@H](N)CC(N)=O ZZXMOQIUIJJOKZ-ZLUOBGJFSA-N 0.000 description 1
- KXFCBAHYSLJCCY-ZLUOBGJFSA-N Asn-Asn-Ser Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CO)C(O)=O KXFCBAHYSLJCCY-ZLUOBGJFSA-N 0.000 description 1
- DMLSCRJBWUEALP-LAEOZQHASA-N Asn-Glu-Val Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](C(C)C)C(O)=O DMLSCRJBWUEALP-LAEOZQHASA-N 0.000 description 1
- HYQYLOSCICEYTR-YUMQZZPRSA-N Asn-Gly-Leu Chemical compound [H]N[C@@H](CC(N)=O)C(=O)NCC(=O)N[C@@H](CC(C)C)C(O)=O HYQYLOSCICEYTR-YUMQZZPRSA-N 0.000 description 1
- XVBDDUPJVQXDSI-PEFMBERDSA-N Asn-Ile-Glu Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCC(=O)O)C(=O)O)NC(=O)[C@H](CC(=O)N)N XVBDDUPJVQXDSI-PEFMBERDSA-N 0.000 description 1
- MHBUWPFQNPJTAS-QAETUUGQSA-N Asn-Leu-Phe-Asp Chemical compound NC(=O)C[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@H](C(=O)N[C@@H](CC(O)=O)C(O)=O)CC1=CC=CC=C1 MHBUWPFQNPJTAS-QAETUUGQSA-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
- KHCNTVRVAYCPQE-CIUDSAMLSA-N Asn-Lys-Asn Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(N)=O)C(O)=O KHCNTVRVAYCPQE-CIUDSAMLSA-N 0.000 description 1
- BCADFFUQHIMQAA-KKHAAJSZSA-N Asn-Thr-Val Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C(C)C)C(O)=O BCADFFUQHIMQAA-KKHAAJSZSA-N 0.000 description 1
- LRCIOEVFVGXZKB-BZSNNMDCSA-N Asn-Tyr-Tyr Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O LRCIOEVFVGXZKB-BZSNNMDCSA-N 0.000 description 1
- XJQRWGXKUSDEFI-ACZMJKKPSA-N Asp-Glu-Asn Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(N)=O)C(O)=O XJQRWGXKUSDEFI-ACZMJKKPSA-N 0.000 description 1
- KYQNAIMCTRZLNP-QSFUFRPTSA-N Asp-Ile-Val Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](C(C)C)C(O)=O KYQNAIMCTRZLNP-QSFUFRPTSA-N 0.000 description 1
- VHUKCUHLFMRHOD-MELADBBJSA-N Asp-Tyr-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CC2=CC=C(C=C2)O)NC(=O)[C@H](CC(=O)O)N)C(=O)O VHUKCUHLFMRHOD-MELADBBJSA-N 0.000 description 1
- 241000432824 Asparagus densiflorus Species 0.000 description 1
- 241000193830 Bacillus <bacterium> Species 0.000 description 1
- 241000193738 Bacillus anthracis Species 0.000 description 1
- 235000002566 Capsicum Nutrition 0.000 description 1
- 235000002567 Capsicum annuum Nutrition 0.000 description 1
- 240000004160 Capsicum annuum Species 0.000 description 1
- 235000008534 Capsicum annuum var annuum Nutrition 0.000 description 1
- 240000008384 Capsicum annuum var. annuum 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
- 241000289763 Dasygaster padockina Species 0.000 description 1
- 108010082495 Dietary Plant Proteins Proteins 0.000 description 1
- 241000305071 Enterobacterales Species 0.000 description 1
- 241000221785 Erysiphales Species 0.000 description 1
- KVYVOGYEMPEXBT-GUBZILKMSA-N Gln-Ala-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](C)NC(=O)[C@@H](N)CCC(N)=O KVYVOGYEMPEXBT-GUBZILKMSA-N 0.000 description 1
- PRBLYKYHAJEABA-SRVKXCTJSA-N Gln-Arg-Leu Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(C)C)C(O)=O PRBLYKYHAJEABA-SRVKXCTJSA-N 0.000 description 1
- RMOCFPBLHAOTDU-ACZMJKKPSA-N Gln-Asn-Ser Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CO)C(O)=O RMOCFPBLHAOTDU-ACZMJKKPSA-N 0.000 description 1
- XWIBVSAEUCAAKF-GVXVVHGQSA-N Gln-His-Val Chemical compound CC(C)[C@@H](C(=O)O)NC(=O)[C@H](CC1=CN=CN1)NC(=O)[C@H](CCC(=O)N)N XWIBVSAEUCAAKF-GVXVVHGQSA-N 0.000 description 1
- MKRDNSWGJWTBKZ-GVXVVHGQSA-N Gln-Val-Lys Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CCCCN)C(=O)O)NC(=O)[C@H](CCC(=O)N)N MKRDNSWGJWTBKZ-GVXVVHGQSA-N 0.000 description 1
- OCJRHJZKGGSPRW-IUCAKERBSA-N Glu-Lys-Gly Chemical compound NCCCC[C@@H](C(=O)NCC(O)=O)NC(=O)[C@@H](N)CCC(O)=O OCJRHJZKGGSPRW-IUCAKERBSA-N 0.000 description 1
- DMYACXMQUABZIQ-NRPADANISA-N Glu-Ser-Val Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](C(C)C)C(O)=O DMYACXMQUABZIQ-NRPADANISA-N 0.000 description 1
- GPSHCSTUYOQPAI-JHEQGTHGSA-N Glu-Thr-Gly Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)NCC(O)=O GPSHCSTUYOQPAI-JHEQGTHGSA-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
- GQGAFTPXAPKSCF-WHFBIAKZSA-N Gly-Ala-Cys Chemical compound NCC(=O)N[C@@H](C)C(=O)N[C@@H](CS)C(=O)O GQGAFTPXAPKSCF-WHFBIAKZSA-N 0.000 description 1
- CIMULJZTTOBOPN-WHFBIAKZSA-N Gly-Asn-Asn Chemical compound NCC(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(N)=O)C(O)=O CIMULJZTTOBOPN-WHFBIAKZSA-N 0.000 description 1
- XRTDOIOIBMAXCT-NKWVEPMBSA-N Gly-Asn-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CC(=O)N)NC(=O)CN)C(=O)O XRTDOIOIBMAXCT-NKWVEPMBSA-N 0.000 description 1
- XBWMTPAIUQIWKA-BYULHYEWSA-N Gly-Asp-Ile Chemical compound CC[C@H](C)[C@@H](C(O)=O)NC(=O)[C@H](CC(O)=O)NC(=O)CN XBWMTPAIUQIWKA-BYULHYEWSA-N 0.000 description 1
- UQJNXZSSGQIPIQ-FBCQKBJTSA-N Gly-Gly-Thr Chemical compound C[C@@H](O)[C@@H](C(O)=O)NC(=O)CNC(=O)CN UQJNXZSSGQIPIQ-FBCQKBJTSA-N 0.000 description 1
- TVUWMSBGMVAHSJ-KBPBESRZSA-N Gly-Leu-Phe Chemical compound NCC(=O)N[C@@H](CC(C)C)C(=O)N[C@H](C(O)=O)CC1=CC=CC=C1 TVUWMSBGMVAHSJ-KBPBESRZSA-N 0.000 description 1
- BMWFDYIYBAFROD-WPRPVWTQSA-N Gly-Pro-Val Chemical compound CC(C)[C@@H](C(O)=O)NC(=O)[C@@H]1CCCN1C(=O)CN BMWFDYIYBAFROD-WPRPVWTQSA-N 0.000 description 1
- 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
- JSLVAHYTAJJEQH-QWRGUYRKSA-N Gly-Ser-Phe Chemical compound NCC(=O)N[C@@H](CO)C(=O)N[C@H](C(O)=O)CC1=CC=CC=C1 JSLVAHYTAJJEQH-QWRGUYRKSA-N 0.000 description 1
- UIQGJYUEQDOODF-KWQFWETISA-N Gly-Tyr-Ala Chemical compound OC(=O)[C@H](C)NC(=O)[C@@H](NC(=O)CN)CC1=CC=C(O)C=C1 UIQGJYUEQDOODF-KWQFWETISA-N 0.000 description 1
- NWOSHVVPKDQKKT-RYUDHWBXSA-N Gly-Tyr-Gln Chemical compound [H]NCC(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CCC(N)=O)C(O)=O NWOSHVVPKDQKKT-RYUDHWBXSA-N 0.000 description 1
- YGHSQRJSHKYUJY-SCZZXKLOSA-N Gly-Val-Pro Chemical compound CC(C)[C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)CN YGHSQRJSHKYUJY-SCZZXKLOSA-N 0.000 description 1
- 241000258937 Hemiptera Species 0.000 description 1
- 241000238631 Hexapoda Species 0.000 description 1
- WJGSTIMGSIWHJX-HVTMNAMFSA-N His-Ile-Gln Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCC(=O)N)C(=O)O)NC(=O)[C@H](CC1=CN=CN1)N WJGSTIMGSIWHJX-HVTMNAMFSA-N 0.000 description 1
- 206010020649 Hyperkeratosis Diseases 0.000 description 1
- 206010020751 Hypersensitivity Diseases 0.000 description 1
- SLQVFYWBGNNOTK-BYULHYEWSA-N Ile-Gly-Asn Chemical compound CC[C@H](C)[C@@H](C(=O)NCC(=O)N[C@@H](CC(=O)N)C(=O)O)N SLQVFYWBGNNOTK-BYULHYEWSA-N 0.000 description 1
