CN109880832A - 水稻叶片叶夹角突变基因pla1及其应用 - Google Patents
水稻叶片叶夹角突变基因pla1及其应用 Download PDFInfo
- Publication number
- CN109880832A CN109880832A CN201910281061.2A CN201910281061A CN109880832A CN 109880832 A CN109880832 A CN 109880832A CN 201910281061 A CN201910281061 A CN 201910281061A CN 109880832 A CN109880832 A CN 109880832A
- Authority
- CN
- China
- Prior art keywords
- pla1
- ala
- rice
- gene
- mutant
- 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.)
- Withdrawn
Links
- 101150059175 PLA1 gene Proteins 0.000 title claims abstract description 68
- 101100058850 Oryza sativa subsp. japonica CYP78A11 gene Proteins 0.000 title claims abstract description 66
- 101100083853 Homo sapiens POU2F3 gene Proteins 0.000 title claims abstract description 63
- 235000007164 Oryza sativa Nutrition 0.000 title claims abstract description 59
- 235000009566 rice Nutrition 0.000 title claims abstract description 57
- 240000007594 Oryza sativa Species 0.000 title description 3
- 241000209094 Oryza Species 0.000 claims abstract description 57
- 108090000623 proteins and genes Proteins 0.000 claims abstract description 32
- 102000004169 proteins and genes Human genes 0.000 claims abstract description 5
- 239000002773 nucleotide Substances 0.000 claims abstract description 4
- 125000003729 nucleotide group Chemical group 0.000 claims abstract description 4
- 238000009395 breeding Methods 0.000 claims description 4
- 230000001488 breeding effect Effects 0.000 claims description 4
- 238000001514 detection method Methods 0.000 claims description 4
- 125000003275 alpha amino acid group Chemical group 0.000 claims 1
- 241000196324 Embryophyta Species 0.000 abstract description 12
- 150000001413 amino acids Chemical group 0.000 abstract description 6
- 101150053185 P450 gene Proteins 0.000 abstract description 3
- ONIBWKKTOPOVIA-UHFFFAOYSA-N Proline Natural products OC(=O)C1CCCN1 ONIBWKKTOPOVIA-UHFFFAOYSA-N 0.000 abstract description 3
- AYFVYJQAPQTCCC-UHFFFAOYSA-N Threonine Natural products CC(O)C(N)C(O)=O AYFVYJQAPQTCCC-UHFFFAOYSA-N 0.000 abstract description 3
- 239000004473 Threonine Substances 0.000 abstract description 3
- 238000005516 engineering process Methods 0.000 abstract description 2
- 206010064571 Gene mutation Diseases 0.000 abstract 1
- 229920003266 Leaf® Polymers 0.000 description 81
- 108020004414 DNA Proteins 0.000 description 42
- 102100026466 POU domain, class 2, transcription factor 3 Human genes 0.000 description 39
- 210000004027 cell Anatomy 0.000 description 17
- 230000014509 gene expression Effects 0.000 description 13
- 239000006228 supernatant Substances 0.000 description 8
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 6
- 238000004458 analytical method Methods 0.000 description 6
- 230000002792 vascular Effects 0.000 description 6
- 101100289385 Arabidopsis thaliana PLA1 gene Proteins 0.000 description 5
- 238000006243 chemical reaction Methods 0.000 description 5
- 239000002299 complementary DNA Substances 0.000 description 5
- 239000007788 liquid Substances 0.000 description 5
- 238000012163 sequencing technique Methods 0.000 description 5
- 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 4
- 101100340782 Oryza sativa subsp. indica ILI1 gene Proteins 0.000 description 4
- 101100340783 Oryza sativa subsp. japonica BHLH154 gene Proteins 0.000 description 4
- 230000015572 biosynthetic process Effects 0.000 description 4
- 238000005119 centrifugation Methods 0.000 description 4
- 238000011161 development Methods 0.000 description 4
- 230000018109 developmental process Effects 0.000 description 4
- VPZXBVLAVMBEQI-UHFFFAOYSA-N glycyl-DL-alpha-alanine Natural products OC(=O)C(C)NC(=O)CN VPZXBVLAVMBEQI-UHFFFAOYSA-N 0.000 description 4
- 239000000047 product Substances 0.000 description 4
- 210000001519 tissue Anatomy 0.000 description 4
- PLUBXMRUUVWRLT-UHFFFAOYSA-N Ethyl methanesulfonate Chemical compound CCOS(C)(=O)=O PLUBXMRUUVWRLT-UHFFFAOYSA-N 0.000 description 3
- 101100125887 Oryza sativa subsp. japonica ILI4 gene Proteins 0.000 description 3
- 102000052708 Recessive Genes Human genes 0.000 description 3
- 108700005079 Recessive Genes Proteins 0.000 description 3
- 102000006382 Ribonucleases Human genes 0.000 description 3
- 108010083644 Ribonucleases Proteins 0.000 description 3
- 230000003321 amplification Effects 0.000 description 3
- 210000000349 chromosome Anatomy 0.000 description 3
- 230000006872 improvement Effects 0.000 description 3
- 238000013507 mapping Methods 0.000 description 3
- 229910052757 nitrogen Inorganic materials 0.000 description 3
- 238000003199 nucleic acid amplification method Methods 0.000 description 3
- 238000012545 processing Methods 0.000 description 3
- 238000011160 research Methods 0.000 description 3
- 230000019491 signal transduction Effects 0.000 description 3
- 238000003786 synthesis reaction Methods 0.000 description 3
- IXVMHGVQKLDRKH-YEJCTVDLSA-N (22s,23s)-epibrassinolide Chemical compound C1OC(=O)[C@H]2C[C@H](O)[C@H](O)C[C@]2(C)[C@H]2CC[C@]3(C)[C@@H]([C@H](C)[C@H](O)[C@@H](O)[C@H](C)C(C)C)CC[C@H]3[C@@H]21 IXVMHGVQKLDRKH-YEJCTVDLSA-N 0.000 description 2
- SVBXIUDNTRTKHE-CIUDSAMLSA-N Ala-Arg-Glu Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(O)=O)C(O)=O SVBXIUDNTRTKHE-CIUDSAMLSA-N 0.000 description 2
- BGNLUHXLSAQYRQ-FXQIFTODSA-N Ala-Glu-Gln Chemical compound C[C@H](N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCC(N)=O)C(O)=O BGNLUHXLSAQYRQ-FXQIFTODSA-N 0.000 description 2
- OEVCHROQUIVQFZ-YTLHQDLWSA-N Ala-Thr-Ala Chemical compound C[C@H](N)C(=O)N[C@@H]([C@H](O)C)C(=O)N[C@@H](C)C(O)=O OEVCHROQUIVQFZ-YTLHQDLWSA-N 0.000 description 2
- IXVMHGVQKLDRKH-VRESXRICSA-N Brassinolide Natural products O=C1OC[C@@H]2[C@@H]3[C@@](C)([C@H]([C@@H]([C@@H](O)[C@H](O)[C@H](C(C)C)C)C)CC3)CC[C@@H]2[C@]2(C)[C@@H]1C[C@H](O)[C@H](O)C2 IXVMHGVQKLDRKH-VRESXRICSA-N 0.000 description 2
- LZZYPRNAOMGNLH-UHFFFAOYSA-M Cetrimonium bromide Chemical compound [Br-].CCCCCCCCCCCCCCCC[N+](C)(C)C LZZYPRNAOMGNLH-UHFFFAOYSA-M 0.000 description 2
- 102000004190 Enzymes Human genes 0.000 description 2
- 108090000790 Enzymes Proteins 0.000 description 2
- 241000588724 Escherichia coli Species 0.000 description 2
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 2
- 108091092584 GDNA Proteins 0.000 description 2
- XOWMDXHFSBCAKQ-SRVKXCTJSA-N Leu-Ser-Leu Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@H](C(O)=O)CC(C)C XOWMDXHFSBCAKQ-SRVKXCTJSA-N 0.000 description 2
- 101710163270 Nuclease Proteins 0.000 description 2
- 240000002582 Oryza sativa Indica Group Species 0.000 description 2
- 238000011529 RT qPCR Methods 0.000 description 2
- 108010078114 alanyl-tryptophyl-alanine Proteins 0.000 description 2
- AVKUERGKIZMTKX-NJBDSQKTSA-N ampicillin Chemical compound C1([C@@H](N)C(=O)N[C@H]2[C@H]3SC([C@@H](N3C2=O)C(O)=O)(C)C)=CC=CC=C1 AVKUERGKIZMTKX-NJBDSQKTSA-N 0.000 description 2
