CN114149994B - 褐飞虱性别决定调控基因Nlfmd2及其在有害生物防治中的用途 - Google Patents
褐飞虱性别决定调控基因Nlfmd2及其在有害生物防治中的用途 Download PDFInfo
- Publication number
- CN114149994B CN114149994B CN202111336896.7A CN202111336896A CN114149994B CN 114149994 B CN114149994 B CN 114149994B CN 202111336896 A CN202111336896 A CN 202111336896A CN 114149994 B CN114149994 B CN 114149994B
- Authority
- CN
- China
- Prior art keywords
- nlfmd2
- brown
- gly
- gene
- brown planthopper
- 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
- 241001556089 Nilaparvata lugens Species 0.000 title claims abstract description 64
- 230000020509 sex determination Effects 0.000 title claims abstract description 13
- 241000607479 Yersinia pestis Species 0.000 title abstract description 8
- 108700005075 Regulator Genes Proteins 0.000 title description 4
- 108090000623 proteins and genes Proteins 0.000 claims abstract description 22
- 230000009368 gene silencing by RNA Effects 0.000 claims abstract description 8
- 238000012228 RNA interference-mediated gene silencing Methods 0.000 claims abstract description 5
- 230000037361 pathway Effects 0.000 claims description 8
- 230000001850 reproductive effect Effects 0.000 abstract description 6
- 241000238631 Hexapoda Species 0.000 abstract description 5
- 108091030071 RNAI Proteins 0.000 abstract description 3
- 238000010353 genetic engineering Methods 0.000 abstract description 3
- 241001465754 Metazoa Species 0.000 abstract description 2
- 210000001161 mammalian embryo Anatomy 0.000 abstract description 2
- 208000009701 Embryo Loss Diseases 0.000 abstract 1
- 241000209094 Oryza Species 0.000 description 22
- 235000007164 Oryza sativa Nutrition 0.000 description 21
- 235000009566 rice Nutrition 0.000 description 21
- 238000011161 development Methods 0.000 description 7
- 230000018109 developmental process Effects 0.000 description 7
- 150000001413 amino acids Chemical class 0.000 description 6
- XKUKSGPZAADMRA-UHFFFAOYSA-N glycyl-glycyl-glycine Chemical compound NCC(=O)NCC(=O)NCC(O)=O XKUKSGPZAADMRA-UHFFFAOYSA-N 0.000 description 6
- 201000010099 disease Diseases 0.000 description 4
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 4
- 108010067216 glycyl-glycyl-glycine Proteins 0.000 description 4
- 108010010096 glycyl-glycyl-tyrosine Proteins 0.000 description 4
- 238000000520 microinjection Methods 0.000 description 4
- 239000002773 nucleotide Substances 0.000 description 4
- 125000003729 nucleotide group Chemical group 0.000 description 4
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 3
- 108091032955 Bacterial small RNA Proteins 0.000 description 3
- HQRHFUYMGCHHJS-LURJTMIESA-N Gly-Gly-Arg Chemical compound NCC(=O)NCC(=O)N[C@H](C(O)=O)CCCN=C(N)N HQRHFUYMGCHHJS-LURJTMIESA-N 0.000 description 3
- HQSKKSLNLSTONK-JTQLQIEISA-N Gly-Tyr-Gly Chemical compound OC(=O)CNC(=O)[C@@H](NC(=O)CN)CC1=CC=C(O)C=C1 HQSKKSLNLSTONK-JTQLQIEISA-N 0.000 description 3
- 241000819999 Nymphes Species 0.000 description 3
- 238000002347 injection Methods 0.000 description 3
- 239000007924 injection Substances 0.000 description 3
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 2
- 235000015001 Cucumis melo var inodorus Nutrition 0.000 description 2
- 240000002495 Cucumis melo var. inodorus Species 0.000 description 2
- INLIXXRWNUKVCF-JTQLQIEISA-N Gly-Gly-Tyr Chemical compound NCC(=O)NCC(=O)N[C@H](C(O)=O)CC1=CC=C(O)C=C1 INLIXXRWNUKVCF-JTQLQIEISA-N 0.000 description 2
- NNCSJUBVFBDDLC-YUMQZZPRSA-N Gly-Leu-Ser Chemical compound NCC(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(O)=O NNCSJUBVFBDDLC-YUMQZZPRSA-N 0.000 description 2
- JQFILXICXLDTRR-FBCQKBJTSA-N Gly-Thr-Gly Chemical compound NCC(=O)N[C@@H]([C@H](O)C)C(=O)NCC(O)=O JQFILXICXLDTRR-FBCQKBJTSA-N 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
- 108010079364 N-glycylalanine Proteins 0.000 description 2
- XALFIVXGQUEGKV-JSGCOSHPSA-N Phe-Val-Gly Chemical compound OC(=O)CNC(=O)[C@H](C(C)C)NC(=O)[C@@H](N)CC1=CC=CC=C1 XALFIVXGQUEGKV-JSGCOSHPSA-N 0.000 description 2
- TZXFLDNBYYGLKA-BZSNNMDCSA-N Tyr-Asp-Tyr Chemical compound C([C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(O)=O)C1=CC=C(O)C=C1 TZXFLDNBYYGLKA-BZSNNMDCSA-N 0.000 description 2
- 241000700605 Viruses Species 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 235000013601 eggs Nutrition 0.000 description 2
- 239000011521 glass Substances 0.000 description 2
- VPZXBVLAVMBEQI-UHFFFAOYSA-N glycyl-DL-alpha-alanine Natural products OC(=O)C(C)NC(=O)CN VPZXBVLAVMBEQI-UHFFFAOYSA-N 0.000 description 2
- XBGGUPMXALFZOT-UHFFFAOYSA-N glycyl-L-tyrosine hemihydrate Natural products NCC(=O)NC(C(O)=O)CC1=CC=C(O)C=C1 XBGGUPMXALFZOT-UHFFFAOYSA-N 0.000 description 2
