CN114836441B - 玉米种子耐贮性相关的ZmRAP2.7基因及其功能标记与应用 - Google Patents
玉米种子耐贮性相关的ZmRAP2.7基因及其功能标记与应用 Download PDFInfo
- Publication number
- CN114836441B CN114836441B CN202210687849.5A CN202210687849A CN114836441B CN 114836441 B CN114836441 B CN 114836441B CN 202210687849 A CN202210687849 A CN 202210687849A CN 114836441 B CN114836441 B CN 114836441B
- Authority
- CN
- China
- Prior art keywords
- gene
- seeds
- seed
- seq
- storability
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- 235000002017 Zea mays subsp mays Nutrition 0.000 title claims abstract description 40
- 101150101112 7 gene Proteins 0.000 title claims abstract description 36
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 title claims abstract description 28
- 235000005822 corn Nutrition 0.000 title claims abstract description 28
- 240000008042 Zea mays Species 0.000 title abstract description 39
- 239000003550 marker Substances 0.000 title description 3
- 108090000623 proteins and genes Proteins 0.000 claims abstract description 155
- 230000035784 germination Effects 0.000 claims abstract description 50
- 230000032683 aging Effects 0.000 claims abstract description 44
- 102000004169 proteins and genes Human genes 0.000 claims abstract description 36
- 230000014509 gene expression Effects 0.000 claims abstract description 30
- 230000002829 reductive effect Effects 0.000 claims abstract description 17
- 230000001105 regulatory effect Effects 0.000 claims description 22
- 238000003860 storage Methods 0.000 claims description 18
- 239000000463 material Substances 0.000 claims description 17
- 239000002299 complementary DNA Substances 0.000 claims description 10
- 230000007226 seed germination Effects 0.000 claims description 7
- 108091028043 Nucleic acid sequence Proteins 0.000 claims description 6
- 230000001276 controlling effect Effects 0.000 claims description 6
- 238000012217 deletion Methods 0.000 claims description 6
- 230000037430 deletion Effects 0.000 claims description 6
- 230000015572 biosynthetic process Effects 0.000 claims description 4
- 230000002103 transcriptional effect Effects 0.000 claims description 3
- 230000002401 inhibitory effect Effects 0.000 claims description 2
- 241000209149 Zea Species 0.000 claims 2
- 230000009456 molecular mechanism Effects 0.000 claims 1
- 102000054766 genetic haplotypes Human genes 0.000 abstract description 20
- 238000003780 insertion Methods 0.000 abstract description 9
- 230000037431 insertion Effects 0.000 abstract description 9
- 240000004808 Saccharomyces cerevisiae Species 0.000 abstract description 7
- 230000027455 binding Effects 0.000 abstract description 6
- 230000000903 blocking effect Effects 0.000 abstract description 6
- 238000009395 breeding Methods 0.000 abstract description 4
- 230000001488 breeding effect Effects 0.000 abstract description 4
- 230000011664 signaling Effects 0.000 abstract description 4
- 238000010356 CRISPR-Cas9 genome editing Methods 0.000 abstract description 3
- 238000012097 association analysis method Methods 0.000 abstract description 2
- 238000010353 genetic engineering Methods 0.000 abstract description 2
- 125000003275 alpha amino acid group Chemical group 0.000 abstract 1
- 230000007812 deficiency Effects 0.000 abstract 1
- 230000004064 dysfunction Effects 0.000 abstract 1
- 238000011222 transcriptome analysis Methods 0.000 abstract 1
- 108020004414 DNA Proteins 0.000 description 37
- 235000018102 proteins Nutrition 0.000 description 31
- JLIDBLDQVAYHNE-YKALOCIXSA-N (+)-Abscisic acid Chemical compound OC(=O)/C=C(/C)\C=C\[C@@]1(O)C(C)=CC(=O)CC1(C)C JLIDBLDQVAYHNE-YKALOCIXSA-N 0.000 description 30
- 238000000034 method Methods 0.000 description 29
- 230000006870 function Effects 0.000 description 18
- 239000002773 nucleotide Substances 0.000 description 16
- 125000003729 nucleotide group Chemical group 0.000 description 16
- 108091023040 Transcription factor Proteins 0.000 description 14
- 102000040945 Transcription factor Human genes 0.000 description 14
- 235000016383 Zea mays subsp huehuetenangensis Nutrition 0.000 description 12
- 235000009973 maize Nutrition 0.000 description 12
- 238000006243 chemical reaction Methods 0.000 description 11
- 230000019491 signal transduction Effects 0.000 description 11
- 241000196324 Embryophyta Species 0.000 description 10
- 150000001413 amino acids Chemical group 0.000 description 10
- 230000000694 effects Effects 0.000 description 10
- 230000035772 mutation Effects 0.000 description 10
- 102000053602 DNA Human genes 0.000 description 9
- 230000017260 vegetative to reproductive phase transition of meristem Effects 0.000 description 9
- 238000012408 PCR amplification Methods 0.000 description 8
- 238000012098 association analyses Methods 0.000 description 8
- 230000008859 change Effects 0.000 description 8
- 239000003153 chemical reaction reagent Substances 0.000 description 8
- 238000012360 testing method Methods 0.000 description 8
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 7
- 101150009243 HAP1 gene Proteins 0.000 description 7
- 240000007594 Oryza sativa Species 0.000 description 7
- 235000007164 Oryza sativa Nutrition 0.000 description 7
- 239000012535 impurity Substances 0.000 description 7
- 230000008569 process Effects 0.000 description 7
- 108010016634 Seed Storage Proteins Proteins 0.000 description 6
- 238000004458 analytical method Methods 0.000 description 6
- 230000005764 inhibitory process Effects 0.000 description 6
- 210000001161 mammalian embryo Anatomy 0.000 description 6
- 239000000203 mixture Substances 0.000 description 6
- 235000009566 rice Nutrition 0.000 description 6
- 241000219195 Arabidopsis thaliana Species 0.000 description 5
- 239000012634 fragment Substances 0.000 description 5
- 239000000499 gel Substances 0.000 description 5
- 239000000047 product Substances 0.000 description 5
- 239000000523 sample Substances 0.000 description 5
- 238000012163 sequencing technique Methods 0.000 description 5
- 239000000243 solution Substances 0.000 description 5
- 238000011144 upstream manufacturing Methods 0.000 description 5
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 4
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 4
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 4
- 108700039691 Genetic Promoter Regions Proteins 0.000 description 4
- 235000001014 amino acid Nutrition 0.000 description 4
- 210000002257 embryonic structure Anatomy 0.000 description 4
- 238000009396 hybridization Methods 0.000 description 4
- 238000007901 in situ hybridization Methods 0.000 description 4
- 230000007246 mechanism Effects 0.000 description 4
- 239000012071 phase Substances 0.000 description 4
- 230000022532 regulation of transcription, DNA-dependent Effects 0.000 description 4
- 230000008439 repair process Effects 0.000 description 4
- 238000006467 substitution reaction Methods 0.000 description 4
- 238000003786 synthesis reaction Methods 0.000 description 4
- 210000001519 tissue Anatomy 0.000 description 4
- 230000009261 transgenic effect Effects 0.000 description 4
- 238000005406 washing Methods 0.000 description 4
- 241000219194 Arabidopsis Species 0.000 description 3
- 229930192334 Auxin Natural products 0.000 description 3
- 108020004705 Codon Proteins 0.000 description 3
- 108700024394 Exon Proteins 0.000 description 3
- 244000068988 Glycine max Species 0.000 description 3
- 235000010469 Glycine max Nutrition 0.000 description 3
- 108700026244 Open Reading Frames Proteins 0.000 description 3
- 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 3
- MUPFEKGTMRGPLJ-RMMQSMQOSA-N Raffinose Natural products O(C[C@H]1[C@@H](O)[C@H](O)[C@@H](O)[C@@H](O[C@@]2(CO)[C@H](O)[C@@H](O)[C@@H](CO)O2)O1)[C@@H]1[C@H](O)[C@@H](O)[C@@H](O)[C@@H](CO)O1 MUPFEKGTMRGPLJ-RMMQSMQOSA-N 0.000 description 3
- 244000098338 Triticum aestivum Species 0.000 description 3
- MUPFEKGTMRGPLJ-UHFFFAOYSA-N UNPD196149 Natural products OC1C(O)C(CO)OC1(CO)OC1C(O)C(O)C(O)C(COC2C(C(O)C(O)C(CO)O2)O)O1 MUPFEKGTMRGPLJ-UHFFFAOYSA-N 0.000 description 3
- 239000002253 acid Substances 0.000 description 3
- 230000009471 action Effects 0.000 description 3
- 230000003321 amplification Effects 0.000 description 3
- 239000002363 auxin Substances 0.000 description 3
- 230000033228 biological regulation Effects 0.000 description 3
- 210000000349 chromosome Anatomy 0.000 description 3
- 238000010367 cloning Methods 0.000 description 3
- 238000013461 design Methods 0.000 description 3
- 238000009826 distribution Methods 0.000 description 3
- 230000006872 improvement Effects 0.000 description 3
- 238000000338 in vitro Methods 0.000 description 3
- SEOVTRFCIGRIMH-UHFFFAOYSA-N indole-3-acetic acid Chemical compound C1=CC=C2C(CC(=O)O)=CNC2=C1 SEOVTRFCIGRIMH-UHFFFAOYSA-N 0.000 description 3
- 239000007788 liquid Substances 0.000 description 3
- 230000004048 modification Effects 0.000 description 3
- 238000012986 modification Methods 0.000 description 3
- 238000003199 nucleic acid amplification method Methods 0.000 description 3
- 108010031719 prolyl-serine Proteins 0.000 description 3
- 230000001681 protective effect Effects 0.000 description 3
- MUPFEKGTMRGPLJ-ZQSKZDJDSA-N raffinose 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[C@@H]2[C@@H]([C@@H](O)[C@@H](O)[C@@H](CO)O2)O)O1 MUPFEKGTMRGPLJ-ZQSKZDJDSA-N 0.000 description 3
- 238000011160 research Methods 0.000 description 3
- ODWSTKXGQGYHSH-FXQIFTODSA-N Ala-Arg-Ala Chemical compound C[C@H](N)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](C)C(O)=O ODWSTKXGQGYHSH-FXQIFTODSA-N 0.000 description 2
- ZVFVBBGVOILKPO-WHFBIAKZSA-N Ala-Gly-Ala Chemical compound C[C@H](N)C(=O)NCC(=O)N[C@@H](C)C(O)=O ZVFVBBGVOILKPO-WHFBIAKZSA-N 0.000 description 2
- 244000045195 Cicer arietinum Species 0.000 description 2
- 235000010523 Cicer arietinum Nutrition 0.000 description 2
- 244000020551 Helianthus annuus Species 0.000 description 2
- 235000003222 Helianthus annuus Nutrition 0.000 description 2
- 240000005979 Hordeum vulgare Species 0.000 description 2
- 235000007340 Hordeum vulgare Nutrition 0.000 description 2
- 102000003820 Lipoxygenases Human genes 0.000 description 2
- 108090000128 Lipoxygenases Proteins 0.000 description 2
- 108010041559 Methionine Sulfoxide Reductases Proteins 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
- 108010002119 Protein D-Aspartate-L-Isoaspartate Methyltransferase Proteins 0.000 description 2
- 238000011529 RT qPCR Methods 0.000 description 2
- 240000006394 Sorghum bicolor Species 0.000 description 2
- 235000011684 Sorghum saccharatum Nutrition 0.000 description 2
- 229910000831 Steel Inorganic materials 0.000 description 2
- YFOCMOVJBQDBCE-NRPADANISA-N Val-Ala-Glu Chemical compound C[C@@H](C(=O)N[C@@H](CCC(=O)O)C(=O)O)NC(=O)[C@H](C(C)C)N YFOCMOVJBQDBCE-NRPADANISA-N 0.000 description 2
- 238000007792 addition Methods 0.000 description 2
- 238000012271 agricultural production Methods 0.000 description 2
- 230000009418 agronomic effect Effects 0.000 description 2
- 108010070783 alanyltyrosine Proteins 0.000 description 2
- 108010050181 aleurone Proteins 0.000 description 2