- KYLIZSDYWQQTFM-PEDHHIEDSA-N Ile-Ile-Arg Chemical compound CC[C@H](C)[C@H](N)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@H](C(O)=O)CCCN=C(N)N KYLIZSDYWQQTFM-PEDHHIEDSA-N 0.000 description 1
- FGBRXCZYVRFNKQ-MXAVVETBSA-N Ile-Phe-Ser Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CO)C(=O)O)N FGBRXCZYVRFNKQ-MXAVVETBSA-N 0.000 description 1
- HZVRQFKRALAMQS-SLBDDTMCSA-N Ile-Trp-Asp Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC1=CNC2=CC=CC=C21)C(=O)N[C@@H](CC(=O)O)C(=O)O)N HZVRQFKRALAMQS-SLBDDTMCSA-N 0.000 description 1
- SITWEMZOJNKJCH-UHFFFAOYSA-N L-alanine-L-arginine Natural products CC(N)C(=O)NC(C(O)=O)CCCNC(N)=N SITWEMZOJNKJCH-UHFFFAOYSA-N 0.000 description 1
- JRJLGNFWYFSJHB-HOCLYGCPSA-N Leu-Gly-Trp Chemical compound [H]N[C@@H](CC(C)C)C(=O)NCC(=O)N[C@@H](CC1=CNC2=C1C=CC=C2)C(O)=O JRJLGNFWYFSJHB-HOCLYGCPSA-N 0.000 description 1
- DPURXCQCHSQPAN-AVGNSLFASA-N Leu-Pro-Pro Chemical compound CC(C)C[C@H](N)C(=O)N1CCC[C@H]1C(=O)N1[C@H](C(O)=O)CCC1 DPURXCQCHSQPAN-AVGNSLFASA-N 0.000 description 1
- BTEMNFBEAAOGBR-BZSNNMDCSA-N Leu-Tyr-Lys Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CC1=CC=C(C=C1)O)C(=O)N[C@@H](CCCCN)C(=O)O)N BTEMNFBEAAOGBR-BZSNNMDCSA-N 0.000 description 1
- MQMIRLVJXQNTRJ-SDDRHHMPSA-N Lys-Gln-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CCC(=O)N)NC(=O)[C@H](CCCCN)N)C(=O)O MQMIRLVJXQNTRJ-SDDRHHMPSA-N 0.000 description 1
- ZMMDPRTXLAEMOD-BZSNNMDCSA-N Lys-His-Phe Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O ZMMDPRTXLAEMOD-BZSNNMDCSA-N 0.000 description 1
- AEIIJFBQVGYVEV-YESZJQIVSA-N Lys-Phe-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CC2=CC=CC=C2)NC(=O)[C@H](CCCCN)N)C(=O)O AEIIJFBQVGYVEV-YESZJQIVSA-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
- SXWQMBGNFXAGAT-FJXKBIBVSA-N Met-Gly-Thr Chemical compound CSCC[C@H](N)C(=O)NCC(=O)N[C@@H]([C@@H](C)O)C(O)=O SXWQMBGNFXAGAT-FJXKBIBVSA-N 0.000 description 1
- WNJXJJSGUXAIQU-UFYCRDLUSA-N Met-Phe-Phe Chemical compound C([C@H](NC(=O)[C@@H](N)CCSC)C(=O)N[C@@H](CC=1C=CC=CC=1)C(O)=O)C1=CC=CC=C1 WNJXJJSGUXAIQU-UFYCRDLUSA-N 0.000 description 1
- QQPMHUCGDRJFQK-RHYQMDGZSA-N Met-Thr-Leu Chemical compound CSCC[C@H](N)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@H](C(O)=O)CC(C)C QQPMHUCGDRJFQK-RHYQMDGZSA-N 0.000 description 1
- AJHCSUXXECOXOY-UHFFFAOYSA-N N-glycyl-L-tryptophan Natural products C1=CC=C2C(CC(NC(=O)CN)C(O)=O)=CNC2=C1 AJHCSUXXECOXOY-UHFFFAOYSA-N 0.000 description 1
- 239000006002 Pepper Substances 0.000 description 1
- 244000046052 Phaseolus vulgaris Species 0.000 description 1
- IWRZUGHCHFZYQZ-UFYCRDLUSA-N Phe-Arg-Tyr Chemical compound C([C@H](N)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(O)=O)C1=CC=CC=C1 IWRZUGHCHFZYQZ-UFYCRDLUSA-N 0.000 description 1
- MQVFHOPCKNTHGT-MELADBBJSA-N Phe-Asp-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CC(=O)O)NC(=O)[C@H](CC2=CC=CC=C2)N)C(=O)O MQVFHOPCKNTHGT-MELADBBJSA-N 0.000 description 1
- INHMISZWLJZQGH-ULQDDVLXSA-N Phe-Leu-Val Chemical compound CC(C)[C@@H](C(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@@H](N)CC1=CC=CC=C1 INHMISZWLJZQGH-ULQDDVLXSA-N 0.000 description 1
- ZUQACJLOHYRVPJ-DKIMLUQUSA-N Phe-Lys-Ile Chemical compound CC[C@H](C)[C@@H](C(O)=O)NC(=O)[C@H](CCCCN)NC(=O)[C@@H](N)CC1=CC=CC=C1 ZUQACJLOHYRVPJ-DKIMLUQUSA-N 0.000 description 1
- RVEVENLSADZUMS-IHRRRGAJSA-N Phe-Pro-Asn Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CC(N)=O)C(O)=O RVEVENLSADZUMS-IHRRRGAJSA-N 0.000 description 1
- ILGCZYGFYQLSDZ-KKUMJFAQSA-N Phe-Ser-His Chemical compound N[C@@H](Cc1ccccc1)C(=O)N[C@@H](CO)C(=O)N[C@@H](Cc1cnc[nH]1)C(O)=O ILGCZYGFYQLSDZ-KKUMJFAQSA-N 0.000 description 1
- GMWNQSGWWGKTSF-LFSVMHDDSA-N Phe-Thr-Ala Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C)C(O)=O GMWNQSGWWGKTSF-LFSVMHDDSA-N 0.000 description 1
- 241000233614 Phytophthora Species 0.000 description 1
- 235000016761 Piper aduncum Nutrition 0.000 description 1
- 240000003889 Piper guineense Species 0.000 description 1
- 235000017804 Piper guineense Nutrition 0.000 description 1
- 235000008184 Piper nigrum Nutrition 0.000 description 1
- QGOZJLYCGRYYRW-KKUMJFAQSA-N Pro-Glu-Tyr Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O QGOZJLYCGRYYRW-KKUMJFAQSA-N 0.000 description 1
- XYSXOCIWCPFOCG-IHRRRGAJSA-N Pro-Leu-Leu Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(C)C)C(O)=O XYSXOCIWCPFOCG-IHRRRGAJSA-N 0.000 description 1
- SEZGGSHLMROBFX-CIUDSAMLSA-N Pro-Ser-Gln Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CO)C(=O)N[C@@H](CCC(N)=O)C(O)=O SEZGGSHLMROBFX-CIUDSAMLSA-N 0.000 description 1
- 101710093543 Probable non-specific lipid-transfer protein Proteins 0.000 description 1
- 238000012300 Sequence Analysis Methods 0.000 description 1
- ZUGXSSFMTXKHJS-ZLUOBGJFSA-N Ser-Ala-Ala Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](C)C(=O)N[C@@H](C)C(O)=O ZUGXSSFMTXKHJS-ZLUOBGJFSA-N 0.000 description 1
- WDXYVIIVDIDOSX-DCAQKATOSA-N Ser-Arg-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@@H](NC(=O)[C@@H](N)CO)CCCN=C(N)N WDXYVIIVDIDOSX-DCAQKATOSA-N 0.000 description 1
- BNFVPSRLHHPQKS-WHFBIAKZSA-N Ser-Asp-Gly Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC(O)=O)C(=O)NCC(O)=O BNFVPSRLHHPQKS-WHFBIAKZSA-N 0.000 description 1
- BGOWRLSWJCVYAQ-CIUDSAMLSA-N Ser-Asp-Leu Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(C)C)C(O)=O BGOWRLSWJCVYAQ-CIUDSAMLSA-N 0.000 description 1
- UBRMZSHOOIVJPW-SRVKXCTJSA-N Ser-Leu-Lys Chemical compound OC[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCCN)C(O)=O UBRMZSHOOIVJPW-SRVKXCTJSA-N 0.000 description 1
- YUJLIIRMIAGMCQ-CIUDSAMLSA-N Ser-Leu-Ser Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(O)=O YUJLIIRMIAGMCQ-CIUDSAMLSA-N 0.000 description 1
- GZGFSPWOMUKKCV-NAKRPEOUSA-N Ser-Pro-Ile Chemical compound CC[C@H](C)[C@@H](C(O)=O)NC(=O)[C@@H]1CCCN1C(=O)[C@@H](N)CO GZGFSPWOMUKKCV-NAKRPEOUSA-N 0.000 description 1
- SNXUIBACCONSOH-BWBBJGPYSA-N Ser-Thr-Ser Chemical compound OC[C@H](N)C(=O)N[C@@H]([C@H](O)C)C(=O)N[C@@H](CO)C(O)=O SNXUIBACCONSOH-BWBBJGPYSA-N 0.000 description 1
- KIEIJCFVGZCUAS-MELADBBJSA-N Ser-Tyr-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CC2=CC=C(C=C2)O)NC(=O)[C@H](CO)N)C(=O)O KIEIJCFVGZCUAS-MELADBBJSA-N 0.000 description 1
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical class [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 description 1
- 244000061458 Solanum melongena Species 0.000 description 1
- 235000002597 Solanum melongena Nutrition 0.000 description 1
- 244000061456 Solanum tuberosum Species 0.000 description 1
