CN102499254A - Molt-inhibiting insecticide and application thereof - Google Patents
Molt-inhibiting insecticide and application thereof Download PDFInfo
- Publication number
- CN102499254A CN102499254A CN2011103354165A CN201110335416A CN102499254A CN 102499254 A CN102499254 A CN 102499254A CN 2011103354165 A CN2011103354165 A CN 2011103354165A CN 201110335416 A CN201110335416 A CN 201110335416A CN 102499254 A CN102499254 A CN 102499254A
- Authority
- CN
- China
- Prior art keywords
- nlecr
- seq
- brown planthopper
- ecr
- gene
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- 239000002917 insecticide Substances 0.000 title claims abstract description 16
- 230000002401 inhibitory effect Effects 0.000 title abstract 4
- 241001556089 Nilaparvata lugens Species 0.000 claims abstract description 46
- 108091032973 (ribonucleotides)n+m Proteins 0.000 claims abstract description 27
- 102000040650 (ribonucleotides)n+m Human genes 0.000 claims abstract description 23
- 230000000692 anti-sense effect Effects 0.000 claims abstract description 9
- 239000003814 drug Substances 0.000 claims abstract description 3
- 239000002773 nucleotide Substances 0.000 claims description 16
- 125000003729 nucleotide group Chemical group 0.000 claims description 16
- 238000005266 casting Methods 0.000 claims description 8
- 238000002360 preparation method Methods 0.000 claims description 3
- 108090000623 proteins and genes Proteins 0.000 abstract description 24
- 238000011161 development Methods 0.000 abstract description 5
- 238000012216 screening Methods 0.000 abstract description 2
- 108091081021 Sense strand Proteins 0.000 abstract 3
- 239000004480 active ingredient Substances 0.000 abstract 1
- 229940079593 drug Drugs 0.000 abstract 1
- 229920002477 rna polymer Polymers 0.000 abstract 1
- 238000013518 transcription Methods 0.000 abstract 1
- 230000035897 transcription Effects 0.000 abstract 1
- 108010057988 ecdysone receptor Proteins 0.000 description 21
- 241000238631 Hexapoda Species 0.000 description 17
- 239000012634 fragment Substances 0.000 description 15
- 230000004083 survival effect Effects 0.000 description 11
- 230000009261 transgenic effect Effects 0.000 description 11
- 241000209094 Oryza Species 0.000 description 10
- 235000007164 Oryza sativa Nutrition 0.000 description 10
- 238000006243 chemical reaction Methods 0.000 description 10
- 235000009566 rice Nutrition 0.000 description 10
- 230000000694 effects Effects 0.000 description 9
- 238000000034 method Methods 0.000 description 9
- 210000003491 skin Anatomy 0.000 description 8
- 230000006870 function Effects 0.000 description 7
- 239000013612 plasmid Substances 0.000 description 7
- 108020004999 messenger RNA Proteins 0.000 description 6
- 235000013399 edible fruits Nutrition 0.000 description 5
- 238000011144 upstream manufacturing Methods 0.000 description 5
- 241001599018 Melanogaster Species 0.000 description 4
- 230000003321 amplification Effects 0.000 description 4
- 230000008859 change Effects 0.000 description 4
- 230000004087 circulation Effects 0.000 description 4
- 238000005859 coupling reaction Methods 0.000 description 4
- 230000018109 developmental process Effects 0.000 description 4
- 230000009368 gene silencing by RNA Effects 0.000 description 4
- 210000004907 gland Anatomy 0.000 description 4
- 239000005556 hormone Substances 0.000 description 4
- 229940088597 hormone Drugs 0.000 description 4
- QCAWEPFNJXQPAN-UHFFFAOYSA-N methoxyfenozide Chemical compound COC1=CC=CC(C(=O)NN(C(=O)C=2C=C(C)C=C(C)C=2)C(C)(C)C)=C1C QCAWEPFNJXQPAN-UHFFFAOYSA-N 0.000 description 4
- 238000003199 nucleic acid amplification method Methods 0.000 description 4
- 230000008569 process Effects 0.000 description 4
- 238000011160 research Methods 0.000 description 4
- 101150076489 B gene Proteins 0.000 description 3
- HPNSNYBUADCFDR-UHFFFAOYSA-N chromafenozide Chemical compound CC1=CC(C)=CC(C(=O)N(NC(=O)C=2C(=C3CCCOC3=CC=2)C)C(C)(C)C)=C1 HPNSNYBUADCFDR-UHFFFAOYSA-N 0.000 description 3
- 230000012447 hatching Effects 0.000 description 3
- 239000000833 heterodimer Substances 0.000 description 3
- 229940042795 hydrazides for tuberculosis treatment Drugs 0.000 description 3
- 230000005764 inhibitory process Effects 0.000 description 3
- 239000007788 liquid Substances 0.000 description 3
- 230000029052 metamorphosis Effects 0.000 description 3