- 229960000723 ampicillin Drugs 0.000 description 2
- 210000001367 artery Anatomy 0.000 description 2
- 230000033228 biological regulation Effects 0.000 description 2
- 230000001413 cellular effect Effects 0.000 description 2
- 235000013339 cereals Nutrition 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 229960000935 dehydrated alcohol Drugs 0.000 description 2
- 230000002255 enzymatic effect Effects 0.000 description 2
- 238000010195 expression analysis Methods 0.000 description 2
- 239000000284 extract Substances 0.000 description 2
- 238000012252 genetic analysis Methods 0.000 description 2
- 108010050848 glycylleucine Proteins 0.000 description 2
- 239000005556 hormone Substances 0.000 description 2
- 229940088597 hormone Drugs 0.000 description 2
- 239000006210 lotion Substances 0.000 description 2
- 238000000034 method Methods 0.000 description 2
- 238000002156 mixing Methods 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 238000002703 mutagenesis Methods 0.000 description 2
- 231100000350 mutagenesis Toxicity 0.000 description 2
- 239000012188 paraffin wax Substances 0.000 description 2
- 238000004321 preservation Methods 0.000 description 2
- 238000011002 quantification Methods 0.000 description 2
- 230000004044 response Effects 0.000 description 2
- 238000010839 reverse transcription Methods 0.000 description 2
- 108010026333 seryl-proline Proteins 0.000 description 2
- 238000005406 washing Methods 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- 101150084750 1 gene Proteins 0.000 description 1
- YYSWCHMLFJLLBJ-ZLUOBGJFSA-N Ala-Ala-Ser Chemical compound C[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@@H](CO)C(O)=O YYSWCHMLFJLLBJ-ZLUOBGJFSA-N 0.000 description 1
- SDMAQFGBPOJFOM-GUBZILKMSA-N Ala-Arg-Arg Chemical compound NC(=N)NCCC[C@H](NC(=O)[C@@H](N)C)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O SDMAQFGBPOJFOM-GUBZILKMSA-N 0.000 description 1
- PBAMJJXWDQXOJA-FXQIFTODSA-N Ala-Asp-Arg Chemical compound C[C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@H](C(O)=O)CCCN=C(N)N PBAMJJXWDQXOJA-FXQIFTODSA-N 0.000 description 1
- GSCLWXDNIMNIJE-ZLUOBGJFSA-N Ala-Asp-Asn Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(N)=O)C(O)=O GSCLWXDNIMNIJE-ZLUOBGJFSA-N 0.000 description 1
- WDIYWDJLXOCGRW-ACZMJKKPSA-N Ala-Asp-Glu Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(O)=O WDIYWDJLXOCGRW-ACZMJKKPSA-N 0.000 description 1
- BTYTYHBSJKQBQA-GCJQMDKQSA-N Ala-Asp-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](CC(=O)O)NC(=O)[C@H](C)N)O BTYTYHBSJKQBQA-GCJQMDKQSA-N 0.000 description 1
- IKKVASZHTMKJIR-ZKWXMUAHSA-N Ala-Asp-Val Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](C(C)C)C(O)=O IKKVASZHTMKJIR-ZKWXMUAHSA-N 0.000 description 1
- KMGOBAQSCKTBGD-DLOVCJGASA-N Ala-His-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@@H](NC(=O)[C@H](C)N)CC1=CN=CN1 KMGOBAQSCKTBGD-DLOVCJGASA-N 0.000 description 1
- LBYMZCVBOKYZNS-CIUDSAMLSA-N Ala-Leu-Asp Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(O)=O)C(O)=O LBYMZCVBOKYZNS-CIUDSAMLSA-N 0.000 description 1
- OYJCVIGKMXUVKB-GARJFASQSA-N Ala-Leu-Pro Chemical compound C[C@@H](C(=O)N[C@@H](CC(C)C)C(=O)N1CCC[C@@H]1C(=O)O)N OYJCVIGKMXUVKB-GARJFASQSA-N 0.000 description 1
- XUCHENWTTBFODJ-FXQIFTODSA-N Ala-Met-Ala Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](C)C(O)=O XUCHENWTTBFODJ-FXQIFTODSA-N 0.000 description 1
- NEBFIUZIGRTIFY-BJDJZHNGSA-N Ala-Met-Ser-Arg Chemical compound CSCC[C@H](NC(=O)[C@H](C)N)C(=O)N[C@@H](CO)C(=O)N[C@H](C(O)=O)CCCNC(N)=N NEBFIUZIGRTIFY-BJDJZHNGSA-N 0.000 description 1
- DRARURMRLANNLS-GUBZILKMSA-N Ala-Met-Val Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](C(C)C)C(O)=O DRARURMRLANNLS-GUBZILKMSA-N 0.000 description 1
- XRUJOVRWNMBAAA-NHCYSSNCSA-N Ala-Phe-Ala Chemical compound OC(=O)[C@H](C)NC(=O)[C@@H](NC(=O)[C@@H](N)C)CC1=CC=CC=C1 XRUJOVRWNMBAAA-NHCYSSNCSA-N 0.000 description 1
- MAZZQZWCCYJQGZ-GUBZILKMSA-N Ala-Pro-Arg Chemical compound [H]N[C@@H](C)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CCCNC(N)=N)C(O)=O MAZZQZWCCYJQGZ-GUBZILKMSA-N 0.000 description 1
- QOIGKCBMXUCDQU-KDXUFGMBSA-N Ala-Thr-Pro Chemical compound C[C@H]([C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](C)N)O QOIGKCBMXUCDQU-KDXUFGMBSA-N 0.000 description 1
- IETUUAHKCHOQHP-KZVJFYERSA-N Ala-Thr-Val Chemical compound CC(C)[C@H](NC(=O)[C@@H](NC(=O)[C@H](C)N)[C@@H](C)O)C(O)=O IETUUAHKCHOQHP-KZVJFYERSA-N 0.000 description 1
- YFWTXMRJJDNTLM-LSJOCFKGSA-N Arg-Ala-His Chemical compound C[C@@H](C(=O)N[C@@H](CC1=CN=CN1)C(=O)O)NC(=O)[C@H](CCCN=C(N)N)N YFWTXMRJJDNTLM-LSJOCFKGSA-N 0.000 description 1
- YUIGJDNAGKJLDO-JYJNAYRXSA-N Arg-Arg-Tyr Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O YUIGJDNAGKJLDO-JYJNAYRXSA-N 0.000 description 1
- PBSOQGZLPFVXPU-YUMQZZPRSA-N Arg-Glu-Gly Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(O)=O)C(=O)NCC(O)=O PBSOQGZLPFVXPU-YUMQZZPRSA-N 0.000 description 1
- UBCPNBUIQNMDNH-NAKRPEOUSA-N Arg-Ile-Ala Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](C)C(O)=O UBCPNBUIQNMDNH-NAKRPEOUSA-N 0.000 description 1
- YBZMTKUDWXZLIX-UWVGGRQHSA-N Arg-Leu-Gly Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(C)C)C(=O)NCC(O)=O YBZMTKUDWXZLIX-UWVGGRQHSA-N 0.000 description 1
- LCBSSOCDWUTQQV-SDDRHHMPSA-N Arg-Met-Pro Chemical compound CSCC[C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](CCCN=C(N)N)N LCBSSOCDWUTQQV-SDDRHHMPSA-N 0.000 description 1
- IOTKDTZEEBZNCM-UGYAYLCHSA-N Asn-Asn-Ile Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O IOTKDTZEEBZNCM-UGYAYLCHSA-N 0.000 description 1
- UYXXMIZGHYKYAT-NHCYSSNCSA-N Asn-His-Val Chemical compound CC(C)[C@@H](C(=O)O)NC(=O)[C@H](CC1=CN=CN1)NC(=O)[C@H](CC(=O)N)N UYXXMIZGHYKYAT-NHCYSSNCSA-N 0.000 description 1
- LANZYLJEHLBUPR-BPUTZDHNSA-N Asn-Met-Trp Chemical compound CSCC[C@@H](C(=O)N[C@@H](CC1=CNC2=CC=CC=C21)C(=O)O)NC(=O)[C@H](CC(=O)N)N LANZYLJEHLBUPR-BPUTZDHNSA-N 0.000 description 1
- KRXIWXCXOARFNT-ZLUOBGJFSA-N Asp-Ala-Ala Chemical compound OC(=O)[C@H](C)NC(=O)[C@H](C)NC(=O)[C@@H](N)CC(O)=O KRXIWXCXOARFNT-ZLUOBGJFSA-N 0.000 description 1
- KVMPVNGOKHTUHZ-GCJQMDKQSA-N Asp-Ala-Thr Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](C)C(=O)N[C@@H]([C@@H](C)O)C(O)=O KVMPVNGOKHTUHZ-GCJQMDKQSA-N 0.000 description 1
- VIRHEUMYXXLCBF-WDSKDSINSA-N Asp-Gly-Glu Chemical compound [H]N[C@@H](CC(O)=O)C(=O)NCC(=O)N[C@@H](CCC(O)=O)C(O)=O VIRHEUMYXXLCBF-WDSKDSINSA-N 0.000 description 1
- SNDBKTFJWVEVPO-WHFBIAKZSA-N Asp-Gly-Ser Chemical compound [H]N[C@@H](CC(O)=O)C(=O)NCC(=O)N[C@@H](CO)C(O)=O SNDBKTFJWVEVPO-WHFBIAKZSA-N 0.000 description 1
- KPNUCOPMVSGRCR-DCAQKATOSA-N Asp-His-Arg Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O KPNUCOPMVSGRCR-DCAQKATOSA-N 0.000 description 1
- GPPIDDWYKJPRES-YDHLFZDLSA-N Asp-Phe-Val Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](C(C)C)C(O)=O GPPIDDWYKJPRES-YDHLFZDLSA-N 0.000 description 1