- 235000002595 Solanum tuberosum Nutrition 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
- XSLXHSYIVPGEER-KZVJFYERSA-N Thr-Ala-Val Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C)C(=O)N[C@@H](C(C)C)C(O)=O XSLXHSYIVPGEER-KZVJFYERSA-N 0.000 description 1
- JKGGPMOUIAAJAA-YEPSODPASA-N Thr-Gly-Val Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)NCC(=O)N[C@@H](C(C)C)C(O)=O JKGGPMOUIAAJAA-YEPSODPASA-N 0.000 description 1
- KPNSNVTUVKSBFL-ZJDVBMNYSA-N Thr-Met-Thr Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CCSC)C(=O)N[C@@H]([C@@H](C)O)C(=O)O)N)O KPNSNVTUVKSBFL-ZJDVBMNYSA-N 0.000 description 1
- WYLAVUAWOUVUCA-XVSYOHENSA-N Thr-Phe-Asp Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CC(O)=O)C(O)=O WYLAVUAWOUVUCA-XVSYOHENSA-N 0.000 description 1
- MUAFDCVOHYAFNG-RCWTZXSCSA-N Thr-Pro-Arg Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CCCNC(N)=N)C(O)=O MUAFDCVOHYAFNG-RCWTZXSCSA-N 0.000 description 1
- OJKVFAWXPGCJMF-BPUTZDHNSA-N Trp-Pro-Ser Chemical compound C1C[C@H](N(C1)C(=O)[C@H](CC2=CNC3=CC=CC=C32)N)C(=O)N[C@@H](CO)C(=O)O OJKVFAWXPGCJMF-BPUTZDHNSA-N 0.000 description 1
- NSOMQRHZMJMZIE-GVARAGBVSA-N Tyr-Ala-Ile Chemical compound CC[C@H](C)[C@@H](C(O)=O)NC(=O)[C@H](C)NC(=O)[C@@H](N)CC1=CC=C(O)C=C1 NSOMQRHZMJMZIE-GVARAGBVSA-N 0.000 description 1
- FGJWNBBFAUHBEP-IHPCNDPISA-N Tyr-Asp-Trp Chemical compound C1=CC=C2C(=C1)C(=CN2)C[C@@H](C(=O)O)NC(=O)[C@H](CC(=O)O)NC(=O)[C@H](CC3=CC=C(C=C3)O)N FGJWNBBFAUHBEP-IHPCNDPISA-N 0.000 description 1
- OLWFDNLLBWQWCP-STQMWFEESA-N Tyr-Gly-Met Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)NCC(=O)N[C@@H](CCSC)C(O)=O OLWFDNLLBWQWCP-STQMWFEESA-N 0.000 description 1
- FWOVTJKVUCGVND-UFYCRDLUSA-N Tyr-Met-Phe Chemical compound CSCC[C@@H](C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)O)NC(=O)[C@H](CC2=CC=C(C=C2)O)N FWOVTJKVUCGVND-UFYCRDLUSA-N 0.000 description 1
- VPEFOFYNHBWFNQ-UFYCRDLUSA-N Tyr-Pro-Tyr Chemical compound C([C@H](N)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(O)=O)C1=CC=C(O)C=C1 VPEFOFYNHBWFNQ-UFYCRDLUSA-N 0.000 description 1
- SLLKXDSRVAOREO-KZVJFYERSA-N Val-Ala-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](C)NC(=O)[C@H](C(C)C)N)O SLLKXDSRVAOREO-KZVJFYERSA-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
- JAKHAONCJJZVHT-DCAQKATOSA-N Val-Lys-Ser Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CO)C(=O)O)N JAKHAONCJJZVHT-DCAQKATOSA-N 0.000 description 1
- MGVYZTPLGXPVQB-CYDGBPFRSA-N Val-Met-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)O)NC(=O)[C@H](CCSC)NC(=O)[C@H](C(C)C)N MGVYZTPLGXPVQB-CYDGBPFRSA-N 0.000 description 1
- GUIYPEKUEMQBIK-JSGCOSHPSA-N Val-Tyr-Gly Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](Cc1ccc(O)cc1)C(=O)NCC(O)=O GUIYPEKUEMQBIK-JSGCOSHPSA-N 0.000 description 1
- DFQZDQPLWBSFEJ-LSJOCFKGSA-N Val-Val-Asn Chemical compound CC(C)[C@@H](C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC(=O)N)C(=O)O)N DFQZDQPLWBSFEJ-LSJOCFKGSA-N 0.000 description 1
- LLJLBRRXKZTTRD-GUBZILKMSA-N Val-Val-Ser Chemical compound CC(C)[C@@H](C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CO)C(=O)O)N LLJLBRRXKZTTRD-GUBZILKMSA-N 0.000 description 1
- 235000005072 Vigna sesquipedalis Nutrition 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 230000002378 acidificating effect Effects 0.000 description 1
- 238000012271 agricultural production Methods 0.000 description 1
- 108010076324 alanyl-glycyl-glycine Proteins 0.000 description 1
- 108010047495 alanylglycine Proteins 0.000 description 1
- 125000000539 amino acid group Chemical group 0.000 description 1
- 229910021529 ammonia Inorganic materials 0.000 description 1
- 230000003321 amplification Effects 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- 108010077245 asparaginyl-proline Proteins 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- 239000003899 bactericide agent Substances 0.000 description 1
- 230000003115 biocidal effect Effects 0.000 description 1
- 230000001488 breeding effect Effects 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- 239000001511 capsicum annuum Substances 0.000 description 1
- 108010089934 carbohydrase Proteins 0.000 description 1
- 230000003197 catalytic effect Effects 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
- 238000005119 centrifugation Methods 0.000 description 1
- 229940124587 cephalosporin Drugs 0.000 description 1
- 150000001780 cephalosporins Chemical class 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
- 230000000052 comparative effect Effects 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 108010069495 cysteinyltyrosine Proteins 0.000 description 1
- 238000006731 degradation reaction Methods 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 238000007598 dipping method Methods 0.000 description 1
- 239000003814 drug Substances 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
- 230000001408 fungistatic effect Effects 0.000 description 1
- 239000000499 gel Substances 0.000 description 1
- 238000001502 gel electrophoresis Methods 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
- 235000013922 glutamic acid Nutrition 0.000 description 1
- 239000004220 glutamic acid Substances 0.000 description 1
- 108010078144 glutaminyl-glycine Proteins 0.000 description 1
- 150000004676 glycans Chemical class 0.000 description 1
- 229930182470 glycoside Natural products 0.000 description 1
- 125000003147 glycosyl group Chemical group 0.000 description 1
- 230000013595 glycosylation Effects 0.000 description 1
- 238000006206 glycosylation reaction Methods 0.000 description 1
- 108010066198 glycyl-leucyl-phenylalanine Proteins 0.000 description 1
- 108010048994 glycyl-tyrosyl-alanine Proteins 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
- 244000038280 herbivores Species 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 239000000411 inducer Substances 0.000 description 1
- 230000006698 induction Effects 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 229960000318 kanamycin Drugs 0.000 description 1
- 229930182823 kanamycin A Natural products 0.000 description 1
- 238000011031 large-scale manufacturing process Methods 0.000 description 1
- 108010073472 leucyl-prolyl-proline Proteins 0.000 description 1
- 238000009630 liquid culture Methods 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 108020004999 messenger RNA Proteins 0.000 description 1
- 230000009456 molecular mechanism Effects 0.000 description 1
