CN108342395A - 刚毛柽柳myb转录因子编码基因及其应用 - Google Patents
刚毛柽柳myb转录因子编码基因及其应用 Download PDFInfo
- Publication number
- CN108342395A CN108342395A CN201810288452.2A CN201810288452A CN108342395A CN 108342395 A CN108342395 A CN 108342395A CN 201810288452 A CN201810288452 A CN 201810288452A CN 108342395 A CN108342395 A CN 108342395A
- Authority
- CN
- China
- Prior art keywords
- thmyb
- genes
- plant
- tamarix hispida
- tamarix
- 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 117
- 241001141153 Tamarix hispida Species 0.000 title claims abstract description 78
- 101000662893 Arabidopsis thaliana Telomere repeat-binding factor 1 Proteins 0.000 title claims abstract description 14
- 101000662890 Arabidopsis thaliana Telomere repeat-binding factor 2 Proteins 0.000 title claims abstract description 14
- 101000662891 Arabidopsis thaliana Telomere repeat-binding factor 3 Proteins 0.000 title claims abstract description 14
- 101000662896 Arabidopsis thaliana Telomere repeat-binding factor 4 Proteins 0.000 title claims abstract description 14
- 101000662897 Arabidopsis thaliana Telomere repeat-binding factor 5 Proteins 0.000 title claims abstract description 14
- 241000196324 Embryophyta Species 0.000 claims abstract description 64
- 150000003839 salts Chemical class 0.000 claims abstract description 29
- 241000219194 Arabidopsis Species 0.000 claims abstract description 19
- 230000015784 hyperosmotic salinity response Effects 0.000 claims abstract description 10
- 238000009395 breeding Methods 0.000 claims abstract description 9
- 230000001488 breeding effect Effects 0.000 claims abstract description 9
- 239000013598 vector Substances 0.000 claims description 20
- 230000009261 transgenic effect Effects 0.000 claims description 11
- 244000234281 Tamarix gallica Species 0.000 claims description 10
- 239000002299 complementary DNA Substances 0.000 claims description 10
- 235000014265 Tamarix gallica Nutrition 0.000 claims description 8
- 241000894006 Bacteria Species 0.000 claims description 4
- 241000124033 Salix Species 0.000 claims description 4
- 244000061176 Nicotiana tabacum Species 0.000 claims description 2
- 235000002637 Nicotiana tabacum Nutrition 0.000 claims description 2
- 230000006798 recombination Effects 0.000 claims description 2
- 238000005215 recombination Methods 0.000 claims description 2
- 235000018185 Betula X alpestris Nutrition 0.000 claims 1
- 235000018212 Betula X uliginosa Nutrition 0.000 claims 1
- 125000003275 alpha amino acid group Chemical group 0.000 claims 1
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 abstract description 42
- 239000011780 sodium chloride Substances 0.000 abstract description 21
- 230000002018 overexpression Effects 0.000 abstract description 13
- 239000013604 expression vector Substances 0.000 abstract description 7
- 230000035784 germination Effects 0.000 abstract description 6
- 230000005764 inhibitory process Effects 0.000 abstract description 5
- 238000003208 gene overexpression Methods 0.000 abstract description 4
- 230000001744 histochemical effect Effects 0.000 abstract description 4
- 238000010353 genetic engineering Methods 0.000 abstract description 3
- 230000035882 stress Effects 0.000 description 33
- 230000014509 gene expression Effects 0.000 description 20
- 108020004414 DNA Proteins 0.000 description 18
- 210000004027 cell Anatomy 0.000 description 13
- 238000006243 chemical reaction Methods 0.000 description 13
- 238000004043 dyeing Methods 0.000 description 12
- WSMYVTOQOOLQHP-UHFFFAOYSA-N Malondialdehyde Chemical compound O=CCC=O WSMYVTOQOOLQHP-UHFFFAOYSA-N 0.000 description 11
- 108091023040 Transcription factor Proteins 0.000 description 11
- 102000040945 Transcription factor Human genes 0.000 description 10
- 239000012634 fragment Substances 0.000 description 10
- 238000004458 analytical method Methods 0.000 description 9
- 238000004040 coloring Methods 0.000 description 9
- 238000000034 method Methods 0.000 description 9
- 108091030071 RNAI Proteins 0.000 description 8
- 230000029087 digestion Effects 0.000 description 8
- 230000009368 gene silencing by RNA Effects 0.000 description 8
- COXVTLYNGOIATD-HVMBLDELSA-N CC1=C(C=CC(=C1)C1=CC(C)=C(C=C1)\N=N\C1=C(O)C2=C(N)C(=CC(=C2C=C1)S(O)(=O)=O)S(O)(=O)=O)\N=N\C1=CC=C2C(=CC(=C(N)C2=C1O)S(O)(=O)=O)S(O)(=O)=O Chemical compound CC1=C(C=CC(=C1)C1=CC(C)=C(C=C1)\N=N\C1=C(O)C2=C(N)C(=CC(=C2C=C1)S(O)(=O)=O)S(O)(=O)=O)\N=N\C1=CC=C2C(=CC(=C(N)C2=C1O)S(O)(=O)=O)S(O)(=O)=O COXVTLYNGOIATD-HVMBLDELSA-N 0.000 description 7
- 229960003699 evans blue Drugs 0.000 description 7
- 241000588724 Escherichia coli Species 0.000 description 6
- 230000003834 intracellular effect Effects 0.000 description 6
- 108091008146 restriction endonucleases Proteins 0.000 description 6
- 238000012795 verification Methods 0.000 description 6
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 6
- 101150078498 MYB gene Proteins 0.000 description 5
- 150000001413 amino acids Chemical class 0.000 description 5
- 239000000969 carrier Substances 0.000 description 5
- 230000012010 growth Effects 0.000 description 5
- 238000005259 measurement Methods 0.000 description 5
- 108700021654 myb Genes Proteins 0.000 description 5
- 102000004169 proteins and genes Human genes 0.000 description 5
- 238000011084 recovery Methods 0.000 description 5
- 238000011160 research Methods 0.000 description 5
- 241000219195 Arabidopsis thaliana Species 0.000 description 4
- 230000004568 DNA-binding Effects 0.000 description 4
- 230000006378 damage Effects 0.000 description 4
- 230000007850 degeneration Effects 0.000 description 4
- 239000003292 glue Substances 0.000 description 4
- 239000007788 liquid Substances 0.000 description 4
- 239000012528 membrane Substances 0.000 description 4
- 239000013612 plasmid Substances 0.000 description 4
- 238000002360 preparation method Methods 0.000 description 4
- 238000012545 processing Methods 0.000 description 4
- 230000001105 regulatory effect Effects 0.000 description 4
- 238000003757 reverse transcription PCR Methods 0.000 description 4
- 239000000243 solution Substances 0.000 description 4
- 238000011144 upstream manufacturing Methods 0.000 description 4
- 230000003321 amplification Effects 0.000 description 3
- 210000000170 cell membrane Anatomy 0.000 description 3
- 230000007613 environmental effect Effects 0.000 description 3
- 230000003993 interaction Effects 0.000 description 3
- 239000000203 mixture Substances 0.000 description 3
- 238000003199 nucleic acid amplification method Methods 0.000 description 3
- 239000002689 soil Substances 0.000 description 3
- 238000013518 transcription Methods 0.000 description 3
- 230000035897 transcription Effects 0.000 description 3
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 2
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 2
- 241000589158 Agrobacterium Species 0.000 description 2
- 101000584287 Arabidopsis thaliana Transcription factor MYB102 Proteins 0.000 description 2
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- 241000193830 Bacillus <bacterium> Species 0.000 description 2
- 108091062157 Cis-regulatory element Proteins 0.000 description 2
- 102000012410 DNA Ligases Human genes 0.000 description 2