- 230000000692 anti-sense effect Effects 0.000 description 2
- 238000007796 conventional method Methods 0.000 description 2
- 238000010219 correlation analysis Methods 0.000 description 2
- 230000003247 decreasing effect Effects 0.000 description 2
- 230000018044 dehydration Effects 0.000 description 2
- 238000006297 dehydration reaction Methods 0.000 description 2
- 239000012154 double-distilled water Substances 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 235000019441 ethanol Nutrition 0.000 description 2
- PHTQWCKDNZKARW-UHFFFAOYSA-N isoamylol Chemical compound CC(C)CCO PHTQWCKDNZKARW-UHFFFAOYSA-N 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 239000012528 membrane Substances 0.000 description 2
- 238000002156 mixing Methods 0.000 description 2
- 239000013642 negative control Substances 0.000 description 2
- 229910052757 nitrogen Inorganic materials 0.000 description 2
- 230000037361 pathway Effects 0.000 description 2
- 108010051242 phenylalanylserine Proteins 0.000 description 2
- 230000010152 pollination Effects 0.000 description 2
- 239000000843 powder Substances 0.000 description 2
- 108010093296 prolyl-prolyl-alanine Proteins 0.000 description 2
- 108010004914 prolylarginine Proteins 0.000 description 2
- 238000010188 recombinant method Methods 0.000 description 2
- 238000006479 redox reaction Methods 0.000 description 2
- 230000004044 response Effects 0.000 description 2
- 230000002441 reversible effect Effects 0.000 description 2
- 238000012216 screening Methods 0.000 description 2
- 230000035939 shock Effects 0.000 description 2
- 238000009331 sowing Methods 0.000 description 2
- 239000010959 steel Substances 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- 238000013518 transcription Methods 0.000 description 2
- 230000035897 transcription Effects 0.000 description 2
- 238000013519 translation Methods 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- 102000049273 1-Cys peroxiredoxin Human genes 0.000 description 1
- 101150017339 ABI5 gene Proteins 0.000 description 1
- 102100039736 Adhesion G protein-coupled receptor L1 Human genes 0.000 description 1
- ZFXQNADNEBRERM-BJDJZHNGSA-N Ala-Ala-Pro-Pro Chemical compound C[C@H](N)C(=O)N[C@@H](C)C(=O)N1CCC[C@H]1C(=O)N1[C@H](C(O)=O)CCC1 ZFXQNADNEBRERM-BJDJZHNGSA-N 0.000 description 1
- CZPAHAKGPDUIPJ-CIUDSAMLSA-N Ala-Gln-Pro Chemical compound C[C@H](N)C(=O)N[C@@H](CCC(N)=O)C(=O)N1CCC[C@H]1C(O)=O CZPAHAKGPDUIPJ-CIUDSAMLSA-N 0.000 description 1
- HXNNRBHASOSVPG-GUBZILKMSA-N Ala-Glu-Leu Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(O)=O HXNNRBHASOSVPG-GUBZILKMSA-N 0.000 description 1
- VWEWCZSUWOEEFM-WDSKDSINSA-N Ala-Gly-Ala-Gly Chemical compound C[C@H](N)C(=O)NCC(=O)N[C@@H](C)C(=O)NCC(O)=O VWEWCZSUWOEEFM-WDSKDSINSA-N 0.000 description 1
- RZZMZYZXNJRPOJ-BJDJZHNGSA-N Ala-Ile-Lys Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCCCN)C(=O)O)NC(=O)[C@H](C)N RZZMZYZXNJRPOJ-BJDJZHNGSA-N 0.000 description 1
- VCSABYLVNWQYQE-UHFFFAOYSA-N Ala-Lys-Lys Natural products NCCCCC(NC(=O)C(N)C)C(=O)NC(CCCCN)C(O)=O VCSABYLVNWQYQE-UHFFFAOYSA-N 0.000 description 1
- IPZQNYYAYVRKKK-FXQIFTODSA-N Ala-Pro-Ala Chemical compound C[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@@H](C)C(O)=O IPZQNYYAYVRKKK-FXQIFTODSA-N 0.000 description 1
- OLVCTPPSXNRGKV-GUBZILKMSA-N Ala-Pro-Pro Chemical compound C[C@H](N)C(=O)N1CCC[C@H]1C(=O)N1[C@H](C(O)=O)CCC1 OLVCTPPSXNRGKV-GUBZILKMSA-N 0.000 description 1
- NCQMBSJGJMYKCK-ZLUOBGJFSA-N Ala-Ser-Ser Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CO)C(=O)N[C@@H](CO)C(O)=O NCQMBSJGJMYKCK-ZLUOBGJFSA-N 0.000 description 1
- KUFVXLQLDHJVOG-SHGPDSBTSA-N Ala-Thr-Thr Chemical compound C[C@H]([C@@H](C(=O)N[C@@H]([C@@H](C)O)C(=O)O)NC(=O)[C@H](C)N)O KUFVXLQLDHJVOG-SHGPDSBTSA-N 0.000 description 1
- RIPMDCIXRYWXSH-KNXALSJPSA-N Ala-Trp-Pro Chemical compound C[C@@H](C(=O)N[C@@H](CC1=CNC2=CC=CC=C21)C(=O)N3CCC[C@@H]3C(=O)O)N RIPMDCIXRYWXSH-KNXALSJPSA-N 0.000 description 1
- 102000005602 Aldo-Keto Reductases Human genes 0.000 description 1
- 108010084469 Aldo-Keto Reductases Proteins 0.000 description 1
- 101100438273 Arabidopsis thaliana CAN1 gene Proteins 0.000 description 1
- 101000942941 Arabidopsis thaliana DNA ligase 6 Proteins 0.000 description 1
- 101100321442 Arabidopsis thaliana ZHD1 gene Proteins 0.000 description 1
- 101100321447 Arabidopsis thaliana ZHD4 gene Proteins 0.000 description 1
- MCYJBCKCAPERSE-FXQIFTODSA-N Arg-Ala-Asp Chemical compound OC(=O)C[C@@H](C(O)=O)NC(=O)[C@H](C)NC(=O)[C@@H](N)CCCN=C(N)N MCYJBCKCAPERSE-FXQIFTODSA-N 0.000 description 1
- OVVUNXXROOFSIM-SDDRHHMPSA-N Arg-Arg-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CCCN=C(N)N)NC(=O)[C@H](CCCN=C(N)N)N)C(=O)O OVVUNXXROOFSIM-SDDRHHMPSA-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
- SQKPKIJVWHAWNF-DCAQKATOSA-N Arg-Asp-Lys Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CCCCN)C(O)=O SQKPKIJVWHAWNF-DCAQKATOSA-N 0.000 description 1
- BEXGZLUHRXTZCC-CIUDSAMLSA-N Arg-Gln-Ser Chemical compound C(C[C@@H](C(=O)N[C@@H](CCC(=O)N)C(=O)N[C@@H](CO)C(=O)O)N)CN=C(N)N BEXGZLUHRXTZCC-CIUDSAMLSA-N 0.000 description 1
- HPKSHFSEXICTLI-CIUDSAMLSA-N Arg-Glu-Ala Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](C)C(O)=O HPKSHFSEXICTLI-CIUDSAMLSA-N 0.000 description 1
- ZATRYQNPUHGXCU-DTWKUNHWSA-N Arg-Gly-Pro Chemical compound C1C[C@@H](N(C1)C(=O)CNC(=O)[C@H](CCCN=C(N)N)N)C(=O)O ZATRYQNPUHGXCU-DTWKUNHWSA-N 0.000 description 1
- WVNFNPGXYADPPO-BQBZGAKWSA-N Arg-Gly-Ser Chemical compound NC(N)=NCCC[C@H](N)C(=O)NCC(=O)N[C@@H](CO)C(O)=O WVNFNPGXYADPPO-BQBZGAKWSA-N 0.000 description 1
- NKNILFJYKKHBKE-WPRPVWTQSA-N Arg-Gly-Val Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)NCC(=O)N[C@@H](C(C)C)C(O)=O NKNILFJYKKHBKE-WPRPVWTQSA-N 0.000 description 1
- PCQXGEUALSFGIA-WDSOQIARSA-N Arg-His-Trp Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](CC1=CNC2=C1C=CC=C2)C(O)=O PCQXGEUALSFGIA-WDSOQIARSA-N 0.000 description 1
- VVJTWSRNMJNDPN-IUCAKERBSA-N Arg-Met-Gly Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCSC)C(=O)NCC(O)=O VVJTWSRNMJNDPN-IUCAKERBSA-N 0.000 description 1
- WKPXXXUSUHAXDE-SRVKXCTJSA-N Arg-Pro-Arg Chemical compound NC(N)=NCCC[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CCCN=C(N)N)C(O)=O WKPXXXUSUHAXDE-SRVKXCTJSA-N 0.000 description 1
- VENMDXUVHSKEIN-GUBZILKMSA-N Arg-Ser-Arg Chemical compound NC(N)=NCCC[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@@H](CCCN=C(N)N)C(O)=O VENMDXUVHSKEIN-GUBZILKMSA-N 0.000 description 1
- BFDDUDQCPJWQRQ-IHRRRGAJSA-N Arg-Tyr-Cys Chemical compound C1=CC(=CC=C1C[C@@H](C(=O)N[C@@H](CS)C(=O)O)NC(=O)[C@H](CCCN=C(N)N)N)O BFDDUDQCPJWQRQ-IHRRRGAJSA-N 0.000 description 1
- DJIMLSXHXKWADV-CIUDSAMLSA-N Asn-Leu-Ser Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@@H](N)CC(N)=O DJIMLSXHXKWADV-CIUDSAMLSA-N 0.000 description 1
- KYQJHBWHRASMKG-ZLUOBGJFSA-N Asn-Ser-Cys Chemical compound NC(=O)C[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@@H](CS)C(O)=O KYQJHBWHRASMKG-ZLUOBGJFSA-N 0.000 description 1
- WLVLIYYBPPONRJ-GCJQMDKQSA-N Asn-Thr-Ala 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(O)=O WLVLIYYBPPONRJ-GCJQMDKQSA-N 0.000 description 1
- NAPNAGZWHQHZLG-ZLUOBGJFSA-N Asp-Asp-Ala Chemical compound C[C@@H](C(=O)O)NC(=O)[C@H](CC(=O)O)NC(=O)[C@H](CC(=O)O)N NAPNAGZWHQHZLG-ZLUOBGJFSA-N 0.000 description 1
- VHWNKSJHQFZJTH-FXQIFTODSA-N Asp-Asp-Met Chemical compound CSCC[C@@H](C(=O)O)NC(=O)[C@H](CC(=O)O)NC(=O)[C@H](CC(=O)O)N VHWNKSJHQFZJTH-FXQIFTODSA-N 0.000 description 1
- XAJRHVUUVUPFQL-ACZMJKKPSA-N Asp-Glu-Asp Chemical compound OC(=O)C[C@H](N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(O)=O)C(O)=O XAJRHVUUVUPFQL-ACZMJKKPSA-N 0.000 description 1
- CLUMZOKVGUWUFD-CIUDSAMLSA-N Asp-Leu-Asn Chemical compound OC(=O)C[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(N)=O)C(O)=O CLUMZOKVGUWUFD-CIUDSAMLSA-N 0.000 description 1
- OEDJQRXNDRUGEU-SRVKXCTJSA-N Asp-Leu-His Chemical compound N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC1=CNC=N1)C(=O)O OEDJQRXNDRUGEU-SRVKXCTJSA-N 0.000 description 1
- CUQDCPXNZPDYFQ-ZLUOBGJFSA-N Asp-Ser-Asp Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(O)=O)C(O)=O CUQDCPXNZPDYFQ-ZLUOBGJFSA-N 0.000 description 1
- BRRPVTUFESPTCP-ACZMJKKPSA-N Asp-Ser-Glu Chemical compound OC(=O)C[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@H](C(O)=O)CCC(O)=O BRRPVTUFESPTCP-ACZMJKKPSA-N 0.000 description 1
- KGHLGJAXYSVNJP-WHFBIAKZSA-N Asp-Ser-Gly Chemical compound OC(=O)C[C@H](N)C(=O)N[C@@H](CO)C(=O)NCC(O)=O KGHLGJAXYSVNJP-WHFBIAKZSA-N 0.000 description 1
- MNQMTYSEKZHIDF-GCJQMDKQSA-N Asp-Thr-Ala Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C)C(O)=O MNQMTYSEKZHIDF-GCJQMDKQSA-N 0.000 description 1
- KNDCWFXCFKSEBM-AVGNSLFASA-N Asp-Tyr-Glu 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](CCC(O)=O)C(O)=O KNDCWFXCFKSEBM-AVGNSLFASA-N 0.000 description 1
- 241000743774 Brachypodium Species 0.000 description 1
- 241000743776 Brachypodium distachyon Species 0.000 description 1
- 108091033409 CRISPR Proteins 0.000 description 1
- 101100342815 Caenorhabditis elegans lec-1 gene Proteins 0.000 description 1
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 1
- 108091062157 Cis-regulatory element Proteins 0.000 description 1
- 108091026890 Coding region Proteins 0.000 description 1
- OIMUAKUQOUEPCZ-WHFBIAKZSA-N Cys-Asn-Gly Chemical compound SC[C@H](N)C(=O)N[C@@H](CC(N)=O)C(=O)NCC(O)=O OIMUAKUQOUEPCZ-WHFBIAKZSA-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
- 108020001738 DNA Glycosylase Proteins 0.000 description 1
- 230000005778 DNA damage Effects 0.000 description 1
- 231100000277 DNA damage Toxicity 0.000 description 1
- 102000028381 DNA glycosylase Human genes 0.000 description 1
- 230000033616 DNA repair Effects 0.000 description 1
- 102100035619 DNA-(apurinic or apyrimidinic site) lyase Human genes 0.000 description 1
- 102000016928 DNA-directed DNA polymerase Human genes 0.000 description 1
- 108010014303 DNA-directed DNA polymerase Proteins 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 101000962067 Gallus gallus Neuroblastoma suppressor of tumorigenicity 1 Proteins 0.000 description 1
- 229930191978 Gibberellin Natural products 0.000 description 1
- POPZASPRNPGIPZ-UHFFFAOYSA-N Gln Gln Ala Pro Chemical compound NC(=O)CCC(N)C(=O)NC(CCC(N)=O)C(=O)NC(C)C(=O)N1CCCC1C(O)=O POPZASPRNPGIPZ-UHFFFAOYSA-N 0.000 description 1
- KVXVVDFOZNYYKZ-DCAQKATOSA-N Gln-Gln-Leu Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(C)C)C(O)=O KVXVVDFOZNYYKZ-DCAQKATOSA-N 0.000 description 1
- PODFFOWWLUPNMN-DCAQKATOSA-N Gln-His-Gln Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](CCC(N)=O)C(O)=O PODFFOWWLUPNMN-DCAQKATOSA-N 0.000 description 1
- XZUUUKNKNWVPHQ-JYJNAYRXSA-N Gln-Phe-Leu Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CC(C)C)C(O)=O XZUUUKNKNWVPHQ-JYJNAYRXSA-N 0.000 description 1
- MXOODARRORARSU-ACZMJKKPSA-N Glu-Ala-Ser Chemical compound C[C@@H](C(=O)N[C@@H](CO)C(=O)O)NC(=O)[C@H](CCC(=O)O)N MXOODARRORARSU-ACZMJKKPSA-N 0.000 description 1