- AHWRSSLYSGLBGD-CIUDSAMLSA-N Asp-Pro-Glu Chemical compound OC(=O)C[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CCC(O)=O)C(O)=O AHWRSSLYSGLBGD-CIUDSAMLSA-N 0.000 description 1
- MGSVBZIBCCKGCY-ZLUOBGJFSA-N Asp-Ser-Ser Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CO)C(O)=O MGSVBZIBCCKGCY-ZLUOBGJFSA-N 0.000 description 1
- RSMZEHCMIOKNMW-GSSVUCPTSA-N Asp-Thr-Thr Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O RSMZEHCMIOKNMW-GSSVUCPTSA-N 0.000 description 1
- XWKBWZXGNXTDKY-ZKWXMUAHSA-N Asp-Val-Ala Chemical compound OC(=O)[C@H](C)NC(=O)[C@H](C(C)C)NC(=O)[C@@H](N)CC(O)=O XWKBWZXGNXTDKY-ZKWXMUAHSA-N 0.000 description 1
- GIKOVDMXBAFXDF-NHCYSSNCSA-N Asp-Val-Leu Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC(C)C)C(O)=O GIKOVDMXBAFXDF-NHCYSSNCSA-N 0.000 description 1
- QOJJMJKTMKNFEF-ZKWXMUAHSA-N Asp-Val-Ser Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@H](C(C)C)NC(=O)[C@@H](N)CC(O)=O QOJJMJKTMKNFEF-ZKWXMUAHSA-N 0.000 description 1
- 101150038693 BRI1 gene Proteins 0.000 description 1
- 101150060196 BZR1 gene Proteins 0.000 description 1
- 102000012410 DNA Ligases Human genes 0.000 description 1
- 108010061982 DNA Ligases Proteins 0.000 description 1
- 238000001712 DNA sequencing Methods 0.000 description 1
- 108010014303 DNA-directed DNA polymerase Proteins 0.000 description 1
- 102000016928 DNA-directed DNA polymerase Human genes 0.000 description 1
- REJJNXODKSHOKA-ACZMJKKPSA-N Gln-Ala-Asp Chemical compound C[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)O)NC(=O)[C@H](CCC(=O)N)N REJJNXODKSHOKA-ACZMJKKPSA-N 0.000 description 1
- BPDVTFBJZNBHEU-HGNGGELXSA-N Glu-Ala-His Chemical compound OC(=O)CC[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@H](C(O)=O)CC1=CN=CN1 BPDVTFBJZNBHEU-HGNGGELXSA-N 0.000 description 1
- LTUVYLVIZHJCOQ-KKUMJFAQSA-N Glu-Arg-Phe Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O LTUVYLVIZHJCOQ-KKUMJFAQSA-N 0.000 description 1
- ZOXBSICWUDAOHX-GUBZILKMSA-N Glu-Asn-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](CC(N)=O)NC(=O)[C@@H](N)CCC(O)=O ZOXBSICWUDAOHX-GUBZILKMSA-N 0.000 description 1
- JPHYJQHPILOKHC-ACZMJKKPSA-N Glu-Asp-Asp Chemical compound OC(=O)CC[C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(O)=O)C(O)=O JPHYJQHPILOKHC-ACZMJKKPSA-N 0.000 description 1
- OAGVHWYIBZMWLA-YFKPBYRVSA-N Glu-Gly-Gly Chemical compound OC(=O)CC[C@H](N)C(=O)NCC(=O)NCC(O)=O OAGVHWYIBZMWLA-YFKPBYRVSA-N 0.000 description 1
- LKOAAMXDJGEYMS-ZPFDUUQYSA-N Glu-Met-Ile Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCSC)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O LKOAAMXDJGEYMS-ZPFDUUQYSA-N 0.000 description 1
- SOEPMWQCTJITPZ-SRVKXCTJSA-N Glu-Met-Lys Chemical compound CSCC[C@@H](C(=O)N[C@@H](CCCCN)C(=O)O)NC(=O)[C@H](CCC(=O)O)N SOEPMWQCTJITPZ-SRVKXCTJSA-N 0.000 description 1
- YQAQQKPWFOBSMU-WDCWCFNPSA-N Glu-Thr-Leu Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(C)C)C(O)=O YQAQQKPWFOBSMU-WDCWCFNPSA-N 0.000 description 1
- GWKBAXRZPLSWJS-QEJZJMRPSA-N Glu-Trp-Cys Chemical compound C1=CC=C2C(=C1)C(=CN2)C[C@@H](C(=O)N[C@@H](CS)C(=O)O)NC(=O)[C@H](CCC(=O)O)N GWKBAXRZPLSWJS-QEJZJMRPSA-N 0.000 description 1
- RJIVPOXLQFJRTG-LURJTMIESA-N Gly-Arg-Gly Chemical compound OC(=O)CNC(=O)[C@@H](NC(=O)CN)CCCN=C(N)N RJIVPOXLQFJRTG-LURJTMIESA-N 0.000 description 1
- UEGIPZAXNBYCCP-NKWVEPMBSA-N Gly-Cys-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CS)NC(=O)CN)C(=O)O UEGIPZAXNBYCCP-NKWVEPMBSA-N 0.000 description 1
- MBOAPAXLTUSMQI-JHEQGTHGSA-N Gly-Glu-Thr Chemical compound [H]NCC(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O MBOAPAXLTUSMQI-JHEQGTHGSA-N 0.000 description 1
- UFPXDFOYHVEIPI-BYPYZUCNSA-N Gly-Gly-Asp Chemical compound NCC(=O)NCC(=O)N[C@H](C(O)=O)CC(O)=O UFPXDFOYHVEIPI-BYPYZUCNSA-N 0.000 description 1
- BXICSAQLIHFDDL-YUMQZZPRSA-N Gly-Lys-Asn Chemical compound [H]NCC(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(N)=O)C(O)=O BXICSAQLIHFDDL-YUMQZZPRSA-N 0.000 description 1
- FJWSJWACLMTDMI-WPRPVWTQSA-N Gly-Met-Val Chemical compound [H]NCC(=O)N[C@@H](CCSC)C(=O)N[C@@H](C(C)C)C(O)=O FJWSJWACLMTDMI-WPRPVWTQSA-N 0.000 description 1
- VNNRLUNBJSWZPF-ZKWXMUAHSA-N Gly-Ser-Ile Chemical compound [H]NCC(=O)N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O VNNRLUNBJSWZPF-ZKWXMUAHSA-N 0.000 description 1
- TVTZEOHWHUVYCG-KYNKHSRBSA-N Gly-Thr-Thr Chemical compound [H]NCC(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O TVTZEOHWHUVYCG-KYNKHSRBSA-N 0.000 description 1
- JBCLFWXMTIKCCB-UHFFFAOYSA-N H-Gly-Phe-OH Natural products NCC(=O)NC(C(O)=O)CC1=CC=CC=C1 JBCLFWXMTIKCCB-UHFFFAOYSA-N 0.000 description 1
- 101000846893 Homo sapiens Fibrillin-1 Proteins 0.000 description 1
- MTFVYKQRLXYAQN-LAEOZQHASA-N Ile-Glu-Gly Chemical compound [H]N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CCC(O)=O)C(=O)NCC(O)=O MTFVYKQRLXYAQN-LAEOZQHASA-N 0.000 description 1
- CAHCWMVNBZJVAW-NAKRPEOUSA-N Ile-Pro-Ser Chemical compound CC[C@H](C)[C@@H](C(=O)N1CCC[C@H]1C(=O)N[C@@H](CO)C(=O)O)N CAHCWMVNBZJVAW-NAKRPEOUSA-N 0.000 description 1
- GMUYXHHJAGQHGB-TUBUOCAGSA-N Ile-Thr-His Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC1=CN=CN1)C(=O)O)N GMUYXHHJAGQHGB-TUBUOCAGSA-N 0.000 description 1
- 102100034343 Integrase Human genes 0.000 description 1
- SITWEMZOJNKJCH-UHFFFAOYSA-N L-alanine-L-arginine Natural products CC(N)C(=O)NC(C(O)=O)CCCNC(N)=N SITWEMZOJNKJCH-UHFFFAOYSA-N 0.000 description 1
- LHSGPCFBGJHPCY-UHFFFAOYSA-N L-leucine-L-tyrosine Natural products CC(C)CC(N)C(=O)NC(C(O)=O)CC1=CC=C(O)C=C1 LHSGPCFBGJHPCY-UHFFFAOYSA-N 0.000 description 1
- 241000880493 Leptailurus serval Species 0.000 description 1
- PBCHMHROGNUXMK-DLOVCJGASA-N Leu-Ala-His Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@H](C(O)=O)CC1=CN=CN1 PBCHMHROGNUXMK-DLOVCJGASA-N 0.000 description 1
- HXWALXSAVBLTPK-NUTKFTJISA-N Leu-Ala-Trp Chemical compound C[C@@H](C(=O)N[C@@H](CC1=CNC2=CC=CC=C21)C(=O)O)NC(=O)[C@H](CC(C)C)N HXWALXSAVBLTPK-NUTKFTJISA-N 0.000 description 1
- NTRAGDHVSGKUSF-AVGNSLFASA-N Leu-Arg-Arg Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CCCN=C(N)N)C(=O)N[C@@H](CCCN=C(N)N)C(O)=O NTRAGDHVSGKUSF-AVGNSLFASA-N 0.000 description 1
- ILJREDZFPHTUIE-GUBZILKMSA-N Leu-Asp-Glu Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(O)=O ILJREDZFPHTUIE-GUBZILKMSA-N 0.000 description 1
- IIKJNQWOQIWWMR-CIUDSAMLSA-N Leu-Cys-Ala Chemical compound C[C@@H](C(=O)O)NC(=O)[C@H](CS)NC(=O)[C@H](CC(C)C)N IIKJNQWOQIWWMR-CIUDSAMLSA-N 0.000 description 1
- VQPPIMUZCZCOIL-GUBZILKMSA-N Leu-Gln-Ala Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](C)C(O)=O VQPPIMUZCZCOIL-GUBZILKMSA-N 0.000 description 1
- OXRLYTYUXAQTHP-YUMQZZPRSA-N Leu-Gly-Ala Chemical compound [H]N[C@@H](CC(C)C)C(=O)NCC(=O)N[C@@H](C)C(O)=O OXRLYTYUXAQTHP-YUMQZZPRSA-N 0.000 description 1
- HYIFFZAQXPUEAU-QWRGUYRKSA-N Leu-Gly-Leu Chemical compound CC(C)C[C@H](N)C(=O)NCC(=O)N[C@H](C(O)=O)CC(C)C HYIFFZAQXPUEAU-QWRGUYRKSA-N 0.000 description 1