- 239000013600 plasmid vector Substances 0.000 description 3
- QYPNKSZPJQQLRK-UHFFFAOYSA-N tebufenozide Chemical compound C1=CC(CC)=CC=C1C(=O)NN(C(C)(C)C)C(=O)C1=CC(C)=CC(C)=C1 QYPNKSZPJQQLRK-UHFFFAOYSA-N 0.000 description 3
- GRRIYLZJLGTQJX-UHFFFAOYSA-N 1,2-dibenzoylhydrazine Chemical class C=1C=CC=CC=1C(=O)NNC(=O)C1=CC=CC=C1 GRRIYLZJLGTQJX-UHFFFAOYSA-N 0.000 description 2
- NKDFYOWSKOHCCO-YPVLXUMRSA-N 20-hydroxyecdysone Chemical compound C1[C@@H](O)[C@@H](O)C[C@]2(C)[C@@H](CC[C@@]3([C@@H]([C@@](C)(O)[C@H](O)CCC(C)(O)C)CC[C@]33O)C)C3=CC(=O)[C@@H]21 NKDFYOWSKOHCCO-YPVLXUMRSA-N 0.000 description 2
- HXWZQRICWSADMH-SEHXZECUSA-N 20-hydroxyecdysone Natural products CC(C)(C)CC[C@@H](O)[C@@](C)(O)[C@H]1CC[C@@]2(O)C3=CC(=O)[C@@H]4C[C@@H](O)[C@@H](O)C[C@]4(C)[C@H]3CC[C@]12C HXWZQRICWSADMH-SEHXZECUSA-N 0.000 description 2
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- 101150103152 ECR gene Proteins 0.000 description 2
- 108010042407 Endonucleases Proteins 0.000 description 2
- 102000004533 Endonucleases Human genes 0.000 description 2
- 108090000790 Enzymes Proteins 0.000 description 2
- 102000004190 Enzymes Human genes 0.000 description 2
- TWRXJAOTZQYOKJ-UHFFFAOYSA-L Magnesium chloride Chemical compound [Mg+2].[Cl-].[Cl-] TWRXJAOTZQYOKJ-UHFFFAOYSA-L 0.000 description 2
- 239000005917 Methoxyfenozide Substances 0.000 description 2
- 241000819999 Nymphes Species 0.000 description 2
- 229910019142 PO4 Inorganic materials 0.000 description 2
- WCUXLLCKKVVCTQ-UHFFFAOYSA-M Potassium chloride Chemical compound [Cl-].[K+] WCUXLLCKKVVCTQ-UHFFFAOYSA-M 0.000 description 2
- 238000012228 RNA interference-mediated gene silencing Methods 0.000 description 2
- 108091030071 RNAI Proteins 0.000 description 2
- 102000034527 Retinoid X Receptors Human genes 0.000 description 2
- 108010038912 Retinoid X Receptors Proteins 0.000 description 2
- 241000254113 Tribolium castaneum Species 0.000 description 2
- 230000008901 benefit Effects 0.000 description 2
- NKDFYOWSKOHCCO-UHFFFAOYSA-N beta-ecdysone Natural products C1C(O)C(O)CC2(C)C(CCC3(C(C(C)(O)C(O)CCC(C)(O)C)CCC33O)C)C3=CC(=O)C21 NKDFYOWSKOHCCO-UHFFFAOYSA-N 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- 239000002299 complementary DNA Substances 0.000 description 2
- 239000012141 concentrate Substances 0.000 description 2
- 238000013461 design Methods 0.000 description 2
- 235000013601 eggs Nutrition 0.000 description 2
- 239000005090 green fluorescent protein Substances 0.000 description 2
- 230000035772 mutation Effects 0.000 description 2
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 2
- 239000010452 phosphate Substances 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 239000000376 reactant Substances 0.000 description 2
- 230000004044 response Effects 0.000 description 2
- 108091008146 restriction endonucleases Proteins 0.000 description 2
- 238000012163 sequencing technique Methods 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 210000001519 tissue Anatomy 0.000 description 2
- QKNYBSVHEMOAJP-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;hydron;chloride Chemical compound Cl.OCC(N)(CO)CO QKNYBSVHEMOAJP-UHFFFAOYSA-N 0.000 description 1
- OPIFSICVWOWJMJ-AEOCFKNESA-N 5-bromo-4-chloro-3-indolyl beta-D-galactoside Chemical compound O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@H]1OC1=CNC2=CC=C(Br)C(Cl)=C12 OPIFSICVWOWJMJ-AEOCFKNESA-N 0.000 description 1
- 108010022579 ATP dependent 26S protease Proteins 0.000 description 1
- 241000589158 Agrobacterium Species 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- 241000238657 Blattella germanica Species 0.000 description 1
- 241000255789 Bombyx mori Species 0.000 description 1
- ZAMOUSCENKQFHK-UHFFFAOYSA-N Chlorine atom Chemical compound [Cl] ZAMOUSCENKQFHK-UHFFFAOYSA-N 0.000 description 1
- 239000005887 Chromafenozide Substances 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
- 241000255581 Drosophila <fruit fly, genus> Species 0.000 description 1
- 241000620209 Escherichia coli DH5[alpha] Species 0.000 description 1
- 108700039691 Genetic Promoter Regions Proteins 0.000 description 1
- 108010043121 Green Fluorescent Proteins Proteins 0.000 description 1
- 102000004144 Green Fluorescent Proteins Human genes 0.000 description 1
- 108700001097 Insect Genes Proteins 0.000 description 1
- 241000258916 Leptinotarsa decemlineata Species 0.000 description 1