- 239000004570 mortar (masonry) Substances 0.000 description 1
- 235000021278 navy bean Nutrition 0.000 description 1
- 238000011017 operating method Methods 0.000 description 1
- 230000036961 partial effect Effects 0.000 description 1
- 230000007170 pathology Effects 0.000 description 1
- 239000008188 pellet Substances 0.000 description 1
- 239000000575 pesticide Substances 0.000 description 1
- 108010064486 phenylalanyl-leucyl-valine Proteins 0.000 description 1
- 108010084572 phenylalanyl-valine Proteins 0.000 description 1
- 230000022589 plant-type hypersensitive response Effects 0.000 description 1
- 229920001282 polysaccharide Polymers 0.000 description 1
- 239000005017 polysaccharide Substances 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 108010090894 prolylleucine Proteins 0.000 description 1
- 238000000751 protein extraction Methods 0.000 description 1
- 230000008929 regeneration Effects 0.000 description 1
- 238000011069 regeneration method Methods 0.000 description 1
- 230000008261 resistance mechanism Effects 0.000 description 1
- 108091008146 restriction endonucleases Proteins 0.000 description 1
- 238000012163 sequencing technique Methods 0.000 description 1
- 108010071207 serylmethionine Proteins 0.000 description 1
- 235000011091 sodium acetates Nutrition 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 238000002798 spectrophotometry method Methods 0.000 description 1
- 230000003068 static effect Effects 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 238000012549 training Methods 0.000 description 1
- 238000011282 treatment Methods 0.000 description 1
- 241001478887 unidentified soil bacteria Species 0.000 description 1
- 238000011144 upstream manufacturing Methods 0.000 description 1
- 108010073969 valyllysine Proteins 0.000 description 1
- 239000005418 vegetable material Substances 0.000 description 1
- 238000012795 verification Methods 0.000 description 1
- 235000020234 walnut Nutrition 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/14—Hydrolases (3)
- C12N9/24—Hydrolases (3) acting on glycosyl compounds (3.2)
- C12N9/2402—Hydrolases (3) acting on glycosyl compounds (3.2) hydrolysing O- and S- glycosyl compounds (3.2.1)
- C12N9/2405—Glucanases
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
- C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
- C12N15/82—Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
- C12N15/8241—Phenotypically and genetically modified plants via recombinant DNA technology
- C12N15/8261—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield
- C12N15/8271—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield for stress resistance, e.g. heavy metal resistance
- C12N15/8279—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield for stress resistance, e.g. heavy metal resistance for biotic stress resistance, pathogen resistance, disease resistance
- C12N15/8282—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield for stress resistance, e.g. heavy metal resistance for biotic stress resistance, pathogen resistance, disease resistance for fungal resistance
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
- C12Y302/00—Hydrolases acting on glycosyl compounds, i.e. glycosylases (3.2)
- C12Y302/01—Glycosidases, i.e. enzymes hydrolysing O- and S-glycosyl compounds (3.2.1)
- C12Y302/01039—Glucan endo-1,3-beta-D-glucosidase (3.2.1.39)
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Genetics & Genomics (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Wood Science & Technology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Zoology (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Biotechnology (AREA)
- Biochemistry (AREA)
- Biomedical Technology (AREA)
- Molecular Biology (AREA)
- Microbiology (AREA)
- Medicinal Chemistry (AREA)
- Cell Biology (AREA)
- Physics & Mathematics (AREA)
- Biophysics (AREA)
- Plant Pathology (AREA)
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
Abstract
The present invention is a kind of glucanase gene PnGlu1 of pseudo-ginseng β 1,3 and its application, discloses a kind of family gene of pathogenesis-related proteins 2 of pseudo-ginsengPnGlu1, its nucleotide sequence such as SEQ IDNO：Described in 1, the dextranases of β 1,3 are encoded；The present invention is confirmed by functional genomics relation technological researchingPnGlu1Gene, which has, improves the antimycotic function of plant, and the present invention is antimycoticPnGlu1It is gene constructed on plant expression vector and overexpression in tobacco is transferred to, transgenic tobacco plant has very strong extracorporeal antifungal activity, overexpressionPnGlu1Growth of the transgene tobacco to four kinds of fungies such as grape seat chamber bacterium, wheel branch sickle-like bacteria, Fusarium solani, colletotrichum gloeosporioides Penzs there is obvious inhibitory action.
Description
Technical field
The present invention relates to molecular biology and genetic engineering relation technological researching field, particularly a kind of pseudo-ginseng β -1,3
Glucanase genePnGlu1And its application.
Background technology
Plant pathogenic fungi refers to parasitize plant and causes the fungi of disease, and the plant pathogenic fungi recorded is up to 8000
More than kind, and more than 30,000 can be caused to plant plant disease, account for the 70%~80% of plant disease sum, belong to the first major class pathogen.At present
Control to crop fungal disease has following several ways：Seed selection simultaneously uses resistant crop varieties；Use chemical sterilization
Agent；Take crop rotation, avoid propagation with soil bacteria and with cause of disease vegetable material etc..But to filter out and resistant make article
Kind needs to spend substantial amounts of manpower and materials and time；Chemical bactericide cost is higher, may produce plant pathogenic fungi anti-
The property of medicine, its residual hazard can produce pollution to environment；Crop rotation can not fundamentally solve the problems, such as the harm of plant pathogenic fungi.In recent years
Come, with the continuous development of Molecular Biology and technology, make people be not only able to deeply recognize plant from molecular level
The mechanism to be interacted with pathogen, but also disease-resistant New Crop Varieties can be quickly and efficiently cultivated by genetic engineering,
It is the new method for improving plant disease-resistant ability.