- 108010061982 DNA Ligases Proteins 0.000 description 2
- 208000005156 Dehydration Diseases 0.000 description 2
- UHPAZODVFFYEEL-QWRGUYRKSA-N Gly-Leu-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)CN UHPAZODVFFYEEL-QWRGUYRKSA-N 0.000 description 2
- 244000081841 Malus domestica Species 0.000 description 2
- 235000011430 Malus pumila Nutrition 0.000 description 2
- KZNQNBZMBZJQJO-UHFFFAOYSA-N N-glycyl-L-proline Natural products NCC(=O)N1CCCC1C(O)=O KZNQNBZMBZJQJO-UHFFFAOYSA-N 0.000 description 2
- 108010077850 Nuclear Localization Signals Proteins 0.000 description 2
- 241001493796 Paraboea crassifolia Species 0.000 description 2
- FKYKZHOKDOPHSA-DCAQKATOSA-N Pro-Leu-Ser Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(O)=O FKYKZHOKDOPHSA-DCAQKATOSA-N 0.000 description 2
- COYHRQWNJDJCNA-NUJDXYNKSA-N Thr-Thr-Thr Chemical compound C[C@@H](O)[C@H](N)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O COYHRQWNJDJCNA-NUJDXYNKSA-N 0.000 description 2
- 206010052428 Wound Diseases 0.000 description 2
- 208000027418 Wounds and injury Diseases 0.000 description 2
- SXEHKFHPFVVDIR-UHFFFAOYSA-N [4-(4-hydrazinylphenyl)phenyl]hydrazine Chemical compound C1=CC(NN)=CC=C1C1=CC=C(NN)C=C1 SXEHKFHPFVVDIR-UHFFFAOYSA-N 0.000 description 2
- 230000002411 adverse Effects 0.000 description 2
- 238000000137 annealing Methods 0.000 description 2
- 108010038633 aspartylglutamate Proteins 0.000 description 2
- 230000033228 biological regulation Effects 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- 239000001045 blue dye Substances 0.000 description 2
- 239000000872 buffer Substances 0.000 description 2
- 238000006555 catalytic reaction Methods 0.000 description 2
- 238000010367 cloning Methods 0.000 description 2
- 238000003501 co-culture Methods 0.000 description 2
- 230000000052 comparative effect Effects 0.000 description 2
- 230000001276 controlling effect Effects 0.000 description 2
- 108010004073 cysteinylcysteine Proteins 0.000 description 2
- 239000008367 deionised water Substances 0.000 description 2
- 229910021641 deionized water Inorganic materials 0.000 description 2
- 238000004925 denaturation Methods 0.000 description 2
- 230000036425 denaturation Effects 0.000 description 2
- 108010054813 diprotin B Proteins 0.000 description 2
- 239000012153 distilled water Substances 0.000 description 2
- 238000001035 drying Methods 0.000 description 2
- 239000000975 dye Substances 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 238000000605 extraction Methods 0.000 description 2
- 230000006698 induction Effects 0.000 description 2
- 238000010630 lipid peroxidation (MDA) assay Methods 0.000 description 2
- 150000002632 lipids Chemical class 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 230000007246 mechanism Effects 0.000 description 2
- 238000002887 multiple sequence alignment Methods 0.000 description 2
- JPXMTWWFLBLUCD-UHFFFAOYSA-N nitro blue tetrazolium(2+) Chemical compound COC1=CC(C=2C=C(OC)C(=CC=2)[N+]=2N(N=C(N=2)C=2C=CC=CC=2)C=2C=CC(=CC=2)[N+]([O-])=O)=CC=C1[N+]1=NC(C=2C=CC=CC=2)=NN1C1=CC=C([N+]([O-])=O)C=C1 JPXMTWWFLBLUCD-UHFFFAOYSA-N 0.000 description 2
- 238000005502 peroxidation Methods 0.000 description 2
- 230000035479 physiological effects, processes and functions Effects 0.000 description 2
- 230000003252 repetitive effect Effects 0.000 description 2
- 230000004044 response Effects 0.000 description 2
- 230000003938 response to stress Effects 0.000 description 2
- 238000012163 sequencing technique Methods 0.000 description 2
- 210000002966 serum Anatomy 0.000 description 2
- 230000002103 transcriptional effect Effects 0.000 description 2
- 229910021642 ultra pure water Inorganic materials 0.000 description 2
- 239000012498 ultrapure water Substances 0.000 description 2
- JBVSSSZFNTXJDX-YTLHQDLWSA-N Ala-Ala-Thr Chemical compound C[C@@H](O)[C@@H](C(O)=O)NC(=O)[C@H](C)NC(=O)[C@H](C)N JBVSSSZFNTXJDX-YTLHQDLWSA-N 0.000 description 1
- QDRGPQWIVZNJQD-CIUDSAMLSA-N Ala-Arg-Gln Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(N)=O)C(O)=O QDRGPQWIVZNJQD-CIUDSAMLSA-N 0.000 description 1
- OKIKVSXTXVVFDV-MMWGEVLESA-N Ala-Ile-Pro Chemical compound CC[C@H](C)[C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](C)N OKIKVSXTXVVFDV-MMWGEVLESA-N 0.000 description 1
- NHWYNIZWLJYZAG-XVYDVKMFSA-N Ala-Ser-His Chemical compound C[C@@H](C(=O)N[C@@H](CO)C(=O)N[C@@H](CC1=CN=CN1)C(=O)O)N NHWYNIZWLJYZAG-XVYDVKMFSA-N 0.000 description 1
- VNFSAYFQLXPHPY-CIQUZCHMSA-N Ala-Thr-Ile Chemical compound [H]N[C@@H](C)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O VNFSAYFQLXPHPY-CIQUZCHMSA-N 0.000 description 1
- NLYYHIKRBRMAJV-AEJSXWLSSA-N Ala-Val-Pro Chemical compound C[C@@H](C(=O)N[C@@H](C(C)C)C(=O)N1CCC[C@@H]1C(=O)O)N NLYYHIKRBRMAJV-AEJSXWLSSA-N 0.000 description 1
- 108700026767 Arabidopsis MYB102 Proteins 0.000 description 1
- JTWOBPNAVBESFW-FXQIFTODSA-N Arg-Cys-Asp Chemical compound C(C[C@@H](C(=O)N[C@@H](CS)C(=O)N[C@@H](CC(=O)O)C(=O)O)N)CN=C(N)N JTWOBPNAVBESFW-FXQIFTODSA-N 0.000 description 1
- YBZMTKUDWXZLIX-UWVGGRQHSA-N Arg-Leu-Gly Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(C)C)C(=O)NCC(O)=O YBZMTKUDWXZLIX-UWVGGRQHSA-N 0.000 description 1
- CQMQJWRCRQSBAF-BPUTZDHNSA-N Asn-Arg-Trp Chemical compound C1=CC=C2C(=C1)C(=CN2)C[C@@H](C(=O)O)NC(=O)[C@H](CCCN=C(N)N)NC(=O)[C@H](CC(=O)N)N CQMQJWRCRQSBAF-BPUTZDHNSA-N 0.000 description 1
- JRCASHGTXZYSPW-XIRDDKMYSA-N Asn-His-Trp Chemical compound C1=CC=C2C(=C1)C(=CN2)C[C@@H](C(=O)O)NC(=O)[C@H](CC3=CN=CN3)NC(=O)[C@H](CC(=O)N)N JRCASHGTXZYSPW-XIRDDKMYSA-N 0.000 description 1
- KMCRKVOLRCOMBG-DJFWLOJKSA-N Asn-Ile-His Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC1=CN=CN1)C(=O)O)NC(=O)[C@H](CC(=O)N)N KMCRKVOLRCOMBG-DJFWLOJKSA-N 0.000 description 1
- HFPXZWPUVFVNLL-GUBZILKMSA-N Asn-Leu-Gln Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(N)=O)C(O)=O HFPXZWPUVFVNLL-GUBZILKMSA-N 0.000 description 1
- NPZJLGMWMDNQDD-GHCJXIJMSA-N Asn-Ser-Ile Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O NPZJLGMWMDNQDD-GHCJXIJMSA-N 0.000 description 1
- SNYCNNPOFYBCEK-ZLUOBGJFSA-N Asn-Ser-Ser Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CO)C(O)=O SNYCNNPOFYBCEK-ZLUOBGJFSA-N 0.000 description 1
- YHXNKGKUDJCAHB-PBCZWWQYSA-N Asn-Thr-His Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CC1=CN=CN1)C(=O)O)NC(=O)[C@H](CC(=O)N)N)O YHXNKGKUDJCAHB-PBCZWWQYSA-N 0.000 description 1
- VPSHHQXIWLGVDD-ZLUOBGJFSA-N Asp-Asp-Asp Chemical compound OC(=O)C[C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(O)=O)C(O)=O VPSHHQXIWLGVDD-ZLUOBGJFSA-N 0.000 description 1
- FANQWNCPNFEPGZ-WHFBIAKZSA-N Asp-Asp-Gly Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(O)=O)C(=O)NCC(O)=O FANQWNCPNFEPGZ-WHFBIAKZSA-N 0.000 description 1
- BKXPJCBEHWFSTF-ACZMJKKPSA-N Asp-Gln-Asp Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(O)=O)C(O)=O BKXPJCBEHWFSTF-ACZMJKKPSA-N 0.000 description 1
- KHBLRHKVXICFMY-GUBZILKMSA-N Asp-Glu-Lys Chemical compound N[C@@H](CC(=O)O)C(=O)N[C@@H](CCC(=O)O)C(=O)N[C@@H](CCCCN)C(=O)O KHBLRHKVXICFMY-GUBZILKMSA-N 0.000 description 1
- VSMYBNPOHYAXSD-GUBZILKMSA-N Asp-Lys-Glu Chemical compound OC(=O)C[C@H](N)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCC(O)=O)C(O)=O VSMYBNPOHYAXSD-GUBZILKMSA-N 0.000 description 1