- 108010062266 glycyl-glycyl-argininal Proteins 0.000 description 2
- 108010026364 glycyl-glycyl-leucine Proteins 0.000 description 2
- 108010089804 glycyl-threonine Proteins 0.000 description 2
- 108010045126 glycyl-tyrosyl-glycine Proteins 0.000 description 2
- 238000005286 illumination Methods 0.000 description 2
- 230000002452 interceptive effect Effects 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000003752 polymerase chain reaction Methods 0.000 description 2
- 238000010839 reverse transcription Methods 0.000 description 2
- 241000894007 species Species 0.000 description 2
- 101150028074 2 gene Proteins 0.000 description 1
- QDRGPQWIVZNJQD-CIUDSAMLSA-N Ala-Arg-Gln Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(N)=O)C(O)=O QDRGPQWIVZNJQD-CIUDSAMLSA-N 0.000 description 1
- NHCPCLJZRSIDHS-ZLUOBGJFSA-N Ala-Asp-Ala Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](C)C(O)=O NHCPCLJZRSIDHS-ZLUOBGJFSA-N 0.000 description 1
- VGPWRRFOPXVGOH-BYPYZUCNSA-N Ala-Gly-Gly Chemical compound C[C@H](N)C(=O)NCC(=O)NCC(O)=O VGPWRRFOPXVGOH-BYPYZUCNSA-N 0.000 description 1
- PGNNQOJOEGFAOR-KWQFWETISA-N Ala-Tyr-Gly Chemical compound OC(=O)CNC(=O)[C@@H](NC(=O)[C@@H](N)C)CC1=CC=C(O)C=C1 PGNNQOJOEGFAOR-KWQFWETISA-N 0.000 description 1
- YFBGNGASPGRWEM-DCAQKATOSA-N Arg-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](CCCN=C(N)N)N YFBGNGASPGRWEM-DCAQKATOSA-N 0.000 description 1
- HQIZDMIGUJOSNI-IUCAKERBSA-N Arg-Gly-Arg Chemical compound N[C@@H](CCCNC(N)=N)C(=O)NCC(=O)N[C@@H](CCCNC(N)=N)C(O)=O HQIZDMIGUJOSNI-IUCAKERBSA-N 0.000 description 1
- CYXCAHZVPFREJD-LURJTMIESA-N Arg-Gly-Gly Chemical compound NC(=N)NCCC[C@H](N)C(=O)NCC(=O)NCC(O)=O CYXCAHZVPFREJD-LURJTMIESA-N 0.000 description 1
- KRQSPVKUISQQFS-FJXKBIBVSA-N Arg-Gly-Thr Chemical compound C[C@@H](O)[C@@H](C(O)=O)NC(=O)CNC(=O)[C@@H](N)CCCN=C(N)N KRQSPVKUISQQFS-FJXKBIBVSA-N 0.000 description 1
- CLICCYPMVFGUOF-IHRRRGAJSA-N Arg-Lys-Leu Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(C)C)C(O)=O CLICCYPMVFGUOF-IHRRRGAJSA-N 0.000 description 1
- BVLIJXXSXBUGEC-SRVKXCTJSA-N Asn-Asn-Tyr Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O BVLIJXXSXBUGEC-SRVKXCTJSA-N 0.000 description 1
- XWFPGQVLOVGSLU-CIUDSAMLSA-N Asn-Gln-Arg Chemical compound NC(=O)C[C@H](N)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@H](C(O)=O)CCCN=C(N)N XWFPGQVLOVGSLU-CIUDSAMLSA-N 0.000 description 1
- ZYPWIUFLYMQZBS-SRVKXCTJSA-N Asn-Lys-Lys Chemical compound C(CCN)C[C@@H](C(=O)N[C@@H](CCCCN)C(=O)O)NC(=O)[C@H](CC(=O)N)N ZYPWIUFLYMQZBS-SRVKXCTJSA-N 0.000 description 1
- YSYTWUMRHSFODC-QWRGUYRKSA-N Asn-Tyr-Gly Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)NCC(O)=O YSYTWUMRHSFODC-QWRGUYRKSA-N 0.000 description 1
- VPSHHQXIWLGVDD-ZLUOBGJFSA-N Asp-Asp-Asp Chemical compound OC(=O)C[C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(O)=O)C(O)=O VPSHHQXIWLGVDD-ZLUOBGJFSA-N 0.000 description 1
- FRSGNOZCTWDVFZ-ACZMJKKPSA-N Asp-Asp-Gln Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CCC(N)=O)C(O)=O FRSGNOZCTWDVFZ-ACZMJKKPSA-N 0.000 description 1
- WCFCYFDBMNFSPA-ACZMJKKPSA-N Asp-Asp-Glu Chemical compound OC(=O)C[C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@H](C(O)=O)CCC(O)=O WCFCYFDBMNFSPA-ACZMJKKPSA-N 0.000 description 1
- RQYMKRMRZWJGHC-BQBZGAKWSA-N Asp-Gly-Met Chemical compound CSCC[C@@H](C(=O)O)NC(=O)CNC(=O)[C@H](CC(=O)O)N RQYMKRMRZWJGHC-BQBZGAKWSA-N 0.000 description 1
- VSMYBNPOHYAXSD-GUBZILKMSA-N Asp-Lys-Glu Chemical compound OC(=O)C[C@H](N)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCC(O)=O)C(O)=O VSMYBNPOHYAXSD-GUBZILKMSA-N 0.000 description 1
- 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
- 241000894006 Bacteria Species 0.000 description 1
- 241001466044 Delphacidae Species 0.000 description 1
- 206010059866 Drug resistance Diseases 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 241001466042 Fulgoromorpha Species 0.000 description 1
- PGPJSRSLQNXBDT-YUMQZZPRSA-N Gln-Arg-Gly Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)NCC(O)=O PGPJSRSLQNXBDT-YUMQZZPRSA-N 0.000 description 1
- JEFZIKRIDLHOIF-BYPYZUCNSA-N Gln-Gly Chemical compound NC(=O)CC[C@H](N)C(=O)NCC(O)=O JEFZIKRIDLHOIF-BYPYZUCNSA-N 0.000 description 1
- SGVGIVDZLSHSEN-RYUDHWBXSA-N Gln-Tyr-Gly Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)NCC(O)=O SGVGIVDZLSHSEN-RYUDHWBXSA-N 0.000 description 1
- CKRUHITYRFNUKW-WDSKDSINSA-N Glu-Asn-Gly Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(N)=O)C(=O)NCC(O)=O CKRUHITYRFNUKW-WDSKDSINSA-N 0.000 description 1