- 241000255908 Manduca sexta Species 0.000 description 1
- 108020005497 Nuclear hormone receptor Proteins 0.000 description 1
- 102000007399 Nuclear hormone receptor Human genes 0.000 description 1
- 108091028043 Nucleic acid sequence Proteins 0.000 description 1
- 239000012807 PCR reagent Substances 0.000 description 1
- 241000382353 Pupa Species 0.000 description 1
- 108010092799 RNA-directed DNA polymerase Proteins 0.000 description 1
- 239000005937 Tebufenozide Substances 0.000 description 1
- 208000002474 Tinea Diseases 0.000 description 1
- 108091023040 Transcription factor Proteins 0.000 description 1
- 102000040945 Transcription factor Human genes 0.000 description 1
- 241000893966 Trichophyton verrucosum Species 0.000 description 1
- 241000251539 Vertebrata <Metazoa> Species 0.000 description 1
- 210000001015 abdomen Anatomy 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 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 1
- 229960000723 ampicillin Drugs 0.000 description 1
- 238000012197 amplification kit Methods 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 238000009395 breeding Methods 0.000 description 1
- 230000001488 breeding effect Effects 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 239000000460 chlorine Substances 0.000 description 1
- 229910052801 chlorine Inorganic materials 0.000 description 1
- 210000001136 chorion Anatomy 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 230000001276 controlling effect Effects 0.000 description 1
- 230000007850 degeneration Effects 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 230000004069 differentiation Effects 0.000 description 1
- 210000002257 embryonic structure Anatomy 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000003912 environmental pollution Methods 0.000 description 1
- 239000002532 enzyme inhibitor Substances 0.000 description 1
- 210000002615 epidermis Anatomy 0.000 description 1
- 230000003203 everyday effect Effects 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 210000002468 fat body Anatomy 0.000 description 1
- 230000035558 fertility Effects 0.000 description 1
- 238000010230 functional analysis Methods 0.000 description 1
- 230000002068 genetic effect Effects 0.000 description 1
- YQOKLYTXVFAUCW-UHFFFAOYSA-N guanidine;isothiocyanic acid Chemical compound N=C=S.NC(N)=N YQOKLYTXVFAUCW-UHFFFAOYSA-N 0.000 description 1
- CNKHSLKYRMDDNQ-UHFFFAOYSA-N halofenozide Chemical compound C=1C=CC=CC=1C(=O)N(C(C)(C)C)NC(=O)C1=CC=C(Cl)C=C1 CNKHSLKYRMDDNQ-UHFFFAOYSA-N 0.000 description 1
- 230000001939 inductive effect Effects 0.000 description 1
- 239000003112 inhibitor Substances 0.000 description 1
- 229910001629 magnesium chloride Inorganic materials 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 108020004017 nuclear receptors Proteins 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 210000001672 ovary Anatomy 0.000 description 1
- 239000001103 potassium chloride Substances 0.000 description 1
- 235000011164 potassium chloride Nutrition 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 239000012264 purified product Substances 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 238000005070 sampling Methods 0.000 description 1
- 230000028327 secretion Effects 0.000 description 1
- 230000019491 signal transduction Effects 0.000 description 1
Images
Landscapes
- Agricultural Chemicals And Associated Chemicals (AREA)
Abstract
The invention discloses molt-inhibiting insecticide and application thereof. Double-stranded RNA (ribonucleic acid) serves as a main active ingredient of the molt-inhibiting insecticide, and a transcription template for the double-stranded RNA is at least one of NlEcR-A, NlEcR-B, and NlEcR-A/B. A sense strand of the NlEcR-A is shown in SEQ (sequence) ID: No.1, while an antisense strand of the NlEcR-A is showed in SEQ ID: No.2. A sense strand of the NlEcR-B is shown in SEQ ID: No.3, while an antisense strand of the NlEcR-B is shown in SEQ ID: No.4. A sense strand of the NlEcR-A/B is shown in SEQ ID: No.5, while an antisense strand of the NlEcR-A/B is shown in SEQ ID: No.6. The double-stranded RNA is obtained by screening important genes related to growth and development of brown plant-hoppers, and is effective in controlling the brown plant-hoppers. Therefore, the molt-inhibiting insecticide can be used for preparing drugs for controlling brown plant-hoppers.