- YFGUZQQCSDZRBN-DCAQKATOSA-N Asp-Pro-Leu Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CC(C)C)C(O)=O YFGUZQQCSDZRBN-DCAQKATOSA-N 0.000 description 1
- ZQFRDAZBTSFGGW-SRVKXCTJSA-N Asp-Ser-Phe Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O ZQFRDAZBTSFGGW-SRVKXCTJSA-N 0.000 description 1
- MGSVBZIBCCKGCY-ZLUOBGJFSA-N Asp-Ser-Ser Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CO)C(O)=O MGSVBZIBCCKGCY-ZLUOBGJFSA-N 0.000 description 1
- DKQCWCQRAMAFLN-UBHSHLNASA-N Asp-Trp-Asp Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CC1=CNC2=C1C=CC=C2)C(=O)N[C@@H](CC(O)=O)C(O)=O DKQCWCQRAMAFLN-UBHSHLNASA-N 0.000 description 1
- NJLLRXWFPQQPHV-SRVKXCTJSA-N Asp-Tyr-Asn Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CC(N)=O)C(O)=O NJLLRXWFPQQPHV-SRVKXCTJSA-N 0.000 description 1
- 235000009109 Betula pendula Nutrition 0.000 description 1
- 244000056139 Brassica cretica Species 0.000 description 1
- 235000003351 Brassica cretica Nutrition 0.000 description 1
- 235000003343 Brassica rupestris Nutrition 0.000 description 1
- LZZYPRNAOMGNLH-UHFFFAOYSA-M Cetrimonium bromide Chemical compound [Br-].CCCCCCCCCCCCCCCC[N+](C)(C)C LZZYPRNAOMGNLH-UHFFFAOYSA-M 0.000 description 1
- 240000002319 Citrus sinensis Species 0.000 description 1
- 235000005976 Citrus sinensis Nutrition 0.000 description 1
- KIQKJXYVGSYDFS-ZLUOBGJFSA-N Cys-Asn-Asn Chemical compound SC[C@H](N)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(N)=O)C(O)=O KIQKJXYVGSYDFS-ZLUOBGJFSA-N 0.000 description 1
- YMBAVNPKBWHDAW-CIUDSAMLSA-N Cys-Asp-Lys Chemical compound C(CCN)C[C@@H](C(=O)O)NC(=O)[C@H](CC(=O)O)NC(=O)[C@H](CS)N YMBAVNPKBWHDAW-CIUDSAMLSA-N 0.000 description 1
- DZSICRGTVPDCRN-YUMQZZPRSA-N Cys-Gly-Lys Chemical compound C(CCN)C[C@@H](C(=O)O)NC(=O)CNC(=O)[C@H](CS)N DZSICRGTVPDCRN-YUMQZZPRSA-N 0.000 description 1
- IZJLAQMWJHCHTN-BPUTZDHNSA-N Cys-Trp-Arg Chemical compound N[C@@H](CS)C(=O)N[C@@H](Cc1c[nH]c2ccccc12)C(=O)N[C@@H](CCCNC(=N)N)C(=O)O IZJLAQMWJHCHTN-BPUTZDHNSA-N 0.000 description 1
- DGQJGBDBFVGLGL-ZKWXMUAHSA-N Cys-Val-Asp Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)O)NC(=O)[C@H](CS)N DGQJGBDBFVGLGL-ZKWXMUAHSA-N 0.000 description 1
- 102000052510 DNA-Binding Proteins Human genes 0.000 description 1
- 101710096438 DNA-binding protein Proteins 0.000 description 1
- 240000000716 Durio zibethinus Species 0.000 description 1
- 235000006025 Durio zibethinus Nutrition 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 1
- 241000206602 Eukaryota Species 0.000 description 1
- ZDJZEGYVKANKED-NRPADANISA-N Gln-Cys-Val Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CS)C(=O)N[C@@H](C(C)C)C(O)=O ZDJZEGYVKANKED-NRPADANISA-N 0.000 description 1
- PNENQZWRFMUZOM-DCAQKATOSA-N Gln-Glu-Leu Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(O)=O PNENQZWRFMUZOM-DCAQKATOSA-N 0.000 description 1
- CAXXTYYGFYTBPV-IUCAKERBSA-N Gln-Leu-Gly Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(C)C)C(=O)NCC(O)=O CAXXTYYGFYTBPV-IUCAKERBSA-N 0.000 description 1
- WBYHRQBKJGEBQJ-CIUDSAMLSA-N Gln-Pro-Cys Chemical compound C1C[C@H](N(C1)C(=O)[C@H](CCC(=O)N)N)C(=O)N[C@@H](CS)C(=O)O WBYHRQBKJGEBQJ-CIUDSAMLSA-N 0.000 description 1
- ARYKRXHBIPLULY-XKBZYTNZSA-N Gln-Thr-Ser Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CO)C(O)=O ARYKRXHBIPLULY-XKBZYTNZSA-N 0.000 description 1
- WIMVKDYAKRAUCG-IHRRRGAJSA-N Gln-Tyr-Glu Chemical compound C1=CC(=CC=C1C[C@@H](C(=O)N[C@@H](CCC(=O)O)C(=O)O)NC(=O)[C@H](CCC(=O)N)N)O WIMVKDYAKRAUCG-IHRRRGAJSA-N 0.000 description 1
- NCWOMXABNYEPLY-NRPADANISA-N Glu-Ala-Val Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](C)C(=O)N[C@@H](C(C)C)C(O)=O NCWOMXABNYEPLY-NRPADANISA-N 0.000 description 1
- NKLRYVLERDYDBI-FXQIFTODSA-N Glu-Glu-Asp Chemical compound OC(=O)CC[C@H](N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(O)=O)C(O)=O NKLRYVLERDYDBI-FXQIFTODSA-N 0.000 description 1
- HNVFSTLPVJWIDV-CIUDSAMLSA-N Glu-Glu-Gln Chemical compound OC(=O)CC[C@H](N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCC(N)=O)C(O)=O HNVFSTLPVJWIDV-CIUDSAMLSA-N 0.000 description 1
- XTZDZAXYPDISRR-MNXVOIDGSA-N Glu-Ile-Lys Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCCCN)C(=O)O)NC(=O)[C@H](CCC(=O)O)N XTZDZAXYPDISRR-MNXVOIDGSA-N 0.000 description 1
- ITVBKCZZLJUUHI-HTUGSXCWSA-N Glu-Phe-Thr Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H]([C@@H](C)O)C(O)=O ITVBKCZZLJUUHI-HTUGSXCWSA-N 0.000 description 1
- RMWAOBGCZZSJHE-UMNHJUIQSA-N Glu-Val-Pro Chemical compound CC(C)[C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](CCC(=O)O)N RMWAOBGCZZSJHE-UMNHJUIQSA-N 0.000 description 1
- VUUOMYFPWDYETE-WDSKDSINSA-N Gly-Gln-Cys Chemical compound C(CC(=O)N)[C@@H](C(=O)N[C@@H](CS)C(=O)O)NC(=O)CN VUUOMYFPWDYETE-WDSKDSINSA-N 0.000 description 1
- NZOAFWHVAFJERA-OALUTQOASA-N Gly-Phe-Trp Chemical compound [H]NCC(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CC1=CNC2=C1C=CC=C2)C(O)=O NZOAFWHVAFJERA-OALUTQOASA-N 0.000 description 1
- WNGHUXFWEWTKAO-YUMQZZPRSA-N Gly-Ser-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](CO)NC(=O)CN WNGHUXFWEWTKAO-YUMQZZPRSA-N 0.000 description 1
- 244000068988 Glycine max Species 0.000 description 1
- 235000010469 Glycine max Nutrition 0.000 description 1
- 244000299507 Gossypium hirsutum Species 0.000 description 1
- 235000009432 Gossypium hirsutum Nutrition 0.000 description 1
- VHHYJBSXXMPQGZ-AVGNSLFASA-N His-Gln-Leu Chemical compound CC(C)C[C@@H](C(=O)O)NC(=O)[C@H](CCC(=O)N)NC(=O)[C@H](CC1=CN=CN1)N VHHYJBSXXMPQGZ-AVGNSLFASA-N 0.000 description 1
- FIMNVXRZGUAGBI-AVGNSLFASA-N His-Glu-Leu Chemical compound [H]N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(O)=O FIMNVXRZGUAGBI-AVGNSLFASA-N 0.000 description 1
- JRHFQUPIZOYKQP-KBIXCLLPSA-N Ile-Ala-Glu Chemical compound CC[C@H](C)[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@H](C(O)=O)CCC(O)=O JRHFQUPIZOYKQP-KBIXCLLPSA-N 0.000 description 1
- PNDMHTTXXPUQJH-RWRJDSDZSA-N Ile-Glu-Thr Chemical compound N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CCC(=O)O)C(=O)N[C@@H]([C@H](O)C)C(=O)O PNDMHTTXXPUQJH-RWRJDSDZSA-N 0.000 description 1
- KEKTTYCXKGBAAL-VGDYDELISA-N Ile-His-Ser Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC1=CN=CN1)C(=O)N[C@@H](CO)C(=O)O)N KEKTTYCXKGBAAL-VGDYDELISA-N 0.000 description 1
- PHRWFSFCNJPWRO-PPCPHDFISA-N Ile-Leu-Thr Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)O)C(=O)O)N PHRWFSFCNJPWRO-PPCPHDFISA-N 0.000 description 1
- ZDNNDIJTUHQCAM-MXAVVETBSA-N Ile-Ser-Phe Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CO)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)O)N ZDNNDIJTUHQCAM-MXAVVETBSA-N 0.000 description 1
- 240000007049 Juglans regia Species 0.000 description 1
- 235000009496 Juglans regia Nutrition 0.000 description 1
- PMGDADKJMCOXHX-UHFFFAOYSA-N L-Arginyl-L-glutamin-acetat Natural products NC(=N)NCCCC(N)C(=O)NC(CCC(N)=O)C(O)=O PMGDADKJMCOXHX-UHFFFAOYSA-N 0.000 description 1
- IBMVEYRWAWIOTN-UHFFFAOYSA-N L-Leucyl-L-Arginyl-L-Proline Natural products CC(C)CC(N)C(=O)NC(CCCN=C(N)N)C(=O)N1CCCC1C(O)=O IBMVEYRWAWIOTN-UHFFFAOYSA-N 0.000 description 1
- SENJXOPIZNYLHU-UHFFFAOYSA-N L-leucyl-L-arginine Natural products CC(C)CC(N)C(=O)NC(C(O)=O)CCCN=C(N)N SENJXOPIZNYLHU-UHFFFAOYSA-N 0.000 description 1