- UMIRPYLZFKOEOH-YVNDNENWSA-N Glu-Gln-Ile Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O UMIRPYLZFKOEOH-YVNDNENWSA-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
- FBEJIDRSQCGFJI-GUBZILKMSA-N Glu-Leu-Ser Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(O)=O FBEJIDRSQCGFJI-GUBZILKMSA-N 0.000 description 1
- GPSHCSTUYOQPAI-JHEQGTHGSA-N Glu-Thr-Gly Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)NCC(O)=O GPSHCSTUYOQPAI-JHEQGTHGSA-N 0.000 description 1
- MXJYXYDREQWUMS-XKBZYTNZSA-N Glu-Thr-Ser Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CO)C(O)=O MXJYXYDREQWUMS-XKBZYTNZSA-N 0.000 description 1
- HHSKZJZWQFPSKN-AVGNSLFASA-N Glu-Tyr-Asp Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CC(O)=O)C(O)=O HHSKZJZWQFPSKN-AVGNSLFASA-N 0.000 description 1
- BRFJMRSRMOMIMU-WHFBIAKZSA-N Gly-Ala-Asn Chemical compound NCC(=O)N[C@@H](C)C(=O)N[C@@H](CC(N)=O)C(O)=O BRFJMRSRMOMIMU-WHFBIAKZSA-N 0.000 description 1
- UGVQELHRNUDMAA-BYPYZUCNSA-N Gly-Ala-Gly Chemical compound [NH3+]CC(=O)N[C@@H](C)C(=O)NCC([O-])=O UGVQELHRNUDMAA-BYPYZUCNSA-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
- GWCRIHNSVMOBEQ-BQBZGAKWSA-N Gly-Arg-Ser Chemical compound [H]NCC(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CO)C(O)=O GWCRIHNSVMOBEQ-BQBZGAKWSA-N 0.000 description 1
- FMNHBTKMRFVGRO-FOHZUACHSA-N Gly-Asn-Thr Chemical compound C[C@@H](O)[C@@H](C(O)=O)NC(=O)[C@H](CC(N)=O)NC(=O)CN FMNHBTKMRFVGRO-FOHZUACHSA-N 0.000 description 1
- LLXVQPKEQQCISF-YUMQZZPRSA-N Gly-Asp-His Chemical compound C1=C(NC=N1)C[C@@H](C(=O)O)NC(=O)[C@H](CC(=O)O)NC(=O)CN LLXVQPKEQQCISF-YUMQZZPRSA-N 0.000 description 1
- DTRUBYPMMVPQPD-YUMQZZPRSA-N Gly-Gln-Arg Chemical compound [H]NCC(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O DTRUBYPMMVPQPD-YUMQZZPRSA-N 0.000 description 1
- BYYNJRSNDARRBX-YFKPBYRVSA-N Gly-Gln-Gly Chemical compound NCC(=O)N[C@@H](CCC(N)=O)C(=O)NCC(O)=O BYYNJRSNDARRBX-YFKPBYRVSA-N 0.000 description 1
- FIQQRCFQXGLOSZ-WDSKDSINSA-N Gly-Glu-Asp Chemical compound [H]NCC(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(O)=O)C(O)=O FIQQRCFQXGLOSZ-WDSKDSINSA-N 0.000 description 1
- LHRXAHLCRMQBGJ-RYUDHWBXSA-N Gly-Glu-Phe Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)O)NC(=O)[C@H](CCC(=O)O)NC(=O)CN LHRXAHLCRMQBGJ-RYUDHWBXSA-N 0.000 description 1
- QITBQGJOXQYMOA-ZETCQYMHSA-N Gly-Gly-Lys Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)CNC(=O)CN QITBQGJOXQYMOA-ZETCQYMHSA-N 0.000 description 1
- GMTXWRIDLGTVFC-IUCAKERBSA-N Gly-Lys-Glu Chemical compound [H]NCC(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCC(O)=O)C(O)=O GMTXWRIDLGTVFC-IUCAKERBSA-N 0.000 description 1
- PDUHNKAFQXQNLH-ZETCQYMHSA-N Gly-Lys-Gly Chemical compound NCCCC[C@H](NC(=O)CN)C(=O)NCC(O)=O PDUHNKAFQXQNLH-ZETCQYMHSA-N 0.000 description 1
- PTIIBFKSLCYQBO-NHCYSSNCSA-N Gly-Lys-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)O)NC(=O)[C@H](CCCCN)NC(=O)CN PTIIBFKSLCYQBO-NHCYSSNCSA-N 0.000 description 1
- WMGHDYWNHNLGBV-ONGXEEELSA-N Gly-Phe-Ala Chemical compound OC(=O)[C@H](C)NC(=O)[C@@H](NC(=O)CN)CC1=CC=CC=C1 WMGHDYWNHNLGBV-ONGXEEELSA-N 0.000 description 1
- QVDGHDFFYHKJPN-QWRGUYRKSA-N Gly-Phe-Cys Chemical compound NCC(=O)N[C@@H](Cc1ccccc1)C(=O)N[C@@H](CS)C(O)=O QVDGHDFFYHKJPN-QWRGUYRKSA-N 0.000 description 1
- 241000258937 Hemiptera Species 0.000 description 1
- UPJODPVSKKWGDQ-KLHWPWHYSA-N His-Thr-Pro Chemical compound C[C@H]([C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](CC2=CN=CN2)N)O UPJODPVSKKWGDQ-KLHWPWHYSA-N 0.000 description 1
- QSPLUJGYOPZINY-ZPFDUUQYSA-N Ile-Asp-Lys Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)N[C@@H](CCCCN)C(=O)O)N QSPLUJGYOPZINY-ZPFDUUQYSA-N 0.000 description 1
- NPROWIBAWYMPAZ-GUDRVLHUSA-N Ile-Asp-Pro Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)N1CCC[C@@H]1C(=O)O)N NPROWIBAWYMPAZ-GUDRVLHUSA-N 0.000 description 1
- SPQWWEZBHXHUJN-KBIXCLLPSA-N Ile-Glu-Ser Chemical compound CC[C@H](C)[C@H](N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CO)C(O)=O SPQWWEZBHXHUJN-KBIXCLLPSA-N 0.000 description 1
- AKOYRLRUFBZOSP-BJDJZHNGSA-N Ile-Lys-Ser Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CO)C(=O)O)N AKOYRLRUFBZOSP-BJDJZHNGSA-N 0.000 description 1
- AUIYHFRUOOKTGX-UKJIMTQDSA-N Ile-Val-Gln Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCC(=O)N)C(=O)O)N AUIYHFRUOOKTGX-UKJIMTQDSA-N 0.000 description 1
- 241000880493 Leptailurus serval Species 0.000 description 1
- OVZLLFONXILPDZ-VOAKCMCISA-N Leu-Lys-Thr Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H]([C@@H](C)O)C(O)=O OVZLLFONXILPDZ-VOAKCMCISA-N 0.000 description 1