- RDPOETHPAQEGDP-ACZMJKKPSA-N Glu-Asp-Ala Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](C)C(O)=O RDPOETHPAQEGDP-ACZMJKKPSA-N 0.000 description 1
- SJPMNHCEWPTRBR-BQBZGAKWSA-N Glu-Glu-Gly Chemical compound OC(=O)CC[C@H](N)C(=O)N[C@@H](CCC(O)=O)C(=O)NCC(O)=O SJPMNHCEWPTRBR-BQBZGAKWSA-N 0.000 description 1
- RAUDKMVXNOWDLS-WDSKDSINSA-N Glu-Gly-Ser Chemical compound OC(=O)CC[C@H](N)C(=O)NCC(=O)N[C@@H](CO)C(O)=O RAUDKMVXNOWDLS-WDSKDSINSA-N 0.000 description 1
- IVGJYOOGJLFKQE-AVGNSLFASA-N Glu-Leu-Lys Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CCCCN)C(=O)O)NC(=O)[C@H](CCC(=O)O)N IVGJYOOGJLFKQE-AVGNSLFASA-N 0.000 description 1
- WNRZUESNGGDCJX-JYJNAYRXSA-N Glu-Leu-Phe Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O WNRZUESNGGDCJX-JYJNAYRXSA-N 0.000 description 1
- ZAPFAWQHBOHWLL-GUBZILKMSA-N Glu-Ser-His Chemical compound C1=C(NC=N1)C[C@@H](C(=O)O)NC(=O)[C@H](CO)NC(=O)[C@H](CCC(=O)O)N ZAPFAWQHBOHWLL-GUBZILKMSA-N 0.000 description 1
- QXPRJQPCFXMCIY-NKWVEPMBSA-N Gly-Ala-Pro Chemical compound C[C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)CN QXPRJQPCFXMCIY-NKWVEPMBSA-N 0.000 description 1
- KKBWDNZXYLGJEY-UHFFFAOYSA-N Gly-Arg-Pro Natural products NCC(=O)NC(CCNC(=N)N)C(=O)N1CCCC1C(=O)O KKBWDNZXYLGJEY-UHFFFAOYSA-N 0.000 description 1
- XZRZILPOZBVTDB-GJZGRUSLSA-N Gly-Arg-Trp Chemical compound C1=CC=C2C(C[C@H](NC(=O)[C@H](CCCNC(N)=N)NC(=O)CN)C(O)=O)=CNC2=C1 XZRZILPOZBVTDB-GJZGRUSLSA-N 0.000 description 1
- YYPFZVIXAVDHIK-IUCAKERBSA-N Gly-Glu-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](CCC(O)=O)NC(=O)CN YYPFZVIXAVDHIK-IUCAKERBSA-N 0.000 description 1
- KAJAOGBVWCYGHZ-JTQLQIEISA-N Gly-Gly-Phe Chemical compound [NH3+]CC(=O)NCC(=O)N[C@H](C([O-])=O)CC1=CC=CC=C1 KAJAOGBVWCYGHZ-JTQLQIEISA-N 0.000 description 1
- TWTPDFFBLQEBOE-IUCAKERBSA-N Gly-Leu-Gln Chemical compound [H]NCC(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(N)=O)C(O)=O TWTPDFFBLQEBOE-IUCAKERBSA-N 0.000 description 1
- 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
- VEPBEGNDJYANCF-QWRGUYRKSA-N Gly-Lys-Lys Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)[C@@H](NC(=O)CN)CCCCN VEPBEGNDJYANCF-QWRGUYRKSA-N 0.000 description 1
- FEUPVVCGQLNXNP-IRXDYDNUSA-N Gly-Phe-Phe Chemical compound C([C@H](NC(=O)CN)C(=O)N[C@@H](CC=1C=CC=CC=1)C(O)=O)C1=CC=CC=C1 FEUPVVCGQLNXNP-IRXDYDNUSA-N 0.000 description 1
- GGAPHLIUUTVYMX-QWRGUYRKSA-N Gly-Phe-Ser Chemical compound OC[C@@H](C([O-])=O)NC(=O)[C@@H](NC(=O)C[NH3+])CC1=CC=CC=C1 GGAPHLIUUTVYMX-QWRGUYRKSA-N 0.000 description 1
- IXHQLZIWBCQBLQ-STQMWFEESA-N Gly-Pro-Phe Chemical compound NCC(=O)N1CCC[C@H]1C(=O)N[C@H](C(O)=O)CC1=CC=CC=C1 IXHQLZIWBCQBLQ-STQMWFEESA-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
- 108010068250 Herpes Simplex Virus Protein Vmw65 Proteins 0.000 description 1
- BIAKMWKJMQLZOJ-ZKWXMUAHSA-N His-Ala-Ala Chemical compound C[C@H](NC(=O)[C@H](C)NC(=O)[C@@H](N)Cc1cnc[nH]1)C(O)=O BIAKMWKJMQLZOJ-ZKWXMUAHSA-N 0.000 description 1
- IDNNYVGVSZMQTK-IHRRRGAJSA-N His-Arg-His Chemical compound C1=C(NC=N1)C[C@@H](C(=O)N[C@@H](CCCN=C(N)N)C(=O)N[C@@H](CC2=CN=CN2)C(=O)O)N IDNNYVGVSZMQTK-IHRRRGAJSA-N 0.000 description 1
- ZYDYEPDFFVCUBI-SRVKXCTJSA-N His-Glu-Met Chemical compound CSCC[C@@H](C(=O)O)NC(=O)[C@H](CCC(=O)O)NC(=O)[C@H](CC1=CN=CN1)N ZYDYEPDFFVCUBI-SRVKXCTJSA-N 0.000 description 1
- SOYCWSKCUVDLMC-AVGNSLFASA-N His-Pro-Arg Chemical compound N[C@@H](Cc1cnc[nH]1)C(=O)N2CCC[C@H]2C(=O)N[C@@H](CCCNC(=N)N)C(=O)O SOYCWSKCUVDLMC-AVGNSLFASA-N 0.000 description 1
- XIGFLVCAVQQGNS-IHRRRGAJSA-N His-Pro-His Chemical compound C([C@H](N)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CC=1NC=NC=1)C(O)=O)C1=CN=CN1 XIGFLVCAVQQGNS-IHRRRGAJSA-N 0.000 description 1
- LNDVNHOSZQPJGI-AVGNSLFASA-N His-Pro-Pro Chemical compound C([C@H](N)C(=O)N1[C@@H](CCC1)C(=O)N1[C@@H](CCC1)C(O)=O)C1=CN=CN1 LNDVNHOSZQPJGI-AVGNSLFASA-N 0.000 description 1
- 101000959588 Homo sapiens Adhesion G protein-coupled receptor L1 Proteins 0.000 description 1
- 101001137256 Homo sapiens DNA-(apurinic or apyrimidinic site) lyase Proteins 0.000 description 1
- NULSANWBUWLTKN-NAKRPEOUSA-N Ile-Arg-Ser Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCCN=C(N)N)C(=O)N[C@@H](CO)C(=O)O)N NULSANWBUWLTKN-NAKRPEOUSA-N 0.000 description 1
- 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
- RQZFWBLDTBDEOF-RNJOBUHISA-N Ile-Val-Pro Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](C(C)C)C(=O)N1CCC[C@@H]1C(=O)O)N RQZFWBLDTBDEOF-RNJOBUHISA-N 0.000 description 1
- 108091092195 Intron Proteins 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
- ROHFNLRQFUQHCH-YFKPBYRVSA-N L-leucine Chemical compound CC(C)C[C@H](N)C(O)=O ROHFNLRQFUQHCH-YFKPBYRVSA-N 0.000 description 1
- FFEARJCKVFRZRR-BYPYZUCNSA-N L-methionine Chemical compound CSCC[C@H](N)C(O)=O FFEARJCKVFRZRR-BYPYZUCNSA-N 0.000 description 1
- QEFRNWWLZKMPFJ-ZXPFJRLXSA-N L-methionine (R)-S-oxide Chemical compound C[S@@](=O)CC[C@H]([NH3+])C([O-])=O QEFRNWWLZKMPFJ-ZXPFJRLXSA-N 0.000 description 1
- QEFRNWWLZKMPFJ-UHFFFAOYSA-N L-methionine sulphoxide Natural products CS(=O)CCC(N)C(O)=O QEFRNWWLZKMPFJ-UHFFFAOYSA-N 0.000 description 1
- WUFYAPWIHCUMLL-CIUDSAMLSA-N Leu-Asn-Ala Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](C)C(O)=O WUFYAPWIHCUMLL-CIUDSAMLSA-N 0.000 description 1
- KAFOIVJDVSZUMD-DCAQKATOSA-N Leu-Gln-Gln Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCC(N)=O)C(O)=O KAFOIVJDVSZUMD-DCAQKATOSA-N 0.000 description 1
- KAFOIVJDVSZUMD-UHFFFAOYSA-N Leu-Gln-Gln Natural products CC(C)CC(N)C(=O)NC(CCC(N)=O)C(=O)NC(CCC(N)=O)C(O)=O KAFOIVJDVSZUMD-UHFFFAOYSA-N 0.000 description 1
- LAGPXKYZCCTSGQ-JYJNAYRXSA-N Leu-Glu-Phe Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O LAGPXKYZCCTSGQ-JYJNAYRXSA-N 0.000 description 1
- AOFYPTOHESIBFZ-KKUMJFAQSA-N Leu-His-His Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](Cc1cnc[nH]1)C(=O)N[C@@H](Cc1cnc[nH]1)C(O)=O AOFYPTOHESIBFZ-KKUMJFAQSA-N 0.000 description 1
- UCBPDSYUVAAHCD-UWVGGRQHSA-N Leu-Pro-Gly Chemical compound CC(C)C[C@H](N)C(=O)N1CCC[C@H]1C(=O)NCC(O)=O UCBPDSYUVAAHCD-UWVGGRQHSA-N 0.000 description 1
- MVHXGBZUJLWZOH-BJDJZHNGSA-N Leu-Ser-Ile Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O MVHXGBZUJLWZOH-BJDJZHNGSA-N 0.000 description 1
- SBANPBVRHYIMRR-GARJFASQSA-N Leu-Ser-Pro Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CO)C(=O)N1CCC[C@@H]1C(=O)O)N SBANPBVRHYIMRR-GARJFASQSA-N 0.000 description 1
- SBANPBVRHYIMRR-UHFFFAOYSA-N Leu-Ser-Pro Natural products CC(C)CC(N)C(=O)NC(CO)C(=O)N1CCCC1C(O)=O SBANPBVRHYIMRR-UHFFFAOYSA-N 0.000 description 1
- ROHFNLRQFUQHCH-UHFFFAOYSA-N Leucine Natural products CC(C)CC(N)C(O)=O ROHFNLRQFUQHCH-UHFFFAOYSA-N 0.000 description 1
- 108060001084 Luciferase Proteins 0.000 description 1
- 239000005089 Luciferase Substances 0.000 description 1
- RLZDUFRBMQNYIJ-YUMQZZPRSA-N Lys-Cys-Gly Chemical compound C(CCN)C[C@@H](C(=O)N[C@@H](CS)C(=O)NCC(=O)O)N RLZDUFRBMQNYIJ-YUMQZZPRSA-N 0.000 description 1
- CKSBRMUOQDNPKZ-SRVKXCTJSA-N Lys-Gln-Met Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCSC)C(O)=O CKSBRMUOQDNPKZ-SRVKXCTJSA-N 0.000 description 1
- PBIPLDMFHAICIP-DCAQKATOSA-N Lys-Glu-Glu Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(O)=O PBIPLDMFHAICIP-DCAQKATOSA-N 0.000 description 1
- WQDKIVRHTQYJSN-DCAQKATOSA-N Lys-Ser-Arg Chemical compound C(CCN)C[C@@H](C(=O)N[C@@H](CO)C(=O)N[C@@H](CCCN=C(N)N)C(=O)O)N WQDKIVRHTQYJSN-DCAQKATOSA-N 0.000 description 1
- WSMYVTOQOOLQHP-UHFFFAOYSA-N Malondialdehyde Chemical compound O=CCC=O WSMYVTOQOOLQHP-UHFFFAOYSA-N 0.000 description 1
- 102100025169 Max-binding protein MNT Human genes 0.000 description 1
- 240000004658 Medicago sativa Species 0.000 description 1
- 235000017587 Medicago sativa ssp. sativa Nutrition 0.000 description 1
- VOOINLQYUZOREH-SRVKXCTJSA-N Met-Gln-Leu Chemical compound CC(C)C[C@@H](C(=O)O)NC(=O)[C@H](CCC(=O)N)NC(=O)[C@H](CCSC)N VOOINLQYUZOREH-SRVKXCTJSA-N 0.000 description 1
- 102000000532 Methionine Sulfoxide Reductases Human genes 0.000 description 1
- 102000016397 Methyltransferase Human genes 0.000 description 1
- 108060004795 Methyltransferase Proteins 0.000 description 1
- YBAFDPFAUTYYRW-UHFFFAOYSA-N N-L-alpha-glutamyl-L-leucine Natural products CC(C)CC(C(O)=O)NC(=O)C(N)CCC(O)=O YBAFDPFAUTYYRW-UHFFFAOYSA-N 0.000 description 1
- KZNQNBZMBZJQJO-UHFFFAOYSA-N N-glycyl-L-proline Natural products NCC(=O)N1CCCC1C(O)=O KZNQNBZMBZJQJO-UHFFFAOYSA-N 0.000 description 1
- 108010047562 NGR peptide Proteins 0.000 description 1
- 101710185569 Peroxiredoxin-6 Proteins 0.000 description 1
- CSDMCMITJLKBAH-SOUVJXGZSA-N Phe-Glu-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CCC(=O)O)NC(=O)[C@H](CC2=CC=CC=C2)N)C(=O)O CSDMCMITJLKBAH-SOUVJXGZSA-N 0.000 description 1
- GRVMHFCZUIYNKQ-UFYCRDLUSA-N Phe-Phe-Val Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](C(C)C)C(O)=O GRVMHFCZUIYNKQ-UFYCRDLUSA-N 0.000 description 1
- JHSRGEODDALISP-XVSYOHENSA-N Phe-Thr-Asn Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(N)=O)C(O)=O JHSRGEODDALISP-XVSYOHENSA-N 0.000 description 1
- FXEKNHAJIMHRFJ-ULQDDVLXSA-N Phe-Val-His Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC1=CN=CN1)C(=O)O)NC(=O)[C@H](CC2=CC=CC=C2)N FXEKNHAJIMHRFJ-ULQDDVLXSA-N 0.000 description 1
- 108700001094 Plant Genes Proteins 0.000 description 1
- 244000061632 Prinsepia uniflora Species 0.000 description 1
- 235000008997 Prinsepia uniflora Nutrition 0.000 description 1
- FYQSMXKJYTZYRP-DCAQKATOSA-N Pro-Ala-Lys Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)[C@H](C)NC(=O)[C@@H]1CCCN1 FYQSMXKJYTZYRP-DCAQKATOSA-N 0.000 description 1
- OOLOTUZJUBOMAX-GUBZILKMSA-N Pro-Ala-Val Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](C)C(=O)N[C@@H](C(C)C)C(O)=O OOLOTUZJUBOMAX-GUBZILKMSA-N 0.000 description 1
- YTWNSIDWAFSEEI-RWMBFGLXSA-N Pro-His-Pro Chemical compound C1C[C@H](NC1)C(=O)N[C@@H](CC2=CN=CN2)C(=O)N3CCC[C@@H]3C(=O)O YTWNSIDWAFSEEI-RWMBFGLXSA-N 0.000 description 1
- KDBHVPXBQADZKY-GUBZILKMSA-N Pro-Pro-Ala Chemical compound OC(=O)[C@H](C)NC(=O)[C@@H]1CCCN1C(=O)[C@H]1NCCC1 KDBHVPXBQADZKY-GUBZILKMSA-N 0.000 description 1
- POQFNPILEQEODH-FXQIFTODSA-N Pro-Ser-Ala Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CO)C(=O)N[C@@H](C)C(O)=O POQFNPILEQEODH-FXQIFTODSA-N 0.000 description 1
- FNGOXVQBBCMFKV-CIUDSAMLSA-N Pro-Ser-Glu Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CO)C(=O)N[C@@H](CCC(O)=O)C(O)=O FNGOXVQBBCMFKV-CIUDSAMLSA-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
- AIOWVDNPESPXRB-YTWAJWBKSA-N Pro-Thr-Pro Chemical compound C[C@H]([C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@@H]2CCCN2)O AIOWVDNPESPXRB-YTWAJWBKSA-N 0.000 description 1
- IMNVAOPEMFDAQD-NHCYSSNCSA-N Pro-Val-Glu Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCC(O)=O)C(O)=O IMNVAOPEMFDAQD-NHCYSSNCSA-N 0.000 description 1
- 102000000538 Protein D-Aspartate-L-Isoaspartate Methyltransferase Human genes 0.000 description 1
- 108010026552 Proteome Proteins 0.000 description 1
- 238000003559 RNA-seq method Methods 0.000 description 1
- BTKUIVBNGBFTTP-WHFBIAKZSA-N Ser-Ala-Gly Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](C)C(=O)NCC(O)=O BTKUIVBNGBFTTP-WHFBIAKZSA-N 0.000 description 1