- IBMVEYRWAWIOTN-RWMBFGLXSA-N Leu-Arg-Pro Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CCCN=C(N)N)C(=O)N1CCC[C@@H]1C(O)=O IBMVEYRWAWIOTN-RWMBFGLXSA-N 0.000 description 1
- XYUBOFCTGPZFSA-WDSOQIARSA-N Leu-Arg-Trp Chemical compound C1=CC=C2C(C[C@H](NC(=O)[C@H](CCCN=C(N)N)NC(=O)[C@@H](N)CC(C)C)C(O)=O)=CNC2=C1 XYUBOFCTGPZFSA-WDSOQIARSA-N 0.000 description 1
- CQGSYZCULZMEDE-SRVKXCTJSA-N Leu-Gln-Pro Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CCC(N)=O)C(=O)N1CCC[C@H]1C(O)=O CQGSYZCULZMEDE-SRVKXCTJSA-N 0.000 description 1
- CQGSYZCULZMEDE-UHFFFAOYSA-N Leu-Gln-Pro Natural products CC(C)CC(N)C(=O)NC(CCC(N)=O)C(=O)N1CCCC1C(O)=O CQGSYZCULZMEDE-UHFFFAOYSA-N 0.000 description 1
- PBGDOSARRIJMEV-DLOVCJGASA-N Leu-His-Ala Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](C)C(O)=O PBGDOSARRIJMEV-DLOVCJGASA-N 0.000 description 1
- USLNHQZCDQJBOV-ZPFDUUQYSA-N Leu-Ile-Asn Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC(N)=O)C(O)=O USLNHQZCDQJBOV-ZPFDUUQYSA-N 0.000 description 1
- JLWZLIQRYCTYBD-IHRRRGAJSA-N Leu-Lys-Arg Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O JLWZLIQRYCTYBD-IHRRRGAJSA-N 0.000 description 1
- KPYAOIVPJKPIOU-KKUMJFAQSA-N Leu-Lys-Lys Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCCCN)C(O)=O KPYAOIVPJKPIOU-KKUMJFAQSA-N 0.000 description 1
- XWEVVRRSIOBJOO-SRVKXCTJSA-N Leu-Pro-Gln Chemical compound [H]N[C@@H](CC(C)C)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CCC(N)=O)C(O)=O XWEVVRRSIOBJOO-SRVKXCTJSA-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
- 102000003960 Ligases Human genes 0.000 description 1
- 108090000364 Ligases Proteins 0.000 description 1
- MYZMQWHPDAYKIE-SRVKXCTJSA-N Lys-Leu-Ala Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](C)C(O)=O MYZMQWHPDAYKIE-SRVKXCTJSA-N 0.000 description 1
- VMTYLUGCXIEDMV-QWRGUYRKSA-N Lys-Leu-Gly Chemical compound OC(=O)CNC(=O)[C@H](CC(C)C)NC(=O)[C@@H](N)CCCCN VMTYLUGCXIEDMV-QWRGUYRKSA-N 0.000 description 1
- YUAXTFMFMOIMAM-QWRGUYRKSA-N Lys-Lys-Gly Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CCCCN)C(=O)NCC(O)=O YUAXTFMFMOIMAM-QWRGUYRKSA-N 0.000 description 1
- HVAUKHLDSDDROB-KKUMJFAQSA-N Lys-Lys-Leu Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(C)C)C(O)=O HVAUKHLDSDDROB-KKUMJFAQSA-N 0.000 description 1
- WBSCNDJQPKSPII-KKUMJFAQSA-N Lys-Lys-Lys Chemical compound NCCCC[C@H](N)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCCCN)C(O)=O WBSCNDJQPKSPII-KKUMJFAQSA-N 0.000 description 1
- 244000141359 Malus pumila Species 0.000 description 1
- VHGIWFGJIHTASW-FXQIFTODSA-N Met-Ala-Asp Chemical compound CSCC[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@H](C(O)=O)CC(O)=O VHGIWFGJIHTASW-FXQIFTODSA-N 0.000 description 1
- FYRUJIJAUPHUNB-IUCAKERBSA-N Met-Gly-Arg Chemical compound CSCC[C@H](N)C(=O)NCC(=O)N[C@H](C(O)=O)CCCNC(N)=N FYRUJIJAUPHUNB-IUCAKERBSA-N 0.000 description 1
- SLQDSYZHHOKQSR-QXEWZRGKSA-N Met-Gly-Ile Chemical compound CC[C@H](C)[C@@H](C(O)=O)NC(=O)CNC(=O)[C@@H](N)CCSC SLQDSYZHHOKQSR-QXEWZRGKSA-N 0.000 description 1
- KPVLLNDCBYXKNV-CYDGBPFRSA-N Met-Val-Ile Chemical compound [H]N[C@@H](CCSC)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O KPVLLNDCBYXKNV-CYDGBPFRSA-N 0.000 description 1
- 108700026244 Open Reading Frames Proteins 0.000 description 1
- 240000007594 Oryza sativa Species 0.000 description 1
- 235000007164 Oryza sativa Nutrition 0.000 description 1
- 238000012408 PCR amplification Methods 0.000 description 1
- UEEVBGHEGJMDDV-AVGNSLFASA-N Phe-Asp-Gln Chemical compound NC(=O)CC[C@@H](C(O)=O)NC(=O)[C@H](CC(O)=O)NC(=O)[C@@H](N)CC1=CC=CC=C1 UEEVBGHEGJMDDV-AVGNSLFASA-N 0.000 description 1
- QPQDWBAJWOGAMJ-IHPCNDPISA-N Phe-Asp-Trp Chemical compound C([C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(O)=O)C1=CC=CC=C1 QPQDWBAJWOGAMJ-IHPCNDPISA-N 0.000 description 1
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 description 1
- 241000218976 Populus trichocarpa Species 0.000 description 1
- HFZNNDWPHBRNPV-KZVJFYERSA-N Pro-Ala-Thr Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](C)C(=O)N[C@@H]([C@@H](C)O)C(O)=O HFZNNDWPHBRNPV-KZVJFYERSA-N 0.000 description 1
- YFNOUBWUIIJQHF-LPEHRKFASA-N Pro-Asp-Pro Chemical compound C1C[C@H](NC1)C(=O)N[C@@H](CC(=O)O)C(=O)N2CCC[C@@H]2C(=O)O YFNOUBWUIIJQHF-LPEHRKFASA-N 0.000 description 1
- JFNPBBOGGNMSRX-CIUDSAMLSA-N Pro-Gln-Ala Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](C)C(O)=O JFNPBBOGGNMSRX-CIUDSAMLSA-N 0.000 description 1
- FISHYTLIMUYTQY-GUBZILKMSA-N Pro-Gln-Gln Chemical compound NC(=O)CC[C@@H](C(O)=O)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@@H]1CCCN1 FISHYTLIMUYTQY-GUBZILKMSA-N 0.000 description 1
- DMKWYMWNEKIPFC-IUCAKERBSA-N Pro-Gly-Arg Chemical compound [H]N1CCC[C@H]1C(=O)NCC(=O)N[C@@H](CCCNC(N)=N)C(O)=O DMKWYMWNEKIPFC-IUCAKERBSA-N 0.000 description 1
- SBVPYBFMIGDIDX-SRVKXCTJSA-N Pro-Pro-Pro Chemical compound OC(=O)[C@@H]1CCCN1C(=O)[C@H]1N(C(=O)[C@H]2NCCC2)CCC1 SBVPYBFMIGDIDX-SRVKXCTJSA-N 0.000 description 1
- IURWWZYKYPEANQ-HJGDQZAQSA-N Pro-Thr-Glu Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCC(O)=O)C(O)=O IURWWZYKYPEANQ-HJGDQZAQSA-N 0.000 description 1
- 239000013614 RNA sample Substances 0.000 description 1
- 238000003559 RNA-seq method Methods 0.000 description 1
- 238000011529 RT qPCR Methods 0.000 description 1
- 240000000528 Ricinus communis Species 0.000 description 1
- 235000004443 Ricinus communis Nutrition 0.000 description 1
- 238000012300 Sequence Analysis Methods 0.000 description 1
- BLPYXIXXCFVIIF-FXQIFTODSA-N Ser-Cys-Arg Chemical compound C(C[C@@H](C(=O)O)NC(=O)[C@H](CS)NC(=O)[C@H](CO)N)CN=C(N)N BLPYXIXXCFVIIF-FXQIFTODSA-N 0.000 description 1
- BQWCDDAISCPDQV-XHNCKOQMSA-N Ser-Gln-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CCC(=O)N)NC(=O)[C@H](CO)N)C(=O)O BQWCDDAISCPDQV-XHNCKOQMSA-N 0.000 description 1
- LRWBCWGEUCKDTN-BJDJZHNGSA-N Ser-Lys-Ile Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O LRWBCWGEUCKDTN-BJDJZHNGSA-N 0.000 description 1
- KQNDIKOYWZTZIX-FXQIFTODSA-N Ser-Ser-Arg Chemical compound OC[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@H](C(O)=O)CCCNC(N)=N KQNDIKOYWZTZIX-FXQIFTODSA-N 0.000 description 1
- XQJCEKXQUJQNNK-ZLUOBGJFSA-N Ser-Ser-Ser Chemical compound OC[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@@H](CO)C(O)=O XQJCEKXQUJQNNK-ZLUOBGJFSA-N 0.000 description 1
- JURQXQBJKUHGJS-UHFFFAOYSA-N Ser-Ser-Ser-Ser Chemical compound OCC(N)C(=O)NC(CO)C(=O)NC(CO)C(=O)NC(CO)C(O)=O JURQXQBJKUHGJS-UHFFFAOYSA-N 0.000 description 1
- PYTKULIABVRXSC-BWBBJGPYSA-N Ser-Ser-Thr Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)O)C(O)=O PYTKULIABVRXSC-BWBBJGPYSA-N 0.000 description 1
- XJDMUQCLVSCRSJ-VZFHVOOUSA-N Ser-Thr-Ala Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C)C(O)=O XJDMUQCLVSCRSJ-VZFHVOOUSA-N 0.000 description 1
- 240000003768 Solanum lycopersicum Species 0.000 description 1
- 235000002560 Solanum lycopersicum Nutrition 0.000 description 1
- 235000010185 Tamarix canariensis Nutrition 0.000 description 1
- 235000010154 Tamarix ramosissima Nutrition 0.000 description 1
- 244000299461 Theobroma cacao Species 0.000 description 1
- 235000009470 Theobroma cacao Nutrition 0.000 description 1