- WSXTWLJHTLRFLW-SRVKXCTJSA-N Lys-Ala-Lys Chemical compound NCCCC[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@@H](CCCCN)C(O)=O WSXTWLJHTLRFLW-SRVKXCTJSA-N 0.000 description 1
- KNKHAVVBVXKOGX-JXUBOQSCSA-N Lys-Ala-Thr Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](C)C(=O)N[C@@H]([C@@H](C)O)C(O)=O KNKHAVVBVXKOGX-JXUBOQSCSA-N 0.000 description 1
- ZUGVARDEGWMMLK-SRVKXCTJSA-N Lys-Ser-Lys Chemical compound NCCCC[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@H](C(O)=O)CCCCN ZUGVARDEGWMMLK-SRVKXCTJSA-N 0.000 description 1
- GIKFNMZSGYAPEJ-HJGDQZAQSA-N Lys-Thr-Asp Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(O)=O)C(O)=O GIKFNMZSGYAPEJ-HJGDQZAQSA-N 0.000 description 1
- VHGIWFGJIHTASW-FXQIFTODSA-N Met-Ala-Asp Chemical compound CSCC[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@H](C(O)=O)CC(O)=O VHGIWFGJIHTASW-FXQIFTODSA-N 0.000 description 1
- ACYHZNZHIZWLQF-BQBZGAKWSA-N Met-Asn-Gly Chemical compound CSCC[C@H](N)C(=O)N[C@@H](CC(N)=O)C(=O)NCC(O)=O ACYHZNZHIZWLQF-BQBZGAKWSA-N 0.000 description 1
- PCTFVQATEGYHJU-FXQIFTODSA-N Met-Ser-Asn Chemical compound [H]N[C@@H](CCSC)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(N)=O)C(O)=O PCTFVQATEGYHJU-FXQIFTODSA-N 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
- XZFYRXDAULDNFX-UHFFFAOYSA-N N-L-cysteinyl-L-phenylalanine Natural products SCC(N)C(=O)NC(C(O)=O)CC1=CC=CC=C1 XZFYRXDAULDNFX-UHFFFAOYSA-N 0.000 description 1
- 108700026244 Open Reading Frames Proteins 0.000 description 1
- 206010033799 Paralysis Diseases 0.000 description 1
- JWQWPTLEOFNCGX-AVGNSLFASA-N Phe-Glu-Ser Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@@H](N)CC1=CC=CC=C1 JWQWPTLEOFNCGX-AVGNSLFASA-N 0.000 description 1
- YYKZDTVQHTUKDW-RYUDHWBXSA-N Phe-Gly-Gln Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)NCC(=O)N[C@@H](CCC(=O)N)C(=O)O)N YYKZDTVQHTUKDW-RYUDHWBXSA-N 0.000 description 1
- ZLGQEBCCANLYRA-RYUDHWBXSA-N Phe-Gly-Glu Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)NCC(=O)N[C@@H](CCC(O)=O)C(O)=O ZLGQEBCCANLYRA-RYUDHWBXSA-N 0.000 description 1
- QPVFUAUFEBPIPT-CDMKHQONSA-N Phe-Gly-Thr Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)NCC(=O)N[C@@H]([C@@H](C)O)C(O)=O QPVFUAUFEBPIPT-CDMKHQONSA-N 0.000 description 1
- BYAIIACBWBOJCU-URLPEUOOSA-N Phe-Ile-Thr Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H]([C@@H](C)O)C(O)=O BYAIIACBWBOJCU-URLPEUOOSA-N 0.000 description 1
- RORUIHAWOLADSH-HJWJTTGWSA-N Phe-Ile-Val Chemical compound CC(C)[C@@H](C(O)=O)NC(=O)[C@H]([C@@H](C)CC)NC(=O)[C@@H](N)CC1=CC=CC=C1 RORUIHAWOLADSH-HJWJTTGWSA-N 0.000 description 1
- SCKXGHWQPPURGT-KKUMJFAQSA-N Phe-Lys-Ser Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CO)C(O)=O SCKXGHWQPPURGT-KKUMJFAQSA-N 0.000 description 1
- SWXSLPHTJVAWDF-VEVYYDQMSA-N Pro-Asn-Thr Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O SWXSLPHTJVAWDF-VEVYYDQMSA-N 0.000 description 1
- UIMCLYYSUCIUJM-UWVGGRQHSA-N Pro-Gly-Lys Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)CNC(=O)[C@@H]1CCCN1 UIMCLYYSUCIUJM-UWVGGRQHSA-N 0.000 description 1
- OFGUOWQVEGTVNU-DCAQKATOSA-N Pro-Lys-Ala Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](C)C(O)=O OFGUOWQVEGTVNU-DCAQKATOSA-N 0.000 description 1
- ULWBBFKQBDNGOY-RWMBFGLXSA-N Pro-Lys-Pro Chemical compound C1C[C@H](NC1)C(=O)N[C@@H](CCCCN)C(=O)N2CCC[C@@H]2C(=O)O ULWBBFKQBDNGOY-RWMBFGLXSA-N 0.000 description 1
- CWZUFLWPEFHWEI-IHRRRGAJSA-N Pro-Tyr-Asp Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CC(O)=O)C(O)=O CWZUFLWPEFHWEI-IHRRRGAJSA-N 0.000 description 1
- SRTCFKGBYBZRHA-ACZMJKKPSA-N Ser-Ala-Glu Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](C)C(=O)N[C@@H](CCC(O)=O)C(O)=O SRTCFKGBYBZRHA-ACZMJKKPSA-N 0.000 description 1
- OQPNSDWGAMFJNU-QWRGUYRKSA-N Ser-Gly-Tyr Chemical compound OC[C@H](N)C(=O)NCC(=O)N[C@H](C(O)=O)CC1=CC=C(O)C=C1 OQPNSDWGAMFJNU-QWRGUYRKSA-N 0.000 description 1
- JIPVNVNKXJLFJF-BJDJZHNGSA-N Ser-Ile-Lys Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCCCN)C(=O)O)NC(=O)[C@H](CO)N JIPVNVNKXJLFJF-BJDJZHNGSA-N 0.000 description 1
- NADLKBTYNKUJEP-KATARQTJSA-N Ser-Thr-Leu Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(C)C)C(O)=O NADLKBTYNKUJEP-KATARQTJSA-N 0.000 description 1
- GSCVDSBEYVGMJQ-SRVKXCTJSA-N Ser-Tyr-Asp Chemical compound C1=CC(=CC=C1C[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)O)NC(=O)[C@H](CO)N)O GSCVDSBEYVGMJQ-SRVKXCTJSA-N 0.000 description 1
- XPNSAQMEAVSQRD-FBCQKBJTSA-N Thr-Gly-Gly Chemical compound C[C@@H](O)[C@H](N)C(=O)NCC(=O)NCC(O)=O XPNSAQMEAVSQRD-FBCQKBJTSA-N 0.000 description 1
- AZGZDDNKFFUDEH-QWRGUYRKSA-N Tyr-Gly-Ser Chemical compound OC[C@@H](C(O)=O)NC(=O)CNC(=O)[C@@H](N)CC1=CC=C(O)C=C1 AZGZDDNKFFUDEH-QWRGUYRKSA-N 0.000 description 1