Description
Technical field
The invention belongs to biological technical field, particularly a kind of casting off a skin suppressed insecticides and application thereof.
Background technology
The too busy to get away molt metamorphosis that grows of insect; The moulting hormone of prothoracic gland secretion is being played the part of important role among this life course; These moulting hormones are generally 20-hydroxyecdysone (20-hydroxyecdysone, 20E) (Perera et al., 1999 among most of insects; Soin et al., 2009).(Ecdysone receptor, EcR) gene is at first identified out (Koelle et al., 1991) by the clone to the ecdysone receptor that coding 20E combines in fruit bat D.melanogaster.EcR combines to form heterodimer (heterodimer) function of competence exertion ecdysone receptor afterwards with ultra valve albumen USP (ultraspicacle); RXR among USP and the vertebrate (retinoid X receptor) is homologue (Perera et al., 1999; Kozlova and Thummel, 2000).The characteristic of EcR and USP also different to some extent (Beatty et al., 2009) among insect not of the same race.Functional EcR can the combining target gene promoter region specific dna sequence; The early stage response gene of a series of encoding transcription factors of activation-inducing (primary response gene) is like the expression of E74, E75 and Broad etc.; These early genes are further regulated growth courses (Cruz et al., 2006) such as secondary replies expression of gene and molt metamorphosis again.Therefore, EcR is for the insect important effect of having grown normally, below mainly make a brief overview with regard to the 26S Proteasome Structure and Function of EcR among the insect.
Independent EcR is the expression that can't regulate and control downstream gene, has only when EcR and USP formation heterodimer, under the effect of 20E, just can exercise the real function of its ecdysone receptor then.Since the gene order of EcR and USP after at first being identified among the fruit bat D.melanogaster, all found gene (Ogura et al., 2005 of this nuclear receptor family among the present increasing insect; Martin et al., 2006; Tan and Palli, 2008).Have now found that EcR among a plurality of tissues such as prothoracic gland, epidermis, fat-body and ovary and developmental stage expressed (Cruz et al. is arranged; 2006); Yet different EcR mainly expresses mainly expression in imaginal discs (imaginal discs) of EcRA with still difference to some extent of the expression of the Worker's Stadium among insect like EcRB1 in fruit bat D.melanogaster in the larva tissue; Find that among lepidopterous insects such as silkworm B.mori and maduca sexta M.sexta EcRB1 mainly expresses among larva; Yet find but that among coleoptera colorado potato beetle Leptinotarsa decemlineata the expression of EcRA among larva is than EcRB1 high a lot (Ogura et al., 2005).This explains that promptly the expression of the same the Worker's Stadium that among different order insects, accounts for the leading EcR that expresses is a difference to some extent.
Except the EcR difference with the Worker's Stadium expression difference, each is also had nothing in common with each other with the function of the Worker's Stadium.In fruit bat D.melanogaster, EcRA, EcRB1 and EcRB2 are stated the death that three gene simultaneous mutations can cause drosophila embryos, but to each gene respectively alone research find that EcRA mainly plays an important role to the adult structure differentiation formation with propupa; Sudden change to EcRB1 can cause and can't normally pupate; Simultaneous mutation to EcRB1 and EcRB2 then can cause stopping of most of larva early developments, and the larva that minority can be grown then can't be accomplished normally pupate (Cruz et al., 2006).In the past among other the non-pattern insect except that fruit bat, owing to lack effective genetic research instrument, the understanding of EcR gene function is stagnated always.After RNAi finds and is widely used in the insect genes functional study, the EcR gene functions of insects such as Groton bug B.germanica and red flour beetle T.castaneum (Cruz et al., 2006 have been carried out studying at present; Tan and Palli, 2008).The EcRA that discovers in Groton bug all has important effect (Cruz et al., 2006) among growth, prothoracic gland disintegration (prothoracic gland degeneration) and the chorion of its larva generate (normal choriogenesis).Research among red flour beetle shows, in the signal transduction of moulting hormone reaction, plays a part leadingly at EcRA among the development by metamorphosis from the larva to the pupa, and weak a lot (Tan and Palli, 2008) are then wanted in the effect of EcRB among this process.