- DSFYPIUSAMSERP-IHRRRGAJSA-N Leu-Leu-Arg Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@H](C(O)=O)CCCN=C(N)N DSFYPIUSAMSERP-IHRRRGAJSA-N 0.000 description 1
- YOKVEHGYYQEQOP-QWRGUYRKSA-N Leu-Leu-Gly Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)NCC(O)=O YOKVEHGYYQEQOP-QWRGUYRKSA-N 0.000 description 1
- VCHVSKNMTXWIIP-SRVKXCTJSA-N Leu-Lys-Ser Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CO)C(O)=O VCHVSKNMTXWIIP-SRVKXCTJSA-N 0.000 description 1
- CPONGMJGVIAWEH-DCAQKATOSA-N Leu-Met-Ala Chemical compound CSCC[C@H](NC(=O)[C@@H](N)CC(C)C)C(=O)N[C@@H](C)C(O)=O CPONGMJGVIAWEH-DCAQKATOSA-N 0.000 description 1
- MUCIDQMDOYQYBR-IHRRRGAJSA-N Leu-Pro-His Chemical compound CC(C)C[C@@H](C(=O)N1CCC[C@H]1C(=O)N[C@@H](CC2=CN=CN2)C(=O)O)N MUCIDQMDOYQYBR-IHRRRGAJSA-N 0.000 description 1
- QESXLSQLQHHTIX-RHYQMDGZSA-N Leu-Val-Thr Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H]([C@@H](C)O)C(O)=O QESXLSQLQHHTIX-RHYQMDGZSA-N 0.000 description 1
- ONGCSGVHCSAATF-CIUDSAMLSA-N Met-Ala-Glu Chemical compound CSCC[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@H](C(O)=O)CCC(O)=O ONGCSGVHCSAATF-CIUDSAMLSA-N 0.000 description 1
- VTKPSXWRUGCOAC-GUBZILKMSA-N Met-Ala-Met Chemical compound CSCC[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@H](C(O)=O)CCSC VTKPSXWRUGCOAC-GUBZILKMSA-N 0.000 description 1
- LRALLISKBZNSKN-BQBZGAKWSA-N Met-Gly-Ser Chemical compound CSCC[C@H](N)C(=O)NCC(=O)N[C@@H](CO)C(O)=O LRALLISKBZNSKN-BQBZGAKWSA-N 0.000 description 1
- DBXMFHGGHMXYHY-DCAQKATOSA-N Met-Leu-Ser Chemical compound CSCC[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(O)=O DBXMFHGGHMXYHY-DCAQKATOSA-N 0.000 description 1
- YGNUDKAPJARTEM-GUBZILKMSA-N Met-Val-Ala Chemical compound CSCC[C@H](N)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](C)C(O)=O YGNUDKAPJARTEM-GUBZILKMSA-N 0.000 description 1
- KZNQNBZMBZJQJO-UHFFFAOYSA-N N-glycyl-L-proline Natural products NCC(=O)N1CCCC1C(O)=O KZNQNBZMBZJQJO-UHFFFAOYSA-N 0.000 description 1
- 108010079364 N-glycylalanine Proteins 0.000 description 1
- 108091028043 Nucleic acid sequence Proteins 0.000 description 1
- 101100466242 Oryza sativa subsp. japonica TUD1 gene Proteins 0.000 description 1
- BBDSZDHUCPSYAC-QEJZJMRPSA-N Phe-Ala-Leu Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](C)C(=O)N[C@@H](CC(C)C)C(O)=O BBDSZDHUCPSYAC-QEJZJMRPSA-N 0.000 description 1
- QMMRHASQEVCJGR-UBHSHLNASA-N Phe-Ala-Pro Chemical compound C([C@H](N)C(=O)N[C@@H](C)C(=O)N1[C@@H](CCC1)C(O)=O)C1=CC=CC=C1 QMMRHASQEVCJGR-UBHSHLNASA-N 0.000 description 1
- CGOMLCQJEMWMCE-STQMWFEESA-N Phe-Arg-Gly Chemical compound NC(N)=NCCC[C@@H](C(=O)NCC(O)=O)NC(=O)[C@@H](N)CC1=CC=CC=C1 CGOMLCQJEMWMCE-STQMWFEESA-N 0.000 description 1
- JOXIIFVCSATTDH-IHPCNDPISA-N Phe-Asn-Trp Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)N[C@@H](CC2=CNC3=CC=CC=C32)C(=O)O)N JOXIIFVCSATTDH-IHPCNDPISA-N 0.000 description 1
- QPQDWBAJWOGAMJ-IHPCNDPISA-N Phe-Asp-Trp Chemical compound C([C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(O)=O)C1=CC=CC=C1 QPQDWBAJWOGAMJ-IHPCNDPISA-N 0.000 description 1
- KJJROSNFBRWPHS-JYJNAYRXSA-N Phe-Glu-Leu Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(O)=O KJJROSNFBRWPHS-JYJNAYRXSA-N 0.000 description 1
- OSBADCBXAMSPQD-YESZJQIVSA-N Phe-Leu-Pro Chemical compound CC(C)C[C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](CC2=CC=CC=C2)N OSBADCBXAMSPQD-YESZJQIVSA-N 0.000 description 1
- MVIJMIZJPHQGEN-IHRRRGAJSA-N Phe-Ser-Val Chemical compound CC(C)[C@@H](C([O-])=O)NC(=O)[C@H](CO)NC(=O)[C@@H]([NH3+])CC1=CC=CC=C1 MVIJMIZJPHQGEN-IHRRRGAJSA-N 0.000 description 1
- HFZNNDWPHBRNPV-KZVJFYERSA-N Pro-Ala-Thr Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](C)C(=O)N[C@@H]([C@@H](C)O)C(O)=O HFZNNDWPHBRNPV-KZVJFYERSA-N 0.000 description 1
- ONPFOYPPPOHMNH-UVBJJODRSA-N Pro-Ala-Trp Chemical compound C[C@@H](C(=O)N[C@@H](CC1=CNC2=CC=CC=C21)C(=O)O)NC(=O)[C@@H]3CCCN3 ONPFOYPPPOHMNH-UVBJJODRSA-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
- BNBBNGZZKQUWCD-IUCAKERBSA-N Pro-Arg-Gly Chemical compound NC(N)=NCCC[C@@H](C(=O)NCC(O)=O)NC(=O)[C@@H]1CCCN1 BNBBNGZZKQUWCD-IUCAKERBSA-N 0.000 description 1
- HAAQQNHQZBOWFO-LURJTMIESA-N Pro-Gly-Gly Chemical compound OC(=O)CNC(=O)CNC(=O)[C@@H]1CCCN1 HAAQQNHQZBOWFO-LURJTMIESA-N 0.000 description 1
- LEIKGVHQTKHOLM-IUCAKERBSA-N Pro-Pro-Gly Chemical compound OC(=O)CNC(=O)[C@@H]1CCCN1C(=O)[C@H]1NCCC1 LEIKGVHQTKHOLM-IUCAKERBSA-N 0.000 description 1
- KBUAPZAZPWNYSW-SRVKXCTJSA-N Pro-Pro-Val Chemical compound CC(C)[C@@H](C(O)=O)NC(=O)[C@@H]1CCCN1C(=O)[C@H]1NCCC1 KBUAPZAZPWNYSW-SRVKXCTJSA-N 0.000 description 1
- QKDIHFHGHBYTKB-IHRRRGAJSA-N Pro-Ser-Phe Chemical compound N([C@@H](CO)C(=O)N[C@@H](CC=1C=CC=CC=1)C(O)=O)C(=O)[C@@H]1CCCN1 QKDIHFHGHBYTKB-IHRRRGAJSA-N 0.000 description 1
- YIPFBJGBRCJJJD-FHWLQOOXSA-N Pro-Trp-Leu Chemical compound CC(C)C[C@@H](C(=O)O)NC(=O)[C@H](CC1=CNC2=CC=CC=C21)NC(=O)[C@@H]3CCCN3 YIPFBJGBRCJJJD-FHWLQOOXSA-N 0.000 description 1
- VDHGTOHMHHQSKG-JYJNAYRXSA-N Pro-Val-Phe Chemical compound CC(C)[C@H](NC(=O)[C@@H]1CCCN1)C(=O)N[C@@H](Cc1ccccc1)C(O)=O VDHGTOHMHHQSKG-JYJNAYRXSA-N 0.000 description 1
- 238000012181 QIAquick gel extraction kit Methods 0.000 description 1
- 238000002123 RNA extraction Methods 0.000 description 1
- 108010092799 RNA-directed DNA polymerase Proteins 0.000 description 1
- 229920000297 Rayon Polymers 0.000 description 1
- HQTKVSCNCDLXSX-BQBZGAKWSA-N Ser-Arg-Gly Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CCCNC(N)=N)C(=O)NCC(O)=O HQTKVSCNCDLXSX-BQBZGAKWSA-N 0.000 description 1
- BNFVPSRLHHPQKS-WHFBIAKZSA-N Ser-Asp-Gly Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC(O)=O)C(=O)NCC(O)=O BNFVPSRLHHPQKS-WHFBIAKZSA-N 0.000 description 1
- SFZKGGOGCNQPJY-CIUDSAMLSA-N Ser-Asp-His Chemical compound C1=C(NC=N1)C[C@@H](C(=O)O)NC(=O)[C@H](CC(=O)O)NC(=O)[C@H](CO)N SFZKGGOGCNQPJY-CIUDSAMLSA-N 0.000 description 1
- SWIQQMYVHIXPEK-FXQIFTODSA-N Ser-Cys-Val Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CS)C(=O)N[C@@H](C(C)C)C(O)=O SWIQQMYVHIXPEK-FXQIFTODSA-N 0.000 description 1
- MIJWOJAXARLEHA-WDSKDSINSA-N Ser-Gly-Glu Chemical compound OC[C@H](N)C(=O)NCC(=O)N[C@H](C(O)=O)CCC(O)=O MIJWOJAXARLEHA-WDSKDSINSA-N 0.000 description 1
- SRSPTFBENMJHMR-WHFBIAKZSA-N Ser-Ser-Gly Chemical compound OC[C@H](N)C(=O)N[C@@H](CO)C(=O)NCC(O)=O SRSPTFBENMJHMR-WHFBIAKZSA-N 0.000 description 1
- XJDMUQCLVSCRSJ-VZFHVOOUSA-N Ser-Thr-Ala Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C)C(O)=O XJDMUQCLVSCRSJ-VZFHVOOUSA-N 0.000 description 1
- DWYAUVCQDTZIJI-VZFHVOOUSA-N Thr-Ala-Ser Chemical compound C[C@@H](O)[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@@H](CO)C(O)=O DWYAUVCQDTZIJI-VZFHVOOUSA-N 0.000 description 1
- ISLDRLHVPXABBC-IEGACIPQSA-N Thr-Leu-Trp Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC1=CNC2=C1C=CC=C2)C(O)=O ISLDRLHVPXABBC-IEGACIPQSA-N 0.000 description 1