- QAYSODICXVZUIA-WLTAIBSBSA-N Tyr-Gly-Thr Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)NCC(=O)N[C@@H]([C@@H](C)O)C(O)=O QAYSODICXVZUIA-WLTAIBSBSA-N 0.000 description 1
- WTTRJMAZPDHPGS-KKXDTOCCSA-N Tyr-Phe-Ala Chemical compound C[C@H](NC(=O)[C@H](Cc1ccccc1)NC(=O)[C@@H](N)Cc1ccc(O)cc1)C(O)=O WTTRJMAZPDHPGS-KKXDTOCCSA-N 0.000 description 1
- RIVVDNTUSRVTQT-IRIUXVKKSA-N Tyr-Thr-Gln Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CCC(=O)N)C(=O)O)NC(=O)[C@H](CC1=CC=C(C=C1)O)N)O RIVVDNTUSRVTQT-IRIUXVKKSA-N 0.000 description 1
- QPZMOUMNTGTEFR-ZKWXMUAHSA-N Val-Asn-Ala Chemical compound C[C@@H](C(=O)O)NC(=O)[C@H](CC(=O)N)NC(=O)[C@H](C(C)C)N QPZMOUMNTGTEFR-ZKWXMUAHSA-N 0.000 description 1
- DBOXBUDEAJVKRE-LSJOCFKGSA-N Val-Asn-Val Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)N[C@@H](C(C)C)C(=O)O)N DBOXBUDEAJVKRE-LSJOCFKGSA-N 0.000 description 1
- ZSZFTYVFQLUWBF-QXEWZRGKSA-N Val-Asp-Met Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)N[C@@H](CCSC)C(=O)O)N ZSZFTYVFQLUWBF-QXEWZRGKSA-N 0.000 description 1
- SCBITHMBEJNRHC-LSJOCFKGSA-N Val-Asp-Val Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)N[C@@H](C(C)C)C(=O)O)N SCBITHMBEJNRHC-LSJOCFKGSA-N 0.000 description 1
- BZMIYHIJVVJPCK-QSFUFRPTSA-N Val-Ile-Asn Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)O)NC(=O)[C@H](C(C)C)N BZMIYHIJVVJPCK-QSFUFRPTSA-N 0.000 description 1
- UVHFONIHVHLDDQ-IFFSRLJSSA-N Val-Thr-Glu Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CCC(=O)O)C(=O)O)NC(=O)[C@H](C(C)C)N)O UVHFONIHVHLDDQ-IFFSRLJSSA-N 0.000 description 1
- 241000746966 Zizania Species 0.000 description 1
- 235000002636 Zizania aquatica Nutrition 0.000 description 1
- 239000011543 agarose gel Substances 0.000 description 1
- 108010044940 alanylglutamine Proteins 0.000 description 1
- 108010047495 alanylglycine Proteins 0.000 description 1
- 108010011559 alanylphenylalanine Proteins 0.000 description 1
- 108010062796 arginyllysine Proteins 0.000 description 1
- 108010093581 aspartyl-proline Proteins 0.000 description 1
- 108010047857 aspartylglycine Proteins 0.000 description 1
- 108010092854 aspartyllysine Proteins 0.000 description 1
- 108010068265 aspartyltyrosine Proteins 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 229910002092 carbon dioxide Inorganic materials 0.000 description 1
- 239000001569 carbon dioxide Substances 0.000 description 1
- 235000013339 cereals Nutrition 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 230000001276 controlling effect Effects 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 239000012154 double-distilled water Substances 0.000 description 1
- 210000002257 embryonic structure Anatomy 0.000 description 1
- 238000004880 explosion Methods 0.000 description 1
- 238000010195 expression analysis Methods 0.000 description 1
- 235000011389 fruit/vegetable juice Nutrition 0.000 description 1
- 108010006664 gamma-glutamyl-glycyl-glycine Proteins 0.000 description 1
- 108010078144 glutaminyl-glycine Proteins 0.000 description 1
- 108010049041 glutamylalanine Proteins 0.000 description 1
- 150000002337 glycosamines Chemical class 0.000 description 1
- 108010001064 glycyl-glycyl-glycyl-glycine Proteins 0.000 description 1
- 108010077435 glycyl-phenylalanyl-glycine Proteins 0.000 description 1
- 108010082286 glycyl-seryl-alanine Proteins 0.000 description 1
- 108010087823 glycyltyrosine Proteins 0.000 description 1
- 238000003306 harvesting Methods 0.000 description 1
- 108010092114 histidylphenylalanine Proteins 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 108010030617 leucyl-phenylalanyl-valine Proteins 0.000 description 1
- 108010003700 lysyl aspartic acid Proteins 0.000 description 1
- 108010064235 lysylglycine Proteins 0.000 description 1
- 238000012423 maintenance Methods 0.000 description 1
- 230000008774 maternal effect Effects 0.000 description 1
- 230000013011 mating Effects 0.000 description 1
- 108010005942 methionylglycine Proteins 0.000 description 1
- 238000000034 method Methods 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 108020004707 nucleic acids Proteins 0.000 description 1
- 150000007523 nucleic acids Chemical class 0.000 description 1
- 102000039446 nucleic acids Human genes 0.000 description 1
- 230000017448 oviposition Effects 0.000 description 1
- 239000013612 plasmid Substances 0.000 description 1
- 108010079317 prolyl-tyrosine Proteins 0.000 description 1
- 210000003296 saliva Anatomy 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 239000000243 solution Substances 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 108010061238 threonyl-glycine Proteins 0.000 description 1