- IGROJMCBGRFRGI-YTLHQDLWSA-N Thr-Ala-Ala Chemical compound C[C@@H](O)[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@@H](C)C(O)=O IGROJMCBGRFRGI-YTLHQDLWSA-N 0.000 description 1
- JVTHIXKSVYEWNI-JRQIVUDYSA-N Thr-Asn-Tyr Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O JVTHIXKSVYEWNI-JRQIVUDYSA-N 0.000 description 1
- MFEBUIFJVPNZLO-OLHMAJIHSA-N Thr-Asp-Asn Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(N)=O)C(O)=O MFEBUIFJVPNZLO-OLHMAJIHSA-N 0.000 description 1
- WDFPMSHYMRBLKM-NKIYYHGXSA-N Thr-Glu-His Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CCC(=O)O)C(=O)N[C@@H](CC1=CN=CN1)C(=O)O)N)O WDFPMSHYMRBLKM-NKIYYHGXSA-N 0.000 description 1
- KRGDDWVBBDLPSJ-CUJWVEQBSA-N Thr-His-Ser Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](CO)C(O)=O KRGDDWVBBDLPSJ-CUJWVEQBSA-N 0.000 description 1
- PCMDGXKXVMBIFP-VEVYYDQMSA-N Thr-Met-Asn Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CC(N)=O)C(O)=O PCMDGXKXVMBIFP-VEVYYDQMSA-N 0.000 description 1
- RVMNUBQWPVOUKH-HEIBUPTGSA-N Thr-Ser-Thr Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)O)C(O)=O RVMNUBQWPVOUKH-HEIBUPTGSA-N 0.000 description 1
- YCQXZDHDSUHUSG-FJHTZYQYSA-N Trp-Thr-Ala Chemical compound C1=CC=C2C(C[C@H](N)C(=O)N[C@@H]([C@H](O)C)C(=O)N[C@@H](C)C(O)=O)=CNC2=C1 YCQXZDHDSUHUSG-FJHTZYQYSA-N 0.000 description 1
- GAYLGYUVTDMLKC-UWJYBYFXSA-N Tyr-Asp-Ala Chemical compound OC(=O)[C@H](C)NC(=O)[C@H](CC(O)=O)NC(=O)[C@@H](N)CC1=CC=C(O)C=C1 GAYLGYUVTDMLKC-UWJYBYFXSA-N 0.000 description 1
- TZXFLDNBYYGLKA-BZSNNMDCSA-N Tyr-Asp-Tyr Chemical compound C([C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(O)=O)C1=CC=C(O)C=C1 TZXFLDNBYYGLKA-BZSNNMDCSA-N 0.000 description 1
- URIRWLJVWHYLET-ONGXEEELSA-N Val-Gly-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)CNC(=O)[C@@H](N)C(C)C URIRWLJVWHYLET-ONGXEEELSA-N 0.000 description 1
- 240000004922 Vigna radiata Species 0.000 description 1
- 235000006582 Vigna radiata Nutrition 0.000 description 1
- 240000006365 Vitis vinifera Species 0.000 description 1
- 235000014787 Vitis vinifera Nutrition 0.000 description 1
- 241000607479 Yersinia pestis Species 0.000 description 1
- 240000000038 Ziziphus mauritiana Species 0.000 description 1
- 235000006545 Ziziphus mauritiana Nutrition 0.000 description 1
- 235000008529 Ziziphus vulgaris Nutrition 0.000 description 1
- 230000036579 abiotic stress Effects 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- 229960000583 acetic acid Drugs 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 108010008685 alanyl-glutamyl-aspartic acid Proteins 0.000 description 1
- 108010005233 alanylglutamic acid Proteins 0.000 description 1
- 108010044940 alanylglutamine Proteins 0.000 description 1
- 108010047495 alanylglycine Proteins 0.000 description 1
- 108010087924 alanylproline Proteins 0.000 description 1
- 229930014669 anthocyanidin Natural products 0.000 description 1
- 150000001452 anthocyanidin derivatives Chemical class 0.000 description 1
- 235000008758 anthocyanidins Nutrition 0.000 description 1
- 235000021016 apples Nutrition 0.000 description 1
- 108010008355 arginyl-glutamine Proteins 0.000 description 1
- 108010043240 arginyl-leucyl-glycine Proteins 0.000 description 1
- 108010093581 aspartyl-proline Proteins 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000003851 biochemical process Effects 0.000 description 1
- 238000007622 bioinformatic analysis Methods 0.000 description 1
- 230000004790 biotic stress Effects 0.000 description 1
- QKSKPIVNLNLAAV-UHFFFAOYSA-N bis(2-chloroethyl) sulfide Chemical compound ClCCSCCCl QKSKPIVNLNLAAV-UHFFFAOYSA-N 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- 210000004899 c-terminal region Anatomy 0.000 description 1
- 238000004364 calculation method Methods 0.000 description 1
- 210000005056 cell body Anatomy 0.000 description 1
- 230000005779 cell damage Effects 0.000 description 1
- 208000037887 cell injury Diseases 0.000 description 1
- 230000001413 cellular effect Effects 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 239000003610 charcoal Substances 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 238000012790 confirmation Methods 0.000 description 1
- 230000007123 defense Effects 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 238000007598 dipping method Methods 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 239000000428 dust Substances 0.000 description 1
- 235000013601 eggs Nutrition 0.000 description 1
- 239000003792 electrolyte Substances 0.000 description 1
- 230000006353 environmental stress Effects 0.000 description 1
- 238000001976 enzyme digestion Methods 0.000 description 1
- 229930003935 flavonoid Natural products 0.000 description 1
- 150000002215 flavonoids Chemical class 0.000 description 1
- 235000017173 flavonoids Nutrition 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- 230000009746 freeze damage Effects 0.000 description 1
- 230000002068 genetic effect Effects 0.000 description 1
- 239000012362 glacial acetic acid Substances 0.000 description 1
- 108010079547 glutamylmethionine Proteins 0.000 description 1
- 108010077515 glycylproline Proteins 0.000 description 1
- 235000002532 grape seed extract Nutrition 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 108010085325 histidylproline Proteins 0.000 description 1
- 229940088597 hormone Drugs 0.000 description 1
- 239000005556 hormone Substances 0.000 description 1
- 230000002209 hydrophobic effect Effects 0.000 description 1
- 230000001771 impaired effect Effects 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 108010077158 leucinyl-arginyl-tryptophan Proteins 0.000 description 1
- 108010044056 leucyl-phenylalanine Proteins 0.000 description 1
- 108010073472 leucyl-prolyl-proline Proteins 0.000 description 1
- 108010000761 leucylarginine Proteins 0.000 description 1
- 230000003859 lipid peroxidation Effects 0.000 description 1
- 108010009298 lysylglutamic acid Proteins 0.000 description 1
- 210000001161 mammalian embryo Anatomy 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 230000010534 mechanism of action Effects 0.000 description 1
- 230000001404 mediated effect Effects 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- 239000002207 metabolite Substances 0.000 description 1
- 238000010369 molecular cloning Methods 0.000 description 1
- 235000010460 mustard Nutrition 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 238000006384 oligomerization reaction Methods 0.000 description 1
- 210000000056 organ Anatomy 0.000 description 1
- 230000003204 osmotic effect Effects 0.000 description 1
- 230000003647 oxidation Effects 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- 239000008363 phosphate buffer Substances 0.000 description 1
- 229910052698 phosphorus Inorganic materials 0.000 description 1
- 239000011574 phosphorus Substances 0.000 description 1
- 230000035790 physiological processes and functions Effects 0.000 description 1
- 230000008121 plant development Effects 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 230000002035 prolonged effect Effects 0.000 description 1
- 238000004064 recycling Methods 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 230000002829 reductive effect Effects 0.000 description 1