- YQHZVYJAGWMHES-ZLUOBGJFSA-N Ser-Ala-Ser Chemical compound OC[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@@H](CO)C(O)=O YQHZVYJAGWMHES-ZLUOBGJFSA-N 0.000 description 1
- WXUBSIDKNMFAGS-IHRRRGAJSA-N Ser-Arg-Tyr Chemical compound NC(N)=NCCC[C@H](NC(=O)[C@H](CO)N)C(=O)N[C@H](C(O)=O)CC1=CC=C(O)C=C1 WXUBSIDKNMFAGS-IHRRRGAJSA-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
- SMIDBHKWSYUBRZ-ACZMJKKPSA-N Ser-Glu-Ala Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](C)C(O)=O SMIDBHKWSYUBRZ-ACZMJKKPSA-N 0.000 description 1
- IOVBCLGAJJXOHK-SRVKXCTJSA-N Ser-His-His Chemical compound C([C@H](NC(=O)[C@H](CO)N)C(=O)N[C@@H](CC=1NC=NC=1)C(O)=O)C1=CN=CN1 IOVBCLGAJJXOHK-SRVKXCTJSA-N 0.000 description 1
- RIAKPZVSNBBNRE-BJDJZHNGSA-N Ser-Ile-Leu Chemical compound OC[C@H](N)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC(C)C)C(O)=O RIAKPZVSNBBNRE-BJDJZHNGSA-N 0.000 description 1
- WNDUPCKKKGSKIQ-CIUDSAMLSA-N Ser-Pro-Gln Chemical compound OC[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CCC(N)=O)C(O)=O WNDUPCKKKGSKIQ-CIUDSAMLSA-N 0.000 description 1
- FZXOPYUEQGDGMS-ACZMJKKPSA-N Ser-Ser-Gln Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CO)C(=O)N[C@@H](CCC(N)=O)C(O)=O FZXOPYUEQGDGMS-ACZMJKKPSA-N 0.000 description 1
- BMKNXTJLHFIAAH-CIUDSAMLSA-N Ser-Ser-Leu Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(O)=O BMKNXTJLHFIAAH-CIUDSAMLSA-N 0.000 description 1
- XQJCEKXQUJQNNK-ZLUOBGJFSA-N Ser-Ser-Ser Chemical compound OC[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@@H](CO)C(O)=O XQJCEKXQUJQNNK-ZLUOBGJFSA-N 0.000 description 1
- VGQVAVQWKJLIRM-FXQIFTODSA-N Ser-Ser-Val Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CO)C(=O)N[C@@H](C(C)C)C(O)=O VGQVAVQWKJLIRM-FXQIFTODSA-N 0.000 description 1
- 240000005498 Setaria italica Species 0.000 description 1
- 235000007226 Setaria italica Nutrition 0.000 description 1
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 1
- 244000062793 Sorghum vulgare Species 0.000 description 1
- 241000592344 Spermatophyta Species 0.000 description 1
- ZTPXSEUVYNNZRB-CDMKHQONSA-N Thr-Gly-Phe Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)NCC(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O ZTPXSEUVYNNZRB-CDMKHQONSA-N 0.000 description 1
- XIULAFZYEKSGAJ-IXOXFDKPSA-N Thr-Leu-His Chemical compound C[C@@H](O)[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@H](C(O)=O)CC1=CNC=N1 XIULAFZYEKSGAJ-IXOXFDKPSA-N 0.000 description 1
- UUSQVWOVUYMLJA-PPCPHDFISA-N Thr-Lys-Ile Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O UUSQVWOVUYMLJA-PPCPHDFISA-N 0.000 description 1
- KERCOYANYUPLHJ-XGEHTFHBSA-N Thr-Pro-Ser Chemical compound C[C@@H](O)[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CO)C(O)=O KERCOYANYUPLHJ-XGEHTFHBSA-N 0.000 description 1
- NDZYTIMDOZMECO-SHGPDSBTSA-N Thr-Thr-Ala Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C)C(O)=O NDZYTIMDOZMECO-SHGPDSBTSA-N 0.000 description 1
- ABCLYRRGTZNIFU-BWAGICSOSA-N Thr-Tyr-His Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CC1=CC=C(C=C1)O)C(=O)N[C@@H](CC2=CN=CN2)C(=O)O)N)O ABCLYRRGTZNIFU-BWAGICSOSA-N 0.000 description 1
- 235000021307 Triticum Nutrition 0.000 description 1
- HJWVPKJHHLZCNH-DVXDUOKCSA-N Trp-Ala-Trp Chemical compound C1=CC=C2C(C[C@H](NC(=O)[C@@H](NC(=O)[C@@H](N)CC=3C4=CC=CC=C4NC=3)C)C(O)=O)=CNC2=C1 HJWVPKJHHLZCNH-DVXDUOKCSA-N 0.000 description 1
- NXJZCPKZIKTYLX-XEGUGMAKSA-N Trp-Glu-Ala Chemical compound C[C@@H](C(=O)O)NC(=O)[C@H](CCC(=O)O)NC(=O)[C@H](CC1=CNC2=CC=CC=C21)N NXJZCPKZIKTYLX-XEGUGMAKSA-N 0.000 description 1
- AKXBNSZMYAOGLS-STQMWFEESA-N Tyr-Arg-Gly Chemical compound NC(N)=NCCC[C@@H](C(=O)NCC(O)=O)NC(=O)[C@@H](N)CC1=CC=C(O)C=C1 AKXBNSZMYAOGLS-STQMWFEESA-N 0.000 description 1
- JFDGVHXRCKEBAU-KKUMJFAQSA-N Tyr-Asp-Lys Chemical compound C1=CC(=CC=C1C[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)N[C@@H](CCCCN)C(=O)O)N)O JFDGVHXRCKEBAU-KKUMJFAQSA-N 0.000 description 1
- YMUQBRQQCPQEQN-CXTHYWKRSA-N Tyr-Ile-Tyr Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC1=CC=C(C=C1)O)C(=O)O)NC(=O)[C@H](CC2=CC=C(C=C2)O)N YMUQBRQQCPQEQN-CXTHYWKRSA-N 0.000 description 1
- FZSPNKUFROZBSG-ZKWXMUAHSA-N Val-Ala-Asp Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@H](C(O)=O)CC(O)=O FZSPNKUFROZBSG-ZKWXMUAHSA-N 0.000 description 1
- TZVUSFMQWPWHON-NHCYSSNCSA-N Val-Asp-Leu Chemical compound CC(C)C[C@@H](C(=O)O)NC(=O)[C@H](CC(=O)O)NC(=O)[C@H](C(C)C)N TZVUSFMQWPWHON-NHCYSSNCSA-N 0.000 description 1
- XGJLNBNZNMVJRS-NRPADANISA-N Val-Glu-Ala Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](C)C(O)=O XGJLNBNZNMVJRS-NRPADANISA-N 0.000 description 1
- OTJMMKPMLUNTQT-AVGNSLFASA-N Val-Leu-Arg Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CCCN=C(N)N)C(=O)O)NC(=O)[C@H](C(C)C)N OTJMMKPMLUNTQT-AVGNSLFASA-N 0.000 description 1
- SSYBNWFXCFNRFN-GUBZILKMSA-N Val-Pro-Ser Chemical compound CC(C)[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CO)C(O)=O SSYBNWFXCFNRFN-GUBZILKMSA-N 0.000 description 1
- MNSSBIHFEUUXNW-RCWTZXSCSA-N Val-Thr-Arg Chemical compound CC(C)[C@H](N)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@H](C(O)=O)CCCN=C(N)N MNSSBIHFEUUXNW-RCWTZXSCSA-N 0.000 description 1
- PQSNETRGCRUOGP-KKHAAJSZSA-N Val-Thr-Asn Chemical compound CC(C)[C@H](N)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@H](C(O)=O)CC(N)=O PQSNETRGCRUOGP-KKHAAJSZSA-N 0.000 description 1
- PDDJTOSAVNRJRH-UNQGMJICSA-N Val-Thr-Phe Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)O)NC(=O)[C@H](C(C)C)N)O PDDJTOSAVNRJRH-UNQGMJICSA-N 0.000 description 1
- JXWGBRRVTRAZQA-ULQDDVLXSA-N Val-Tyr-Leu Chemical compound CC(C)C[C@@H](C(=O)O)NC(=O)[C@H](CC1=CC=C(C=C1)O)NC(=O)[C@H](C(C)C)N JXWGBRRVTRAZQA-ULQDDVLXSA-N 0.000 description 1
- 241000607479 Yersinia pestis Species 0.000 description 1
- 235000007244 Zea mays Nutrition 0.000 description 1
- HLVPIMVSSMJFPS-UHFFFAOYSA-N abscisic acid beta-D-glucopyranosyl ester Natural products O1C(CO)C(O)C(O)C(O)C1OC(=O)C=C(C)C=CC1(O)C(C)=CC(=O)CC1(C)C HLVPIMVSSMJFPS-UHFFFAOYSA-N 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- 230000006978 adaptation Effects 0.000 description 1
- 108010076324 alanyl-glycyl-glycine Proteins 0.000 description 1
- 108010086434 alanyl-seryl-glycine Proteins 0.000 description 1
- 108010005233 alanylglutamic acid Proteins 0.000 description 1
- 108010070944 alanylhistidine Proteins 0.000 description 1
- 108010087924 alanylproline Proteins 0.000 description 1
- 125000000539 amino acid group Chemical group 0.000 description 1
- 238000000137 annealing Methods 0.000 description 1
- 230000006907 apoptotic process Effects 0.000 description 1
- 108010013835 arginine glutamate Proteins 0.000 description 1
- 108010008355 arginyl-glutamine Proteins 0.000 description 1
- 108010091092 arginyl-glycyl-proline Proteins 0.000 description 1
- 108010069926 arginyl-glycyl-serine Proteins 0.000 description 1
- 108010038633 aspartylglutamate Proteins 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 230000037429 base substitution Effects 0.000 description 1
- 238000003287 bathing Methods 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 102000023732 binding proteins Human genes 0.000 description 1
- 108091008324 binding proteins Proteins 0.000 description 1
- 210000004899 c-terminal region Anatomy 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- 150000001728 carbonyl compounds Chemical class 0.000 description 1
- 210000004027 cell Anatomy 0.000 description 1
- 230000010307 cell transformation Effects 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 125000000151 cysteine group Chemical group N[C@@H](CS)C(=O)* 0.000 description 1
- 108010060199 cysteinylproline Proteins 0.000 description 1
- GVJHHUAWPYXKBD-UHFFFAOYSA-N d-alpha-tocopherol Natural products OC1=C(C)C(C)=C2OC(CCCC(C)CCCC(C)CCCC(C)C)(C)CCC2=C1C GVJHHUAWPYXKBD-UHFFFAOYSA-N 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- FCRACOPGPMPSHN-UHFFFAOYSA-N desoxyabscisic acid Natural products OC(=O)C=C(C)C=CC1C(C)=CC(=O)CC1(C)C FCRACOPGPMPSHN-UHFFFAOYSA-N 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 230000006866 deterioration Effects 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- FSXRLASFHBWESK-UHFFFAOYSA-N dipeptide phenylalanyl-tyrosine Natural products C=1C=C(O)C=CC=1CC(C(O)=O)NC(=O)C(N)CC1=CC=CC=C1 FSXRLASFHBWESK-UHFFFAOYSA-N 0.000 description 1
- 230000013020 embryo development Effects 0.000 description 1
- 230000002255 enzymatic effect Effects 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 238000010195 expression analysis Methods 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- 239000003517 fume Substances 0.000 description 1
- 238000010230 functional analysis Methods 0.000 description 1
- 108010063718 gamma-glutamylaspartic acid Proteins 0.000 description 1
- 238000012215 gene cloning Methods 0.000 description 1
- 230000008303 genetic mechanism Effects 0.000 description 1
- IXORZMNAPKEEDV-UHFFFAOYSA-N gibberellic acid GA3 Natural products OC(=O)C1C2(C3)CC(=C)C3(O)CCC2C2(C=CC3O)C1C3(C)C(=O)O2 IXORZMNAPKEEDV-UHFFFAOYSA-N 0.000 description 1
- 239000003448 gibberellin Substances 0.000 description 1
- 108010079547 glutamylmethionine Proteins 0.000 description 1
- JYPCXBJRLBHWME-UHFFFAOYSA-N glycyl-L-prolyl-L-arginine Natural products NCC(=O)N1CCCC1C(=O)NC(CCCN=C(N)N)C(O)=O JYPCXBJRLBHWME-UHFFFAOYSA-N 0.000 description 1
- 108010066198 glycyl-leucyl-phenylalanine Proteins 0.000 description 1
- 108010050848 glycylleucine Proteins 0.000 description 1
- 108010037850 glycylvaline Proteins 0.000 description 1
- 230000012010 growth Effects 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 108010040030 histidinoalanine Proteins 0.000 description 1
- 108010045383 histidyl-glycyl-glutamic acid Proteins 0.000 description 1
- 108010028295 histidylhistidine Proteins 0.000 description 1
- 230000001632 homeopathic effect Effects 0.000 description 1
- 230000009001 hormonal pathway Effects 0.000 description 1
- 230000015784 hyperosmotic salinity response Effects 0.000 description 1
- 238000005213 imbibition Methods 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 108010053037 kyotorphin Proteins 0.000 description 1
- 108010044311 leucyl-glycyl-glycine Proteins 0.000 description 1
- 108010057821 leucylproline Proteins 0.000 description 1
- 230000000670 limiting effect Effects 0.000 description 1
- 230000003859 lipid peroxidation Effects 0.000 description 1
- 150000002632 lipids Chemical class 0.000 description 1
- 238000003670 luciferase enzyme activity assay Methods 0.000 description 1
- 230000014759 maintenance of location Effects 0.000 description 1
- 229940118019 malondialdehyde Drugs 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 229930182817 methionine Natural products 0.000 description 1
- 108010020410 methionine sulfoxide reductase Proteins 0.000 description 1
- 108010005942 methionylglycine Proteins 0.000 description 1
- 235000019713 millet Nutrition 0.000 description 1
- 238000007857 nested PCR Methods 0.000 description 1
- 238000007899 nucleic acid hybridization Methods 0.000 description 1
- 229920001542 oligosaccharide Polymers 0.000 description 1