Inhibitor is developed the control that is used for insect because the important function of EcR among insect growth is grown, many non-hormones are casted off a skin.These are mainly dibenzoyl hydrazine class (dibenzoylhydrazine; DBH) compound, that on market, sells at present comprises four big types: worm hydrazides (tebufenozide) RH-5992, methoxyfenozide (methoxyfenozide) RH-2485, ring worm hydrazides (chromafenozide) ANS-118 and chlorine worm hydrazides (Halofenozide) RH-0345.Important advantage of these insecticides is exactly environmentally safe close friend, but these insecticides are most of only to lepidopterous insects effective (comprising RH-5992, RH-2485 and ANS-118) at present, and few part also has effect (RH-0345) to coleopteron.This has limited the range of application of this insecticides greatly, the mechanism of growing based on the insect growth of EcR is furtherd investigate will help us to search out new cast off a skin inhibition insecticides (Nakagawa, 2005; Soin et al., 2009).
Summary of the invention
The shortcoming that primary and foremost purpose of the present invention is to overcome prior art provides a kind of inhibition insecticides of casting off a skin with not enough.
Another object of the present invention is to provide described casting off a skin to suppress the application of insecticides.
The object of the invention is realized through following technical proposals: a kind of type of casting off a skin suppresses insecticide, and main active is a double-stranded RNA, and the template of transcribing of this double-stranded RNA is at least a among NlEcR-A, NlEcR-B or the NlEcR-A/B;
The nucleotide sequence of the positive-sense strand of NlEcR-A (216bp) as follows:
5’-GCTAGTAAAGCGAGAACAGTTGGACGACACCACCCCGTTACGGGGGGGTTCCCCAAGGGGCAGCCCGACCCCCCAGGGGGGCCTGAGGGGCTCCTCGTGGCCCCCCTCGCCAAGGGACATGACTCCCTCCTATGGCGGGGGTGGCACTCCCCTAAATGGCTACCCTTCTCCCACCATGTCATCGCAGCACAGCAACTATGACAGCTGCCTCAGTCC-3’；
The nucleotide sequence of the antisense strand of NlEcR-A (216bp) as follows:
5’-GGACTGAGGCAGCTGTCATAGTTGCTGTGCTGCGATGACATGGTGGGAGAAGGGTAGCCATTTAGGGGAGTGCCACCCCCGCCATAGGAGGGAGTCATGTCCCTTGGCGAGGGGGGCCACGAGGAGCCCCTCAGGCCCCCCTGGGGGGTCGGGCTGCCCCTTGGGGAACCCCCCCGTAACGGGGTGGTGTCGTCCAACTGTTCTCGCTTTACTAGC-3’；
The nucleotide sequence of the positive-sense strand of NlEcR-B (223bp) as follows:
5’-GAGGAGGTGGAGGAGTAGTGATGGGTCATCATCTGCATCAGGCCGGCCTGGCAATGCTGCAGCAGCGGATGATGATGAGTCAGCATGGATTTCATCAGCAGAGTCAGCATCAGGGTGGGCTGCATCATGGTGGGACTACAACACTGGTGCGAGCGGCGGTTGCACATGTTTCGAGCAATGTGTCGGATAGCGGATCTGTGTCAGGACGCGAAGACCTGTCACC-3’；
The nucleotide sequence of the antisense strand of NlEcR-B (223bp) as follows:
5’-GGTGACAGGTCTTCGCGTCCTGACACAGATCCGCTATCCGACACATTGCTCGAAACATGTGCAACCGCCGCTCGCACCAGTGTTGTAGTCCCACCATGATGCAGCCCACCCTGATGCTGACTCTGCTGATGAAATCCATGCTGACTCATCATCATCCGCTGCTGCAGCATTGCCAGGCCGGCCTGATGCAGATGATGACCCATCACTACTCCTCCACCTCCTC-3’；
The nucleotide sequence of the positive-sense strand of NlEcR-A/B (360bp) as follows:
5’-ACTTCCAGAACGAATTCGAGCACCCGAGCGAGGAGGACCTGAAGAGGATTGGATGTTTGAATCTACCCAGTCAGGTGGCCCAAGACCAGCAGGCGGAGAGCGACATGAGATTCCGTCACATAACAGAAATCACCATTTTGACAGTGCAGCTGATTGTCGAATTCGCCAAACGACTGCCCGGCTTCGACAAACTACTCAGAGAGGACCAGATTGTACTGCTCAAGGCATGTTCCAGCGAAGTGATGATGCTTCGCACGGCCAGAAAGTACGACGTGAACACAGACTCTATCCTGTTCGCCAACAACCAGCCCTACACGAGGGACTCCTACACGCTGGCGGGCATGGGTTATGTGGTGGAGG-3’；
The nucleotide sequence of the antisense strand of NlEcR-A/B (360bp) as follows:
5’-CCTCCACCACATAACCCATGCCCGCCAGCGTGTAGGAGTCCCTCGTGTAGGGCTGGTTGTTGGCGAACAGGATAGAGTCTGTGTTCACGTCGTACTTTCTGGCCGTGCGAAGCATCATCACTTCGCTGGAACATGCCTTGAGCAGTACAATCTGGTCCTCTCTGAGTAGTTTGTCGAAGCCGGGCAGTCGTTTGGCGAATTCGACAATCAGCTGCACTGTCAAAATGGTGATTTCTGTTATGTGACGGAATCTCATGTCGCTCTCCGCCTGCTGGTCTTGGGCCACCTGACTGGGTAGATTCAAACATCCAATCCTCTTCAGGTCCTCCTCGCTCGGGTGCTCGAATTCGTTCTGGAAGT-3’；
The described inhibition insecticide of casting off a skin is applied to prepare the medicine of preventing and treating brown planthopper.