- 238000010839 reverse transcription Methods 0.000 description 1
- 235000009566 rice Nutrition 0.000 description 1
- 230000010496 root system development Effects 0.000 description 1
- 239000000523 sample Substances 0.000 description 1
- 230000002000 scavenging effect Effects 0.000 description 1
- 230000024053 secondary metabolic process Effects 0.000 description 1
- 230000019491 signal transduction Effects 0.000 description 1
- UIIMBOGNXHQVGW-UHFFFAOYSA-M sodium bicarbonate Substances [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 description 1
- 229910000030 sodium bicarbonate Inorganic materials 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 239000008223 sterile water Substances 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
- -1 superoxide ion Chemical class 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 239000008399 tap water Substances 0.000 description 1
- 235000020679 tap water Nutrition 0.000 description 1
- 230000010474 transient expression Effects 0.000 description 1
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 description 1
- 108010084932 tryptophyl-proline Proteins 0.000 description 1
- 239000005418 vegetable material Substances 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
- 240000004494 yellow birch Species 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
- C12N15/8273—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield for stress resistance, e.g. heavy metal resistance for drought, cold, salt resistance
Landscapes
- Health & Medical Sciences (AREA)
- 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)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
Abstract
本发明属于植物基因工程育种技术领域，尤其涉及刚毛柽柳MYB转录因子编码基因及其应用。本发明提供的刚毛柽柳ThMYB基因可用于提高植物耐盐性或选育耐盐性转基因植物。分别构建ThMYB基因的过表达和抑制表达载体，通过瞬时侵染获得转基因刚毛柽柳，经组织化学染色和生理指标测定证实ThMYB基因过表达株系具有明显的耐盐能力；将ThMYB基因转入拟南芥，转基因T3代拟南芥种子在150mM NaCl胁迫后平均发芽率是野生型的6.21倍，根长是野生型的1.41倍，鲜重是野生型的1.66倍。ThMYB基因的同源和异源过表达结果均表明该基因能够明显提高转基因植物的耐盐能力，是用于耐盐基因工程育种的优良基因。
Description
技术领域
本发明属于植物基因工程育种技术领域，尤其涉及刚毛柽柳MYB转录因子编码基因及其应用。
背景技术
植物逆境胁迫包括生物胁迫和非生物胁迫。生物胁迫主要为病虫害、杂草等；非生物胁迫主要为干旱、高盐碱、水涝、高温及冻害等。在胁迫刺激下，一些转录因子过量表达，将信号传递和放大，调控相应的下游功能基因的表达，从而提高植物的抗逆能力。由于转录因子能调控多个下游基因的表达，因此，在植物基因工程育种中，与转入一个功能基因相比，转入一个转录因子更能提高植物的抗逆能力。
转录因子(transcription factor,TF)，亦称反式作用因子，是一类能与真核生物基因启动子的顺式作用元件发生特异性相互作用的DNA结合蛋白，通过它们之间及与其他相关蛋白之间的相互作用，激活或抑制转录。根据蛋白质的结构，转录因子主要具有4个功能域，为DNA结合域(DNA-binding domain)，转录调控域(transcription regulationdomain，包括激活域和抑制域)，寡聚化位点(oligomerization site)和核定位信号(nuclear localization signal,NLS)。转录因子通过这些功能域，在特定的时间进入细胞核与启动子顺式作用元件或其他转录因子的功能域相互作用来调控基因的转录表达。
MYB类转录因子是转录因子家族中数量最大，功能最多的转录因子家族之一。MYB蛋白的N端都有一个由1-3个重复序列组成的MYB结构域(MYB结构域具体由51-52个氨基酸残基采用螺旋-转角-螺旋的形式与DNA结合组成)。每个重复序列通常含有3个间隔分布的色氨酸残基，形成疏水中心，识别特异的DNA结合位点。由于识别过程具有灵活性，作用机制多变，这就意味着MYB蛋白可以通过改变DNA结合域识别不同的目标基因，从而拥有完全不同的生理功能。MYB蛋白的C端是一个激活或抑制结构域。和保守的MYB结构域不同的是，其它区域是高度可变的。
已有研究表明MYB蛋白在植物中参与很多生理和生化过程，如调节次生代谢，参与类黄酮和花青素的合成，控制细胞形态发生，调节胚的形成以及花和种子的发育，控制细胞周期以及光和激素的信号途径等。
同时，也发现MYB蛋白参与了多种防御反应和胁迫反应。如水稻的OsMYB4基因，它是一个冷胁迫诱导的基因，OsMYB4基因过表达引起了254个基因的上调表达，这些上调表达基因大多数不仅使植物的抗寒能力提高，而且使植物的耐干旱以及耐高盐等抗逆境胁迫能力提高。将OsMYB4基因转入苹果内，结果显示提高了转基因苹果的抗干旱和低温能力。GmMYB基因能通过调控抗逆响应基因而在大豆的抗逆胁迫中起着不同的作用。拟南芥的AtMYB68基因能够抵抗高温胁迫，AtMYB2基因能够使拟南芥的耐寒能力显著增强。
随着研究的不断深入，科研人员发现MYB蛋白的功能越来越多，呈现多样化的特点。结构相似的MYB蛋白在功能上可能存在诸多相似性，但是它们在功能上不是绝对的冗余，更可能是功能上的重叠。同时还发现，在不同的物种甚至相同的器官中，结构相似的MYB蛋白在作用方式和功能上也可能存在显著的差异。如从抗旱植物厚叶旋蒴苣苔(Boeacrassifolia)中分离了一个BcMYB1基因，该基因序列与拟南芥AtMYB102基因高度相似，两个基因虽然都参与了水分胁迫响应。但是两个基因的表达模式差别非常大。AtMYB102能被外源ABA胁迫所诱导，而BcMYB1基因对ABA处理不敏感。此外，AtMYB102的表达同时依赖参与创伤和水分胁迫，而BcMYB1几乎不被创伤信号所诱导。
刚毛柽柳(Tamarix hispida)是一种抗逆能力非常优良的木本盐生植物，能在含盐量达1％的土壤中形成天然林，因此是研究耐盐机理和进行耐盐基因克隆的理想材料。目前对刚毛柽柳的MYB转录因子的耐盐功能了解较少。一些研究仅表明MYB基因的表达受盐胁迫诱导，可能参与植物的耐盐调控，但还不能深入认识其耐盐机理，无法利用刚毛柽柳的MYB转录因子编码基因培育出耐盐能力优良的木本植物。
发明内容
为解决上述现有技术的不足，本发明提供了刚毛柽柳MYB转录因子编码基因及其应用。
本发明的技术方案：
刚毛柽柳MYB转录因子编码基因，即刚毛柽柳ThMYB基因，所述刚毛柽柳ThMYB基因的cDNA序列如SEQ ID No：1所示。
进一步的，所述刚毛柽柳ThMYB基因的cDNA序列编码的氨基酸序列如SEQ ID NO：2所示。
一种含有所述刚毛柽柳ThMYB基因的植物过表达载体。
一种含有所述植物过表达载体的重组基因工程菌。
刚毛柽柳ThMYB基因在提高植物耐盐性或选育耐盐性转基因植物中的应用。
进一步的，所述应用包括构建含有刚毛柽柳ThMYB基因的植物过表达载体，将所构建的植物过表达载体转化到木本植物中，培育得到耐盐性转基因木本植物。
进一步的，所述木本植物为杨树或白桦树。
进一步的，所述应用包括构建含有刚毛柽柳ThMYB基因的植物过表达载体，将所构建的植物过表达载体转化到草本植物中，培育得到耐盐性转基因草本植物。
进一步的，所述草本植物为拟南芥或烟草。
本发明的有益效果：
本发明提供的刚毛柽柳MYB转录因子编码基因，即刚毛柽柳ThMYB基因可用于提高植物耐盐性或选育耐盐性转基因植物。分别构建刚毛柽柳ThMYB基因的过表达和抑制表达载体，通过瞬时侵染获得转基因刚毛柽柳，经组织化学染色和生理指标测定等试验证实刚毛柽柳ThMYB基因过表达株系具有明显的耐盐能力，且由于空白对照和抑制表达株系；同时将刚毛柽柳ThMYB基因转入模式植物拟南芥中，转基因T3代拟南芥种子在150mM NaCl胁迫后平均发芽率是野生型的6.21倍，根长是野生型的1.41倍，鲜重是野生型的1.66倍。刚毛柽柳ThMYB基因的同源和异源过表达结果均表明该基因能够明显提高转基因植物的耐盐能力，是用于林木耐盐基因工程育种的优良基因。
附图说明
图1为ThMYB基因保守区预测图；
图2为刚毛柽柳ThMYB与其它13种植物MYB蛋白多序列比对图；
图3为NaCl胁迫下ThMYB基因表达模式分析图；
图4为NaCl胁迫下转ThMYB基因和对照刚毛柽柳组织化学染色分析图；
图5为NaCl胁迫下转基因刚毛柽柳H2O2含量的测定分析图；
图6为NaCl胁迫下转基因刚毛柽柳MDA含量的测定分析图；
图7为NaCl胁迫下转基因刚毛柽柳相对电导率含量的测定分析图；
图8为ThMYB转基因和野生型(WT)拟南芥在盐胁迫后种子萌发表型观察比较图；
图9为ThMYB转基因和野生型(WT)拟南芥在盐胁迫后萌发率比较图；
图10为ThMYB转基因和野生型(WT)拟南芥在盐胁迫后根系发育表型观察比较图；
图11为ThMYB转基因和野生型(WT)拟南芥在盐胁迫后根长和鲜重比较图；
图12为ThMYB转基因和野生型(WT)拟南芥在盐胁迫后植株发育表型观察比较图。
具体实施方式
下面结合实施例对本发明的技术方案做进一步的说明，但并不局限于此，凡是对本发明技术方案进行修改或者等同替换，而不脱离本发明技术方案的精神和范围，均应涵盖在本发明的保护范围中。
实施例1：刚毛柽柳ThMYB基因的克隆、生物信息学分析及结构域验证
1、刚毛柽柳ThMYB基因的克隆
1.1提取刚毛柽柳总RNA
刚毛柽柳种子播种于炭土与沙比例为2:1的人工土泥中，置于相对湿度为65–75％，光强为400μmol·m-2·s-1，平均温度为22±2℃温室中生长。待生长2个月后，用0.3mol·L-1NaHCO3溶液浇灌根部，进行胁迫处理，分别在处理0h(胁迫处理前)、12h、24h和48h后，取其叶和根放入液氮中速冻用于RNA的提取。CTAB法提取各处理时期的RNA，将提取的RNA样品送至深圳华大基因科技有限公司进行转录组文库的构建和测序。
1.2克隆刚毛柽柳ThMYB基因
将获得的各处理时期的RNA测序结果进行对比拼接，对测序比对拼接后的结果用“MYB transcription factor”作为关键词进行查找，对查找后的序列进一步利用BLASTX软件进行比对确认，结合ORF founder程序(http://www.ncbi.nlm.nih.gov/gorf.html)确定是否具有完整的开放读码框。选择具有完整开放读码框的MYB基因。
根据ThMYB基因特征，设计了如SEQ ID NO：3和SEQ ID NO：4所示的特异性上下游引物(ThMYB-F和ThMYB-R)，以柽柳cDNA为模板，进行ThMYB基因RT-PCR克隆。
RT-PCR反应体系为20μL，其中包括模板2μL，ThMYB-F和ThMYB-R引物(10μmol/L)各1μL，dNTP Mix(10mmol/L)0.4μL，10×LA Taq PCR buffer 2.0μL，LA Taq(5U/μL)0.25μL。反应程序为94℃预变性3min；94℃变性30s，58℃退火30s，72℃延伸1min，35个循环；72℃延伸10min；使用胶回收试剂盒对扩增目的片段进行回收。
1.3测序验证刚毛柽柳ThMYB基因
将胶回收得到的目的基因连接到pMD18-T载体，将连接产物转化到大肠杆菌感受态细胞，挑取大肠杆菌感受态单克隆分别用基因引物ThMYB-F和ThMYB-R和如SEQ ID NO：5和SEQ ID NO：6所示的载体引物pMD18-T-F和pMD18-T-R进行PCR，将得到的特异性片段进行测序验证，最终获得如SEQ ID No：1所示的刚毛柽柳ThMYB基因的全长cDNA序列。
2、利用生物信息学软件和在线网络资源对ThMYB基因进行序列分析及结构域验证
本实施例获得的刚毛柽柳ThMYB基因的cDNA序列全长1014bp，编码如SEQ ID NO：2所示的337个氨基酸。利用网址为(http://au.expasy.org/tools/protparam.html)的ProtParam软件计算推导刚毛柽柳ThMYB基因编码的蛋白质分子量及理论等电点，结果表明该基因编码蛋白分子量为37.5，理论等电点为5.05。
通过网址为(http://blast.ncbi.nlm.nih.gov/Blast.cgi？PROGRAM＝blastx&BLAST_PROG RAMS＝blastx&PAGE_TYPE＝BlastSearch&SHOW_DEFAULTS＝on&LINK_LOC＝blasthom e)的BLASTx数据库对刚毛柽柳ThMYB基因进行相似性分析，比对结果如图1所示，本实施例获得的刚毛柽柳ThMYB基因含有保守的MYB结构域。
利用网址为(http://www.ncbi.nlm.nih.gov/BLAST/)的Blast程序进行序列同源性搜索，选择与其相似程度高的其它13种不同植物的MYB基因氨基酸序列，通过BioEdit软件进行多序列比对。
所选择的13种植物的MYB基因分别为：
Ziziphus jujuba(XP_015901922.1)、Gossypium hirsutum(XP_016723612.1)、
Vitis vinifera(XP_010660298.1)、Durio zibethinus(XP_022769989.1)、
Theobroma cacao(EOY03898.1)、Populus trichocarpa(XP_006372565.1)、
Solanum lycopersicum(NP_001234300.2)、Malus domestica(XP_008343128.1)、
Citrus sinensis(XP_006482113.1)、Ricinus communis(XP_015582549.1)、
Juglans regia(XP_018811302.1)、Vigna radiata(XP_014503716.1)、
Arabidopsis thaliana(AT4G12350)。
进一步通过Clustal软件对该基因编码蛋白序列与其他相似程度高的13种植物的MYB基因的序列同源性进行比较，结果如图2所示，刚毛柽柳MYB蛋白与其他各植物中MYB蛋白的同源性并不高，但都具有保守的MYB结构域。
实施例2：构建刚毛柽柳ThMYB基因的过表达载体