- 150000002482 oligosaccharides Chemical class 0.000 description 1
- 210000000056 organ Anatomy 0.000 description 1
- 230000002018 overexpression Effects 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- 230000001590 oxidative effect Effects 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 239000008188 pellet Substances 0.000 description 1
- 229920001184 polypeptide Polymers 0.000 description 1
- 230000029279 positive regulation of transcription, DNA-dependent Effects 0.000 description 1
- 239000002244 precipitate Substances 0.000 description 1
- 230000001376 precipitating effect Effects 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 238000004321 preservation Methods 0.000 description 1
- 108090000765 processed proteins & peptides Proteins 0.000 description 1
- 102000004196 processed proteins & peptides Human genes 0.000 description 1
- 108010016009 procolipase activation peptide Proteins 0.000 description 1
- 230000002062 proliferating effect Effects 0.000 description 1
- 230000002035 prolonged effect Effects 0.000 description 1
- 108010077112 prolyl-proline Proteins 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 230000004853 protein function Effects 0.000 description 1
- 230000005451 protein repair Effects 0.000 description 1
- 108010063589 raffinose synthase Proteins 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 238000006722 reduction reaction Methods 0.000 description 1
- 230000002786 root growth Effects 0.000 description 1
- 230000008117 seed development Effects 0.000 description 1
- 230000005562 seed maturation Effects 0.000 description 1
- 230000008054 signal transmission Effects 0.000 description 1
- 239000000741 silica gel Substances 0.000 description 1
- 229910002027 silica gel Inorganic materials 0.000 description 1
- 238000002791 soaking Methods 0.000 description 1
- 239000007790 solid phase Substances 0.000 description 1
- 238000009987 spinning Methods 0.000 description 1
- 238000010561 standard procedure Methods 0.000 description 1
- 230000035882 stress Effects 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 229910052717 sulfur Inorganic materials 0.000 description 1
- 238000001308 synthesis method Methods 0.000 description 1
- 108010061238 threonyl-glycine Proteins 0.000 description 1
- 229960001295 tocopherol Drugs 0.000 description 1
- 229930003799 tocopherol Natural products 0.000 description 1
- 235000010384 tocopherol Nutrition 0.000 description 1
- 239000011732 tocopherol Substances 0.000 description 1
- 108091006106 transcriptional activators Proteins 0.000 description 1
- 230000037426 transcriptional repression Effects 0.000 description 1
- 108091006107 transcriptional repressors Proteins 0.000 description 1
- 230000001052 transient effect Effects 0.000 description 1
- 108700004896 tripeptide FEG Proteins 0.000 description 1
- 235000021122 unsaturated fatty acids Nutrition 0.000 description 1
- 150000004670 unsaturated fatty acids Chemical class 0.000 description 1
- 238000012795 verification Methods 0.000 description 1
- 239000011782 vitamin Substances 0.000 description 1
- 229940088594 vitamin Drugs 0.000 description 1
- 229930003231 vitamin Natural products 0.000 description 1
- 235000013343 vitamin Nutrition 0.000 description 1
- 150000003722 vitamin derivatives Chemical class 0.000 description 1
- 238000012070 whole genome sequencing analysis Methods 0.000 description 1
- 239000012224 working solution Substances 0.000 description 1
- GVJHHUAWPYXKBD-IEOSBIPESA-N α-tocopherol Chemical compound OC1=C(C)C(C)=C2O[C@@](CCC[C@H](C)CCC[C@H](C)CCCC(C)C)(C)CCC2=C1C GVJHHUAWPYXKBD-IEOSBIPESA-N 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
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
- C12Q1/6888—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms
- C12Q1/6895—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms for plants, fungi or algae
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q2600/00—Oligonucleotides characterized by their use
- C12Q2600/13—Plant traits
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q2600/00—Oligonucleotides characterized by their use
- C12Q2600/156—Polymorphic or mutational markers
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Health & Medical Sciences (AREA)
- Analytical Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Biophysics (AREA)
- Botany (AREA)
- Zoology (AREA)
- Molecular Biology (AREA)
- General Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Wood Science & Technology (AREA)
- Biotechnology (AREA)
- Genetics & Genomics (AREA)
- Mycology (AREA)
- Physics & Mathematics (AREA)
- Immunology (AREA)
- Microbiology (AREA)
- Gastroenterology & Hepatology (AREA)
- Medicinal Chemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
本发明涉及作物分子育种、分子生物学和基因工程领域,特别涉及了一种玉米种子耐贮性相关的ZmRAP2.7基因克隆鉴定及在提高种子耐贮性中的应用。我们采用全基因组关联分析法,获得与种子人工加速老化发芽率显著关联的4个SNP,落在ZmRAP2.7基因上。该基因的Mu***突变体和CRISPR‑Cas9敲除系的耐贮性显著下降。转录组分析发现该基因功能缺失导致ABA信号传导功能异常;酵母单杂和凝胶阻滞证实该蛋白能结合到ZCN9和3个ABA信号传导基因启动子上转录抑制基因表达。四个显著关联SNP造成该蛋白氨基酸序列改变,进而影响与ZCN9等启动子的结合活性,导致不同单倍型自交系的耐贮性差异。
Description
技术领域
本发明属于作物分子育种、分子生物学和基因工程领域,涉及一种玉米种子耐贮性相关的ZmRAP2.7基因及其编码蛋白序列,具体涉及一个控制玉米种子人工加速老化萌发和耐贮性相关基因ZmRAP2.7的克隆及应用。
背景技术
种子是基本的农业生产资料,高活力种子具有明显的生长优势和生产潜力,是保证作物高产稳产的基础。作物种子需要经历一段时间的贮藏才能用于播种,而贮藏过程中种子会逐步发生劣变,导致活力下降,甚至造成种子报废或播后无法出苗的农业生产事故。挖掘耐贮藏基因,并应用到育种中,是提高种子耐贮藏特性的有效方法,对提高种子田间出苗质量和作物高产稳产具有重要意义。
种子耐贮性的本质是种子在经过一定时间的贮藏后,种子萌发能力的保持能力,因此耐贮性评价最直接的方法是将种子在特定环境中贮藏一定时间后进行发芽能力测定,根据发芽率来评价种子耐贮性。但这种方法耗时长、效率低。人工加速老化发芽法是采用高温高湿的条件来处理种子,一般经过几天的处理后,种子的劣变程度可以达到贮藏几年的效果,因此被广泛用来评价种子的耐贮性。
近年来,利用人工加速老化的方法,在拟南芥、大豆、鹰嘴豆、向日葵和水稻中,通过不同的正向遗传学和反向遗传学手段,克隆了一些耐贮藏相关基因,促进了对种子耐贮性遗传机理的理解。对于目前已克隆的大部分基因,根据其发挥功能的方式不同,大致可分为保护类基因和修复类基因。保护类基因主要与种皮成分、非还原性糖含量、热激响应因子、氧化还原反应相关。其中以氧化还原反应相关的基因报道最多,如拟南芥中维生素E编码基因VITAMIN E1(VTE1)和VTE2可限制贮藏过程中非酶脂质氧化的物质增加,1-cysperoxiredoxin(PER1)基因可消除半胱氨酸残基产生的活性氧,水稻Aldo-ketoreductase1(AKR1)基因有助于降低脂质过氧化过程产生的丙二醛等活性羰基化合物,lipoxygenase(LOX)基因可有助于减少不饱和脂肪酸的氧化反应。修复类基因主要与DNA和蛋白的修复过程相关。L-异天冬氨酰甲基转移酶编码基因Protein L-isoaspartyl methyltransferase(PIMT),在拟南芥、水稻、鹰嘴豆中被报道可有效减少L-异天冬氨酸残基在种子中的积累,促进种子寿命和发芽活力的提高。苜蓿中蛋氨酸亚砜还原酶编码基因methioninesulfoxide reductases(MSR),可有效阻止蛋氨酸氧化成蛋氨酸亚砜,延长种子寿命。此外,拟南芥中DNA连接酶VI,DNA糖基化酶OGG1等被报道在种子老化过程中参与DNA损伤的修复过程。
值得注意的是,转录因子由于可同时调控多个下游基因,常常位于保护类和修复类基因的上游,可从多个层面调控种子的耐贮性。其中以脱落酸ABA、赤霉素GA和生长素三个激素途径中的转录因子报道最多。ABA途径中的ABSCISIC ACID INSENSITIVE3(ABI3)基因属于植物特有的B3转录因子家族,其与LEAFY COTYLEDON 1(LEC1)、LEC2和FUS3FUSCA3(FUS3)三个转录因子共同调节种子的成熟过程,突变后造成种子耐贮藏能力下降。保护类基因non-yellow coloring 1(nyc1)和Tonoplast intrinsic proteins 3(TIP3),是ABI3的下游调控功能基因。ABI5编码碱性亮氨酸拉链类型的转录因子,通过调节棉子糖家族低聚糖代谢和晚期胚胎丰富蛋白Late embryogenesis abundant(LEA)来调控耐贮藏性状,突变后可造成LEA蛋白减少、生育酚增加,导致种子中细胞性程序死亡增加,蛋白质发生错误折叠,最终降低种子耐贮性。拟南芥转录因子ARABIDOPSIS THALIANA HOMEOBOX25(AtHB25),编码同源异型框型转录因子,其过表达可增加种子中的GA含量,从而提高种子的耐贮性。而同时敲除HB25和同源基因HB22、HB31后,种子寿命迅速下降。向日葵中,生长素途径的转录抑制子AUXIN-RESPONSIVE PROTEIN 27(HaIAA27)可与热激响应转录因子HaHSFA9互作,调控种子耐贮性。此外,一些MADS,MYB家族转录因子,也被报道参与调控种子的贮藏性。
与拟南芥、水稻相比,玉米种子由于种胚大且含油量高,更易发生老化变质,不耐贮藏。但鉴于玉米基因组复杂度高且全基因组测序完成时间稍晚,耐贮藏相关基因克隆相对缓慢,目前仅有两个基因的功能研究比较清楚,即ZmRS和ZmDREB2A。ZmRS编码棉子糖合成酶Raffinose Synthase,是棉子糖合成的关键酶,可改变种子中的碳分配,从而调控玉米种子活力及耐老化能力。ZmDREB2A基因编码脱水反应性元素结合蛋白家族转录因子,通过调控下游ZmGH3.2和ZmRAFS基因的表达,影响生长素和棉子糖含量,进而调控玉米种子耐贮性,突变后会导致种子耐贮藏能力显著下降。
除了上述两个基因,当前耐贮性基因克隆的工作主要集中在QTL初定位上。如Wang等在45℃/85%相对湿度下人工加速老化处理玉米种子2、3和4天,从F2群体和重组自交系群体中分别检测到21和28个QTL位点。Liu等利用F2:3群体和重组自交系玉米群体,人工老化5天后,鉴定出13个QTL位点。与传统的连锁分析相比,全基因组关联分析花费少时间短,仅用自然群体就可以完成,近年来在抗旱性、耐盐性、耐脱水性等复杂农艺性状研究中已取得了重要进展,但在种子耐贮性研究中还未有报道。
本发明利用全基因组关联分析的方法,从自然群体中挖掘到一个调控玉米种子耐贮性的重要转录因子ZmRAP2.7,其突变后种子耐贮藏能力显著下降。重测序结果表明,不同单倍型与下游基因启动子的结合强弱是造成不同关联群体材料耐贮性差异的原因;转录组、酵母单杂和凝胶阻滞等技术发现该基因通过转录抑制下游基因ZmABI5、ZmPP2C、ZmPYL3和ZCN9,最终影响ABA信号转导过程和种子贮藏特性。在此基础上,细致研究该基因的作用机理和调控网络,研究结果不仅有助于解析玉米种子耐贮性的作用机制及其进化机理,而且具有潜在的应用价值,对于耐贮藏玉米新品种培育具有重要意义。
发明内容
本发明的目的之一在于提供一种新的影响种子ABA信号通路导致种子耐贮性变化的基因ZmRAP2.7的DNA序列、cDAN序列和该基因所编码的功能蛋白的氨基酸序列,所述基因编码蛋白是一种植物组织内普遍存在并且十分重要的AP2蛋白。该功能的缺失导致玉米ABA信号转导缺陷,从而导致玉米种子耐贮性降低。
本发明的第一个发明目的是:提供一种调控玉米耐贮性的新基因ZmRAP2.7,其特征在于,是选自如下1)的DNA分子或2)的cDNA分子:
SEQ ID NO.1所示的DNA分子(克隆自玉米自交系B73的基因组DNA):该DNA的分子由2742个核苷酸组成,翻译起始于1bp,终止于2742bp。ZmRAP2.7基因包含9个外显子,8个内含子(图4A)。
SEQ ID NO.2所示的DNA分子(克隆自玉米自交系B73的cDNA):该DNA的分子由1413个核苷酸组成,翻译起始于1bp,终止与1413bp,是SEQ ID NO.1转录出来的cDNA序列。
在SEQ ID NO.1基础之上经过一至数个碱基替换所形成的能影响种子耐贮性的DNA分子;
在0.1×SSPE(或0.1×SSC)、0.1%(w/v)SDS的溶液中,65℃条件下杂交并洗膜,能够与SEQ ID NO.2的DNA分子杂交且编码植物种子耐贮性相关蛋白的DNA分子;
与SEQ ID NO.2的DNA分子具有85%以上的同源性且编码植物种子耐贮性相关蛋白的DNA分子;
本发明的第二个发明目的是:提供上述基因所编码的调控种子耐贮性的相关蛋白质。
在一个实施方案中,所述基因编码如下1)或2)所述的蛋白质;
1)序列表的SEQ ID NO.3所示的氨基酸序列组成的蛋白质;
2)将序列表的SEQ ID NO.3经过一个或数个氨基酸残基的替换成具有影响植物功能的相关蛋白质。
本发明的第三个发明目的是:提供在植物种子器官中调节下游基因序列转录表达的启动子及顺势作用元件。
在一个实施方案中,所述启动子序列如SEQ ID NO.4、SEQ ID NO.5、SEQ ID NO.6、SEQ ID NO.7所示DNA序列,该序列能够被SEQ ID NO.3所述蛋白转录抑制。
序列表的SEQ ID NO.4为所述ZmABI5启动子的序列,总长1028个核苷酸,包含SEQID NO.3转录调控的重要元件。
序列表的SEQ ID NO.5为所述ZmPP2C启动子的序列,总长1270个核苷酸,包含SEQID NO.3转录调控的重要元件。
序列表的SEQ ID NO.6为所述ZmPYL3启动子的序列,总长1111个核苷酸,包含SEQID NO.3转录调控的重要元件。
序列表的SEQ ID NO.7为所述ZCN9启动子的序列,总长1145个核苷酸,包含SEQ IDNO.3转录调控的重要元件。
本发明的第四个发明目的是:提供上述基因在用于转基因改良作物的用途。
在一个实施方案中,所述基因用于抑制作物种子ABA信号转导,以便导入外源基因以获得优质的转基因作物。
在一具体实施方案中,所述改良包括产量提高、品质提高、抗病虫害、抗逆、抗倒伏等农艺性状的改良。
在另一具体实施方案中,所述作物是自花授粉或者异花授粉作物。
在一个更加具体的实施方案中,所述作物包括但不仅限于玉米、水稻、高粱、小麦。
本发明的第五个发明目的是:RAP2.7及其同源基因在种子耐贮藏性状中的作用未见报道,通过研究RAP2.7及其下游靶基因调控玉米种子耐贮性的功能,解析RAP2.7的作用机制和调控网络。
本发明通过全基因组关联分析挖掘到ZmRAP2.7,并通过突变体和敲除系证实了功能,通过表达分析、酵母单杂和凝胶阻滞等技术证实该基因可转录抑制3个ABA信号通路基因和1个ZCN基因。这些结果可为玉米种子耐贮性研究和分子设计育种提供新基因和新调控网络。
术语定义
术语“影响种子ABA信号通路导致种子耐贮性变化的基因”指一段具有编码蛋白能力的核苷酸序列,该序列特异编码具有转录激活活性的转录因子,如SEQ ID NO.2的第1-1413位核苷酸序列及其简并序列。
术语“简并序列”是指,位于SEQ ID NO.2的编码框第1-1413位核苷酸序列中,有一个或多个密码子被编码相同氨基酸的简并密码子所取代后而产生的序列。由于密码子的简并性,许多与SEQ ID NO.2的第1-1413位核苷酸序列同源性低至70%的简并序列也能编码出SEQ ID NO.2所编码的氨基酸序列。
所述“影响种子ABA信号通路导致种子耐贮性变化的基因”,还包括在中度严格条件下,更加高度严格条件也可以与SEQ ID NO.2的核苷酸序列杂交的核苷酸序列。其中,中等严格条件下可以是0.1×SSPE(或0.1×SSC)、0.1%(w/v)SDS的溶液中,65℃条件下杂交并洗膜的条件。
所述“影响种子ABA信号通路导致种子耐贮性变化的基因”,还包括与SEQ ID NO.2从1-1413位的核苷酸序的同源性至少70%,较佳的80%、82%、85%、86%、88%、89%同源性,更佳的具有90%、91%、92%、93%、94%同源性,最佳的具有至少95%、96%、97%、98%、99%同源性的核苷酸序列。
所述“影响种子ABA信号通路导致种子耐贮性变化的基因”,还包括能编码具有天然的调控玉米种子耐贮性基因ZmRAP2.7相同功能蛋白的SEQ ID NO.2开放阅读框序列的变异形式。这些变异形式包括(但并不限于):1个或几个核苷酸的缺失、***和/或取代,以及5’或3’端添加几个(通常为60个以内,较佳的为30个以内,更佳的为10个以内,最佳的为5个以内)的核苷酸。在本领域中,用性能相近或相似的氨基酸进行取代时,通常不会改变蛋白质的功能;在C末端和/或N末端添加一个或数个氨基酸通常也不会改变蛋白质的功能。
另外,所述“影响种子ABA信号通路导致种子耐贮性变化的基因”的核苷酸全长序列或其片段通常可以用PCR扩增法、重组法或人工合成的方法获得。对于PCR扩增法,可根据本实施例公开的有关核苷酸序列,尤其是开放阅读框序列来设计引物,并用市售的cDNA文库或按本领域技术人员已知的常规方法所制备的cDNA文库作为模板,扩增到有关序列。当序列较长时,通常需要两次或者多次的巢式PCR扩增,然后将各次PCR扩增产物按正确的顺序拼接在一起。一旦获得了有关序列,可以用重组法来大批量的获得有关序列。通常是将其克隆入载体,然后通过细胞转化等常规方法从增殖的宿主细胞中分离得到有关序列。此外,还可通过化学合成将突变引入实施例蛋白序列中。除了用重组法产生之外,实施例蛋白的片段还可用固相技术,通过直接合成多肽而加以生产。在体外合成蛋白质可以用人工或者自动进行,可以分别化学合成实施例蛋白的各片段,然后用化学方法加以连接,以产生全长的蛋白质分子。
附图说明
为了更清楚地说明本发明实施例或现有技术中的技术方案,下面将对实施例或现有技术描述中所需要使用的附图作简单地介绍。
图1(A)2011、2016、2017三个年份玉米种子的人工加速老化发芽率密度分布图,横坐标为老化发芽率,纵坐标为密度;(B)三个年份人工加速老化发芽率的相关性分析;(C-E)以三年老化发芽率平均值作为表型数据,全基因组关联分析的QQ plot图(C)、曼哈顿图(D)和局部曼哈顿图(E);(F)最显著SNP(Lead SNP)位点上下游100kb所包括的基因结构示意图;AAG,accelerated aging germination,人工加速老化发芽;
图2(a)ZmRAP2.7基因的Mu***突变体和对照系种子的人工加速老化发芽表型;(b)ZmRAP2.7基因的敲除系KO#1和对照系种子的人工加速老化表型;(c)和(d)分别为(a)和(b)图对应的发芽率统计结果,其中NAG,AG3和AG6分别表示未老化,老化3天和老化6天的发芽率;(e)ZmRAP2.7基因在不同组织中的表达量,纵坐标为TPM值,数据来自Chen J.,etal.2014;(f)ZmRAP2.7基因在种子萌发过程中的相对表达量变化;