- 238000013518 transcription Methods 0.000 description 1
- 230000035897 transcription Effects 0.000 description 1
- 230000009261 transgenic effect Effects 0.000 description 1
- 108010073969 valyllysine Proteins 0.000 description 1
- 230000002792 vascular Effects 0.000 description 1
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/43504—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from invertebrates
- C07K14/43563—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from invertebrates from insects
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K67/00—Rearing or breeding animals, not otherwise provided for; New or modified breeds of animals
- A01K67/033—Rearing or breeding invertebrates; New breeds of invertebrates
- A01K67/0333—Genetically modified invertebrates, e.g. transgenic, polyploid
- A01K67/0337—Genetically modified Arthropods
- A01K67/0339—Genetically modified insects, e.g. Drosophila melanogaster, medfly
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/87—Introduction of foreign genetic material using processes not otherwise provided for, e.g. co-transformation
- C12N15/89—Introduction of foreign genetic material using processes not otherwise provided for, e.g. co-transformation using microinjection
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K2217/00—Genetically modified animals
- A01K2217/07—Animals genetically altered by homologous recombination
- A01K2217/075—Animals genetically altered by homologous recombination inducing loss of function, i.e. knock out
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K2227/00—Animals characterised by species
- A01K2227/70—Invertebrates
- A01K2227/706—Insects, e.g. Drosophila melanogaster, medfly
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Zoology (AREA)
- Genetics & Genomics (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Environmental Sciences (AREA)
- Molecular Biology (AREA)
- Wood Science & Technology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Biochemistry (AREA)
- Biophysics (AREA)
- General Health & Medical Sciences (AREA)
- Biodiversity & Conservation Biology (AREA)
- Animal Husbandry (AREA)
- Insects & Arthropods (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Animal Behavior & Ethology (AREA)
- Physics & Mathematics (AREA)
- Medicinal Chemistry (AREA)
- Gastroenterology & Hepatology (AREA)
- Microbiology (AREA)
- Toxicology (AREA)
- Plant Pathology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
Abstract
本发明属于动物基因工程技术领域，本发明提供了褐飞虱性别调控基因Nlfmd2及其在有害生物防治中的用途。所述褐飞虱性别调控基因Nlfmd2的序列如SEQ ID No.1所示。通过RNA干扰技术在褐飞虱体内敲除Nlfmd2后，雌性褐飞虱出现向雄虫转变的嵌合体性状，并失去生殖能力，雄性褐飞虱精子失去活力，从而丧失生殖能力。通过母本RNAi敲低基因Nlfmd2在下一代胚胎中的表达量，则引起胚胎死亡。这表明褐飞虱基因Nlfmd2在褐飞虱性别决定中起到重要作用，鉴于参与昆虫性别决定的基因具有高度特异性，可将该基因用于褐飞虱生物防治中，具有良好的应用场景。
Description
技术领域
本发明属于动物基因工程技术领域，具体涉及一种褐飞虱性别决定调控基因Nlfmd2及其在有害生物防治中的用途。
背景技术
褐飞虱(Nilaparvata lugens(Stal))属于半翅目飞虱科(Hemiptera:Delphacidae)，是一种可以远距离迁飞危害的水稻害虫，对环境有极强的适应性，是目前危害我国和许多东南亚等国家水稻的首要害虫。褐飞虱对寄主植物选择单一，只在水稻或野生稻上取食和产卵。褐飞虱主要通过刺吸寄主水稻韧皮部汁液给水稻生长发育造成严重影响，褐飞虱取食时形成的唾液鞘会阻塞维管束，而褐飞虱在取食后又分泌大量的“蜜露”，“蜜露”中含有大量的氨基酸和糖，可以使寄主水稻感染细菌，受褐飞虱危害严重的水稻会引起瘫痪倒伏，引起水稻“虱烧”现象，导致大田水稻严重减产甚至绝收。同时除了褐飞虱自身需求对水稻植株造成影响外，也作为媒介传播水稻病毒病如草状丛矮病和齿叶矮缩病，水稻病毒病会给水稻生产产量带来灾难性后果。20世纪80年代开始，我国的褐飞虱发生面积约1330-2000万hm2，约为我国种植水稻面积的二分之一，给我国水稻生产造成严重损失，在近三十年来看，褐飞虱危害我国水稻的主要特征有三个：爆发频率增加、危害范围扩大、危害程度加大。
长期以来，大量不合理使用化学防治，已经在成褐飞虱的抗药性激增，并造成褐飞虱大发生，严重威胁我国水稻产区粮食安全。随着生物技术，尤其是基因工程技术的发展，运用生物学手段进行褐飞虱防治已经成为新植物保护的重要手段。昆虫性别决定并保证了个体的发育和物种的延续，不同种类昆虫性别决定通路被报道各不相同，性别通路基因具有成为生物防治的靶标基因的潜力。
发明内容
本发明是针对水稻害虫褐飞虱的生物学防治，提供了影响褐飞虱性别决定通路基因Nlfmd2，及其功能和表达分析。
一方面，本发明提供了一种褐飞虱性别决定通路基因Nlfmd2，该基因是从水稻褐飞虱克隆得到的基因，其核苷酸序列如下(图1)：
ATGGCTGATTCCACTCTGGAATACGACATGAGTAACGGAGAATTTCAACAGGGCGGCGAGGATTACACCCAGGACGATCAAATGAACGGATCTGGCGAAAATGGAGGAGCTAACGAAGGAGGCGACAGTGGAAGCGCCGAAGCACCAGGCAAAGATGACGAAAGAAAACTGTTTGTTGGTGGACTCAGCTGGGAAACCTCTGACAAGGAACTCCGTGATCATTTCGGTACATTTGGTGAAATCGAAAGCGTAAATGTGAAGACTGATCCAAATACTGGCAGATCAAGAGGATTCGCGTTCATTGTTTTCAAGTCTGCGGATGCTATCGATAAGGTAGTCAACGCTGGAGACCATGTGATCAACAACAAGAAAATTGACCCAAAGAAAGCCAAGGCCAGACAAGGCAAGATTTTCGTCGGAGGATTGTCGGCTGAATTATCCGATGATGATATCAAGTCATACTTTGCTCAGTATGGAACCATTGTTCAAGTTGATATGCCGTACGACAAGTCAAAGAACCAGAGGAAGGGATTCTGCTTCATCACCTTCGAATCAGAACAGATTGTCACTGAAGTTCTTAAGACACCCAAGGCTTCAATCAAAGGCAAAGAAGTTGACGTGAGGAAAGCGACGCCGAAGCCTGATGGCATGGGCAACACGAGGGGAGGTAGAGGAGCCGGCCGTGGCACTCGAGGCAGGGGCCGTGGAGGATTTGGTCAGGGAGGCTATGGACAGGGAGGTTACGGTAACTACGGAGCTGGTGGCTATGGCACTGGCTACGATTACTATGGCAGTGGTGGCTACGGAGGTGGATACGGCGGAGGCTACGGAACCTACGACTACTCGGGATACGGCGGCTATGGTAGCTATGATGGATATGGAGCTTATGGCAACAACTATGAGACAGGATATGGCACTGGTGGAGGTGGACGAGGAGGCCGAGGAAAAGGTACCGGAGGCGGCTATGGTGGTGGCGGCAAGCAGAGGGGCGGCGGCCGAGGCCAGAGGCACACTCCCTACTGA(SEQ NO.1)，或经取代、缺失和/或增加核苷酸与所述核苷酸序列具有至少70％同源性，优选至少80％同源性，更优选至少90％同源性。