- DEGCBBCMYWNJNA-RHYQMDGZSA-N Thr-Pro-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@@H]1CCCN1C(=O)[C@@H](N)[C@@H](C)O DEGCBBCMYWNJNA-RHYQMDGZSA-N 0.000 description 1
- 102100032120 Toll/interleukin-1 receptor domain-containing adapter protein Human genes 0.000 description 1
- -1 Trizol kit Proteins 0.000 description 1
- NIWAGRRZHCMPOY-GMVOTWDCSA-N Trp-Ala-Tyr Chemical compound C[C@@H](C(=O)N[C@@H](CC1=CC=C(C=C1)O)C(=O)O)NC(=O)[C@H](CC2=CNC3=CC=CC=C32)N NIWAGRRZHCMPOY-GMVOTWDCSA-N 0.000 description 1
- NRFTYDWKWGJLAR-MELADBBJSA-N Tyr-Asp-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CC(=O)O)NC(=O)[C@H](CC2=CC=C(C=C2)O)N)C(=O)O NRFTYDWKWGJLAR-MELADBBJSA-N 0.000 description 1
- AKRHKDCELJLTMD-BVSLBCMMSA-N Tyr-Trp-Arg Chemical compound C1=CC=C2C(=C1)C(=CN2)C[C@@H](C(=O)N[C@@H](CCCN=C(N)N)C(=O)O)NC(=O)[C@H](CC3=CC=C(C=C3)O)N AKRHKDCELJLTMD-BVSLBCMMSA-N 0.000 description 1
- DDRBQONWVBDQOY-GUBZILKMSA-N Val-Ala-Arg Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@@H](CCCN=C(N)N)C(O)=O DDRBQONWVBDQOY-GUBZILKMSA-N 0.000 description 1
- COYSIHFOCOMGCF-WPRPVWTQSA-N Val-Arg-Gly Chemical compound CC(C)[C@H](N)C(=O)N[C@H](C(=O)NCC(O)=O)CCCN=C(N)N COYSIHFOCOMGCF-WPRPVWTQSA-N 0.000 description 1
- COYSIHFOCOMGCF-UHFFFAOYSA-N Val-Arg-Gly Natural products CC(C)C(N)C(=O)NC(C(=O)NCC(O)=O)CCCN=C(N)N COYSIHFOCOMGCF-UHFFFAOYSA-N 0.000 description 1
- FEFZWCSXEMVSPO-LSJOCFKGSA-N Val-His-Ala Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](Cc1cnc[nH]1)C(=O)N[C@@H](C)C(O)=O FEFZWCSXEMVSPO-LSJOCFKGSA-N 0.000 description 1
- AEMPCGRFEZTWIF-IHRRRGAJSA-N Val-Leu-Lys Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCCN)C(O)=O AEMPCGRFEZTWIF-IHRRRGAJSA-N 0.000 description 1
- RFKJNTRMXGCKFE-FHWLQOOXSA-N Val-Leu-Trp Chemical compound C1=CC=C2C(C[C@H](NC(=O)[C@@H](NC(=O)[C@@H](N)C(C)C)CC(C)C)C(O)=O)=CNC2=C1 RFKJNTRMXGCKFE-FHWLQOOXSA-N 0.000 description 1
- CKTMJBPRVQWPHU-JSGCOSHPSA-N Val-Phe-Gly Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)NCC(=O)O)N CKTMJBPRVQWPHU-JSGCOSHPSA-N 0.000 description 1
- XBJKAZATRJBDCU-GUBZILKMSA-N Val-Pro-Ala Chemical compound CC(C)[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@@H](C)C(O)=O XBJKAZATRJBDCU-GUBZILKMSA-N 0.000 description 1
- PZTZYZUTCPZWJH-FXQIFTODSA-N Val-Ser-Ser Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CO)C(=O)N[C@@H](CO)C(=O)O)N PZTZYZUTCPZWJH-FXQIFTODSA-N 0.000 description 1
- IRAUYEAFPFPVND-UVBJJODRSA-N Val-Trp-Ala Chemical compound C1=CC=C2C(C[C@H](NC(=O)[C@@H](N)C(C)C)C(=O)N[C@@H](C)C(O)=O)=CNC2=C1 IRAUYEAFPFPVND-UVBJJODRSA-N 0.000 description 1
- RTJPAGFXOWEBAI-SRVKXCTJSA-N Val-Val-Arg Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](C(C)C)C(=O)N[C@H](C(O)=O)CCCN=C(N)N RTJPAGFXOWEBAI-SRVKXCTJSA-N 0.000 description 1
- AOILQMZPNLUXCM-AVGNSLFASA-N Val-Val-Lys Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](C(C)C)C(=O)N[C@H](C(O)=O)CCCCN AOILQMZPNLUXCM-AVGNSLFASA-N 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- 238000007605 air drying Methods 0.000 description 1
- 108010045350 alanyl-tyrosyl-alanine Proteins 0.000 description 1
- 108010047495 alanylglycine Proteins 0.000 description 1
- 108010087924 alanylproline Proteins 0.000 description 1
- 150000001298 alcohols Chemical class 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- 108010043240 arginyl-leucyl-glycine Proteins 0.000 description 1
- 238000009412 basement excavation Methods 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 230000032823 cell division Effects 0.000 description 1
- 238000000546 chi-square test Methods 0.000 description 1
- 238000010367 cloning Methods 0.000 description 1
- 230000002380 cytological effect Effects 0.000 description 1
- 230000009089 cytolysis Effects 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000010828 elution Methods 0.000 description 1
- 238000006911 enzymatic reaction Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 239000000706 filtrate Substances 0.000 description 1
- 238000011010 flushing procedure Methods 0.000 description 1
- 239000012634 fragment Substances 0.000 description 1
- 108010006664 gamma-glutamyl-glycyl-glycine Proteins 0.000 description 1
- 238000012214 genetic breeding Methods 0.000 description 1
- 108010049041 glutamylalanine 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
- 108010072405 glycyl-aspartyl-glycine Proteins 0.000 description 1
- 108010051307 glycyl-glycyl-proline Proteins 0.000 description 1
- 108010025801 glycyl-prolyl-arginine Proteins 0.000 description 1
- 108010089804 glycyl-threonine Proteins 0.000 description 1
- 108010081551 glycylphenylalanine Proteins 0.000 description 1
- 108010025306 histidylleucine Proteins 0.000 description 1
- 238000005286 illumination Methods 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 150000002596 lactones Chemical class 0.000 description 1
- 230000011890 leaf development Effects 0.000 description 1
- 108010012058 leucyltyrosine Proteins 0.000 description 1
- 239000006166 lysate Substances 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 230000035800 maturation Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000010369 molecular cloning Methods 0.000 description 1
- 230000035772 mutation Effects 0.000 description 1
- 108020004707 nucleic acids Proteins 0.000 description 1
- 102000039446 nucleic acids Human genes 0.000 description 1
- 150000007523 nucleic acids Chemical class 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 230000035764 nutrition Effects 0.000 description 1
- 230000037361 pathway Effects 0.000 description 1
- 108010024607 phenylalanylalanine Proteins 0.000 description 1
- 230000000243 photosynthetic effect Effects 0.000 description 1
- 239000013612 plasmid Substances 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 230000001376 precipitating effect Effects 0.000 description 1
- 108010087846 prolyl-prolyl-glycine Proteins 0.000 description 1
- 108010079317 prolyl-tyrosine Proteins 0.000 description 1
- 108010015796 prolylisoleucine Proteins 0.000 description 1
- 238000003753 real-time PCR Methods 0.000 description 1
- 238000011084 recovery Methods 0.000 description 1
- 238000002310 reflectometry Methods 0.000 description 1
- 230000003252 repetitive effect Effects 0.000 description 1
- 238000005204 segregation Methods 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 108010071207 serylmethionine Proteins 0.000 description 1
- 239000002689 soil Substances 0.000 description 1
- 238000009331 sowing Methods 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 238000010998 test method Methods 0.000 description 1
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 description 1
- 238000012795 verification Methods 0.000 description 1
Landscapes
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