(g)ZmRAP2.7基因在授粉后18天的种子中的原位杂交图,上面为反义探针杂交结果,下面为正义探针作为负对照的杂交结果,bar=1mm;
图3(A)ZmRAP2.7敲除系与对照系的种子,人工老化3天萌发24小时后的差异表达基因统计图,蓝色为下调基因,红色为上调基因;(B)差异表达基因的GO功能分析;(C)ZmRAP2.7敲除系和对照系人工加速老化3天后萌发不同时间点种胚的ABA含量测定;G0,G3和G6分别代表未萌发,萌发3小时,萌发6小时的种胚;(D)和(E)分别为ZmRAP2.7敲除系KO#1和对照系的老化3天萌发24小时的种胚中ZmPYL3、ZmPP2C、ZmABI5(D)以及ZCN9、ZCN10(E)的相对表达量;(F)ZmRAP2.7蛋白与ZmPYL3、ZmPP2C、ZmABI5以及ZCN9、ZCN10基因启动子序列的酵母单杂结果,AD为空载体为负对照,AD-ZmRAP2.7为ZmRAP2.7基因连入pB42AD载体;(G)ZmRAP2.7蛋白与ZmPYL3、ZmPP2C、ZmABI5以及ZCN9基因启动子的凝胶阻滞试验结果;
图4(A)ZmRAP2.7基因的候选基因关联分析显示不同标记之间的连锁关系;(B)基于显著关联的基因区的4个SNP可将关联群体材料分为四种单倍型;(C)-(E)分别为+MITE-Hap1、+MITE-Hap2、-MITE-Hap2三组单倍型材料的吐丝期(C)、散粉期(D)和人工加速老化发芽率(E)的比较,MITE为前人报道的ZmRAP2.7基因上游35kb处的一个转座子***,远程调控ZmRAP2.7基因表达,具有调控玉米开花的功能;(F)ZmRAP2.7基因在Hap1和Hap2单倍型自交系中的表达量比较;(G)单倍型Hap1和Hap2的ZmRAP2.7基因启动子的荧光素酶瞬时试验结果,GAL4-LUC为报告载体,不同单倍型连入GAL4-DB载体中,并采用与转录激活因子VP16融合来增加本底表达水平。
具体实施方式
本发明公开了玉米种子耐贮性相关的ZmRAP2.7基因及其功能标记与应用,本领域技术人员可以借鉴本文内容,适当改进工艺参数实现。特别需要指出的是,所有类似的替换和改动对本领域技术人员来说是显而易见的,它们都被视为包括在本发明。本发明的方法及应用已经通过较佳实施例进行了描述,相关人员明显能在不脱离本发明内容、精神和范围内对本文所述的方法和应用进行改动或适当变更与组合,来实现和应用本发明技术。
以下的实施例便于更好的理解本发明,但不限定本发明的应用范围。下属实施例中的所有技术和科学术语,如无特殊说明,均为本发明所属领域普通技术人员通常理解的相同含义。除非有相反证明,本发明所使用或者提及的技术均为本领域普通技术人员公认的标准技术。所述试验材料,如无特别注明,均为本发明领域通用的试验材料。下属实施例中所用的试验试剂,如无特殊说明,均为常规生化试剂商店购买得到的。材料、方法和实施例仅作阐述用,而非加以限制。
本发明包括利用序列表锁住序列调控种子耐贮性、即利用本发明提供的基因序列在基因组、和/或转录组、和/或蛋白组水平影响其它植物中相同或同源基因的功能来达到控制种子耐贮性的目的。例如,下述方法但不限于下述方法:通过天然序列的变异导致基因表达抑制或蛋白质功能的丧失、通过向植物中转入所述基因的反义序列或引入发卡结构、或将所述基因与其它序列(DNA或RNA)相结合产生新的具有功能活性的DNA或者RNA链,来影响或改变植物基因的功能,或其它本领域技术人员已知的可用于影响种子耐贮性的技术方法中的任何一种技术方法。
本发明包括我们利用368关联群体三年的人工加速老化发芽率关联到ZmRAP2.7基因。利用该基因的突变体和CRISPR-Cas9敲除系,确认ZmRAP2.7基因对种子耐贮性的影响。该基因位于玉米8号染色体。
本发明通过对ZmRAP2.7基因重测序,进行候选基因的关联分析,进一步将变异位点锁定在编码区的4个SNP,并以此为依据分为四种单倍型,其中两组只有2个和1个材料,属于极端稀有材料。另外两种主要单倍型中,Hap1单倍型的人工加速老化发芽率显著低于Hap2单倍型。
本发明通过对RAP2.7转录因子的顺式作用元件分析,同时结合RAP2.7的转录组数据,找到了一个可能的下游作用因子,影响种子的萌发。
该基因序列及其同源序列可从任何种子植物中获得,包括但不限于玉米(Zeamays)、水稻(Oryza sativa)、高粱(Sorghum bicolor)、普通小麦(Triticum aestivum)、谷子(Setaria italica)、大麦(Hordeum vulgare)、短柄草(Brachypodium distachyon)、拟南芥(Arabidopsis thaliana)、大豆(Glycine max)等。获得方法包括但不限于:通过玉米ZmRAP2.7基因序列利用blastx、blastn或通过氨基酸序列利用blastp从其它植物的基因组序列数据库、和/或cDNA序列数据库、和/或蛋白质序列数据库中调取;以ZmRAP2.7基因的DNA或cDNA或RNA序列为参考序列设计引物,直接从其它植物的基因组DNA或cDNA或RNA中利用PCR的方法直接获得;以玉米ZmRAP2.7的基因序列设计探针,利用核酸杂交的方法从基因组文库中分离含有同源基因序列的DNA或cDNA或RNA片段。
下文通过说明和阐述提供了更为详细的描述,但这并非意欲对本发明的范围加以限制。
实施例1:利用全基因组关联分析法克隆ZmRAP2.7
本发明利用三个不同年份收获的368份不同来源的自交系,在玉米种子萌发最佳条件(25℃,标准发芽条件)下发芽7天,统计发芽率,2011、2016和2017年分别有273、318和290份种子的标准发芽率高于85%,说明这些种子具备正常的萌发能力。然后对这些材料进行人工老化发芽实验(45℃和95%湿度下处理3天),发现老化发芽率呈现偏正态分布(图1A),不同种子批的均值在68%-87%之间,两两之间的相关性为0.26-0.41(图1B)。
我们以三年数据的平均值作为表型数据,结合该群体的已发表的55万个SNP标记,采用混合线性模型,进行全基因组关联分析,得到4个显著SNP位点,全部位于8号染色体的GRMZM2G700665(ZmRAP2.7)基因上,跨越1.8kb的染色体区段(图1C-1E),同时对该区域上下游100kb的区域进行搜索,发现该区域只有2个编码基因(图1F)。根据maizeGDB网站(www.maizegdb.org)信息,另一个基因在种子中没有表达,而ZmRAP2.7在种子中高表达,因此将ZmRAP2.7选为候选基因进行功能验证。
实施例2:ZmRAP2.7敲除和突变后导致种子耐贮藏能力显著降低
早在2008年就报道ZmRAP2.7是一个玉米花期的负调控因子,随后在2019年详细报道了该基因调控花期和气生根生长的功能,但它在种子活力中的功能未见报道。我们获得了发表文献中创制的ZmRAP2.7转基因敲除系和Mu***突变体,进行标准发芽试验,发现敲除系和突变体的标准发芽率与对照没有明显差异(图2a-2d)。但人工加速老化3天和6天后,敲除系的发芽率分别下降到19.3%和2.6%,而对照为56.7%和10.3%(图2c);Mu***系的发芽率分别降到46.7%和16.7%,而对照依然维持在97.3%和75.0%(图2d),说明ZmRAP2.7基因功能缺失降低了玉米种子的人工老化发芽率。
我们利用公共数据库对ZmRAP2.7基因的表达模式进行了分析,发现该基因是组成型表达的基因,但在萌发时的种子、根和叶片中的表达量较高(图2e)。我们利用qPCR技术发现,发现随着种子萌发的进行,该基因呈现先上升后下降的表达变化趋势,在萌发12h的表达量最高(图2f)。因此我们对萌发12h的种子进行ZmRAP2.7基因的原位杂交,但因种子太硬而未能获得高质量的组织切片,考虑基因在种子发育和萌发阶段的组织表达特性一般不会改变,因此改用授粉后18天的种子进行原位杂交,发现ZmRAP2.7基因主要在种胚和糊粉层中表达,在胚乳中几乎观察不到表达信号(图2g),这一表达部位与过去认为的种子耐贮性主要与胚和糊粉层的活性相关的生理结论相吻合。
综上所述,ZmRAP2.7基因的Mu***系和Crispr敲除系的老化发芽率均显著下降,说明该基因具有调控玉米种子耐贮性的功能。
实施例3:ZmRAP2.7转录调控下游基因筛选
利用ZmRAP2.7 KO#1转基因株系和对照系,收集人工老化3天萌发24小时的种胚,进行转录组测序分析,获得1750个差异基因,其中上调708个,下调1042个(图3A)。对这些差异基因进行GO分析,发现功能聚类主要与ABA信号响应相关(图3B)。此外,对人工加速老化三天的种子进行萌发不同时间段的ABA含量测定,发现吸胀3小时后,敲除系的ABA含量显著高于对照系(图3C)。由于ABA具有抑制种子萌发的功能,这些结果说明人工加速老化后,ZmRAP2.7功能缺失很可能导致无法解除(或部分解除)ABA对萌发的抑制,从而导致萌发率下降。
GO聚类的ABA信号途径组中有18个ABA信号通路相关基因。同时,根据文献报道推测,ZmRAP2.7还可能直接调控ZCN基因。我们逐个扩增这些基因约1kb的启动子序列,进行酵母单杂测试,发现ABA信号通路基因ZmABI5、ZmPP2C和ZmPYL3,以及开花基因ZCN8的同源基因ZCN9和ZCN10基因的与启动子能够与ZmRAP2.7蛋白互作,产生单杂信号(图3F)。
进一步对这些基因启动子进行缺失分析(即将启动子分为多个小片段),通过酵母单杂筛选获得能与ZmRAP2.7蛋白结合的核心序列。然后合成这些核心序列的标记探针,利用凝胶阻滞(EMSA)试验,体外验证这些核心基序与ZmRAP2.7蛋白在体外互作,证实ZmRAP2.7能与ZmABI5、ZmPP2C、ZmPYL3、ZCN9的启动子核心基序结合(图3G)。
同时,我们对ZmABI5、ZmPP2C、ZmPYL3和ZCN9等基因在ZmRAP2.7敲除系中的表达进行分析,发现这些基因在敲除系和对照系的干种子中表达量差异不大,但在老化3天萌发24小时的种子中,这些基因在敲除系中的表达量显著高于对照种子(图3D和3E),说明ZmRAP2.7功能缺失导致ZmABI5、ZmPP2C、ZmPYL3和ZCN9基因在人工加速老化种子萌发过程中的表达量无法正常下降,因而无法减弱ABA信号对种子萌发的抑制,导致发芽率下降。
总之,基因表达、原位杂交、酵母单杂和凝胶阻滞等结果说明ZmRAP2.7蛋白能够转录抑制ABA信号通路基因ZmABI5、ZmPP2C和ZmPYL3,以及ZCN8的同源基因ZCN9的表达,抑制人工加速老化种子的ABA信号通路,进而促进萌发。
实施例4:不同单倍型ZmRAP2.7对下游基因的调控能力具有显著差异
对ZmRAP2.7的基因组序列(包含3428bp的基因区序列和1473bp启动子序列)进行重测序,结合网上发布的368份玉米自交系的基因组重测序序列数据,发现该基因区存在4个InDel变异和98个SNP变异,其中在启动子区存在2个大的***序列,即***长度为1551bp和3562bp。利用所有多态性位点,结合人工加速老化发芽表型数据,进行候选基因关联分析,发现最初GWAS定位到的基因外显子上的4个SNP位点的关联显著性依然最高,而启动子区的两个***/缺失与种子人工加速老化发芽率表型不存在显著关联性(图2a)。此外,前期报ZmRAP2.7上游35kb处存在一个与花期调控密切相关的MITE***,这个***与种子人工加速老化发芽率表型不相关。这些结果说明ZmRAP2.7的功能变异不在启动子区,很有可能是基因外显子上的4个SNP。
利用Primer5.0软件进行引物设计,开发出2对分子辅助标记(如表1):利用表1引物扩增368份材料的ZmRAP2.7基因组序列。通过测序对扩增的基因序列进行验证。所有PCR扩增均使用KOD FX DNA polymerase(TOYOBO CO.,LTD.Life Science Department,Osaka,Japan),并按照产品说明的反应体系和条件,在杭州朗基A300快速梯度PCR仪进行扩增。PCR产物送往上海生工生物工程公司进行测序。测序后结果使用DNAMAN生物软件进行序列拼接后比对分析,发现ZmRAP2.7不同材料存在多个SNP差异,同时验证了关联分析获得的4个显著SNP:SNP190、SNP1772、SNP1789、SNP1825,并且利用这4个SNP能够将368份自交系分成4组,其中两组只有2个和1个材料,属于极端稀有材料,另外两种主要单倍型中,Hap1(序列SEQ ID NO.8和SEQ ID NO.10)有19个系,Hap2(序列SEQ ID NO.9和SEQ ID NO.11)有346个系(图4B)。
表1.引物信息
对实施例2中开发的2个分子辅助标记进行验证,具体操作如下。其中,玉米种子DNA提取方法如下。1)取野生型和敲除系萌发3天的种子,于冰盒中剥去果皮,放入事先写好编号的2.0ml离心管中(管底提前放1钢珠)。2)按顺序将已放入样品和钢珠的离心管摆放在打样仪专用的离心管架(8×5)上,整体浸泡在盛有液氮的保鲜盒中1-2min(注:液氮以刚没过离心管架少许为宜)。3)将冷冻好的离心管架放入打样仪(MiniG-1600)的卡槽中,拧紧,关盖,1200转/min的速度打样30s。4)加入800μLCTAB提取液(65℃预热),65℃水浴30min,中间取出颠倒3次。5)加入800μL氯仿∶异戊醇(24∶1)萃取液,盖紧盖子,上下颠倒混匀(注意标记不要被擦掉;氯仿为腐蚀性试剂,需要带PE一次性手套在通风橱中操作)。
6)12,000rpm离心10min。吸取上清液300μL,转入新的1.5mL微量离心管中,加入等量预冷的无水乙醇,弃枪头,盖紧盖子,写好编号并检查无误,上下颠倒混匀。-20℃冰箱中沉淀60min以上。7)12,000rpm离心30min至沉淀附于离心管底部,小心倒去上清液(注:沉淀后的DNA呈白色粉末状于离心管底部,容易随液体倒出,此步骤小心缓慢操作)。8)于离心管中加入1ml 70%乙醇,盖上管盖上下轻轻颠倒洗涤,离心5min,弃掉乙醇,重复洗涤2遍,将1.5mL微量离心管倒置(下面铺好吸水纸),自然干燥。9)加入300μL的ddH2O,Votex溶解沉淀(如不溶,可以借助加热50℃以下溶解)。10)样品置于-20℃冰箱中保存备用。
利用设计的引物对所获得的DNA进行PCR扩增,方法和程序如下:1)先打开制冰机的水源开关,再打开制冰机的电源开关,制冰。2)将下列试剂从-20℃冰箱中取出解冻:2×PCR master mix(擎科生物科技有限公司)、正反向引物溶液以及模板DNA(注:当某一试剂完全解冻,即放置于冰上。经常用的ddH2O、引物工作液(10μM)、模板DNA可放于4℃冰箱中)。3)所有试剂解冻完毕后,5,000rpm离心数秒,放回冰上待用。4)配制PCR反应的混合液,按照表2依次加入各试剂(反应体系为10μL)。下表2同时列出了1个反应(1R)、10个反应(10R)、50个反应(50R)的混合液配方。配好后将所有试剂放回4℃或-20℃冰箱。5)混匀,5,000rpm离心数秒。6)将混合液分装至96孔PCR反应板(每孔200μL最大体积)中,再加入1μL模板DNA,做好标记,盖96孔硅胶垫封口。7)将PCR反应板嵌入PCR扩增仪的200μL孔中,合上热盖,旋紧。8)打开PCR扩增仪电源开关,选择预设的反应程序(如表3),开始扩增。9)扩增完后,退出反应程序,回到主菜单,关闭电源开关。打开热盖,取出PCR反应板置于PCR管架上,放入4℃冰箱中,待用。
表2.PCR反应体系
表3.PCR扩增条件
注:不同引物的退火温度不同,一般都在56-60℃之间,延伸时间根据PCR产物大小有变动,产物1kb以下均延伸30s,1kb以上可适当延长至60s。
为了进一步确认MITE元件和显著性最高的4个SNP位点与花期及耐贮性的关系,我们将所有材料分为+MITE-Hap1、+MITE-Hap2和-MITE-Hap2三种类型(Hap1型的19个系全部含有MITE元件),进行多重比较,发现MITE元件仅与花期相关,与耐贮性无关,而显著性最高的4个SNP位点,仅影响种子耐贮性,与花期无关(图4C-4E)。我们还对18个Hap1和随机选择的53个Hap2单倍型的种胚进行qPCR分析,发现ZmRAP2.7基因表达量在两个单倍型材料间无差异(图4F)。这些结果说明4个SNP位点不影响ZmRAP2.7基因表达量和玉米花期,这与启动子区不存在显著关联位点相一致,进一步说明功能变异和基因表达量无关。为了探究不同单倍型造成种子人工加速老化发芽率差异的原因,我们首先对Hap1和Hap2单倍型的氨基酸序列进行分析,发现4个显著SNP位点,全部导致氨基酸序列发生改变(A>V、S>R、P>S和L>V),这些关键氨基酸改变可能影响转录因子对下游基因的结合活性。因此,我们利用荧光素酶活性检测***检测两种单倍型与下游基因启动子的结合能力差异,发现Hap2单倍型对下游基因的抑制作用更强(图4G),可以推测Hap2单倍型的强结合能力可能导致对下游ABA信号基因和ZCN基因的强抑制效果,从而具有更强的抑制ABA信号的功效和促进种子人工加速老化后的萌发率的功能。
总之,本实验室对三个年份收获的368份玉米自交系群体的3个批次种子进行人工加速老化发芽试验,全基因组关联分析获得4个显著关联SNP位点,均位于ZmRAP2.7基因外显子上。该基因的Mu转座子***突变体和CRISPR-Cas9敲除系的种子人工加速老化发芽率显著下降,确认ZmRAP2.7基因对种子耐贮性的影响。将ZmRAP2.7基因的敲除系和野生型老化3d萌发24h的种子胚进行RNA-Seq分析,发现差异表达基因主要聚类到ABA信号途径,酵母单杂和EMSA试验发现ZmRAP2.7能直接结合到ABA信号传导基因ZmABI5、ZmPP2C和ZmPYL3,以及ZCN9的启动子上,转录抑制这些基因表达,进而影响种子的人工加速老化发芽率。并且找到了ZmRAP2.7的功能变异位点是基因编码区上的4个SNP,导致氨基酸的非同义突变,最终造成ZmRAP2.7对下游基因启动子调控强度的变化,引起了种子耐贮性的差异。
以上所述仅是本发明的优选实施方式,应当指出,对于本技术领域的普通技术人员来说,在不脱离本发明原理的前提下,还可以做出若干改进和润饰,这些改进和润饰也应视为本发明的保护范围。
序列表
<110> 中国农业大学
<120> 玉米种子耐贮性相关的ZmRAP2.7基因及其功能标记与应用
<130> MP22000538Z
<160> 15
<170> SIPOSequenceListing 1.0
<210> 1
<211> 2742
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 1
atgcagttgg atctgaacgt ggccgaggcg ccgccgccgg tggagatgga ggcgagcgac 60
tcggggtcgt cggtgctgaa cgcgtcggaa gcggcgtcgg cgggcggcgc gcccgcgccg 120
gcggaggagg gatctagctc aacgccggcc gtgctggagt tcagcatcct catccggagc 180
gatagcgacg cggccggcgc ggacgaggac gaggacgcca cgccatcgcc tcctcctcgc 240
caccgccacc agcaccagca gcagctcgtg acccgcgagc tgttcccggc cggcgccggt 300
ccgccggccc cgacgccgcg gcattgggcc gagctcggct tcttccgcgc cgacctgcag 360
cagcaacagg cgccgggccc caggatcgtg ccgcacccac acgccgcgcc gccgccggcc 420
aagaagagcc gccgcggccc gcgctcccgc agctcgcagt accgcggcgt caccttctac 480
cgccgcacag gccgctggga gtcccacatc tggtcagtac taccactgtc tacaactagc 540
cacaccacac cgattgcttc cgactctcat taatttctga cacaaactct ccgtcttcct 600
cctcttctcc cgcgacgcag ggattgcggc aagcaggtgt acctaggtga gcaagagcag 660
atctcttttg cgttcccaaa gatttttccc cttttagttc cttatcccat cccatctcga 720
atggcctagc taaccgattc agtggtggtc cggctgctgg ccgatatacg caggtggatt 780
cgacaccgct cacgccgctg caaggcacgc actggactgg acgcccagaa ttcttcgtca 840
tgtgagtctc tgaccgaatt gattgattaa cgagtctctg gctcctggaa ctcgcagggc 900
gtacgaccgg gcggcgatca agttccgcgg cgtcgacgcc gacatcaact tcaacctcag 960
cgactacgag gacgacatga agcagatggg gagcctgtcc aaggaggagt tcgtgcacgt 1020
cctgcgccgt cagagcaccg gcttctcgag aggcagctcc aggtacagag gcgtcaccct 1080
gcacaagtgc ggccgctggg aggcgcgcat ggggcagttc ctcggcaaga agtaagaacc 1140
aaccaacgct tcttttcttt ttcttttttt tatagcatgc agtgatgatt caaccttagt 1200
tgtgcctttc ctcctaatcc tatatatgta ggatttagta ctggttgact atataagtat 1260
atatgtattg ttcagtaaaa gtatacatag gttagctgca tgtttatgta tgtagctggt 1320
tgtttcaatc agaagataaa aaaaaaggga agtagtggct agggaattcc tccaatcctc 1380
accggtggga acgccgtgct tgggtgcagg ggaacgccgt gcttgggtgc aggtacatat 1440
accttgggct attcgacagc gaagtagagg ctgcaaggtt cttcatcttg gattctgccg 1500
ttcatatatg cataatcatg tcttttaatt tccaaagggt tgagtaccga ctcgattcct 1560
cttcgtgtct tttttctttc tttcttcgaa atccagagcc tacgacaagg ccgccatcaa 1620
atgcaatggc agagaggccg tgacgaactt cgagccgagc acgtatcacg gggagctgcc 1680
gactgaaggt acgtattttc tttctgcata tatatatctt caggtattat tggctattaa 1740
actgcttgga tcttactgct tcttctgcag ttgctgatgt cgatctgaac ctgagcatat 1800
ctcagccgag cccccaaaga gacaagaaca gctgcctagg tctgcagctc caccacggac 1860
cattcgaggg ctccgaactg aagaaaacca aggcaagcgc taacgataga tataccttga 1920
caagctagta tcaaacaaaa ccagtaaaaa aagtttactt tcttgtcgaa tttcattgcc 1980
tacctgatgt acgtacttgt gcttctgcac aaaataacga aatccttttg ccctctgatg 2040
atgatgcaga tcgacgatgc tccctctgag ctaccgggcc gccctcgtca gctgtctcct 2100
ctcgtggctg agcatccgcc ggcctggcct gcgcagccgc ctcacccctt cttcgtcttc 2160
acaaaccatg aggttaggtg acagctactg atcgagatgc agcagcagtt caaacctgtc 2220
tgttccaagg acctttaggc cggattacca aatcatcggt caactgtcct gtctgttata 2280
tatttatgtg ttaatttata atacaagtgt gactattttt caaaccttcc ttcaaaatgc 2340
atgaaaagag ttttttttaa cgaaaggcga aaagaaaata tgatacttgg gacaggagca 2400
agcttggatc atcagaaagt attattaatt aggatcactg agctgttcat tttgttcttg 2460
agtcaatcct aatcgtacta tgtcagtgaa tgaacttgtg ttgcaccaat gcagatgagt 2520
gcatcaggag atctccacag gaggcctgca ggggctgttc ccagctgggc atggcaggtg 2580
gcagcagcag ctcctcctcc tgccgccctg ccgtcgtccg ctgcagcatc atcaggattc 2640