一方面，本发明提供了一种由所述核苷酸序列(SEQ NO.1)翻译的氨基酸序列Nlfmd2氨基酸序列，其序列如下(图1)：
MADSTLEYDMSNGEFQQGGEDYTQDDQMNGSGENGGANEGGDSGSAEAPGKDDERKLFVGGLSWETSDKELRDHFGTFGEIESVNVKTDPNTGRSRGFAFIVFKSADAIDKVVNAGDHVINNKKIDPKKAKARQGKIFVGGLSAELSDDDIKSYFAQYGTIVQVDMPYDKSKNQRKGFCFITFESEQIVTEVLKTPKASIKGKEVDVRKATPKPDGMGNTRGGRGAGRGTRGRGRGGFGQGGYGQGGYGNYGAGGYGTGYDYYGSGGYGGGYGGGYGTYDYSGYGGYGSYDGYGAYGNNYETGYGTGGGGRGGRGKGTGGGYGGGGKQRGGGRGQRHTPY(SEQ NO.2)或经取代、缺失和/或增加氨基酸与所述氨基酸序列具有至少70％同源性，优选至少80％同源性，更优选至少90％同源性。
本发明提供了褐飞虱性别决定通路基因Nlfmd2在生物学防治褐飞虱中的应用。优选的，用于成虫，若虫和卵期褐飞虱的防治。
本发明提供了Nlfmd2在褐飞虱性别特异性发育中的作用，具体为在褐飞虱三龄若虫期使用RNAi敲除Nlfmd2后，雌性褐飞虱产卵瓣缩短，卵巢不发育，失去生殖能力，与此同时雄性褐飞虱精子发育受到影响，失去活力，从而也失去繁殖能力；在雌成虫第一天使用显微注射将干扰双链注射进褐飞虱体内后，其产下的卵中Nlfmd2也被敲除，同时造成褐飞虱胚胎不发育，没有若虫孵化。
本发明提供的Nlfmd2 RNAi在褐飞虱若虫期造成褐飞虱雌虫和雄虫失去生殖能力，在雌性褐飞虱成虫初期造成后代胚胎期死亡。这表明褐飞虱性别决定通路基因Nlfmd2在褐飞虱性别特异性发育中起到了关键性作用，同时在生物学防治褐飞虱中(转基因水稻等)具有重要的价值，具有良好的应用前景。
附图说明
图1为Nlfmd2基因的核酸和氨基酸序列。
图2基因Nlfmd2在褐飞虱整个发育历期都表达。
图3在褐飞虱2龄期敲除Nlfmd2后，褐飞虱出现明显死亡。
图4在褐飞虱雌虫体内敲除基因Nlfmd2后，没有后代孵化并且产卵量明显下降。
图5为在3龄期使用小RNA干扰敲除基因Nlfmd2后，褐飞虱雌虫出现雄性化现象。
图6为在3龄期使用小RNA干扰敲除基因Nlfmd2后，褐飞虱雌虫卵巢不发育。
具体实施方式
下面通过具体实施例对本发明进行说明，实施例中未作详细描述的技术手段属于本领域专业技术人员熟知的常规技术。实施例只用于说明本发明，但不限制本发明的范围，任何本领域的技术人员在不付出创造性劳动的情况下，以本发明的实施例为基础所获得的其他实施例均属于本发明的保护范围。
实施例1：基因Nlfmd2全长的获得以及干扰RNA双链的合成。
褐飞虱种群在秀水128水稻苗上饲养，养虫室温度设定为26±0.5摄氏度，光照设定为12小时光照，12小时黑暗。总的RNA从10头雌虫和15头雄虫中提取，总量为1微克的RNA被用来反转录，反转录体系为10微升，将反应液稀释10倍(取9.8微升稀释到89.2微升的双蒸水中)，被用来作为模板。完整的开放阅读框序列通过特异性引物Nlfmd2-S：ACCCACTATTGCTGCTATTT(SEQ NO.3)，Nlfmd2-S：CAACACCCACTGCTAAAGA(SEQ NO.4)，经聚合酶链式反应(Polymerase Chain Reaction，PCR)得到，并连接PMD-19T载体上，进行复制筛选，并测序。干扰双链是使用N端带有T7启动子的双向特异性引物dsNlfmd2-S:GATCACTAATACGACTCACTATAGGGAGAATCTTTAGCAGTGGGTGT(SEQ NO.5)，dsNlfmd2-AS:GATCACTAATACGACTCACTATAGGGAGATGGTTCGGTGATATTCA(SEQ NO.6)与带有特异性靶序列的PMD-19T质粒的PCR产物为模板，使用MEGAscript T7体外转录试剂盒，合成的双链使用1％的琼脂糖凝胶进行检测，然后稀释1倍，放-80摄氏度冰箱保存。
实施例2：褐飞虱若虫期干扰和母本干扰。
使用拉针仪P-97，使用程序heat＝800，pull＝150，vel＝150，time＝80，将毛细管拉成适合显微注射的玻璃针，并将双链注入玻璃针中。选取2龄或3龄褐飞虱幼虫或刚羽化的雌性褐飞虱，使用二氧化碳麻醉1分钟，将褐飞虱置于体视镜下，使用显微注射仪进行注射，其中显微注射仪推荐参数为注射压力(injection pressure，Pi)1000hPa，压力维持时间(injection time，ti)0.5s，补偿压力(compensation pressure，pc)20hpa。
实施例3：死亡率统计和现象记录。
将完成注射的褐飞虱若虫饲养在水稻幼苗上，每24小时统计存活的若虫，并统计现象在表1中。
表1
结果表明：褐飞虱3龄期使用小RNA干扰敲除基因Nlfmd2后，褐飞虱雄虫失去生殖能力，与对照雌虫交配后，没有后代孵化。
以上所述为本发明较佳的具体实施方式，但在其基础上可以进行一些改进或修改，这对任何熟悉本领域的技术人员而言是显而易见的。因此，在本发明基础上所作的这些修改和改进，均属于本发明要求保护的范围。
序列表
<110> 宁波大学
<120> 褐飞虱性别决定调控基因Nlfmd2及其在有害生物防治中的用途
<130> NKY-001
<160> 6
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1023
<212> DNA
<213> 人工序列()
<400> 1
atggctgatt ccactctgga atacgacatg agtaacggag aatttcaaca gggcggcgag 60
gattacaccc aggacgatca aatgaacgga tctggcgaaa atggaggagc taacgaagga 120
ggcgacagtg gaagcgccga agcaccaggc aaagatgacg aaagaaaact gtttgttggt 180
ggactcagct gggaaacctc tgacaaggaa ctccgtgatc atttcggtac atttggtgaa 240
atcgaaagcg taaatgtgaa gactgatcca aatactggca gatcaagagg attcgcgttc 300
attgttttca agtctgcgga tgctatcgat aaggtagtca acgctggaga ccatgtgatc 360
aacaacaaga aaattgaccc aaagaaagcc aaggccagac aaggcaagat tttcgtcgga 420
ggattgtcgg ctgaattatc cgatgatgat atcaagtcat actttgctca gtatggaacc 480
attgttcaag ttgatatgcc gtacgacaag tcaaagaacc agaggaaggg attctgcttc 540
atcaccttcg aatcagaaca gattgtcact gaagttctta agacacccaa ggcttcaatc 600
aaaggcaaag aagttgacgt gaggaaagcg acgccgaagc ctgatggcat gggcaacacg 660
aggggaggta gaggagccgg ccgtggcact cgaggcaggg gccgtggagg atttggtcag 720
ggaggctatg gacagggagg ttacggtaac tacggagctg gtggctatgg cactggctac 780
gattactatg gcagtggtgg ctacggaggt ggatacggcg gaggctacgg aacctacgac 840
tactcgggat acggcggcta tggtagctat gatggatatg gagcttatgg caacaactat 900
gagacaggat atggcactgg tggaggtgga cgaggaggcc gaggaaaagg taccggaggc 960
ggctatggtg gtggcggcaa gcagaggggc ggcggccgag gccagaggca cactccctac 1020
tga 1023
<210> 2
<211> 340
<212> PRT
<213> 人工序列()
<400> 2
Met Ala Asp Ser Thr Leu Glu Tyr Asp Met Ser Asn Gly Glu Phe Gln
1 5 10 15
Gln Gly Gly Glu Asp Tyr Thr Gln Asp Asp Gln Met Asn Gly Ser Gly
20 25 30
Glu Asn Gly Gly Ala Asn Glu Gly Gly Asp Ser Gly Ser Ala Glu Ala
35 40 45
Pro Gly Lys Asp Asp Glu Arg Lys Leu Phe Val Gly Gly Leu Ser Trp
50 55 60
Glu Thr Ser Asp Lys Glu Leu Arg Asp His Phe Gly Thr Phe Gly Glu
65 70 75 80
Ile Glu Ser Val Asn Val Lys Thr Asp Pro Asn Thr Gly Arg Ser Arg
85 90 95
Gly Phe Ala Phe Ile Val Phe Lys Ser Ala Asp Ala Ile Asp Lys Val
100 105 110
Val Asn Ala Gly Asp His Val Ile Asn Asn Lys Lys Ile Asp Pro Lys
115 120 125
Lys Ala Lys Ala Arg Gln Gly Lys Ile Phe Val Gly Gly Leu Ser Ala
130 135 140
Glu Leu Ser Asp Asp Asp Ile Lys Ser Tyr Phe Ala Gln Tyr Gly Thr
145 150 155 160