The present invention has following advantage and effect with respect to prior art:
The main method of control brown planthopper has cultural control, chemical control and biological control at present.Because popularity rate is difficult to improve problems such as the resistance effect that brings with chemicals and environmental pollution, makes the brown planthopper preventing and controlling be difficult to obtain sustainable development.The present invention is through utilizing brown planthopper self the relevant important gene of growing, and screening obtains double-stranded RNA, and in order to the brown planthopper control, effect is remarkable.
Description of drawings
Fig. 1 is the survival rate figure that feeds brown planthopper behind the 0.1 μ g/ μ l dsEcRAs.
Fig. 2 is the survival rate figure that feeds brown planthopper behind the 0.5 μ g/ μ l dsEcRAs.
Fig. 3 is the survival rate figure that feeds brown planthopper behind the 0.1 μ g/ μ l dsEcRBs.
Fig. 4 is the survival rate figure that feeds brown planthopper behind the 0.5 μ g/ μ l dsEcRBs.
Fig. 5 is the survival rate figure that feeds brown planthopper behind the 0.1 μ g/ μ l dsEcRc.
Fig. 6 is the survival rate figure that feeds brown planthopper behind the 0.5 μ g/ μ l dsEcRc.
Fig. 7 is the phenotype variation diagram of brown planthopper behind the feeding dsRNA.
Fig. 8 is the 0.1 hatching progeny size figure of μ g/ μ l dsEcRc after 10 days that feed.
Fig. 9 feeds that the mRNA of brown planthopper expresses spirogram behind the 0.1 μ g/ μ l dsEcRAs.
Figure 10 feeds that the mRNA of brown planthopper expresses spirogram behind the 0.5 μ g/ μ l dsEcRAs.
Figure 11 feeds that the mRNA of brown planthopper expresses spirogram behind the 0.1 μ g/ μ l dsEcRc.
Figure 12 feeds that the mRNA of brown planthopper expresses spirogram behind the 0.5 μ g/ μ l dsEcRc.
Figure 13 is the survival rate figure that feeds brown planthopper behind the dsEcRc transgenic paddy rice.
Figure 14 is the hatching progeny size figure that feeds brown planthopper behind the dsEcRc transgenic paddy rice.
Figure 15 is the hatching progeny size figure that feeds brown planthopper behind the dsEcRc transgenic paddy rice.
Embodiment
Below in conjunction with embodiment and accompanying drawing the present invention is described in further detail, but embodiment of the present invention is not limited thereto.
(1) chooses brown planthopper nymph (J.Chen et al. in each in the length of time; 2010.Feeding-based RNA interference of a trhalose phosphate synthase gene in the brown planthopper, Nilaparvata lugens.Insect Molecular Biology.19 (6): 777-786).
(2) extraction of total RNA: get 20 nymphs; Grind in the liquid nitrogen; Extract total RNA (J.Chen et al. with guanidine isothiocyanate method; 2010.Feeding-based RNA interference of a trhalose phosphate synthase gene in the brown planthopper, Nilaparvata lugens.Insect Molecular Biology.19 (6): 777-786).
(3) cDNA first chain is synthetic: according to the RACE of CLONTECH company kit (Clontech BD SMART
TMRACE cDNA Amplification Kit, Takara, Japan) specification carries out, and the total RNA of 10 μ g adds reactant liquor 20 μ l.42 ℃ of reaction 90min, 72 ℃ of 10min cessation reactions.Per 20 μ l reactant liquors are formed: 50mmol potassium chloride, 3mmol magnesium chloride, 10mmol Tris-HCl pH8.3,1mmol DTT, 5 μ mold NTP, 25 RNA of unit enzyme inhibitors, 8 AMV of unit revertases.
(4) following is that example is described the preparation process with the NlEcR-A specific fragment, the same NlEcR-A of preparation process of NlECR-B specific fragment and the common conservative fragments of NlEcR-A/B.
1. PCR reagent (TransTaq archaeal dna polymerase HiFi, Beijing Quanshijin Biotechnology Co., Ltd) and reaction condition:
At first in the PCR pipe, add following reagent:
The concrete steps of above-mentioned PCR are following:
With 94 ℃ of sex change 4min of above-mentioned mixed liquor, get into following circulation then: 94 ℃ of 30s, 65 ℃ of 30s, 72 ℃ of 30s carry out 35 circulations altogether, and last 72 ℃ are extended 10min.