根据植物过表达载体pROKII的多克隆位点及ThMYB基因特征，通过在ThMYB基因5’端和3’端分别引入XbaI和KpnI限制性内切酶位点设计了如SEQ ID NO：9和SEQ ID NO：10所示的基因特异性上下游引物pROKⅡ-ThMYB-F和pROKⅡ-ThMYB-R，以柽柳cDNA为模板，进行ThMYB基因RT-PCR克隆。
RT-PCR反应体系为20μL，其中包括模板2μL，pROKⅡ-ThMYB-F和pROKⅡ-ThMYB-R特异性引物(10μmol/L)各1μL，dNTP Mix(10mmol/L)0.4μL，10×LA Taq PCR buffer 2.0μL，LA Taq(5U/μL)0.25μL。反应程序为94℃预变性3min；94℃变性30s，58℃退火30s，72℃延伸1min，35个循环；72℃延伸7min。
使用胶回收试剂盒对扩增目的片段进行回收。用KpnI和XbaI酶分别对胶回收ThMYB基因产物和pROKII质粒进行双酶切，分别回收后用T4连接酶进行连接。将连接产物转化到大肠杆菌感受态细胞，挑取大肠杆菌感受态单克隆，利用如SEQ ID NO：9、SEQ ID NO：10所示的基因特异性上下游引物pROKⅡ-ThMYB-F、pROKⅡ-ThMYB-R和SEQ ID NO：7、SEQ IDNO：8所示的过表达载体pROKII的上下游引物pROKⅡ-F、pROKⅡ-R进行PCR验证，并进一步测序验证。序列正确的载体即为ThMYB基因过表达载体，命名为pROKⅡ-ThMYB，转化农杆菌菌株EHA105。将构建好的EHA105(pROKⅡ-ThMYB)保存在-80℃超低温冰箱备用。
对比例1：构建刚毛柽柳ThMYB基因的抑制表达载体
1、构建pFGC5941-ThMYB-Cis
根据植物抑制表达载体pFGC5941的多克隆位点，及ThMYB基因的特征，通过在ThMYB基因5’端和3’端分别引入NcoⅠ、AscⅠ限制性内切酶位点设计引物。
以pROKII-ThMYB质粒为模板，如SEQ ID NO：11和SEQ ID NO：12所示的pFGC5941-ThMYB-Cis-F/R为引物进行PCR，扩增带有AscI、NcoI限制性内切酶位点的目的基因。用胶回收试剂盒对PCR产物ThMYB-Cis目的片段进行回收纯化。用AscI、NcoI限制性内切酶分别双酶切ThMYB-Cis目的基因片段和植物干扰载体pFGC5941质粒。37℃温育4h后，将获得的酶切产物分别进行回收纯化，用T4DNA ligase将纯化好的ThMYB-Cis双酶切目的片段与植物干扰载体pFGC5941双酶切大片段进行16℃催化连接12～16h，重组为载体pFGC5941-ThMYB-Cis，进而转化大肠杆菌感受态细胞，检测完毕，保存备用。
2、构建pFGC5941-ThMYB-Anti
以pROKII-ThMYB质粒为模板，如SEQ ID NO：13和SEQ ID NO：14所示的pFGC5941-ThMYB-Anti-F/R为引物进行PCR扩增带有XbaI、BamHI限制性内切酶位点的目的基因。同上，胶回收后用XbaI、BamHI限制性内切酶分别双酶切ThMYB-Anti目的基因片段和重组载体pFGC5941-ThMYB-Cis。将获得的酶切产物分别进行回收纯化，用T4DNA ligase将纯化好的ThMYB-Anti双酶切目的片段与pFGC5941-ThMYB-Cis双酶切大片段进行16℃催化连接12～16h。将连接产物转化到大肠杆菌感受态细胞，挑取单克隆进行菌液PCR检测阳性克隆，并进一步测序验证。获得ThMYB基因抑制表达载体，命名为pFGC5941-ThMYB，转农杆菌菌株EHA105。将构建好的EHA105(pFGC5941-ThMYB)保存在-80℃超低温冰箱备用。
实施例3：刚毛柽柳ThMYB基因的瞬时转化
本实施例利用瞬时转化技术将实施例2构建完成的过表达载体pROKII-ThMYB(OE)和对比例1构建完成的抑制表达载体pFGC5941-ThMYB(SE)分别转入刚毛柽柳中，使ThMYB基因瞬时过表达或抑制表达。同时，将空pROKII载体转入刚毛柽柳作为对照(CON)。
具体方法如下：
一、选取继代培养30-35天的刚毛柽柳组培苗，株高为2.5-3.0cm，将选取的刚毛柽柳组培苗放入高渗透处理液中浸泡5-10min；
二、将步骤一高渗处理过的刚毛柽柳组培苗迅速转移至加有含目的基因的根癌农杆菌的侵染液中，其中侵染液OD600为0.6-0.8，置于真空泵中对其抽真空5-10min，然后将其置于恒温振荡器24～26℃，100～130rpm侵染4～5h；
三、取出步骤二中侵染完成后的组培苗，无菌水洗涤3-4次，用无菌滤纸吸干水分，***共培养固体培养基，共培养32～38h。
为了进一步分析ThMYB基因在瞬时过表达和抑制表达刚毛柽柳中的表达情况，提取150mM NaCl胁迫12h、24h和36h后瞬时侵染刚毛柽柳总RNA，反转录为cDNA，利用qRT-PCR对ThMYB基因的表达量进行分析，结果如图3所示，与CON相比，ThMYB在OE中的表达量明显增高，而在RNAi抑制表达株系中明显减少，这表明本实施例成功获得了ThMYB基因瞬时转化刚毛柽柳。
实施例4：NaCl胁迫下瞬时转基因刚毛柽柳苗的组织化学染色分析
本实施例主要通过对转基因刚毛柽柳苗的组织进行NBT、DAB和Evans Blue染色来分析植物体内O2 -和H2O2的含量，以分析植物体细胞膜的完整性情况。
硝基四氮唑蓝(Nitroblue Tetrazolium，NBT)还原法可研究细胞内超氧离子水平，材料蓝色的深浅反映细胞内O2 -的多少。本实施例中1mg/mL NBT染色液的具体配制方法为：称取50mgNBT溶于50mL磷酸缓冲液(pH7.0)。
二氨基联苯胺(Diaminobenzidine，DAB)染色通过颜色的深浅，可以反映胞内H2O2含量的多少。本实施例中1mg/mL DAB染色液的具体配制方法为：称取50mg DAB溶于50mL磷酸缓冲液(pH7.0)。
伊文思蓝(Evans Blue)染色可通过染色深浅判断植物细胞膜是否完整。正常的活细胞，胞膜结构完整，能够排斥伊文思蓝染色液，使之不能够进入胞内。而丧失活性或细胞膜不完整的细胞，细胞膜的通透性增加，可被伊文思蓝染成蓝色。并且丧失活性或细胞膜不完整的细胞数量越多，伊文思蓝染色结果越深。本实施例中1mg/mL Evans blue染色液的具体配制方法为：称取50mg Evans blue溶于50mL蒸馏水。
本实施例染色处理的转基因植株为瞬时转化过表达载体pROKII-ThMYB(OE)的刚毛柽柳植株、瞬时转化抑制表达载体pFGC5941-ThMYB(RNAi)的刚毛柽柳植株和瞬时转化空pROKII载体的空白(CON)对照刚毛柽柳植株。
本实施例的染色方法为：分别将150mM NaCl胁迫0h、2h后的上述三种转基因刚毛柽柳置于10mL离心管中，分别加入不同染色液使叶片全部浸没在染色液中，室温放置适当时间，待染色结果明显后，用脱色液(冰乙酸：酒精＝1:3)脱色。
染色结果如图4所示，未胁迫时，三种瞬时表达株系着色程度基本相同。胁迫2h后，3种转基因株系着色都加深。但NBT染色中SE株系着色最深，OE株系着色最浅，表明ThMYB基因的过表达能有效清除细胞体内O2 -含量，而抑制表达则相反，导致胞内O2 -的积累增多；同样，DAB染色中，OE株系着色最浅，SE株系着色比CON更深，表明胁迫后OE株系H2O2含量最少，SE株系H2O2含量最多。进一步的Evans blue染色结果中OE株系着色最浅，SE株系着色比CON更深，表明ThMYB基因的过表达能明显改善NaCl胁迫后细胞膜受损情况。综上结果表明盐胁迫后ThMYB基因的过表达能提高植物的ROS清除能力，从而减轻胁迫对细胞的损伤。
实施例5：NaCl胁迫下瞬时转基因刚毛柽柳的生理生化指标分析
本实施例将瞬时侵染后的刚毛柽柳置于150mM NaCl分别胁迫12和24h后测其生理指标，分析比较了3种转基因刚毛柽柳的H2O2含量、丙二醛(MDA)含量、电导率情况。
本实施例染色处理的转基因植株为瞬时转化过表达载体pROKII-ThMYB(OE)的刚毛柽柳植株、瞬时转化抑制表达载体pFGC5941-ThMYB(RNAi)的刚毛柽柳植株和瞬时转化空pROKII载体的空白(CON)对照刚毛柽柳植株。
本实施例中H2O2含量，MDA含量的测量采用了南京建成生物工程研究所生产的H2O2含量，MDA含量测定试剂盒；
相对电导率测定具体方法如下：1)50ml玻璃三角瓶的准备：用去离子水洗净，灭菌，烘干，用灭菌的去离子水平衡24h，在烘箱内烘干备用；2)将植物材料用自来水洗去表面的灰尘，再用双蒸水和超纯水分别冲洗3次，用洁净滤纸吸净表面的水分。取面积相等的叶片分别放入烧杯内，加超纯水50ml，用抽真空仪器抽15min，用电导仪测其电导值，记录S1；3)然后放入90℃水浴中20min，冷却至室温，测其电导率，记录S2；4)计算公式：相对电导率＝S1/S2。
H2O2是细胞内的氧化代谢产物，植物抵御逆境胁迫能力越差，细胞的受损程度越严重，H2O2含量越高。图5显示了150mM NaCl分别胁迫0h、12和24h后三种转基因刚毛柽柳的H2O2含量测定结果，结果显示150mM NaCl胁迫下，OE株系在胁迫12h和24h后的H2O2含量均保持在较低水平，特别是胁迫24h时ThMYB的过表达株系OE的H2O2含量是CON的0.81倍，是RNAi的0.77倍。
植物在逆境条件下，细胞会发生膜脂过氧化作用并产生丙二醛(MDA)。MDA通常作为脂质过氧化指标，表示细胞膜脂过氧化程度和植物对逆境条件反应的强弱。图6显示了150mM NaCl分别胁迫0h、12和24h后三种转基因刚毛柽柳的MDA含量测定结果，结果显示与0h的MDA含量相比，胁迫后3个转基因刚毛柽柳株系的MDA含量均不同程度的增大且在各个胁迫时间点均为RNAi的MDA含量最高，CON次之，OE的MDA含量最低。由此可知RNAi株系中细胞受损程度高和死亡数量大，OE株系中细胞受损程度低和死亡数量较少。
相对电导率也是表明细胞膜受损的一个生理指标，胞膜受损导致电解质外渗增加。图7显示了150mM NaCl分别胁迫0h、12和24h后三种转基因刚毛柽柳的相对电导率，结果显示ThMYB基因的过表达株系OE的相对电导率低于CON和RNAi株系，说明OE植株的耐盐能力高于CON和RNAi株系。
实施例6：转ThMYB基因拟南芥的获得和耐盐性鉴定
利用农杆菌介导的浸花法转化野生型拟南芥Columbia生态型，获得T3代转基因纯合过表达株系。为了分析过表达ThMYB基因对拟南芥耐盐能力的影响，将转ThMYB基因拟南芥T3代过表达株系(OE1、OE2)和野生型种子进行消毒，然后播种于1/2MS和含有150mM NaCl的1/2MS培养基上，10～15d，统计萌发率。结果如图8和图9所示，正常生长条件下，转基因拟南芥与野生型拟南芥生长及表型没有差别，表明ThMYB基因对植物的生长及表型没有明显的干扰。而150mM NaCl胁迫条件下，转基因拟南芥株系的平均发芽率是野生型株系发芽率的6.21倍。
同时为了测定盐胁迫对转基因和非转基因(野生型)拟南芥生长的影响，将在正常培养基上生长3d大的拟南芥小苗分别移栽到1/2MS和含150mM NaCl的1/2MS培养基上，生长14d后统计根长和鲜重。结果如图10和图11显示，盐胁迫后转基因株系的平均根长是野生型的1.41倍，鲜重是野生型的1.66倍(图9)。
对正常生长三周大的转基因和非转基因(野生型)拟南芥土培苗进行盐胁迫，结果如图12所示，在非胁迫条件下(control)，OE1和OE2两个不同转基因株系以及WT的生长状态没有明显的差别；150mM NaCl胁迫5-7天后，OE1和OE2两个转基因株系以及WT的生长状态都发生了变化，ThMYB基因的过表达株系OE1和OE2的生长状态明显好于WT，野生型wt株系叶面变黄，萎蔫状况比OE1和OE2两个转基因株系严重，这说明OE1和OE2植株的抗逆能力高于WT。
综上，通过对转基因刚毛柽柳和拟南芥分析，从同源和异源过表达ThMYB基因两个方面，全面地分析了刚毛柽柳ThMYB基因的耐盐功能，结果表明刚毛柽柳ThMYB基因明显提高了转基因植物的耐盐能力，因此将该基因用于制备转基因植物尤其是转基因林木具有非常重要的应用前景。
以上实施例中未作具体说明的分子生物学实验方法，均参照《分子克隆实验指南》(第三版，J.萨姆布鲁克)一书中所列的具体方法进行，或者按照试剂盒产品说明书进行操作。
SEQUENCE LISTING
<110> 东北林业大学
<120> 刚毛柽柳MYB转录因子编码基因及其应用
<130> 1
<160> 18
<170> PatentIn version 3.3
<210> 1
<211> 1014
<212> DNA
<213> 刚毛柽柳MYB转录因子编码基因的cDNA序列
<400> 1
atgggaaggc aaccgtgttg tgataaagta ggattgaaga aaggaccctg gactgccgaa 60
gaagacaaga agcttatcag cttcatcctt acaaacggcc agtgctgctg gagagctgtt 120
cctaaactcg cagggttgct gaggtgtggg aagagctgca ggttgagatg gacaaactat 180
ttaaggccag atctcaagag gggtctgctt gatgaaaaag aagagcagat ggtcattgat 240
cttcatgcta aacttggcaa cagatggtcc aagattgcct cccacctccc aggcagaaca 300
gataacgaga tcaagaatca ttggaacacc cacatcaaga agaagctcaa gaaaatgggc 360
atcgatcccc tcactcacag ccctctccca cctcctgcca ccgaccaaga tcctcaagct 420
gcccggcagc caccgacaga aacagaacat ccgccgccgc cgcagcagca gacgtctagc 480
tcctcacgcg acgacggcat cgagacgacg atgaacgcga ccatttccat cgccgaggac 540
aaggaacagt atgagatggc cgacgcagcg acggaccatc aactggaggc cgtggacagc 600
ttctgtgtag acgaagtgcc gctgcttcag ccacacgagc ttgattataa cttagctgct 660
attcctgctc cgacggcagc cacgtccacc agttcaactt cttcatcgtc gagcacggct 720
cgtttgggag gttctctctc acagccacag gaattattcg actggcctta cgattactgt 780
aataacaaca gcagcatcca cagcgacagc agcagcagta gcagcactac tactaccact 840
accaacttgc aacgctgtga cgtgggattc tgggatgatg atgagttcac aaactccatt 900
gattgggact tgttgatcaa taacattcat tatgacgcac agctgggccc tgatcctctt 960
ccactcagcc aatgtgtgcc actgtctttt gatcaactac ctcagaactt ttag 1014
<210> 2
<211> 337
<212> PRT
<213> 刚毛柽柳MYB转录因子编码基因的cDNA序列编码的氨基酸序列
<400> 2
Met Gly Arg Gln Pro Cys Cys Asp Lys Val Gly Leu Lys Lys Gly Pro
1 5 10 15
Trp Thr Ala Glu Glu Asp Lys Lys Leu Ile Ser Phe Ile Leu Thr Asn
20 25 30
Gly Gln Cys Cys Trp Arg Ala Val Pro Lys Leu Ala Gly Leu Leu Arg
35 40 45
Cys Gly Lys Ser Cys Arg Leu Arg Trp Thr Asn Tyr Leu Arg Pro Asp
50 55 60
Leu Lys Arg Gly Leu Leu Asp Glu Lys Glu Glu Gln Met Val Ile Asp
65 70 75 80
Leu His Ala Lys Leu Gly Asn Arg Trp Ser Lys Ile Ala Ser His Leu
85 90 95
Pro Gly Arg Thr Asp Asn Glu Ile Lys Asn His Trp Asn Thr His Ile