本发明属于基因工程技术领域，具体涉及水稻叶片叶夹角突变基因PLA1及其应用，所述突变基因PLA1的核苷酸序列如SEQ ID No.1所示，其编码蛋白质的氨基酸序列如SEQ ID No.2所示，与野生型相比，突变基因PLA1在P450基因(LOC_Os10g26340)的第2外显子的第453位碱基由C变为了A，使得编码的氨基酸由脯氨酸变成了苏氨酸。该基因突变后的水稻表现为植株矮化、叶片叶夹角增大，能够将该基因用于水稻品种选育，选育出具有观赏价值的观赏型水稻。
Description
技术领域
本发明属于基因工程技术领域，具体涉及水稻叶片叶夹角突变基因PLA1及其应用。
背景技术
水稻是极其重要的农作物，在我国作为单产最高、总产量最高以及消费人群最多的粮食作物，在保障我国粮食安全方面有着重要意义。因此，水稻高产一直是育种家们在品种选育中最重要的目标。籽粒、穗型和株型是影响水稻产量的三大因素，是遗传育种中最重要的选择性状。水稻的株型改良对于合理密植、提高光能利用率有重要意义，紧凑的水稻株型有利于水稻高产的形成。水稻叶片挺立、夹角变小导致对光的反射率较小，可增加叶片双面受光，提高冠层的光合效率，从而使得干物质积累增加，同时也能改善群体的下层光照,有利于根系活力的提高，进而达到提升水稻产量的目的。因此发掘和克隆新的调控水稻叶夹角的基因对于提高水稻产量有着重要的理论意义和指导意义。此外研究者一直都在不断的发掘水稻独特的观赏性，力寻收集一批在叶色、穗型、株高、株型、籽粒等不同变异的水稻。叶夹角增大、叶片发育周期提前造成茎秆着生的叶片增多且对称生长提高了水稻株型的独特性，水稻叶色的变异也增加了水稻的观赏性，为培育“彩色水稻”提供丰富的种质资源。
发明内容
本发明的一个目的在于提供水稻叶片叶夹角突变基因PLA1及应用。
本发明的再一个目的在于提供用于检测水稻叶片叶夹角突变基因PLA1的引物对以及含有该引物对的试剂盒。
为了实现上述目的，本发明利用甲基磺酸乙酯(EMS)诱变籼型水稻保持系西大1B获得了一个遗传稳定的水稻叶夹角增大突变体(命名为PLA1)。遗传分析发现该突变性状受隐性单基因(命名为PLA1)控制。分子标记定位结果显示，该基因位于第10号染色体RM25367和XJ-1之间86.7kb的区间内。
在上述基因定位的基础上，本发明通过候选基因预测、同源搜索及基因序列差异比较，确定了PLA1的核苷酸序列如SEQ ID No.1所示，编码的蛋白质的氨基酸序列如SEQ IDNo.2所示。与野生型相比，PLA1基因在P450基因(LOC_Os10g26340)的第2外显子的第453位碱基由C变为了A，使得编码的氨基酸由脯氨酸变成了苏氨酸。然后，本发明继续对该基因的表达模式和激素响应进行了研究，初步确定PLA1基因可能通过影响BU1和ILI1等BR信号传导途径中上游基因调控叶枕近轴面细胞的发育从而影响水稻叶夹角的大小。
本发明还提供用于检测检测水稻叶片叶夹角突变基因PLA1的引物对，包括包括正向引物：5′-CATGGCTCGCCCACCTCTACG-3′，反向引物(5′-3′)：5′-CTCCCAGAGGACGGCGACCAT-3′。
本发明还提供还有上述引物对的检测试剂盒。
本发明还提供了水稻叶片叶夹角突变基因PLA1在水稻选育中的应用，应用该基因选育出极具观赏性的水稻品种。
本发明的有益效果为：本发明提供了水稻叶片叶夹角突变基因PLA1，并通过实验初步验证，该基因可能通过BR信号传导途径调控水稻叶夹角的发育，为改良水稻株型提供了方向。
附图说明
图1为野生型和突变体PLA1叶夹角的组织学观察；其中，A为苗期野生型和突变体PLA1三片功能叶的叶夹角大小统计图；B为分蘖期野生型和突变体PLA1三片功能叶的叶夹角大小统计图；C为成熟期野生型和突变体PLA1三片功能叶的叶夹角大小统计图；D为成熟期野生型和突变体单株分蘖；E为野生型和突变体倒一叶叶夹角；F为野生型与突变体倒二叶叶夹角；G为野生型与突变体倒三叶叶夹角；H为野生型倒二叶叶枕细胞；I为突变体倒二叶叶枕细胞；a为图H中a区域放大图；b为图I中b区域放大图；c为图H中c区域放大图；d为图I中d区域放大图；J为野生型近轴面与远轴面细胞长度；K为突变体近轴面与远轴面细胞宽度；
图2为野生型(WT)和突变体(PLA1)叶片的组织学观察；其中，A为苗期野生型和突变体叶片；B为苗期野生型和突变体叶片中部放大图；C为野生型叶片横切图；D为突变体叶片横切图；E为野生型叶片中脉细胞；F为突变体叶片中脉细胞；a为图C中a区域放大图；b为图C中b区域放大图；c为图D中c区域的放大图；d为图D中d区域的放大图；G为野生型和突变体大维管束数量统计图；H为野生型和突变体小维管束数量统计图；I为野生型和突变体大维管束长度和宽度统计图；J为野生型和突变体小维管束长度和宽度统计图；
图3为野生型和突变体茎秆的组织学观察；其中，A为成熟期的野生型和突变体植株；B为野生型和突变体的穗长、倒一节间至倒四节间长；C为野生型倒二节节间的横切图；D为突变体倒二节节间的横切图；E为野生型倒二节节间横切图的放大图；F为突变体倒二节节间横切图的放大图；G为野生型倒二节节间纵切图；H为突变体倒二节节间纵切图；I为野生型和突变体茎秆横切图中维管束数量统计图；J为野生型和突变体茎秆纵切图中细胞层数统计图；K为野生型和突变体纵切图中细胞长度统计图；L为野生型和突变体纵切图中细胞宽度统计图；
图4为PLA1基因的分子定位图；其中，A为PLA1的分子定位；B为野生型和突变体的碱基变化；
图5为PLA1基因在野生型中的表达模式；
图6为突变体PLA1对油菜素内酯BR的响应及相关基因分析结果；其中，A为不同BR处理时长下的野生型和突变体PLA1中的qPCR分析结果；B为野生型和突变体叶夹角对外源BR的敏感性检测结果；C为野生型和突变体PLA1外源BR处理前后的叶夹角变化结果；D为BR生物信号传导和合成相关基因的表达分析结果。
具体实施方式
下面通过具体实施例对本发明进行清楚、完整地描述，显然，所描述的实施例仅仅是本发明的一部分实施例，而不是全部的实施例。基于本发明的实施例，本领域普通技术人员所获得的所有其他实施例，都属于本发明的保护范围。
实施例中未注明具体条件的实验方法，通常按照常规条件，例如分子克隆实验指南(第三版，J.萨姆布鲁克等著，黄培堂等译，科学出版社，2002年)中所述的条件，或按照制造厂商所建议的条件。
实施例中使用的材料：野生型西大1B和水稻叶片叶夹角增大突变体PLA1由西南大学水稻研究所提供；M-MLV逆转录酶、高保真DNA聚合酶PFU、T4 DNA连接酶、Trizol试剂盒、DNA凝胶回收试剂盒、质粒提取试剂盒购自TaKaRa公司；氨苄青霉素(Ampicillin,Amp)为Sigma公司产品；引物合成和DNA测序由上海英俊生物技术有限公司完成；其它化学试剂购自北京鼎国生物技术有限责任公司；大肠杆菌DH5α由西南大学水稻研究所提供。
实施例1
西南大学水稻研究所利用EMS诱变籼型水稻保持系西大1B获得了一个遗传稳定的水稻叶夹角增大突变体(命名为PLA1)，表现为植株矮化、叶片叶夹角增大。观察发现，植株矮化是由于PLA1穗长、倒1-4节节间长均变短，且达到显著水平，见图3B，“*”表示在0.05水平上差异显著，“**”表示在0.01水平上差异显著；田间种植条件下分别统计苗期、分蘖期、成熟期野生型和突变体的叶夹角，发现三片功能叶的叶夹角在各时期大于野生型，见图1A-C,其中，1st表示倒一叶叶夹角，2nd表示倒二叶叶夹角，3rd表示倒三叶叶夹角。除此之外突变体还表现为窄叶和茎秆变细。
为了进一步分析突变体叶夹角增大、叶片变窄、茎秆变细的原因，利用石蜡切片进行细胞学观察，观察叶枕部位的细胞结构发现突变体近轴面细胞相对于野生型变长，达到极显著水平，突变体近轴面细胞宽度极显著短于野生型；远轴面的细胞长度和宽度差异均不显著。即突变体叶夹角增大是由于近轴面细胞细长造成，见图1D-K，其中，ad表示近轴面，ab表示远轴面；苗期开始突变体PLA1的叶片与野生型相比严重变窄，且这种性状一直持续到成熟期，石蜡切片结果显示，PLA1叶片变窄是维管束数量变少引起,见图2；统计野生型和突变体PLA1茎秆横切的维管束数目、细胞长度和宽度发现突变体PLA1维管束数目极显著减少，而细胞长度和宽度无显著变化，且相同放大倍数下，突变体PLA1茎秆横切的结构图小于野生型，统计茎秆纵切的细胞层数发现突变体PLA1的细胞层数显著少于野生型，即茎秆变细主要受细胞分裂影响，见图3。
实施例2水稻叶片叶夹角增大突变基因PLA1的定位、测序及功能验证
1、水稻叶片叶夹角突变基因PLA1的定位
用表型正常的缙恢10号与突变体杂交，F1代表型正常，说明该突变体受隐性基因控制。F2代群体中出现明显的分离，分别表现双亲性状。经卡方测验，正常株﹕突变株符合3∶1分离比，表明突变体受隐性单基因(命名为PLA1)控制，利用F2隐性群体进行基因定位，共获得947个突变株。按照CTAB法提取亲本基因池和突变基因池的DNA；
(1)将10株亲本水稻叶片和突变体水稻叶片剪碎后分别装在2ml的离心管内速冻于液氮中，利用组织研磨机将其打碎成粉末，加入500μl的60℃预热的CTAB液体后65℃温浴30min，期间10min左右摇匀一次；
(2)加入500μl氯仿，上下颠倒混匀，室温放置2min后5000rpm离心5min，将上清液吸取到新的1.5ml的离心管中，加入两倍上清液体积冰冻后的无水乙醇轻轻摇匀直至絮状沉淀抱团；
(3)12000rpm离心2min后倒掉上清液；
(4)600μl无水乙醇洗涤一次，12000rpm离心2min后倒掉上清液；
(5)风干至无液体残留后加入200μl的ddH2O,-20℃保存备用。
选用400对均匀分布于12条染色体上的SSR标记筛选出缙恢10号和突变体PLA1之间的多态性标记，利用多态性标记对突变基因池和正常基因池进行分析(参照硕士学位论文：水稻叶缘白化突变体mal的遗传分析与基因定位，2014年，22页)，结果第10号染色体上的XYD-3和XYD-5与PLA1表现连锁。在两标记间进一步设计引物，引物序列祥见表1，最终将PLA1定位在RM25367和XJ-1之间，物理距离为246kb，见图4A。
表1基因精细定位所用引物及其序列
2、水稻叶片叶夹角突变基因PLA1的测序
在对PLA1精细定位的基础上，通过候选基因在线预测(http://mendel.cs.rhul.ac.uk)和Gramene数据库(http://ensembl.gramene.org/genome-browser/index.html)给出的基因序列，下载候选基因的基因组DNA序列，使用BLAST在线比对和VectorNTI10软件设计测序引物：正向引物序列(5′-3′)为：5′-CATGGCTCGCCCACCTCTACG-3′，反向引物序列(5′-3′)为：5′-CTCCCAGAGGACGGCGACCAT-3′。分别用野生型和突变体DNA模板扩增目的片段，扩增体系为：SYBRPremix Ex TaqⅡ 10μl，PCRForward Primer 0.6μl，PCR Reverse Primer 0.6μl，cDNA模板2μl，RNase Free dd H2O6.8μl，总反应体系20μl。将扩增到的单一的目的条带胶块切下利用胶回收试剂盒回收目的片段，然后将片段16℃过夜连接到19-T载体上，转化大肠杆菌DH5α，待37℃培养一天后挑选单一阳性克隆，送测序公司测序，将测序结果在VectorNTI10软件上比对差异。结果显示PLA1的核苷酸序列如SEQ ID No.1所示，其编码蛋白质的氨基酸序列如SEQ ID No.2所示。与野生型相比，PLA1基因在P450基因(LOC_Os10g26340)的第2外显子的第453位碱基由C变为了A，使得编码的氨基酸由脯氨酸变成了苏氨酸，见图4B。
实施例3水稻叶片叶夹角增大突变基因PLA1的表达模式
为了解水稻叶片叶夹角增大突变基因的表达模式，通过实时荧光定量qPCR对野生型进行表达量分析发现，(参照博士学位论文：水稻小穗发育相关基因MFS1的图位克隆与功能分析，2013年，29页)水稻叶片叶夹角增大突变基因在各个组织部位都有表达，且在叶片中表达最高，其次是根和鞘，在茎中表达量最低。见图5，其中，Ro代表根，St代表茎，Le代表叶，Sh代表鞘，Sp-1代表长度为0-2cm的穗，Sp-2代表长度为2-5cm的穗。
实施例4水稻叶片叶夹角突变基因PLA1的激素响应实验分析
(1)将突变体PLA1和野生型种子催芽，选取露白一致的种子播种在营养土中，放置于恒温培养箱中培养，待突变体PLA1和野生型长至三叶一心期时用10μmol L–1的BR(油菜素内酯)分别处理野生型和突变体PLA1，且处理时长为0h、0.5h、1h、2h、4h、6h。另外利用不同浓度的24-eBL处理野生型和突变体PLA1，结果表明随着外源24-eBL浓度的增加，野生型和突变体PLA1的叶夹角不断增大，当浓度增大为0.01μmol L–1时野生型的叶夹角达到最大，随着浓度升高叶夹角趋于稳定，而突变体叶夹角不断增大，10μmol L–1时叶夹角达到最大值见图6B；叶夹角的变化量由野生型(△B)的7°增加到突变体PLA1(△S)的56.5°，上升了87.6％，达到极显著差异水平见图6C。取各处理下的野生型和突变体PLA1提取RNA，并反转录成cDNA，保存备用。
(2)定量PCR分析
取10μmol L–1的BR(油菜素内酯)处理时长不同的野生型和突变体PLA1的地上部分于2.0ml的无核酸酶的离心管内，立刻置于液氮中。按照以下步骤提取RNA(参照天根植物总RNA提取试剂盒说明书)：
a将速冻于液氮后的离心管置于组织研磨机45HZ，60s完全打碎；
b加入RNA裂解液，用移液枪混匀后室温放置3-5min后加入等量的稀释液；
c组织裂解混匀后室温12000rpm离心5min，小心吸取上清液；
d加入0.5倍上清液体积的无水乙醇吹打混匀后全部转移至离心柱中，12000rpm离心1min之后倒掉上清液；
e加入600μlRNA洗液，12000rpm离心45s后弃滤液，加入50μlDNA酶Ⅰ(10╳DNA酶Ⅰ缓冲液5μl，DNA酶Ⅰ5μl，无核酸酶氺40μl)到吸附膜中央，室温放置15min；
f加入600μlRNA洗液12000rpm离心45s，倒掉上清液，重复操作一次后将离心管重新放置于收集管上，12000rpm离心2min；
g将离心柱转移到洗脱管上，在膜中央加入50-200μl无核酸氺，室温静置2min，12000rpm离心1min即可得到RNA。
RNA提取后按照下面体系反转录成cDNA(参照TaKaRa发转录试剂盒说明书)：5╳gDNA Ereasar Buffer 2μl，gDNA Ereasar 1μl，RNA 7μl，42℃反应2min后4℃保存1min；然后在上述10μl体系中加入Primefxcript Enzyme MixⅠ1μl，RT Primer Mix 1μl，5╳Primescript Buffer 4μl，RNase Free ddH2O 4μl，37℃反应15min，85℃5s后4℃保存1min所得产物即为cDNA产物。
按照以下组分配制PCR反应体系分析野生型和突变体PLA1中的表达量：SYBRPremix Ex TaqⅡ10μl，PCR Forward Primer 0.6μl，PCR Reverse Primer 0.6μl，cDNA模板2μl，RNase Free dd H2O 6.8μl，总反应体系20μl，扩增在Real-Time System定量PCR仪上进行。结果显示当处理时长达到1h时野生型表达量急剧升高，升高量约为0h时的120倍，随后表达量骤减，与0h时相当且后期趋于稳定，见图6A；同时取各浓度处理后的野生型和突变体PLA1，提取RNA，反转录成cDNA，进行qPCR分析，结果表明,与野生型相比，BR信号传导途径基因BRI1、BZR1、BU1、ILI1、TUD1在突变体PLA1中的表达量均上调，其中ILI1在突变体PLA1中的表达量为野生型的76.5倍，且8倍；但BR合成途径调控基因D4、D2和D11表达量变化不似信号传导途径基因变化明显，见图6D，相关定量PCR引物序列见表2。因此初步确定水稻叶片叶夹角增大突变基因PLA1可能通过影响BU1和ILI1等BR信号传导途径中上游基因调控叶枕近轴面细胞的发育从而影响水稻叶夹角的大小。