Wherein, the sequence of the upstream primer EcR-AsF of amplification NlEcR-A specific fragment is: 5 '-GCTAGTAAAGCGAGAACAGTTGG-3 '; The sequence of downstream primer EcR-AsR is: 5 '-GGACTGAGGCAGCTGTCATAG-3 ';
The sequence of the upstream primer EcR-BsF of amplification NlEcR-B specific fragment is: 5 '-GAGGAGGTGGAGGAGTAGTGATG-3 '; The sequence of downstream primer EcR-BsR is: 5 '-GGTGACAGGTCTTCGCGTC-3 ';
The sequence of the upstream primer EcR-CF of the common conservative region of amplification NlEcR-A/B is: 5 '-ACTTCCAGAACGAATTCGAGC-3 '; The sequence of downstream primer EcR-CR is: 5 '-CCTCCACCACATAACCCATG-3 '.
2. amplified production purifying: utilize OMEGA purification kit purifying pcr amplified fragment.
3. intermediate carrier obtains: with purified product according to following coupled reaction system; Be cloned into pMD18-T carrier (TAKARA Company products); Transformed into escherichia coli DH5 α cell (Beijing Quanshijin Biotechnology Co., Ltd's product), containing ampicillin (100 mcg/ml) and 5-and smelling-the LB flat board of 4-chloro-3-indoles-β-D-galactoside (X-gal 0.2 mcg/ml) on incubated overnight.
The coupled reaction system:
4. the purifying of plasmid: the picking hickie, be inoculated in the LB liquid nutrient medium, collect bacterium liquid after 37 ℃ of shaking table incubated overnight, press the OMEGA kit and extract plasmid.
5. sequencing and homology retrieval:, obtain following sequence through the sequencing result analysis:
The nucleotide sequence of NlECR-A gene (long 216bp):
GCTAGTAAAGCGAGAACAGTTGGACGACACCACCCCGTTACGGGGGGGTTCCCCAAGGGGCAGCCCGACCCCCCAGGGGGGCCTGAGGGGCTCCTCGTGGCCCCCCTCGCCAAGGGACATGACTCCCTCCTATGGCGGGGGTGGCACTCCCCTAAATGGCTACCCTTCTCCCACCATGTCATCGCAGCACAGCAACTATGACAGCTGCCTCAGTCC；
The nucleotide sequence of NlECR-B gene (long 223bp):
GAGGAGGTGGAGGAGTAGTGATGGGTCATCATCTGCATCAGGCCGGCCTGGCAATGCTGCAGCAGCGGATGATGATGAGTCAGCATGGATTTCATCAGCAGAGTCAGCATCAGGGTGGGCTGCATCATGGTGGGACTACAACACTGGTGCGAGCGGCGGTTGCACATGTTTCGAGCAATGTGTCGGATAGCGGATCTGTGTCAGGACGCGAAGACCTGTCACC；
The nucleotide sequence of NlECR-A/B gene (long 360bp):
ACTTCCAGAACGAATTCGAGCACCCGAGCGAGGAGGACCTGAAGAGGATTGGATGTTTGAATCTACCCAGTCAGGTGGCCCAAGACCAGCAGGCGGAGAGCGACATGAGATTCCGTCACATAACAGAAATCACCATTTTGACAGTGCAGCTGATTGTCGAATTCGCCAAACGACTGCCCGGCTTCGACAAACTACTCAGAGAGGACCAGATTGTACTGCTCAAGGCATGTTCCAGCGAAGTGATGATGCTTCGCACGGCCAGAAAGTACGACGTGAACACAGACTCTATCCTGTTCGCCAACAACCAGCCCTACACGAGGGACTCCTACACGCTGGCGGGCATGGGTTATGTGGTGGAGG。
The plasmid called after pMDR-EcRAs that contains the NlECR-A gene; The plasmid that contains the NlECR-B gene, called after pMDT-EcRBs; The plasmid called after pMDT-EcRc that contains the NlECR-A/B gene.
(1) design has the Auele Specific Primer that T7 starts, and the dsRNA that carries out three sequence fragments is external synthetic, also designs the primer of green fluorescence protein gene simultaneously, as the conduct contrast.