100 105 110
Lys Lys Lys Leu Lys Lys Met Gly Ile Asp Pro Leu Thr His Ser Pro
115 120 125
Leu Pro Pro Pro Ala Thr Asp Gln Asp Pro Gln Ala Ala Arg Gln Pro
130 135 140
Pro Thr Glu Thr Glu His Pro Pro Pro Pro Gln Gln Gln Thr Ser Ser
145 150 155 160
Ser Ser Arg Asp Asp Gly Ile Glu Thr Thr Met Asn Ala Thr Ile Ser
165 170 175
Ile Ala Glu Asp Lys Glu Gln Tyr Glu Met Ala Asp Ala Ala Thr Asp
180 185 190
His Gln Leu Glu Ala Val Asp Ser Phe Cys Val Asp Glu Val Pro Leu
195 200 205
Leu Gln Pro His Glu Leu Asp Tyr Asn Leu Ala Ala Ile Pro Ala Pro
210 215 220
Thr Ala Ala Thr Ser Thr Ser Ser Thr Ser Ser Ser Ser Ser Thr Ala
225 230 235 240
Arg Leu Gly Gly Ser Leu Ser Gln Pro Gln Glu Leu Phe Asp Trp Pro
245 250 255
Tyr Asp Tyr Cys Asn Asn Asn Ser Ser Ile His Ser Asp Ser Ser Ser
260 265 270
Ser Ser Ser Thr Thr Thr Thr Thr Thr Asn Leu Gln Arg Cys Asp Val
275 280 285
Gly Phe Trp Asp Asp Asp Glu Phe Thr Asn Ser Ile Asp Trp Asp Leu
290 295 300
Leu Ile Asn Asn Ile His Tyr Asp Ala Gln Leu Gly Pro Asp Pro Leu
305 310 315 320
Pro Leu Ser Gln Cys Val Pro Leu Ser Phe Asp Gln Leu Pro Gln Asn
325 330 335
Phe
<210> 3
<211> 30
<212> DNA
<213> 人工序列
<400> 3
gctctagaat gggaaggcaa ccgtgttgtg 30
<210> 4
<211> 30
<212> DNA
<213> 人工序列
<400> 4
cggggtaccc taaaagttct gaggtagttg 30
<210> 5
<211> 24
<212> DNA
<213> 人工序列
<400> 5
agcggataac aatttcacac agga 24
<210> 6
<211> 24
<212> DNA
<213> 人工序列
<400> 6
cgccagggtt ttcccagtca cgac 24
<210> 7
<211> 22
<212> DNA
<213> 人工序列
<400> 7
acaggtttcc cgactggaaa gc 22
<210> 8
<211> 20
<212> DNA
<213> 人工序列
<400> 8
ggcgaacgtg gcgagaaagg 20
<210> 9
<211> 30
<212> DNA
<213> 人工序列
<400> 9
gctctagaat gggaaggcaa ccgtgttgtg 30
<210> 10
<211> 30
<212> DNA
<213> 人工序列
<400> 10
cggggtaccc taaaagttct gaggtagttg 30
<210> 11
<211> 29
<212> DNA
<213> 人工序列
<400> 11
catgccatgg gacgatgaac gcgaccatt 29
<210> 12
<211> 31
<212> DNA
<213> 人工序列
<400> 12
ttggcgcgcc acagtaatcg taaggccagt c 31
<210> 13
<211> 27
<212> DNA
<213> 人工序列
<400> 13
gctctagaga cgatgaacgc gaccatt 27
<210> 14
<211> 29
<212> DNA
<213> 人工序列
<400> 14
cgggatccac agtaatcgta aggccagtc 29
<210> 15
<211> 22
<212> DNA
<213> 人工序列
<400> 15
ataaggaagt tcatttcatt tg 22
<210> 16
<211> 20
<212> DNA
<213> 人工序列
<400> 16
caatcaaatg aagagccaat 20
<210> 17
<211> 22
<212> DNA
<213> 人工序列
<400> 17
cttacttaca cttgccttgg ag 22
<210> 18
<211> 21
<212> DNA
<213> 人工序列
<400> 18
atctgagcta cacatgctca g 21
Claims (9)
1.刚毛柽柳MYB转录因子编码基因，即刚毛柽柳ThMYB基因，其特征在于所述刚毛柽柳ThMYB基因的cDNA序列如SEQ ID No：1所示。
2.如权利要求1所述刚毛柽柳ThMYB基因，其特征在于所述cDNA序列编码的氨基酸序列如SEQ ID NO：2所示。
3.一种含有权利要求1所述刚毛柽柳ThMYB基因的植物过表达载体。
4.一种含有权利要求4所述植物过表达载体的重组基因工程菌。
5.如权利要求1所述的刚毛柽柳ThMYB基因在提高植物耐盐性或选育耐盐性转基因植物中的应用。
6.如权利要求5所述刚毛柽柳ThMYB基因的应用，其特征在于所述应用包括构建含有刚毛柽柳ThMYB基因的植物过表达载体，将所构建的植物过表达载体转化到木本植物中，培育得到耐盐性转基因木本植物。
7.如权利要求6所述刚毛柽柳ThMYB基因的应用，其特征在于所述木本植物为杨树或白桦树。
8.如权利要求5所述刚毛柽柳ThMYB基因的应用，其特征在于所述应用包括构建含有刚毛柽柳ThMYB基因的植物过表达载体，将所构建的植物过表达载体转化到草本植物中，培育得到耐盐性转基因草本植物。
9.如权利要求8所述刚毛柽柳ThMYB基因的应用，其特征在于所述草本植物为拟南芥或烟草。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810288452.2A CN108342395B (zh) | 2018-03-30 | 2018-03-30 | 刚毛柽柳myb转录因子编码基因及其应用 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810288452.2A CN108342395B (zh) | 2018-03-30 | 2018-03-30 | 刚毛柽柳myb转录因子编码基因及其应用 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN108342395A true CN108342395A (zh) | 2018-07-31 |
| CN108342395B CN108342395B (zh) | 2019-11-05 |
Family
ID=62957817
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201810288452.2A Expired - Fee Related CN108342395B (zh) | 2018-03-30 | 2018-03-30 | 刚毛柽柳myb转录因子编码基因及其应用 |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN108342395B (zh) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110951754A (zh) * | 2020-01-06 | 2020-04-03 | 东北林业大学 | 刚毛柽柳col转录因子编码基因及其应用 |
| CN111118036A (zh) * | 2020-03-02 | 2020-05-08 | 东北林业大学 | 刚毛柽柳phd3转录因子编码基因及其应用 |
| CN112725356A (zh) * | 2021-02-08 | 2021-04-30 | 南京林业大学 | 一种鹅掌楸转录因子LcbHLH16421基因及其应用 |
| CN116891856A (zh) * | 2023-07-17 | 2023-10-17 | 江西省科学院生物资源研究所 | 赣彤1号CcMYB10_LIKE基因及其表达蛋白和应用 |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1624132A (zh) * | 2004-11-10 | 2005-06-08 | 东北林业大学 | 多枝柽柳Myb转录因子基因 |
-
2018
- 2018-03-30 CN CN201810288452.2A patent/CN108342395B/zh not_active Expired - Fee Related
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1624132A (zh) * | 2004-11-10 | 2005-06-08 | 东北林业大学 | 多枝柽柳Myb转录因子基因 |
Non-Patent Citations (2)
| Title |
|---|
| 张凤娇等: "7个柽柳ThMYB基因的鉴定及表达", 《东北林业大学学报》 * |
| 杨桂燕等: "ThMYB3 和 ThDof2 对 ThVHAc1 基因表达的调控", 《北京林业大学学报》 * |
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110951754A (zh) * | 2020-01-06 | 2020-04-03 | 东北林业大学 | 刚毛柽柳col转录因子编码基因及其应用 |
| CN110951754B (zh) * | 2020-01-06 | 2022-05-10 | 东北林业大学 | 刚毛柽柳col转录因子编码基因及其应用 |
| CN111118036A (zh) * | 2020-03-02 | 2020-05-08 | 东北林业大学 | 刚毛柽柳phd3转录因子编码基因及其应用 |
| CN111118036B (zh) * | 2020-03-02 | 2023-01-24 | 东北林业大学 | 刚毛柽柳phd3转录因子编码基因及其应用 |
| CN112725356A (zh) * | 2021-02-08 | 2021-04-30 | 南京林业大学 | 一种鹅掌楸转录因子LcbHLH16421基因及其应用 |
| CN112725356B (zh) * | 2021-02-08 | 2022-02-01 | 南京林业大学 | 一种鹅掌楸转录因子LcbHLH16421基因及其应用 |
| CN116891856A (zh) * | 2023-07-17 | 2023-10-17 | 江西省科学院生物资源研究所 | 赣彤1号CcMYB10_LIKE基因及其表达蛋白和应用 |
Also Published As
| Publication number | Publication date |
|---|---|
| CN108342395B (zh) | 2019-11-05 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Divya et al. | Constitutive expression of mustard annexin, AnnBj1 enhances abiotic stress tolerance and fiber quality in cotton under stress | |
| CN101265294B (zh) | 一种抗病相关的小麦myb蛋白及其编码基因与应用 | |
| CN108342395B (zh) | 刚毛柽柳myb转录因子编码基因及其应用 | |
| CN106244607A (zh) | 棉花细胞色素p450 cyp94c1基因在抗黄萎病中的应用 | |
| Zhang et al. | Identification of pepper CaSBP08 gene in defense response against Phytophthora capsici infection | |
| CN114921474A (zh) | 一种提高白桦耐盐能力的BpbHLH10基因及其编码的蛋白 | |
| CN110923250B (zh) | 棉花黄萎病抗性相关基因GhSDH1-1的应用 | |
| CN108676804A (zh) | 拟南芥at5g49330基因在盐胁迫反应方面的应用 | |
| CN106191001A (zh) | 磷脂酶PLDζ1基因在提高植物耐盐性中的应用 | |
| CN114790449B (zh) | 钙依赖蛋白激酶基因GhCPK4在植物抗黄萎病中的应用 | |
| Li et al. | Overexpression of MpbHLH transcription factor, an encoding ICE1-like protein, enhances Foc TR4-resistance of Cavendish banana | |
| CN115838734A (zh) | C2h2型锌指蛋白基因hstl在调控水稻耐盐性中的应用 | |
| CN104561044B (zh) | 一种提高白桦抗逆性的基因及其编码的蛋白 | |
| CN104561045B (zh) | 一种提高白桦抗逆性的转录因子及其编码的蛋白 | |
| CN111235160B (zh) | 一种密罗木基因MfWRKY70及其应用 | |
| CN111118024B (zh) | 一种密罗木基因MfbHLH44及其应用 | |
| Meir et al. | Transforming a NEP1 toxin gene into two Fusarium spp. to enhance mycoherbicide activity on Orobanche—failure and success | |
| Wijekoon et al. | The role of a putative peroxisomal-targeted epoxide hydrolase of Nicotiana benthamiana in interactions with Colletotrichum destructivum, C. orbiculare or Pseudomonas syringae pv. tabaci | |
| KR102127184B1 (ko) | 벼 유래의 ak102606 유전자의 항산화능, 환경 스트레스 및 수확량 조절자로서의 용도 | |
| Yuan et al. | Ectopic expression of BrIQD35 promotes drought stress tolerance in Nicotiana benthamiana | |
| CN106811448B (zh) | 棉花酪氨酸蛋白磷酸酶GhPTP1及其编码基因和应用 | |
| CN101686642A (zh) | 增强植物外伤耐受性的方法 | |
| CN114736279B (zh) | 一种植物抗逆相关蛋白PvNAC52及其编码基因和应用 | |
| CN101111595A (zh) | 新型高等植物的制作方法以及高等植物的生长促进方法 | |
| KR101460743B1 (ko) | 병저항성 관련 키티네이즈 유전자 CaChitIV 및 이를 이용한 형질전환 식물체 |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20191105 |