表2定量PCR引物序列
以上所述仅为本发明的较佳实施例而已，并不用以限制本发明，凡在本发明的精神和原则之内，所有的任何修改、等同替换、改进等，均应包含在本发明的保护范围之内。
序列表
<110> 西南大学
<120> 水稻叶片叶夹角突变基因PLA1及其应用
<130> 2019
<160> 38
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1754
<212> DNA
<213> Oryza sativa
<400> 1
atggcaatgg ccaccgccac cgcctcctcc tgcgtcgacg ccacgtggtg ggcgtacgcc 60
ctcccggcgc tcctcggcgc cgacaccctc tgcgcccacc cggcgctgct cgccggcgcc 120
gtcctcctgg ccttcgccac cgccgcggtg ctcgcctggg ccgcgtcccc cggcgggccg 180
gcgtgggcgc acggccgcgg ccgcctcggc gcgacgccca tcgaggggcc ccgggggctc 240
cccgtgttcg gcagcatctt cgcgctctcc cggggcctcc cgcaccgcgc gctcgacgcg 300
atgtcgcgcg acgcggcggc gccacgggcg agggagctca tggcgttctc cgtcggggag 360
acgccggcgg tggtgtcgtc gtgcccggcg acggcgaggg aggtgctcgc gcacccgtcg 420
ttcgccgacc gcccgctgaa gcgctcggcg cgggagctgc tgttcgcgcg cgccatcggg 480
ttcgccccca gcggcgagta ctggcgcctc ctccgccgca tcgcctccac ccacctcttc 540
tcccctcgcc gcgtcgccgc gcacgagccg gggcgccagg ccgacgccac ggcgatgctg 600
tccgccatgg ccgccgagca gtccgccacc ggcgccgtcg tgctccgccc ccacctccag 660
gccgccgcgc tcaacaacat catgggcagc gtgttcggcc ggcgctacga cgtctcctcc 720
tcctccggcg ccgccgccga cgaggccgag cagctcaaga gcatggtgcg cgaggggttc 780
gagctcctcg gcgcgttcaa ctggtccgac cacctcccat ggctcgccca cctctacgac 840
cccaaccacg tcgcccgccg ctgcgccgcg ctcgtccccc gcgtccaggc gttcgtccgc 900
ggcgtcatcc gcgaccaccg cctccgccgc gactcctcct ccaccgccgc cgacaatgcc 960
gacttcgtcg acgtcctcct ctccctcgag gcccacgaga acctcgccga ggacgacatg 1020
gtcgccgtcc tctgggtaaa aaaaaaaaaa aaaaaacaaa ttctactcaa acatttcaaa 1080
ctcaaatgtt tttttaaaaa tgtttttgtg tattttggca ggagatgata tttcgtggga 1140
cggacacgac ggcgttggtg acggagtggt gcatggcgga ggtggtgagg aacccggcgg 1200
tgcaggcgag gctgagggcg gaggtggacg cggcggtggg cggcgacggg tgtcccagcg 1260
acggcgacgt ggcgcggatg ccgtacctgc aggcggtggt gaaggagacg ctgagggcgc 1320
acccgccggg gccgctgctg agctgggcgc ggctggccac cgccgacgtg gggctcgcca 1380
acggcatggt ggtgccggcg ggcacgacgg cgatggtgaa catgtgggcc atcacccacg 1440
acggcgaggt gtgggccgac ccggaggcgt tcgcgccgga gcggttcatc ccgtcggagg 1500
gcggcgccga cgtcgacgtc cgcggcggcg acctccgcct ggcgccgttc ggcgccgggc 1560
gccgcgtctg ccccggcaag aacctcggcc tcgccaccgt caccctctgg gtcgcccgcc 1620
tcgtccacgc cttcgactgg ttcctccccg acggctcgcc gccggtgtcc ctcgacgagg 1680
tcctcaagct ctccctcgag atgaagaccc ctctcgccgc cgccgccacc ccccgccgcc 1740
gccgcgccgc ctga 1754
<210> 2
<211> 555
<212> PRT
<213> Oryza sativa
<400> 2
Met Ala Met Ala Thr Ala Thr Ala Ser Ser Cys Val Asp Ala Thr Trp
1 5 10 15
Trp Ala Tyr Ala Leu Pro Ala Leu Leu Gly Ala Asp Thr Leu Cys Ala
20 25 30
His Pro Ala Leu Leu Ala Gly Ala Val Leu Leu Ala Phe Ala Thr Ala
35 40 45
Ala Val Leu Ala Trp Ala Ala Ser Pro Gly Gly Pro Ala Trp Ala His
50 55 60
Gly Arg Gly Arg Leu Gly Ala Thr Pro Ile Glu Gly Pro Arg Gly Leu
65 70 75 80
Pro Val Phe Gly Ser Ile Phe Ala Leu Ser Arg Gly Leu Pro His Arg
85 90 95
Ala Leu Asp Ala Met Ser Arg Asp Ala Ala Ala Pro Arg Ala Arg Glu
100 105 110
Leu Met Ala Phe Ser Val Gly Glu Thr Pro Ala Val Val Ser Ser Cys
115 120 125
Pro Ala Thr Ala Arg Glu Val Leu Ala His Pro Ser Phe Ala Asp Arg
130 135 140
Pro Leu Lys Arg Ser Ala Arg Glu Leu Leu Phe Ala Arg Ala Ile Gly
145 150 155 160
Phe Ala Pro Ser Gly Glu Tyr Trp Arg Leu Leu Arg Arg Ile Ala Ser
165 170 175
Thr His Leu Phe Ser Pro Arg Arg Val Ala Ala His Glu Pro Gly Arg
180 185 190
Gln Ala Asp Ala Thr Ala Met Leu Ser Ala Met Ala Ala Glu Gln Ser
195 200 205
Ala Thr Gly Ala Val Val Leu Arg Pro His Leu Gln Ala Ala Ala Leu
210 215 220
Asn Asn Ile Met Gly Ser Val Phe Gly Arg Arg Tyr Asp Val Ser Ser
225 230 235 240
Ser Ser Gly Ala Ala Ala Asp Glu Ala Glu Gln Leu Lys Ser Met Val
245 250 255
Arg Glu Gly Phe Glu Leu Leu Gly Ala Phe Asn Trp Ser Asp His Leu
260 265 270
Pro Trp Leu Ala His Leu Tyr Asp Pro Asn His Val Ala Arg Arg Cys
275 280 285
Ala Ala Leu Val Pro Arg Val Gln Ala Phe Val Arg Gly Val Ile Arg
290 295 300
Asp His Arg Leu Arg Arg Asp Ser Ser Ser Thr Ala Ala Asp Asn Ala
305 310 315 320
Asp Phe Val Asp Val Leu Leu Ser Leu Glu Ala His Glu Asn Leu Ala
325 330 335
Glu Asp Asp Met Val Ala Val Leu Trp Glu Met Ile Phe Arg Gly Thr
340 345 350
Asp Thr Thr Ala Leu Val Thr Glu Trp Cys Met Ala Glu Val Val Arg
355 360 365
Asn Pro Ala Val Gln Ala Arg Leu Arg Ala Glu Val Asp Ala Ala Val
370 375 380
Gly Gly Asp Gly Cys Pro Ser Asp Gly Asp Val Ala Arg Met Pro Tyr
385 390 395 400
Leu Gln Ala Val Val Lys Glu Thr Leu Arg Ala His Pro Pro Gly Pro
405 410 415
Leu Leu Ser Trp Ala Arg Leu Ala Thr Ala Asp Val Gly Leu Ala Asn
420 425 430
Gly Met Val Val Pro Ala Gly Thr Thr Ala Met Val Asn Met Trp Ala
435 440 445
Ile Thr His Asp Gly Glu Val Trp Ala Asp Pro Glu Ala Phe Ala Pro
450 455 460
Glu Arg Phe Ile Pro Ser Glu Gly Gly Ala Asp Val Asp Val Arg Gly
465 470 475 480
Gly Asp Leu Arg Leu Ala Pro Phe Gly Ala Gly Arg Arg Val Cys Pro
485 490 495
Gly Lys Asn Leu Gly Leu Ala Thr Val Thr Leu Trp Val Ala Arg Leu
500 505 510
Val His Ala Phe Asp Trp Phe Leu Pro Asp Gly Ser Pro Pro Val Ser
515 520 525
Leu Asp Glu Val Leu Lys Leu Ser Leu Glu Met Lys Thr Pro Leu Ala
530 535 540
Ala Ala Ala Thr Pro Arg Arg Arg Arg Ala Ala
545 550 555
<210> 3
<211> 19
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 3
gttctgcgtc accgacttg 19
<210> 4
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 4
ccaaacaaag acctcctggt aag 23
<210> 5
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 5
tgcgtttccc acatacatgg 20
<210> 6
<211> 25
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 6
caatcacatt ctactaactt ctgcc 25
<210> 7
<211> 21
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 7
ggcctgtatg tgcaacagtt a 21
<210> 8
<211> 25
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 8
acatttattg atttgtggtg agttt 25
<210> 9
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 9
ttaagtcctg tagtaggtca cacc 24
<210> 10
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 10
cgatgagtca gattgaagta gc 22
<210> 11
<211> 21
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 11
catggctcgc ccacctctac g 21
<210> 12
<211> 21
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 12
ctcccagagg acggcgacca t 21
<210> 13
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 13
cttgcggtgc ttgacctgtc gtat 24
<210> 14
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 14
tggctcttcg gaaatgtggc aag 23
<210> 15
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 15
acaacaacga ggtgctcaag 20
<210> 16
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 16
ttacatccct tgcggtaggt 20
<210> 17
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 17
gtagccagct tgatctcatc tc 22
<210> 18
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 18
gggacgactc tactgcatca 20
<210> 19
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 19
gttcgaagtc tcctgcttcc 20
<210> 20
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 20
tgttcccaac agattcctca 20
<210> 21
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 21