tccaacaccg ccacgacagc tgccaccacc gccccatcgg cctcctccct ccggtactgc 2700
ccgccgccgc cgccgccgtc gagccatcac catccccgct ga 2742
<210> 2
<211> 1413
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 2
atgcagttgg atctgaacgt ggccgaggcg ccgccgccgg tggagatgga ggcgagcgac 60
tcggggtcgt cggtgctgaa cgcgtcggaa gcggcgtcgg cgggcggcgc gcccgcgccg 120
gcggaggagg gatctagctc aacgccggcc gtgctggagt tcagcatcct catccggagc 180
gatagcgacg cggccggcgc ggacgaggac gaggacgcca cgccatcgcc tcctcctcgc 240
caccgccacc agcaccagca gcagctcgtg acccgcgagc tgttcccggc cggcgccggt 300
ccgccggccc cgacgccgcg gcattgggcc gagctcggct tcttccgcgc cgacctgcag 360
cagcaacagg cgccgggccc caggatcgtg ccgcacccac acgccgcgcc gccgccggcc 420
aagaagagcc gccgcggccc gcgctcccgc agctcgcagt accgcggcgt caccttctac 480
cgccgcacag gccgctggga gtcccacatc tgggattgcg gcaagcaggt gtacctaggt 540
ggattcgaca ccgctcacgc cgctgcaagg gcgtacgacc gggcggcgat caagttccgc 600
ggcgtcgacg ccgacatcaa cttcaacctc agcgactacg aggacgacat gaagcagatg 660
gggagcctgt ccaaggagga gttcgtgcac gtcctgcgcc gtcagagcac cggcttctcg 720
agaggcagct ccaggtacag aggcgtcacc ctgcacaagt gcggccgctg ggaggcgcgc 780
atggggcagt tcctcggcaa gaagtacata taccttgggc tattcgacag cgaagtagag 840
gctgcaagag cctacgacaa ggccgccatc aaatgcaatg gcagagaggc cgtgacgaac 900
ttcgagccga gcacgtatca cggggagctg ccgactgaag ttgctgatgt cgatctgaac 960
ctgagcatat ctcagccgag cccccaaaga gacaagaaca gctgcctagg tctgcagctc 1020
caccacggac cattcgaggg ctccgaactg aagaaaacca agatcgacga tgctccctct 1080
gagctaccgg gccgccctcg tcagctgtct cctctcgtgg ctgagcatcc gccggcctgg 1140
cctgcgcagc cgcctcaccc cttcttcgtc ttcacaaacc atgagatgag tgcatcagga 1200
gatctccaca ggaggcctgc aggggctgtt cccagctggg catggcaggt ggcagcagca 1260
gctcctcctc ctgccgccct gccgtcgtcc gctgcagcat catcaggatt ctccaacacc 1320
gccacgacag ctgccaccac cgccccatcg gcctcctccc tccggtactg cccgccgccg 1380
ccgccgccgt cgagccatca ccatccccgc tga 1413
<210> 3
<211> 470
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 3
Met Gln Leu Asp Leu Asn Val Ala Glu Ala Pro Pro Pro Val Glu Met
1 5 10 15
Glu Ala Ser Asp Ser Gly Ser Ser Val Leu Asn Ala Ser Glu Ala Ala
20 25 30
Ser Ala Gly Gly Ala Pro Ala Pro Ala Glu Glu Gly Ser Ser Ser Thr
35 40 45
Pro Ala Val Leu Glu Phe Ser Ile Leu Ile Arg Ser Asp Ser Asp Ala
50 55 60
Ala Gly Ala Asp Glu Asp Glu Asp Ala Thr Pro Ser Pro Pro Pro Arg
65 70 75 80
His Arg His Gln His Gln Gln Gln Leu Val Thr Arg Glu Leu Phe Pro
85 90 95
Ala Gly Ala Gly Pro Pro Ala Pro Thr Pro Arg His Trp Ala Glu Leu
100 105 110
Gly Phe Phe Arg Ala Asp Leu Gln Gln Gln Gln Ala Pro Gly Pro Arg
115 120 125
Ile Val Pro His Pro His Ala Ala Pro Pro Pro Ala Lys Lys Ser Arg
130 135 140
Arg Gly Pro Arg Ser Arg Ser Ser Gln Tyr Arg Gly Val Thr Phe Tyr
145 150 155 160
Arg Arg Thr Gly Arg Trp Glu Ser His Ile Trp Asp Cys Gly Lys Gln
165 170 175
Val Tyr Leu Gly Gly Phe Asp Thr Ala His Ala Ala Ala Arg Ala Tyr
180 185 190
Asp Arg Ala Ala Ile Lys Phe Arg Gly Val Asp Ala Asp Ile Asn Phe
195 200 205
Asn Leu Ser Asp Tyr Glu Asp Asp Met Lys Gln Met Gly Ser Leu Ser
210 215 220
Lys Glu Glu Phe Val His Val Leu Arg Arg Gln Ser Thr Gly Phe Ser
225 230 235 240
Arg Gly Ser Ser Arg Tyr Arg Gly Val Thr Leu His Lys Cys Gly Arg
245 250 255
Trp Glu Ala Arg Met Gly Gln Phe Leu Gly Lys Lys Tyr Ile Tyr Leu
260 265 270
Gly Leu Phe Asp Ser Glu Val Glu Ala Ala Arg Ala Tyr Asp Lys Ala
275 280 285
Ala Ile Lys Cys Asn Gly Arg Glu Ala Val Thr Asn Phe Glu Pro Ser
290 295 300
Thr Tyr His Gly Glu Leu Pro Thr Glu Val Ala Asp Val Asp Leu Asn
305 310 315 320
Leu Ser Ile Ser Gln Pro Ser Pro Gln Arg Asp Lys Asn Ser Cys Leu
325 330 335
Gly Leu Gln Leu His His Gly Pro Phe Glu Gly Ser Glu Leu Lys Lys
340 345 350
Thr Lys Ile Asp Asp Ala Pro Ser Glu Leu Pro Gly Arg Pro Arg Gln
355 360 365
Leu Ser Pro Leu Val Ala Glu His Pro Pro Ala Trp Pro Ala Gln Pro
370 375 380
Pro His Pro Phe Phe Val Phe Thr Asn His Glu Met Ser Ala Ser Gly
385 390 395 400
Asp Leu His Arg Arg Pro Ala Gly Ala Val Pro Ser Trp Ala Trp Gln
405 410 415
Val Ala Ala Ala Ala Pro Pro Pro Ala Ala Leu Pro Ser Ser Ala Ala
420 425 430
Ala Ser Ser Gly Phe Ser Asn Thr Ala Thr Thr Ala Ala Thr Thr Ala
435 440 445
Pro Ser Ala Ser Ser Leu Arg Tyr Cys Pro Pro Pro Pro Pro Pro Ser
450 455 460
Ser His His His Pro Arg
465 470
<210> 4
<211> 1028
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 4
gatttaaatc ccaaacaaat caaaaatttt gttattttat ttcaatctta tttaatccaa 60
atgtattggc aataaacaaa caaggcaaac caaacaccct tctacactgt acccggacaa 120
gtacgtcatg cagatacaaa ttccgcgagg cacgccggac gcgccattcg agtttgaatg 180
aataggagta gacgatcacg ctacgtcggt aacatacact gaggtgtctt ggctgcagcc 240
gagctagatg tcaacagcgt gctcgtgctc gtcgggaagc cgtaggaaac ccggttcaga 300
tggtcacgcc tcctcgcggt ccatcagcag tgacctgtcc cgtttaggaa acggccggag 360
agcaacgtca cactcacacg cgctaatttt ggcctctgcc aggcaacgga acagagcacg 420
ccgagctcac tggtatataa aaaaataatt aatacataac agaaccgggg aaaaggagcc 480
cgaggtttcg tcgtatactc ttttgtgtaa caacaccgct ccctgtatct atctgatctc 540
gtgcggcggt ggagaagtcg acaacaagca cgtgctgaat gttcgtcagg gtcctgcact 600
ggggtgcgta gattattctg tcaggataag tactgcgctc tggcctctgc gcaacatgcg 660
acgccggaga gcgcagaaga tcctctgcaa gaagctgctg gtaattcact tctgaacaaa 720
gatccaaaca gaggaagcgt ctatccacgc atgctagaga cgatgccgat cagtcatcag 780
agcctgaccg aagcgcagca aactgcggct gcgcgcgtaa tttggactgg ctgtttgtta 840
ttgatgacga tgaagcccct acccggcacc gcacctgggt tttctcgatg catgcaccaa 900
tgttttctga tcaccacgcc tggctttcgc atgcaggcta taagcacgcg tcgcgttcaa 960
caacgcatca gcagagagtg tcctaaaagg agccagtctc ataggtcgac tgagagatgg 1020
cgtccgag 1028
<210> 5
<211> 1270
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 5
ctaatagtta gtaatgttat aaaaaatatt tctatcttat taaatttaag aaatacactc 60
acaatgtact atcttatttt aaataaagtg atttctcaca ccaaacaatc taaaatctaa 120
ggtgattcat ctggacgcgt ggtcgcagca ccgtggatgc gcgcggcggc ccgagtttcc 180
cccatgccga gttgccgagt cgacgagcgg cgactcagtc ccagccgccc catggccgcc 240
gccgctcggc acccacagcg tgcttgcagc ggcagatctc ggcacgggac ggcgcggctt 300
ctctctccgt gcgcatcgct tttccccagg tgcacctccc ctcaacacca acgtggccac 360
gccagcgcgc aaccaccgcc tcccgcccgc ttccccaacg caagccttct acacgtgtct 420
ccctccgtgc cgcgaccgac cgcccacggc cctcccacag aatgcgaccg ccccaaccac 480
gtgtcctttc tccatgacta cacccgttct ctcacgctcc cgccattgag attcttctct 540
ctggcgggtg gacagcgctc caaccagtcc cacatgtcat cgggacgaat cctgtgggct 600
tcaacaaaga cgggtgaaat agagcccaga tattttaggc agcccaagag acagagccca 660
tgtggtgggg accacactag gcatcaataa atcagctgcg gaggcggact gccccaccgg 720
ggagcgccac gtgtgtgcgg cacggggatt aggcgataac accggcgtcg tggtcgccgt 780
taccatcaac ctcctcatgg gcccacgcac cttgcccacc tggtaggcca tggacaccac 840
gtgtccaccg cgccgagcaa gagagcaaca cgcgtccaga tcctgccggg acggacgggc 900
ccgatgcgcc ggaccaggac cacgtcgcaa caactgtacc tgtccaggcg ctgcccgtta 960
cggcgacgga cacctgcccc tctgcccctg cccgcctcgc ttccttcctc ttcttccctc 1020
tatctgtatt gccccgaaac cttcgtgccc gtttcgcctc gcgcctgcgt tgctctgcta 1080
tttccatttc cattgcaccc atttccgggg tcgcctgcgt tgcttcgcgt gggagcgtga 1140
ttcgtttagg cctctcgcca atttcggagg gcgacggcga cgtgctttcg gcgggcgggc 1200
agagaaaatc tagagtcttg ggtttgccag acgcgccttt gtgggagaca gacacttgct 1260
ctcgcctcca 1270
<210> 6
<211> 1111
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 6
tacgacagca acgatgcaaa aggtggtcgg ttgctgtcca gccaggcctg ggcagatatt 60
tgaccaccaa agcaccgtga agtccttgct ggacacggga gacgcgacga acacaaccgc 120
acggacggat catggatcga tcactcgcgc tgggtgccac aggtcctccg gtgtggggca 180
tcattcacgc ggcgccgaac caccacacca gctgtgccaa aaacgacgag cattaccgtc 240
gcctcccctc gccgagctac aagtggccgg ccggactggc cattcttccg gcgagccata 300
ccatgtctgg catggcgttc tagatacacc accaactttt tgtcatccca ccccacatcc 360
acatgtgcgt gcagtggacc tttttctttc tttctttcca tcacataccc tttttggttg 420
gcgcggcaca tactagcaat ggggggtgaa aatcggatcg ctttggacaa acaagtgaca 480
atccgcgtga ctgattgtaa catctgctcc cacaagttgg ctcataaaca agatcaacca 540
gccttaacaa caaataaatc gacccggccc cccacaatgg gcgtcgcttt cctccaattc 600
taatttgacc acacccagtt gcccccaacc accaccctag tccggccagc tttcctccca 660
cgcccaccca ttggaagctc aaggccggaa cccaaaccgc catctttcta ctgagaggaa 720
actatggccg agacaacaac tcgcccgagt gaccctaaac tagtccctgc aacttgcgag 780
gccgcagtaa aagtaatggc aagtgtgtgc tgctgctgcc ctggcgattt tcgtcgcgac 840
gagtgaccat gaccggcacc acctccccag ttgtttgttc cagttccagg tatataagcg 900
aagcgcccgc ctctctcctc cagctccacc accaccagct cccaccccca cccatccctt 960
gcccgttcgt cctcctccct cccctctacg gaacagaaca gccctgccca accaaagaac 1020
cttgagagcc ttgaggtggg gtagggtgtg tctgcctggc acagagagta gcagagcagt 1080
gcagagcaaa gcagtcaggc aagcagcagc a 1111
<210> 7
<211> 1145
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 7
tgctctgaaa cattttaata ataccaaaac atgagtttat attcatactt cacccaacaa 60
ccataactca taaccatagt tcaccaccaa cgatgttcaa gtacacaagc aaaatcacaa 120
cacataacca tagttcatca aacaactacg atacatgaat tcatatcacc atgacatacc 180
aagttcaatt catcagacac catgacacta atcatgaccc acaatctaaa aattctcacc 240
aacgaaattc gaaaaccacg atagcttcca atcgaatggt agattcatga aattaaagca 300
tgggcatcat taggatgcct aaccaagtgc tgatagacaa tatttttttc ttattatttc 360
tgtcttggca gtaaaatcta gatattatta ttatgaatta actttttatt atttgtggaa 420
tgtagatttt tttgtaatat caatctctac tatatttaat tgtgagtttc ttcacctctc 480
cacgctcacg cactactatt agcgaatttt gttgtaaaaa aatgtatcag ccagggatta 540
aacccctgcc ccttataaga aggagacaaa caaatttaac cattgcaccg tttaatttta 600
tgttacatat ataagagata ttatatatga tatagtagca atattaatgt gtatgagcac 660
tagaagtaga tattttattt tgtcgaacgg ccgtccggag gcagtagata tcaatggact 720
gaaatgtgga cattatccca cctggtccag atcatcacgg gtacacgtgc cacgctgccg 780
accgatggcc cagctctgaa gctaacgtgt caagtacgcc cgcccccctg cattgcgtcg 840
tgccaatagt ccaagtccca tggcagaacc cagcgctttg tcccgtcccg cctgtgcaca 900
gctagcgccc tctcggaacc gtgtacgtca tcgatcgtgt cacgcgcgcg acggaagaag 960
ttaagcacgc tgcagcaacg cggcagagca catccgtagt gtgtgcatgc atcacagtca 1020
cacacacaca gcagaagaag aagaaaccga acgagggttt agctagcaaa ataaacagaa 1080
gcaagcaagc tagctagagc taaggatcga gatcgagatc gaccgaccga cgacgatcag 1140
ctagc 1145
<210> 8
<211> 1520
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 8
caccagttcg ccaggtagtt agcagcccag ttgcgactgg ggatcggtgg cgggctgccg 60
cttgcgagtt gtaagcttgg aggggagggg agcaggagca ggagatgcag ttggatctga 120
acgtggccga ggcgccgccg ccggtggaga tggaggcgag cgactcgggg tcgtcggtgc 180
tgaacgcgtc ggaagcggcg tcggcgggcg gcgcgcccgc gccggcggag gagggatcta 240
gctcaacgcc ggccgtgctg gagttcagca tcctcatccg gagcgatagc gacgcggccg 300
gcgcggacga ggacgaggac gccacgccat cgcctcctcc tcgccaccgc caccagcacc 360
agcagcagct cgtgacccgc gagctgttcc cggccggcgc cggtccgccg gccccgacgc 420
cgcggcattg ggccgagctc ggcttcttcc gcgccgacct gcagcagcaa caggcgccgg 480
gccccaggat cgtgccgcac ccacacgccg cgccgccgcc ggccaagaag agccgccgcg 540
gcccgcgctc ccgcagctcg cagtaccgcg gcgtcacctt ctaccgccgc acaggccgct 600
gggagtccca catctggtca gtactaccac tgtctacaac tagccacacc acaccgattg 660
cttccgactc tcattaattt ctgacacaaa ctctccgtct tcctcctctt ctcccgcgac 720
gcagggattg cggcaagcag gtgtacctag gtgagcaaga gcagatctct tttgcgttcc 780
caaagatttt tcccctttta gttccttatc ccatcccatc tcgaatggcc tagctaaccg 840
attcagtggt ggtccggctg ctggccgata tacgcaggtg gattcgacac cgctcacgcc 900
gctgcaaggc acgcactgga ctggacgccc agaattcttc gtcatgtgag tctctgaccg 960