The family member of pathogenesis-related proteins second of beta-1,3-glucanase（van LOON L C, van STRIEN E
A. The families of pathogenesis related proteins, their activities, and
comparative an analysis of PR1 type proteins. Physiol Mol Plant Pathol, 1999,
55: 85-97.）.β -1,3 dextranases are widely distributed in higher plant, including pollen tube wall, fine cell wall, screen casing end wall
Etc. having presence in structure, they play a significant role in plant normal growth development.In addition, β -1,3 dextranases are being planted
Key player is also play in the resistance mechanism of thing（Ou Yangbo, Li Han rosy clouds plants β-1,3-dextranase and its gene
Chinese biological engineering magazine, 2002,22 (6):18-23.）.During the external pathogen of plant resistant, β -1, glucan
Enzyme is the important hydrolase of synthesis in plant hypersensitive response (Hypersensitive Response, HR).Plant resistant germ
The approach of infringement is varied, including reinforces cell membrane, produces antibacterial material, and its preferred mode is then HR.According to gene
Evolutionary relationship, protein features it is similar with amino acid sequence, β -1,3 dextranases can be divided into three types：It is positioned at liquid
The alkaline type of bubble, extracellular positioning acid type, also have some other beta-1,3 glucanase in sequence with both the above class
Type differs greatly, can be separately as one kind.
Known β -1,3 dextranases belong to the family of glycosyl hydrolase the 17th, and its member has common sequence knot
Structure：(LIVM) the X 2G of 2 X 2 (LIVMFYW) 32 (STAG), 22 W of E 2 (ST) 2 G, 2 P 2 (ST) 2, such as exist
Just there is VNVVVSESGWPSDG structure in the beta-1,3 glucanase of pea（Chang M M, Culley D E, Hadwiger
L A. Nucleotide sequence of a pea (Pisum sativum L. ) β-1,3 glucanase gene.
Plant Physiol, 1993, 101: 1121-1122）, several conservative glutamic acid and asparagus fern ammonia in the enzyme amino acid sequence
Sour residue may be relevant with the hydrolysis of glycoside bond that beta-1,3 glucanase is catalyzed.Alkaline beta-1,3 glucanase generally has a liquid
Carboxyl terminal polypeptide (Carboxyl Terminal Polypeptide, CTPP) structure of positioning is steeped, is often contained in CTPP
Glycosylation site and CTPP excision signal amino acid structures, it is extracellular that CTPP shortage make it that dextranase is secreted into.Benzene in CTPP
The presence of Ala-Gly dipeptides is probably the signal of CTPP excisions.Therefore, CTPP presence or absence turns into β -1, and 3 Portugals gather
The important evidence of carbohydrase classification.In addition, β -1, also containing several relatively conservative sections in 3 dextranase mature peptides, particularly
There is a highly conserved catalytic site.Glucanase gene such as tobacco, Kidney bean, pea and soybean is all quite protected containing two
The peptide chain kept, VVSESGWPS and FAMFDEN.
The structural core of fungal cell wall is the compound that β -1,3 glucans of β -1,3 branches and chitin are cross-linked to form
（Adams DJ. Fungal cell wall chitinases and glucanases. Microbiology, 2004.
150: 2029-2035.）.β -1,3 dextranase energy catalyzing hydrolysis β -1,3 glucans, so as to suppress the growth of fungi, propagation,
The product oligosaccharides of hydrolysis can turn into the important motivating factor of plant defense response（Ham K S, Wu S C, Darvill A
G, et al. Fungal pathogens secrete an inhibitor protein that distinguishes
isoforms of plant pathogenesis-related endo-β-1,3-glucanases. Plant Journal,
1997, 11(2): 169-179.）.Bacteriostatic test plate shows, β -1,3 dextranases to phytophthora (Phytophthora capsici) mycelial growth is inhibited（Kim Y J, Hwang B K. Isolation of a basic 34
kiloDalton β-1,3-glucanase with inhibitory activity against Phytophthora capsici from pepper stems. Physiological & Molecular Plant Pathology, 1997,
50(2): 103-115.）.Soybean (Glycine max) in, it has been found that glucan exciton associated proteins GEBP
(Glucan Elicitor Binding Protein, GEBP), is positioned on the plasma membrane of soybean root cells, being capable of specificity
Ground combination β -1, the oligosaccharides exciton discharged in 3 glucan degradation processes, induce the defense response of plant（Ham K S,
Wu S C, Darvill A G, et al. Fungal pathogens secrete an inhibitor protein
that distinguishes isoforms of plant pathogenesis-related endo-β-1,3-
glucanases. Plant Journal, 1997, 11(2): 169-179.）.Beta-1,3 glucanase is common with chitinase
Fungistatic effect becomes apparent during effect.The oligosaccharides discharged in hydrolytic process by fungal cell wall can be used as plant a variety of
The motivating factor of disease resistance response, induce comprehensive disease resistance response of plant（Klarzynski O, Plesse B, Joubert J
M, et al. Linear beta-1,3 glucans are elicitors of defense responses in
tobacco. Plant Physiol, 2000. 124(3): 1027-1038.）.
Pseudo-ginseng (Panax notoginseng) main product in Yunnan Wenshan Prefecture Yanshan County, Maguan, Xichou, Guangnan, Malipo,
Also there is plantation on the ground such as Funing, Qiu north etc., another Guangxi Tianyang County, Jingxi, Tiandong County, Debao.The pseudo-ginseng of Yunnan Wenshan Prefecture is with a long history, production
Amount is big, quality is good, practises and claims " literary pseudo-ginseng ", " pseudo-ginseng ", is famous genunie medicinal materials.Pseudo-ginseng is grown in the moon and covered in environment throughout the year, disease
Insect pest generation is more serious, according to statistics, the pest and disease damage occurred on pseudo-ginseng kind about more than 20.Wherein, it is main to have root rot, black
Pinta, Northern leaf spot, epidemic disease, powdery mildew, slug, cutworm, aphid, scale insect, looper etc., pseudo-ginseng is every year all because of the harm of disease pest
Cause extremely serious loss.It is to ensure a major measure of pseudo-ginseng Yield and quality to strengthen the pseudo-ginseng prevention and control of plant diseases, pest control.In addition,
Recognize the molecular mechanism of pseudo-ginseng defense responses, and clone and be particularly important to using related disease-resistant gene.
The content of the invention
The purpose of the present invention is the full-length gene that clone obtains the beta-1,3 glucanase with antifungal activity from pseudo-ginsengPnGlu1；Pseudo-ginseng beta-1,3 glucanase genePnGlu1Nucleotide sequence such as SEQ IDNO：Shown in 1, the gene cDNA total length sequence
1417bp is classified as, 5 ' non-translational regions of ORFs, 63bp comprising a 1140bp, 214bp 3 ' non-translational regions, coding
Such as SEQ IDNO：The protein of amino acid sequence shown in 2.
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 mediated method, by entering one
Whether the step experimental verification gene has antimycotic activity, is resisted for the later-stage utilization improvement of genes tobacco and other plant
The ability of fungal disease lays the foundation.This unnamed gene is by inventorPnGlu1。
Plant β -1,3 dextranases can be by pathogen, herbivore and other biological or abiotic inducer, mechanical treatments
Or induction is stimulated to produce.Chitin and β -1,3 glucans are covered in cell wall as fungi cell wall main component, β -1,3 glucans
Outermost layer, dextranase have hydrolysis to the polysaccharide that these expose.Beta-1,3 glucanase hydrolyzes the cell membrane of fungi, and makes
Pathogen mycelia apical cell's wall is thinning, and then imbalance occurs for turgescence and cell wall tension, so as to cause mycelium top swollen
It is swollen crush, it is final dead.What is more important, the oligosaccharides discharged in hydrolytic process by fungal cell wall being capable of conduct
The motivating factor of a variety of disease resistance responses of plant, induce comprehensive disease resistance response of plant.
The present invention relates to separation to includePnGlu1DNA fragmentation and identify its function, to the gene carry out sequence homology
Analysis, findPnGlu1The protein sequence and walnut of coding（Juglans regia）, cocoa（Theobroma cacao）, it is peppery
Green pepper（Capsicum annuum）Dextranase protein similarities be respectively 67%, 66% and 63%.PnGlu1Gene code one
The protein being made up of 379 amino acid residues, its molecular mass are 41.8KD, isoelectric point（pI）For 7.75.Protein sequence
In include 29 acidic amino acids（D, E）, 34 basic amino acids（K,R,H）.Sequence shown in overexpression sequence table SEQ ID
Can strengthen tobacco to grape seat chamber bacterium (Botryosphaeria dothidea), wheel branch sickle-like bacteria (Fusarium verticillioide), Fusarium solani (F. solani), colletotrichum gloeosporioides Penz (Colletotrichum gloeosporioides) resistance.