Ile Val Gln Val Asp Met Pro Tyr Asp Lys Ser Lys Asn Gln Arg Lys
165 170 175
Gly Phe Cys Phe Ile Thr Phe Glu Ser Glu Gln Ile Val Thr Glu Val
180 185 190
Leu Lys Thr Pro Lys Ala Ser Ile Lys Gly Lys Glu Val Asp Val Arg
195 200 205
Lys Ala Thr Pro Lys Pro Asp Gly Met Gly Asn Thr Arg Gly Gly Arg
210 215 220
Gly Ala Gly Arg Gly Thr Arg Gly Arg Gly Arg Gly Gly Phe Gly Gln
225 230 235 240
Gly Gly Tyr Gly Gln Gly Gly Tyr Gly Asn Tyr Gly Ala Gly Gly Tyr
245 250 255
Gly Thr Gly Tyr Asp Tyr Tyr Gly Ser Gly Gly Tyr Gly Gly Gly Tyr
260 265 270
Gly Gly Gly Tyr Gly Thr Tyr Asp Tyr Ser Gly Tyr Gly Gly Tyr Gly
275 280 285
Ser Tyr Asp Gly Tyr Gly Ala Tyr Gly Asn Asn Tyr Glu Thr Gly Tyr
290 295 300
Gly Thr Gly Gly Gly Gly Arg Gly Gly Arg Gly Lys Gly Thr Gly Gly
305 310 315 320
Gly Tyr Gly Gly Gly Gly Lys Gln Arg Gly Gly Gly Arg Gly Gln Arg
325 330 335
His Thr Pro Tyr
340
<210> 3
<211> 20
<212> DNA
<213> 人工序列()
<400> 3
acccactatt gctgctattt 20
<210> 4
<211> 19
<212> DNA
<213> 人工序列()
<400> 4
caacacccac tgctaaaga 19
<210> 5
<211> 47
<212> DNA
<213> 人工序列()
<400> 5
gatcactaat acgactcact atagggagaa tctttagcag tgggtgt 47
<210> 6
<211> 46
<212> DNA
<213> 人工序列()
<400> 6
gatcactaat acgactcact atagggagat ggttcggtga tattca 46
Claims (3)
1.一种褐飞虱性别决定通路基因Nlfmd2在生物学防治褐飞虱中的用途，其特征在于：所述褐飞虱性别决定通路基因Nlfmd2序列为SEQ ID NO.1。
2.根据权利要求1所述的用途，其特征在于：其中所述褐飞虱为成虫，若虫和卵期的褐飞虱。
3.权利要求1或2所述的用途，其特征在于：通过RNA干扰技术在褐飞虱体内敲除Nlfmd2后实现。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111336896.7A CN114149994B (zh) | 2021-11-12 | 2021-11-12 | 褐飞虱性别决定调控基因Nlfmd2及其在有害生物防治中的用途 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111336896.7A CN114149994B (zh) | 2021-11-12 | 2021-11-12 | 褐飞虱性别决定调控基因Nlfmd2及其在有害生物防治中的用途 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN114149994A CN114149994A (zh) | 2022-03-08 |
| CN114149994B true CN114149994B (zh) | 2023-05-09 |
Family
ID=80459992
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202111336896.7A Active CN114149994B (zh) | 2021-11-12 | 2021-11-12 | 褐飞虱性别决定调控基因Nlfmd2及其在有害生物防治中的用途 |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN114149994B (zh) |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104232651A (zh) * | 2014-07-18 | 2014-12-24 | 中国计量学院 | 褐飞虱Nlcdc2基因及其编码产物与应用 |
| CN104593391A (zh) * | 2015-01-14 | 2015-05-06 | 中国计量学院 | 褐飞虱生存及生长发育相关的NlPIK3R1基因、编码蛋白及其应用 |
| CN110066798A (zh) * | 2019-04-16 | 2019-07-30 | 杭州师范大学 | 褐飞虱NlInR基因作为靶点在制备防治褐飞虱药物中的应用 |
| CN110684095A (zh) * | 2019-10-25 | 2020-01-14 | 中国水稻研究所 | 褐飞虱雄虫特异表达的蛋白磷酸基因NlPPP1-Y及其应用 |
-
2021
- 2021-11-12 CN CN202111336896.7A patent/CN114149994B/zh active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104232651A (zh) * | 2014-07-18 | 2014-12-24 | 中国计量学院 | 褐飞虱Nlcdc2基因及其编码产物与应用 |
| CN104593391A (zh) * | 2015-01-14 | 2015-05-06 | 中国计量学院 | 褐飞虱生存及生长发育相关的NlPIK3R1基因、编码蛋白及其应用 |
| CN110066798A (zh) * | 2019-04-16 | 2019-07-30 | 杭州师范大学 | 褐飞虱NlInR基因作为靶点在制备防治褐飞虱药物中的应用 |
| CN110684095A (zh) * | 2019-10-25 | 2020-01-14 | 中国水稻研究所 | 褐飞虱雄虫特异表达的蛋白磷酸基因NlPPP1-Y及其应用 |
Non-Patent Citations (1)
| Title |
|---|
| Ji-Chong Zhuo等.A feminizing switch in a hemimetabolous insect.SCIENCE ADVANCES.2021,第7卷第1-13页. * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN114149994A (zh) | 2022-03-08 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN104480117B (zh) | 花生nbs‑lrr基因及其在烟草抗青枯病中的应用 | |
| AU2014350744C1 (en) | Method for controlling pests | |
| CN103718896A (zh) | 控制害虫的方法 | |
| CN1308345C (zh) | 一种海岛棉脂质转移蛋白及其编码基因与应用 | |
| CN110195049B (zh) | 褐飞虱眼色基因NlGCHI及其编码蛋白、应用 | |
| CN113248586B (zh) | 褐飞虱pib14蛋白及其编码基因在调控植物抗褐飞虱中的应用 | |
| CN114149994B (zh) | 褐飞虱性别决定调控基因Nlfmd2及其在有害生物防治中的用途 | |
| CN113584077A (zh) | 沉默桑实杯盘菌关键致病基因CsGPA1的沉默载体及其应用和方法 | |
| CN102851297B (zh) | 一种烟蚜hunchback基因cDNA及其应用 | |
| CN109468336B (zh) | 褐飞虱蛋白磷酸基因NlPP1、蛋白及其dsRNA和应用 | |
| CN111826364B (zh) | 一种抗病虫害相关基因及其应用 | |
| CN102533811A (zh) | 枳促分裂原活化蛋白激酶基因PtrMAPK克隆及用于提高植物抗旱能力 | |
| CN114525303B (zh) | CaM2基因作为调控因子在提高植物对虫害胁迫抗性中的应用 | |
| CN114989283A (zh) | Tcp19蛋白在调控水稻抗纹枯病中的应用 | |
| Balakrishnan et al. | Rice physical defenses and their role against insect herbivores | |
| CN108456683A (zh) | 一个调控水稻抽穗期基因sid1的功能及应用 | |
| CN111411113B (zh) | 梨保卫细胞钾离子吸收通道基因PbrKAT1及其应用 | |
| CN108822210B (zh) | 蝶蛹金小蜂毒液Kazal-type丝氨酸蛋白酶抑制剂PpSPI24蛋白及应用 | |
| CN112266919A (zh) | 水稻源抗虫相关基因OsIDP1及其编码产物与应用 | |
| CN112062825B (zh) | 一种灰飞虱基因LsECP1及其编码产物与应用 | |
| CN106701780B (zh) | 调控石榴花胚珠发育的PgAG基因及其用途 | |
| CN114807156B (zh) | 褐飞虱ncs2基因或编码的蛋白作为靶点在制备防治褐飞虱药物中的应用 | |
| CN114317549B (zh) | 毒蕈型c型乙酰胆碱受体在飞蝗防治中的应用 | |
| KR102397634B1 (ko) | 진딧물의 ERR유전자에 특이적인 dsRNA 및 이를 이용한 진딧물 방제용 조성물 과 방제방법 | |
| CN104255771B (zh) | 杀虫蛋白的用途 |
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 |