aattgattga ttaacgagtc tctggctcct ggaactcgca gggcgtacga ccgggcggcg 1020
atcaagttcc gcggcgtcga cgccgacatc aacttcaacc tcagcgacta cgaggacgac 1080
atgaagcaga tggggagcct gtccaaggag gagttcgtgc acgtcctgcg ccgtcagagc 1140
accggcttct cgagaggcag ctccaggtac agaggcgtca ccctgcacaa gtgcggccgc 1200
tgggaggcgc gcatggggca gttcctcggc aagaagtaag aaccaaccaa cgcttctttt 1260
ctttttcttt tttttatagc atgcagtgat gattcaacct tagttgtgcc tttcctccta 1320
atcctatata tgtaggattt agtactggtt gactatataa gtatatatgt attgttcagt 1380
aaaagtatac ataggttagc tgcatgttta tgtatgtagc tggttgtttc aatcagaaga 1440
taaaaaaaaa gggaagtagt ggctagggaa ttcctccaat cctcaccggt gggaacgccg 1500
tgcttgggtg caggtacata 1520
<210> 9
<211> 1527
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 9
caccagttcg ccaggtagtt agcagcccag ttgcgactgg ggatcggtgg cgggctgccg 60
cttgcgagtt gtaagcttgg aggggagggg agcaggagca ggagatgcag ctggatctga 120
acgtggccga ggcgccgccg ccggtggaga tggaggcgag cgactcgggg tcgtcggtgc 180
tgaacgcgtc ggaagcggcg tcggcgggcg gcgcgcccgc gccggcggag gaggggtcca 240
gctcaacgcc ggccgcgctg gagttcagca tcctcatccg gagcgacagc gacgtggccg 300
gcgcggacga ggacgaggac gccacgccgt cgcctcctcc tcgccaccgc caccagcacc 360
agcagcagct cgtgacccgc gagctgttcc cggctggcgc cggcccgccg gccccggcgc 420
cgcggcattg ggccgagctc ggcttcttcc gcgccgacct gcagcagcaa caggcgccgg 480
gccccaggat cgtgccgcac ccacacgccg cgccgccgcc ggccaagaag agccgccgcg 540
gcccgcgctc ccgcagctcg cagtaccgcg gcgtcacctt ctaccgccgc acaggccgct 600
gggagtccca catctggtca gtactaccac tgtctacaac tagccacacc acaccgattg 660
cttccgactc tcattaattt ctgacacaaa ctctccgtct tcctcctctt cttccacgac 720
gcagggattg cggcaagcag gtgtacctag gtgagcaaga gcagatctct tttgcgttcc 780
caaagatttt tcccctttta gctcctcatc ccatcccatc tcgaatggcc tagctaaccg 840
attcagtggt ggtccggctg ctggccgata tacgcaggtg gattcgacac cgctcacgcc 900
gctgcaaggc acgcactgga ctggacgccc agaattcttc gtcatgtgag tctctgaccg 960
gattagttga ttaacgagtc tctgggtcct ggaactcgca gggcgtacga ccgggcggcg 1020
atcaagttcc gcggcgtgga cgccgacatc aacttcaacc tcagcgacta cgaggacgac 1080
atgaagcaga tggggagcct gtccaaggag gagttcgtgc acgtcctgcg ccgccagagc 1140
accggcttct cgagaggcag ctccaggtac agaggcgtca ccctgcacaa gtgcggccgc 1200
tgggaggcgc gcatggggca gttcctcggc aagaagtaag aaccaacaac caacgcttct 1260
tttctttttc tttttttata gcatgcagat gatgattcaa ccttagttgt gcctctcctc 1320
ctaatcctat gtaggattta gtactggttg actacatatc tactattgtt atatatgtat 1380
tgttcagtaa aagtatacat aggttagctg catgtttatg tatgtagctg gttgtttcaa 1440
tcagaagata aaaaaaaggg aagtagtggc tagggaattc ctccaatcct caccggtggg 1500
aacgccgtgc ttgggtgcag gtacata 1527
<210> 10
<211> 1615
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 10
gtggctaggg aattcctcca atcctcaccg gtgggaacgc cgtgcttggg tgcaggtaca 60
tataccttgg gctattcgac agcgaagtag aggctgcaag gttcttcatc ttggattctg 120
ccgttcatat atgcataatc atgtctttta atttccaaag ggttgagtac cgactcgatt 180
cctcttcgtg tcttttttct ttctttcttc gaaatccaga gcctacgaca aggccgccat 240
caaatgcaat ggcagagagg ccgtgacgaa cttcgagccg agcacgtatc acggggagct 300
gccgactgaa ggtacgtatt ttctttctgc atatatatat cttcaggtat tattggctat 360
taaactgctt ggatcttact gcttcttctg cagttgctga tgtcgatctg aacctgagca 420
tatctcagcc gagcccccaa agagacaaga acagctgcct aggtctgcag ctccaccacg 480
gaccattcga gggctccgaa ctgaagaaaa ccaaggcaag cgctaacgat agatatacct 540
tgacaagcta gtatcaaaca aaaccagtaa aaaaagttta ctttcttgtc gaatttcatt 600
gcctacctga tgtacgtact tgtgcttctg cacaaaataa cgaaatcctt ttgccctctg 660
atgatgatgc agatcgacga tgctccctct gagctaccgg gccgccctcg tcagctgtct 720
cctctcgtgg ctgagcatcc gccggcctgg cctgcgcagc cgcctcaccc cttcttcgtc 780
ttcacaaacc atgaggttag gtgacagcta ctgatcgaga tgcagcagca gttcaaacct 840
gtctgttcca aggaccttta ggccggatta ccaaatcatc ggtcaactgt cctgtctgtt 900
atatatttat gtgttaattt ataatacaag tgtgactatt tttcaaacct tccttcaaaa 960
tgcatgaaaa gagttttttt taacgaaagg cgaaaagaaa atatgatact tgggacagga 1020
gcaagcttgg atcatcagaa agtattatta attaggatca ctgagctgtt cattttgttc 1080
ttgagtcaat cctaatcgta ctatgtcagt gaatgaactt gtgttgcacc aatgcagatg 1140
agtgcatcag gagatctcca caggaggcct gcaggggctg ttcccagctg ggcatggcag 1200
gtggcagcag cagctcctcc tcctgccgcc ctgccgtcgt ccgctgcagc atcatcagga 1260
ttctccaaca ccgccacgac agctgccacc accgccccat cggcctcctc cctccggtac 1320
tgcccgccgc cgccgccgcc gtcgagccat caccatcccc gctgagagaa tcaagaagcc 1380
gcactgtaaa tctgccggga agctagcatt ttccccccgg cccctccccc tctccgggcg 1440
ttgcgacttt ttcagttttg cgccgccggc cggggtggtg gtttcttgta gccgatcgat 1500
tggattcctc gtattactgc tgcttacact cccaattaag tgaaaaaaaa acgctcctct 1560
actctttaca ctacacacac tgttagctga tcgattggac gtacttgcta gctgc 1615
<210> 11
<211> 1618
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 11
gtggctaggg aattcctcca atcctcaccg gtgggaacgc cgtgcttggg tgcaggtaca 60
tataccttgg gctattcgac agcgaagtag aggctgcaag gttcttcatc ttggattctg 120
ccgttcatat atgcataatc atgtctttta atttccaaag ggttgagtac cgactcgatt 180
cctcttcgtg tcttttttct ttctttcttc gaaatccaga gcctacgaca aggccgccat 240
caaatgcaat ggcagagagg ccgtgacgaa cttcgagccg agcacgtatc acggggagct 300
gccgactgaa ggtacgtttt tctttctgca tatatataat cttcaggtat tattattggc 360
tattaaactg cttggatttt actgcttctt ctgcagttgc tgatgtcgat ctgaacctga 420
gaatatctca gccgagctcc caaagagaca agaacagctg cctaggtctg caggtccacc 480
acggaccatt cgagggctcc gaactgaaga aaaccaaggc aagcgctaac gatagatata 540
ccttgacaag ctagtatcaa acaaaaccag taaaaaaagt ttactttctt gtcgaatttc 600
attgcctacc tgatgtacgt acttgtgctt ctgcacaaaa taacgaaatc cttttgccct 660
ctgatgatga tgcagatcga cgatgctccc tctgagctac cgggccgccc tcgtcagctg 720
tctcctctcg tggctgagca tccgccggcc tggcctgcgc agccgcctca ccccttcttc 780
gtcttcacaa accatgaggt taggtgacag ctactgatcg agatgcagca gcagttcaaa 840
cctgtctgtt ccaaggacct ttaggccgga ttaccaaatc atcggtcaac tgtcctgtct 900
gttatatatt tatgtgttaa tttataatac aagtgtgact atttttcaaa ccttccttca 960
aaatgcatga aaagagtttt ttttaacgaa aggcgaaaag aaaatatgat acttgggaca 1020
ggagcaagct tggatcatca gaaagtatta ttaattagga tcactgagct gttcattttg 1080
ttcttgagtc aatcctaatc gtactatgtc agtgaatgaa cttgtgttgc accaatgcag 1140
atgagtgcat caggagatct ccacaggagg cctgcagggg ctgttcccag ctgggcatgg 1200
caggtggcag cagcagctcc tcctcctgcc gccctgccgt cgtccgctgc agcatcatca 1260
ggattctcca acaccgccac gacagctgcc accaccgccc catcggcctc ctccctccgg 1320
tactgcccgc cgccgccgcc gccgtcgagc catcaccatc cccgctgaga gaatcaagaa 1380
gccgcactgt aaatctgccg ggaagctagc attttccccc cggcccctcc ccctctccgg 1440
gcgttgcgac tttttcagtt ttgcgccgcc ggccggggtg gtggtttctt gtagccgatc 1500
gattggattc ctcgtattac tgctgcttac actcccaatt aagtgaaaaa aaaacgctcc 1560
tctactcttt acactacaca cactgttagc tgatcgattg gacgtacttg ctagctgc 1618
<210> 12
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 12
caccagttcg ccaggtagtt 20
<210> 13
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 13
tatgtacctg cacccaagca 20
<210> 14
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 14
gtggctaggg aattcctcca at 22
<210> 15
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 15
gcagctagca agtacgtcca 20
Claims (3)
1.ZmRAP2.7基因在调控玉米种子人工加速老化萌发率和种子耐贮性中的应用,其特征在于:种子经过模拟长时间贮藏的人工加速老化处理后,进行种子萌发时,缺失该基因的功能,会降低种子的萌发率;所述ZmRAP2.7的基因是如SEQ ID NO:1所示的DNA序列或如SEQID NO:2所示的cDNA序列;所述ZmRAP2.7基因的编码蛋白是如SEQ ID NO:3所示的蛋白序列。
2.根据权利要求1所述应用,其特征在于:在种子经过人工加速老化处理后,进行萌发时,缺失该基因则不能产生蛋白来抑制下游基因ZmABI5、ZmPP2C、ZmPYL3和ZCN9的转录表达,种子无法降低ABA信号并开始正常萌发,因而最终降低种子萌发能力;所述ZmABI5的启动子如SEQ ID NO:4所示;所述ZmPP2C的启动子如SEQ ID NO:5所示;所述ZmPYL3的启动子如SEQ ID NO:6所示;所述ZCN9的启动子如SEQ ID NO:7所示。
3.根据权利要求1所述应用,其特征在于:抑制所述基因ZmRAP2.7的表达,获得人工加速老化发芽率降低的材料,用于挖掘更多与种子萌发相关的生物合成途径,以及挖掘其分子机理,为高品质玉米的选育奠定基础。
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202210687849.5A CN114836441B (zh) | 2022-06-17 | 2022-06-17 | 玉米种子耐贮性相关的ZmRAP2.7基因及其功能标记与应用 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202210687849.5A CN114836441B (zh) | 2022-06-17 | 2022-06-17 | 玉米种子耐贮性相关的ZmRAP2.7基因及其功能标记与应用 |
Publications (2)
Publication Number | Publication Date |
---|---|
CN114836441A CN114836441A (zh) | 2022-08-02 |
CN114836441B true CN114836441B (zh) | 2024-05-07 |
Family
ID=82575152
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202210687849.5A Active CN114836441B (zh) | 2022-06-17 | 2022-06-17 | 玉米种子耐贮性相关的ZmRAP2.7基因及其功能标记与应用 |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN114836441B (zh) |
Families Citing this family (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN115992149B (zh) * | 2022-11-17 | 2024-04-23 | 西北农林科技大学 | 玉米ZmRAFS基因用于提高植物耐涝性的应用 |
Family Cites Families (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CA2575388C (en) * | 2004-07-29 | 2013-04-30 | Pioneer Hi-Bred International, Inc. | Methods and compositions for modulating flowering and maturity in plants |
-
2022
- 2022-06-17 CN CN202210687849.5A patent/CN114836441B/zh active Active
Also Published As
Publication number | Publication date |
---|---|
CN114836441A (zh) | 2022-08-02 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
Brocard et al. | Regulation and role of the Arabidopsis abscisic acid-insensitive 5 gene in abscisic acid, sugar, and stress response | |
Zhang et al. | Differential gene expression analysis of Yunnan red pear, Pyrus pyrifolia, during fruit skin coloration | |
Chinnusamy et al. | ICE1: a regulator of cold-induced transcriptome and freezing tolerance in Arabidopsis | |
Li et al. | TAC4 controls tiller angle by regulating the endogenous auxin content and distribution in rice | |
Li et al. | Characterization and fine mapping of a novel rice narrow leaf mutant nal9 | |
Grasser et al. | Transcript elongation factor TFIIS is involved in Arabidopsis seed dormancy | |
US20120317676A1 (en) | Method of producing plants having enhanced transpiration efficiency and plants produced therefrom | |
Ueda et al. | qUVR-10, a major quantitative trait locus for ultraviolet-B resistance in rice, encodes cyclobutane pyrimidine dimer photolyase | |
CN107418957B (zh) | 控制水稻耐冷基因cold2及其应用 | |
CN110982826B (zh) | 一种控制玉米籽粒发育基因ZmDek605的克隆、功能研究及标记挖掘 | |
CN114836441B (zh) | 玉米种子耐贮性相关的ZmRAP2.7基因及其功能标记与应用 | |
Zheng et al. | Mutation types of CYP71P1 cause different phenotypes of mosaic spot lesion and premature leaf senescence in rice | |
CN112592393B (zh) | 水稻早衰控制基因es1-3及其应用 | |
Way et al. | Identification of differentially expressed genes in wheat undergoing gradual water deficit stress using a subtractive hybridisation approach | |
CN108570474A (zh) | 水稻花发育基因eh1及其应用 | |
CN113186195A (zh) | 玉米籽粒穗发芽相关突变基因ZmVp#5的功能标记及其应用 | |
US6635811B1 (en) | Pre-harvest sprouting | |
CN111153980B (zh) | 一种植物粒型相关蛋白OsSDSG及其编码基因与应用 | |
CN109121420B (zh) | 耐寒植物 | |
Przybecki et al. | The isolation of cDNA clones from cucumber (Cucumis sativus L.) floral buds coming from plants differing in sex | |
Lechner et al. | A gene trap Dissociation insertion line, associated with a RING-H2 finger gene, shows tissue specific and developmental regulated expression of the gene in Arabidopsis | |
CN113151323A (zh) | 一种控制玉米籽粒发育基因ZmRH4的克隆、功能研究及标记挖掘 | |
Ming-Li et al. | Cloning and SNP analysis of TT1 gene in Brassica juncea | |
Lv et al. | BOS1 is a basic helix–loop–helix transcription factor involved in regulating panicle development in rice | |
CN111218457A (zh) | 一种水稻mit2基因及其编码蛋白与应用 |
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 |