The present invention is by pseudo-ginseng beta-1,3 glucanase genePnGlu1Apply and improving tobacco to grape seat chamber bacterium, wheel branch
In sickle-like bacteria, Fusarium solani, colletotrichum gloeosporioides Penz resistance, concrete operations are as follows：
（1）With Fusarium solani be inoculated with Panax notoginseng root, take inoculation after 24 h Roots of Panax Notoginseng extraction total serum IgE, using the total serum IgE of extraction as
Template, with oligo (dT) 18 for reverse transcription primer, pass through reverse transcriptase chain reaction (reverse
Transcription-polymerase chain reaction, RT-PCR) amplifyPnGlu1Code area, then by it
It is connected on pMD-18T carriers, the clone with target gene is obtained through sequencing；
（2）Use restriction enzymeBamHI andEcoRI digestions pMD-18T-PnGlu1, target gene piece is obtained by glue reclaim
Section, with same digestion with restriction enzyme plant expression vector pCAMBIA2300s, carrier large fragment needed for glue reclaim acquisition,
It will be obtained againPnGlu1Genetic fragment is connected with pCAMBIA2300s fragments, plant overexpression vector is built, afterwards by institute's structure
The recombinant vector built is 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 to fungi, is finally filtered 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, and simple to operate, is readily available Gao Kangcai
Material.The present invention is from pseudo-ginsengPnGlu1Gene can strengthen the resistance of plant against fungal, by the channel genes tobacco, can produce
Raw new varieties and new material with fungus resistant.The disease brought using technique for gene engineering reduction fungi has obvious excellent
Gesture and the importance do not replaced.It can be that the offers such as large-scale production crop, flowers are convenient, a large amount of reduction chemical pesticides
Use, can also be that agricultural production is cost-effective, reduce environmental pollution and raise the management level, therefore the present invention is with wide
Market application foreground.
Brief description of the drawings
Fig. 1 is the present inventionPnGlu1The PCR testing result schematic diagrames of transgene tobacco genomic DNA, in figure：Marker is
DL2000 DNA Marker (the precious biology in Dalian)；Positive control is plasmid pMD-18T-PnGlu1Product is tied for the PCR of template；
WT is the product that non-transgenic tobacco (wild type) STb gene is template PCR；
Fig. 2 is of the invention positivePnGlu1In transgene tobaccoPnGlu1The 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 primer；Positive control is plasmid pMD-18T-PnGlu1For the PCR primer of template；
Fig. 3 is the present inventionPnGlu1Transgene tobacco In Vitro Bacteriostatic design sketch；A, b, c, d are to glue spore anthrax respectively in figure
Bacterium, wheel branch sickle-like bacteria, grape seat chamber bacterium, Fusarium solani；WT is the total protein of wild-type tobacco；Buffer is blank control,
I.e. without protein control (being used for the buffer solution for extracting 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：PnGlu1Full-length gene is cloned and sequence analysis
The conidiospore suspension of Fusarium solani is prepared, root dipping is inoculated with the annual min of pseudo-ginseng 30, takes the pseudo-ginseng of 24 h after inoculation
Extract total serum IgE in root.With liquid nitrogen by Roots of Panax Notoginseng grind into powder, then it is transferred in centrifuge tube, is extracted using guanidine isothiocyanate method
Total serum IgE, reverse transcriptase M-MLV (promega) is used using total serum IgE as the chains of templated synthesis cDNA first, reaction system and operated
Cheng Wei：5 μ g Total RNA are taken, sequentially add 50 ng oligo（dT）, 2 μ L dNTP（2.5mM each）, DEPC water extremely
Reaction volume is 14.5 μ L；After mixing, 4 are then sequentially added in the min of cooled on ice 5 rapidly after 70 DEG C of min of heat denatured 5
μL 5×First-stand buffer、0.5 μL RNasin（200U）、1 μL M-MLV（200U）, mix and centrifuge in short-term,
42 DEG C of h of warm bath 1.5,70 DEG C of 10 min of heating, terminating reaction after taking-up.It is standby that -20 DEG C of preservations are placed in after the synthesis of the chains of cDNA first
With.
Using the first chain cDNA of synthesis as template, amplifying target genesPnGlu1, upstream and downstream primer sequence used is respectively
5 ' TCATCAGTACGTAGTTCTCTTACTTC3 ' and 5 ' AGCTAAGCTAGTTACATGTCACTCT3 '.Using AdvantageTM
2 PCR Enzyme（Clontech）Amplify target gene；PCR reaction conditions：95℃4 min；95 DEG C of 30 s, 54 DEG C 30
S, 72 DEG C of 80 s, 30 circulations；72℃ 10 min；Reaction system（20 μL）For 1 μ L cDNA, 2 10 × Advantage of μ L
2 PCR Buffer、1.8μL 50×dNTP Mix （10mM each）, 0.2 μ L forward primers（10 μM）, 0.2 μ L reversely draw
Thing（10 μM）、0.2 μL Advantage 2 PCR Polymerase Mix、14.6μL PCR-Grade water；PCR is tied
Shu Hou, 5 μ L are taken to be used for agarose gel electrophoresis, to detect the specificity of amplified production and size.
Resulting PCR primer only has a DNA band, therefore directly carries out TA clones to PCR primer, 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 primers are taken, are sequentially added
1 μL pMD18-T vector（50 ng/μL）With 2.5 μ L 2 × Ligation solution I, 16 DEG C of mistakes are placed in after mixing
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 amplificationPnGlu1Special primer identify multiple cloning sites insertionPnGlu1Clone, the clone identified is sequenced, finally
ObtainPnGlu1Full-length cDNA is 1417bp, passes through NCBI ORF finder（http://
www.ncbi.nlm.nih.gov/gorf/gorf.html）Analysis finds that it includes 1140bp opening code-reading frame（See sequence
List）,PnGlu1One protein PnGlu1 containing 379 amino acid of coding, its protein is 41.8 k D, etc.
Electric point (pI) is 7.75, and it is basic protein to show the albumen.
Embodiment 2：Plant overexpression vector is built
Using a small amount of extraction agent boxes of SanPrep pillar DNAs（Give birth to work in Shanghai）Extraction insertionPnGlu1Escherichia coli matter
Grain pMD-18T-PnGlu1And plant expression vector pCAMBIA2300s plasmid, take 1 μ L be used for agarose gel electrophoresis with
The integrality and concentration level of plasmid are extracted in detection；Use restriction enzymeBamHI（TaKaRa）WithEcoRI（TaKaRa）Point
It is other to plasmid pMD-18T-PnGlu1Double digestion is carried out with pCAMBIA2300s（100 μ L systems）, reaction system and operating process
For：Take 20 μ L pMD-18T-PnGlu1With pCAMBIA2300s plasmids, sequentially add 10 μ L10 × H buffer, 4 μ LBamHI、6μLEcoRI、60μL ddH2O, centrifuged in short-term after mixing, be placed in 37 DEG C of reaction overnights；By all digestion products points in
Electrophoresis is carried out in Ago-Gel, it is then rightPnGlu1Fragment and pCAMBIA2300s carriers large fragment carry out glue reclaim respectively,
Whole process uses SanPrep pillar DNA glue reclaim kits（Give birth to work in Shanghai）；1 μ L recovery products are taken to pass through Ago-Gel
Electrophoresis detection reclaims the size and concentration of fragment, is placed in -20 DEG C and saves backup.
Utilize T4 DNA Ligase（TaKaRa）, by recoveryPnGlu1 DNA fragmentation and pCAMBIA2300s carrier-pellets
Section connects, reaction system（20 μL）It is with operating process：Take 10 μ LPnGlu1 DNA 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, centrifuged 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 solutionPnGlu1Special primer enter performing PCR, pick outPnGlu1With
The clone that pCAMBIA2300s is successfully connected, the bacterial strain detected are placed in -80 DEG C and saved backup if the positive, addition glycerine.