ccggtctagg ctctgttctc 20
<210> 22
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 22
acacttgctc cttcagcctt 20
<210> 23
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 23
ttctctccaa gcttcaggcc ct 22
<210> 24
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 24
tgcacgtctc ctgcaaaacc ct 22
<210> 25
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 25
gacccaactc tggtcaatcc 20
<210> 26
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 26
caagttgtgg gaccaaagtg 20
<210> 27
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 27
cacaagagaa tgcctccaga 20
<210> 28
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 28
attgggctct cgtagctcat 20
<210> 29
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 29
cttgcagcct tcctctctct 20
<210> 30
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 30
gcgtggagag aaactgaaca 20
<210> 31
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 31
tatgtgccaa caaaggagga 20
<210> 32
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 32
cctctggcct cctacatcat 20
<210> 33
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 33
caaatcctgg acctcttgct 20
<210> 34
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 34
tcctccctca gttcttggac 20
<210> 35
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 35
gaggtggaag gagaaggaca 20
<210> 36
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 36
ctggtgacca agtggtgaag 20
<210> 37
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 37
tcacctccac aaagctcaag 20
<210> 38
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 38
gcaaactttc ttgcctcctc 20
Claims (6)
1.水稻叶片叶夹角突变基因PLA1，其特征在于，所述突变基因PLA1的核苷酸序列如SEQID No.1所示。
2.权利要求1所述的水稻叶片叶夹角突变基因PLA1编码的蛋白质，其特征在于，所述蛋白质的氨基酸序列如SEQ ID No.2所示。
3.用于检测权利要求1所述水稻叶片叶夹角突变基因PLA1的引物对，其特征在于，包括正向引物：5′-CATGGCTCGCCCACCTCTACG-3′，反向引物(5′-3′)：5′-CTCCCAGAGGACGGCGACCAT-3′。
4.包括权利要求3所述引物对的检测试剂盒。
5.权利要求1所述的水稻叶片叶夹角突变基因PLA1在水稻品种选育中的应用。
6.权要求1所述的水稻叶片叶夹角突变基因PLA1在观赏型水稻品种选育中的应用。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910281061.2A CN109880832A (zh) | 2019-04-09 | 2019-04-09 | 水稻叶片叶夹角突变基因pla1及其应用 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910281061.2A CN109880832A (zh) | 2019-04-09 | 2019-04-09 | 水稻叶片叶夹角突变基因pla1及其应用 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN109880832A true CN109880832A (zh) | 2019-06-14 |
Family
ID=66936540
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201910281061.2A Withdrawn CN109880832A (zh) | 2019-04-09 | 2019-04-09 | 水稻叶片叶夹角突变基因pla1及其应用 |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN109880832A (zh) |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2005070195A1 (ja) * | 2004-01-26 | 2005-08-04 | Japan As Represented By The President Of National Institute Of Genetics | 種子重量の増加したトランスジェニック植物とその利用 |
| EP2695944A2 (en) * | 2008-08-20 | 2014-02-12 | BASF Plant Science GmbH | Transgenic plants with increased yield |
| CN106191080A (zh) * | 2015-05-07 | 2016-12-07 | 杭州瑞丰生物科技有限公司 | Cyp78a基因在增加玉米株高和增强植株长势中的应用 |
| CN109207508A (zh) * | 2018-08-03 | 2019-01-15 | 浙江大学 | 一种提高农作物产量的方法 |
-
2019
- 2019-04-09 CN CN201910281061.2A patent/CN109880832A/zh not_active Withdrawn
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2005070195A1 (ja) * | 2004-01-26 | 2005-08-04 | Japan As Represented By The President Of National Institute Of Genetics | 種子重量の増加したトランスジェニック植物とその利用 |
| EP2695944A2 (en) * | 2008-08-20 | 2014-02-12 | BASF Plant Science GmbH | Transgenic plants with increased yield |
| CN106191080A (zh) * | 2015-05-07 | 2016-12-07 | 杭州瑞丰生物科技有限公司 | Cyp78a基因在增加玉米株高和增强植株长势中的应用 |
| CN109207508A (zh) * | 2018-08-03 | 2019-01-15 | 浙江大学 | 一种提高农作物产量的方法 |
Non-Patent Citations (4)
| Title |
|---|
| KAZUMARU MIYOSHI等: ""PLASTOCHRON1，a timekeper of leaf initiation in rice,encodes cytochrome P45O"", 《PNAS》 * |
| MCCOMBIE,W.R.等: ""Genomic sequence for Oryza sativa，Nipponbare strain,clone OSJNBa0044A10,from chromosome 10,complete sequence"", 《GENBANK DATABASE》 * |
| 封功能等: ""水稻类树状突变体pla1-5的鉴定与基因克隆"", 《中国水稻科学》 * |
| 张晓琼等: ""LAZY1通过油菜素内酯途径调控水稻叶夹角的发育"", 《作物学报》 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Schwarz et al. | Cytokinin regulates the activity of the inflorescence meristem and components of seed yield in oilseed rape | |
| Sun et al. | Comparative transcriptome analysis reveals resistance-related genes and pathways in Musa acuminata banana'Guijiao 9'in response to Fusarium wilt | |
| Kaur et al. | Recent advances in cucumber (Cucumis sativus L.) | |
| Sharma et al. | A single NLR gene confers resistance to leaf and stripe rust in wheat | |
| US9745596B2 (en) | Identification and use of KRP mutants in wheat | |
| Braun et al. | Gene expression profiling and fine mapping identifies a gibberellin 2-oxidase gene co-segregating with the dominant dwarfing gene Ddw1 in rye (Secale cereale L.) | |
| Li et al. | Transcriptomic and biochemical analysis of upland cotton (Gossypium hirsutum) and a chromosome segment substitution line from G. hirsutum× G. barbadense in response to Verticillium dahliae infection | |
| CN116096901B (zh) | 植物病原体效应子和疾病抗性基因鉴定、组合物和使用方法 | |
| US20180244733A1 (en) | Isolated polypeptides and polynucleotides encoding same for generating plants with increased cuticlar water permeability | |
| CN116656867B (zh) | 用于检测西瓜全缘叶形的snp位点及应用 | |
| CN105985965A (zh) | 控制水稻粒型、外观品质和产量的基因gw7及其应用 | |
| Nguyen et al. | Ribosomal DNA locus variation and REMAP analysis of the diploid and triploid complexes of Lilium lancifolium | |
| CN107974457A (zh) | 果实大小增大的植物 | |
| Ferreira | Molecular analysis of genebanks for sustainable conservation and increased use of crop genetic resources. | |
| CN107058335A (zh) | 一种产生PVY抗性的NteIF4E‑1基因突变体及其应用 | |
| Bisi et al. | Molecular characterization of the S-alleles and compatibility among hybrid pear tree cultivars for subtropical regions | |
| Seguin et al. | Rubber tree (Hevea brasiliensis) | |
| CN105925587B (zh) | 一个受低温响应的早期水稻叶绿体发育基因及其检测方法和应用 | |
| Jairin et al. | Application of marker-assisted breeding to improve biotic stress resistance for rainfed lowland rice in Northeastern Thailand. | |
| CN109880832A (zh) | 水稻叶片叶夹角突变基因pla1及其应用 | |
| Nguyen | Molecular dissection of drought resistance in crop plants: from traits to genes | |
| Markussen et al. | Positioning of sex-correlated markers for Populus in a AFLP-and SSR-marker based genetic map of Populus tremula x tremuloides | |
| CN115074380B (zh) | 抽穗期调控基因Ehd5及其应用 | |
| Dong et al. | Fine mapping of the dominant glandless Gene Gl 2 e in Sea-island cotton (Gossypium barbadense L.) | |
| Sathyan et al. | Genes induced by WDS are differentially expressed in two populations of aleppo pine (Pinus halepensis) |
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 | ||
| WW01 | Invention patent application withdrawn after publication | ||
| WW01 | Invention patent application withdrawn after publication |
Application publication date: 20190614 |