Using SanPrep pillar plasmid extraction kits（Give birth to work in Shanghai）Extract and purify in above-mentioned Escherichia coli
pCAMBIA2300s-PnGlu1Plasmid.Frozen-thawed method is then used by the plant expression vector pCAMBIA2300s- of above-mentioned structurePnGlu1It is transferred in agrobacterium tumefaciens lba4404 competent cell.Operating procedure is：Take 2 μ g pCAMBIA2300s-PnGlu1
Plasmid is added in the centrifuge tube containing 200 μ L competent cells, the min of ice bath 5 after gently mixing, is then continued in liquid nitrogen and is freezed
1 min, 37 DEG C of min of water-bath 5 are then immediately placed in, the min of ice bath 2,800 μ L LB Liquid Cultures of addition are based on 28 immediately afterwards
DEG C h of shaken cultivation 4.Agrobacterium after activation is applied on the LB solid mediums containing 50 mg/L Km, 28 DEG C of static trainings
Support.Picking individual colonies shake bacterium, then with amplificationPnGlu1Specific primer enter performing PCR, detect pCAMBIA2300s-PnGlu1It is
It is no to be transferred in Agrobacterium, it is placed in -80 DEG C for positive colony, after addition glycerine and saves 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-pickled 30s, with being used after sterile water washing
0.1% HgCl2Soak 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, 16 h/d illumination）, monthly use 1/2MS culture mediums subculture once later.
That preservation is taken out from -80 DEG C of refrigerators contains pCAMBIA2300s-PnGlu1The 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.The muddy bacterium solutions of 1 mL are drawn to the LB solid mediums containing 50 mg/L Km, 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, it 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 into co-cultivation base
Upper carry out incubated at room temperature, the co-cultivation base of Transformation of tobacco is MS+0.02mg/L 6-BA+2.1 mg/L NAA+30g/L
Sucrose+6g/L agar, co-culture 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 be 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
200 mg/L Cef MS culture medium squamous subcultures, because tobacco callus differentiation rate is higher, therefore need to enter regeneration plant traveling one
Step screening, tobacco regrowth, which is moved on the MS culture mediums containing 50 mg/L Km, makes it take root, finally preferable from taking root
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 plantPnGlu1
Special primer enter performing PCR, after PCR terminates, take 8 μ L products be used for agarose gel electrophoresis to detect positive transgenic plant,
The amplification of Partial Tobacco transfer-gen plant as shown in figure 1,PnGlu1Transgene tobacco screens 47 plants of positive transgenics altogether
Plant.
Embodiment 4：In transgene tobaccoPnGlu1Expression analysis and transfer-gen plant antifungal activity analysis
Take positive transgenic individual plant and non-transgenic tobacco（Wild type）Tender leaf extraction total serum IgE, reverse transcription generation cDNA the
One chain, and expanded as templatePnGlu1Special primer enter performing PCR, according in each transgenosis individual plant of PCR interpretations of resultPnGlu1The expression of transcriptional level, Total RNAs extraction and RT-PCR method 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 individual plant as shown in Fig. 2 detect in 31 transgenosis individual plants altogetherPnGlu1In transcriptional level great expression, the numbering of these individual plants is 1~31.
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, albumen is added when colony growth to diameter is about 2 ~ 3cm, analyze transfer-gen plant body
Outer antifungal activity.
In order to prevent albumen that other living contaminantses are extracted, whole vegetable protein extraction process is sterile working, first
First take 1 g transgene tobacco individual plants（Numbering is respectively 1,6,13,21）And wild-type leaves are put into mortar, 1 mL albumen is added
Extract solution（1M NaCl, 0.1M sodium acetates, 1% PVP, pH6）, it is fully ground；It is transferred in 1.5 mL centrifuge tubes, 4 DEG C after mixing
Stand overnight, 4 DEG C of centrifugation 30min（12,000g/min）, supernatant is taken in 1.5 new mL centrifuge tubes, and is taken in right amount with ultraviolet
Spectrophotometric determination total protein concentration.The total protein concentration of transgenosis and WT lines is adjusted to 0.2 μ g/ μ L, then
20 μ L drops are taken respectively on the aseptic filter paper of each fungi culture medium, except adding different transgenosis on the flat board of each fungi
The total protein of tobacco plant, while the total protein of parallel addition wild-type tobacco and blank control（Extract molten used in albumen
Liquid）, the situation of each processing fungi growth is after a few days observed in 28 DEG C of cultures, and is evaluated accordinglyPnGlu1Transgene tobacco it is external
Antifungal activity, as a result as shown in figure 3,PnGlu1Transgene tobacco albumen is to grape seat chamber bacterium, wheel branch sickle-like bacteria, eggplant corruption reaping hook
The growth of bacterium, colletotrichum gloeosporioides Penz has very strong inhibitory action.
Sequence table
<110>Kunming University of Science and Technology
<120>A kind of pseudo-ginseng beta-1,3 glucanase gene PnGlu1 and its application
<160> 4
<170> PatentIn version 3.3
<210> 1
<211> 1417
<212> DNA
<213> Panax notoginseng
<220>
<221> mRNA
<222> (1)..(1417)
<220>
<221> 5'UTR
<222> (1)..(63)
<220>
<221> CDS
<222> (64)..(1203)
<220>
<221> 3'UTR
<222> (1204)..(1417)
<400> 1
gcggggaata tcatacattt catcagtacg tagttctctt acttctgatt gctttatata 60
tatatgttct tcaccatgac tatattttcc agaagaacta ataacagttt cctcgtaatg 120
acacctatac tgcttcttct ggggtttatg attgcaagct ttaagattac aggggtagaa 180
tctgttggcg cgtgttatgg aatgctagga aacaatctcc cacctgcatc agaagttgta 240
aatctataca aatcatacaa ccttgatcga atgagactct acgatccaaa tcgagccgct 300
atacaagctc tacaaaactc taatatcgaa gttatgattg gtgtcccaaa ctcagacctc 360
cagcgtctgg ccaatgatcc tggctatgca tacgactggg tgtacggaaa tcttgtagat 420
tacccacaag tcaaatttcg gtacatagcc gtaggaaatg aagtgagtcc cattaacggt 480
ggcacagcct ggctagctcc gttcgtctta ccagccatgc aacacatcca aacggcagta 540
ctttcggcag ctcgactggc aaacacggtt aaggtgtcaa ccgcaataga tatgacatta 600
ataggaaact cttatccccc ttcacaaggt agctttaggg gagatattag ggcatatttt 660
gatccgatta ttcggtttct tgtcaacaac aatgcgccct tgctagctaa tgtgtacccg 720
tattttagcc acattgggaa tccgcgtgat atttctttgt cttacgcaat tttcactgct 780
ccggggccag tgatatggga caatggcctt ggttaccaga atcttttcga tgcaatgatg 840
gatgctttat atgcggctgt tgagagggcc ggaggtggct cgttgaaggt ggtggtatca 900
gaaactggat ggccgtctgc cggaggagtg gcgacaactt ttgataatgc gcgtaattat 960
tactctagat tgattcaaca tgtggaaaag ggaaccccta ggaggccggg gagactagag 1020
acctacatgt ttgcgacgtt tgatgaaaat aataaaaatc cagaatatga gaagcatttt 1080
ggattgtttt tcccaaataa gcagcccaag tttccactca gaatttccat gggtactgga 1140
tctggggata tcgtttctga tggaaactct actagtttgg gttgggttaa gagtgacatg 1200
taactagctt agctagaggg tttgtaatat aataatattg catgatttgc ctttacatgc 1260
atgctttgag tttgggttga ataagtgtaa gcgatcatga tatgaatatt gatgtttgat 1320
ttaatttctt tgtatttaat ttgtaagttt ttgataagtg taaacaagga cgtttcatgt 1380
ttgttttgta caaaaaaaaa aaaaaaaaaa aaaaaaa 1417
<210> 2
<211> 379
<212> PRT
<213> Panax notoginseng
<400> 2
Met Phe Phe Thr Met Thr Ile Phe Ser Arg Arg Thr Asn Asn Ser Phe
1 5 10 15
Leu Val Met Thr Pro Ile Leu Leu Leu Leu Gly Phe Met Ile Ala Ser
20 25 30
Phe Lys Ile Thr Gly Val Glu Ser Val Gly Ala Cys Tyr Gly Met Leu
35 40 45
Gly Asn Asn Leu Pro Pro Ala Ser Glu Val Val Asn Leu Tyr Lys Ser
50 55 60
Tyr Asn Leu Asp Arg Met Arg Leu Tyr Asp Pro Asn Arg Ala Ala Ile
65 70 75 80
Gln Ala Leu Gln Asn Ser Asn Ile Glu Val Met Ile Gly Val Pro Asn
85 90 95
Ser Asp Leu Gln Arg Leu Ala Asn Asp Pro Gly Tyr Ala Tyr Asp Trp
100 105 110
Val Tyr Gly Asn Leu Val Asp Tyr Pro Gln Val Lys Phe Arg Tyr Ile
115 120 125
Ala Val Gly Asn Glu Val Ser Pro Ile Asn Gly Gly Thr Ala Trp Leu
130 135 140
Ala Pro Phe Val Leu Pro Ala Met Gln His Ile Gln Thr Ala Val Leu
145 150 155 160
Ser Ala Ala Arg Leu Ala Asn Thr Val Lys Val Ser Thr Ala Ile Asp
165 170 175
Met Thr Leu Ile Gly Asn Ser Tyr Pro Pro Ser Gln Gly Ser Phe Arg
180 185 190
Gly Asp Ile Arg Ala Tyr Phe Asp Pro Ile Ile Arg Phe Leu Val Asn
195 200 205
Asn Asn Ala Pro Leu Leu Ala Asn Val Tyr Pro Tyr Phe Ser His Ile
210 215 220
Gly Asn Pro Arg Asp Ile Ser Leu Ser Tyr Ala Ile Phe Thr Ala Pro
225 230 235 240
Gly Pro Val Ile Trp Asp Asn Gly Leu Gly Tyr Gln Asn Leu Phe Asp
245 250 255
Ala Met Met Asp Ala Leu Tyr Ala Ala Val Glu Arg Ala Gly Gly Gly
260 265 270
Ser Leu Lys Val Val Val Ser Glu Thr Gly Trp Pro Ser Ala Gly Gly
275 280 285
Val Ala Thr Thr Phe Asp Asn Ala Arg Asn Tyr Tyr Ser Arg Leu Ile
290 295 300
Gln His Val Glu Lys Gly Thr Pro Arg Arg Pro Gly Arg Leu Glu Thr
305 310 315 320
Tyr Met Phe Ala Thr Phe Asp Glu Asn Asn Lys Asn Pro Glu Tyr Glu
325 330 335
Lys His Phe Gly Leu Phe Phe Pro Asn Lys Gln Pro Lys Phe Pro Leu
340 345 350
Arg Ile Ser Met Gly Thr Gly Ser Gly Asp Ile Val Ser Asp Gly Asn
355 360 365
Ser Thr Ser Leu Gly Trp Val Lys Ser Asp Met
370 375
<210> 3
<211> 26
<212> DNA
<213>Artificial sequence
<400> 3
tcatcagtac gtagttctct tacttc 26
<210> 4
<211> 25
<212> DNA
<213>Artificial sequence
<400> 4
agctaagcta gttacatgtc actct 25
Claims (2)
- A kind of 1. pseudo-ginseng beta-1,3 glucanase genePnGlu1, its nucleotide sequence such as SEQ IDNO：Shown in 1, coding such as SEQ IDNO：The protein of amino acid sequence shown in 2.
- 2. the pseudo-ginseng beta-1,3 glucanase gene described in claim 1PnGlu1Tobacco is being improved to grape seat chamber bacterium (Botryosphaeria dothidea), wheel branch sickle-like bacteria (Fusarium verticillioide), Fusarium solani (F. solani), colletotrichum gloeosporioides Penz (Colletotrichum gloeosporioides) application in resistance.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710540033.9A CN107354166B (en) | 2017-07-05 | 2017-07-05 | Panax notoginseng β -1,3 glucanase gene PnGlu1 and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710540033.9A CN107354166B (en) | 2017-07-05 | 2017-07-05 | Panax notoginseng β -1,3 glucanase gene PnGlu1 and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN107354166A true CN107354166A (en) | 2017-11-17 |
| CN107354166B CN107354166B (en) | 2020-05-15 |
Family
ID=60293123
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710540033.9A Active CN107354166B (en) | 2017-07-05 | 2017-07-05 | Panax notoginseng β -1,3 glucanase gene PnGlu1 and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN107354166B (en) |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102174547A (en) * | 2011-01-14 | 2011-09-07 | 昆明理工大学 | A pear beta-1, 3-glucanase gene PpGlu and its application |
| CN105861517A (en) * | 2016-04-20 | 2016-08-17 | 昆明理工大学 | Panax notoginseng antimicrobial peptide gene PnSN1 and application thereof |
-
2017
- 2017-07-05 CN CN201710540033.9A patent/CN107354166B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102174547A (en) * | 2011-01-14 | 2011-09-07 | 昆明理工大学 | A pear beta-1, 3-glucanase gene PpGlu and its application |
| CN105861517A (en) * | 2016-04-20 | 2016-08-17 | 昆明理工大学 | Panax notoginseng antimicrobial peptide gene PnSN1 and application thereof |
Non-Patent Citations (1)
| Title |
|---|
| DA SILVA C ET AL.: "Vitis vinifera class I beta-1,3-glucanase (LOC100232986), mRNA", 《GENBANK：NM_001280967》 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN107354166B (en) | 2020-05-15 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN110818783B (en) | Lilium regale WRKY transcription factor gene LrWRKY2 and application thereof | |
| CN105861517B (en) | A kind of Radix Notoginseng antibacterial peptide gene PnSN1 and its application | |
| CN106244599B (en) | A kind of 1 family gene PnPR1-2 of Radix Notoginseng pathogenesis-related proteins and application | |
| CN105755020B (en) | Radix Notoginseng mitogen-activated protein kinase kinase gene PnMAPKK1 and its application | |
| CN101736024B (en) | Thaumatin-like protein gene PpTLP from pyrus pyrifolia nakai with antifungal activity and application | |
| Jung et al. | Pepper gene encoding a basic β-1, 3-glucanase is differentially expressed in pepper tissues upon pathogen infection and ethephon or methyl jasmonate treatment | |
| CN104878019B (en) | Yangbi bulla walnut class sprouts fibroin gene JsGLP1 and application | |
| CN107267526B (en) | Radix Notoginseng myb transcription factor gene PnMYB2 and its application | |
| AU718274B2 (en) | Antifungal proteins, DNA coding therefore, and hosts incorporating same | |
| CN104131015A (en) | Application of lilium regale wilson pathogenesis-related protein 10 gene LrPR10-5 | |
| CN104878028B (en) | Yangbi bulla walnut chitinase gene JsCHI1 and application | |
| CN112359049B (en) | Lilium regale chitinase gene LrCHI2 and application thereof | |
| CN104878041B (en) | Yangbi bulla walnut transcription factor gene JsWRKY1 application | |
| CN107964547B (en) | A kind of 10 gene PnPR10-3 of Radix Notoginseng pathogenesis-related proteins and application | |
| CN103320448B (en) | Lilium regle bZIP transcription factor LrbZIP1 and application | |
| CN107267525B (en) | Application of panax notoginseng polygalacturonase inhibitor protein gene PnPGIP | |
| CN102174547B (en) | Beta-1,3-glucanase gene (i)PpGlu(/i) of Pyrus pyrifolia Nakai and application thereof | |
| CN104878027B (en) | Yangbi bulla walnut ribonuclease gene JsRNase and application | |
| CN107365794A (en) | Pseudo-ginseng chitinase gene PnCHI1 application | |
| CN104774847B (en) | Yangbi bulla walnut Pro-rich GFP JsPRP1 and application | |
| CN109295068B (en) | Pseudo-ginseng sweet protein gene PnTLP2 and application | |
| CN107354166A (en) | A kind of glucanase gene PnGlu1 of pseudo-ginseng β 1,3 and its application | |
| CN104152465A (en) | Lilium regale cytochrome b5 gene LrCyt-b5 and application thereof | |
| CN108707611B (en) | Pseudo-ginseng reverse osmosis protein genePnOLP1And applications | |
| CN104774855B (en) | The gene JsPR10 1 of Yangbi bulla walnut pathogenesis-related proteins 10 application |
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 |