NlEcR-As dsRNA amplimer:
EcR-AsF：5′-GCTAGTAAAGCGAGAACAGTTGG-3′；
EcR-AsR：5′-GGACTGAGGCAGCTGTCATAG-3′；
EcR-AsT7F：5′-TAATACGACTCACTATAGGGGCTAGTAAAGCGAGAACAGTTGG-3′；
EcR-AsT7R：5′-TAATACGACTCACTATAGGGGGACTGAGGCAGCTGTCATAG-3′；
NlEcR-Bs dsRNA amplimer:
EcR-BsF：5′-GAGGAGGTGGAGGAGTAGTGATG-3′；
EcR-BsR：5′-GGTGACAGGTCTTCGCGTC-3′；
EcR-BsT7F：5′-TAATACGACTCACTATAGGGGAGGAGGTGGAGGAGTAGTGATG-3′；
EcR-BsT7R：5′-TAATACGACTCACTATAGGGGGTGACAGGTCTTCGCGTC-3′；
NlEcR-c dsRNA amplimer:
EcR-cF：5′-ACTTCCAGAACGAATTCGAGC-3′；
EcR-cR：5′-CCTCCACCACATAACCCATG-3′；
EcR-cT7F：5′-TAATACGACTCACTATAGGGACTTCCAGAACGAATTCGAGC-3′；
EcR-cT7R：5′-TAATACGACTCACTATAGGGCCTCCACCACATAACCCATG-3′；
GFP dsRNA amplimer:
GFP-F：5′-GCACCATCTTCTTCAAGGACG-3′；
GFP-R：5′-ACTCCAGCAGGACCATGTGAT-3′；
GFP-T7F：5′-TAATACGACTCACTATAGGGGCACCATCTTCTTCAAGGACG-3′；
GFP-T7R：5′-TAATACGACTCACTATAGGGACTCCAGCAGGACCATGTGAT-3′。
(2) plasmid that the name that obtains with embodiment 1 respectively is called pMDT-EcRAs, pMDT-EcRBs and pMDT-EcRc is a template, uses step (1) designed primer to increase according to following reaction condition, obtains A fragment, B fragment and C fragment respectively.Through dsRNA synthetic agent box (T7RiboMAXTMExpress RNAi System, Promega, USA) method; Synthetic these three corresponding dsRNA of fragment; Called after dsEcRAs, dsEcRBs and dsEcRc are mixed in brown planthopper artificial feed (Fu et al. separately; 2001) in, brown planthopper is fed.Adopting dsRNA final concentration in artificial feed is 0.1 μ g/ μ l and two gradients of 0.5 μ g/ μ l, continued to feed 10 days, with the double-stranded RNA dsGFP of green fluorescent protein as contrast (the lucky right bio tech ltd in Shanghai, pEGFP-N1 plasmid).
The PCR reaction condition: 94 ℃ of sex change 4min get into following circulation then: 94 ℃ of 30s, and 50 ℃ of 30s, 72 ℃ of 30s carry out 35 circulations altogether, and last 72 ℃ are extended 10min.
(3) in the middle of the process of in step (2), mentioning of feeding; Add up the brown planthopper survival rate and the variation of observation phenotype of every day; No matter be under 0.1 μ g/ μ l or 0.5 μ g/ μ l concentration; The dsRNA of three fragments is to the appreciable impact of having grown of brown planthopper, and all there is significant difference (shown in Fig. 1～6) in its survival rate with control group, and brown planthopper Dysecdysis phenotype (as shown in Figure 7) occurs.Among Fig. 7, the Dysecdysis that is caused by dsEcRAs mainly concentrates on head and the back of brown planthopper, the Dysecdysis that is caused by dsEcRBs mainly concentrates on belly and the foot of brown planthopper, and the Dysecdysis phenotype that is caused by dsEcRc has then contained above having been mentioned.After choosing the dsEcRc that fed in addition; The female Dan Xiong of brachypterism type list that sprouted wings one day matches, after laying eggs 15 days, and the back algebraically that statistics hatches out; Compare with contrast; There is significance to reduce (as shown in Figure 8), explains, hindered normal development and the breeding of brown planthopper through the dsRNA of these three fragments that brown planthopper continued feed.
(4) the same step of method (2); In the middle of the feeding dsRNA process 2 days, 4 days, 8 days; Sampling in 10 days; The mRNA relative expression quantity that carries out brown planthopper ecdysone receptor gene through the method for quantitative fluorescent PCR changes, and no matter dsEcRAs and dsEcRc are under 0.1 μ g/ μ l and 0.5 μ g/ μ l concentration, and NlEcR gene mRNA expression amount is all significantly reduced (shown in Fig. 9～12).Presentation of results can effectively disturb the expression of target gene through the dsRNA of brown planthopper is continued feed dsEcRAs and dsEcRc fragment.
(5) on above functional analysis basis; We choose fragment three is that the conservative nucleotide sequence (long 360bp) of NlECR-A/B gene carries out the transgenic paddy rice structure; Obtain forward sequence (BamHI/SacI) and reverse sequence (HindIII/SalI) by having added corresponding restriction enzyme site primer amplification, amplimer is distinguished as follows:
Forward sequence upstream primer (BamHI): 5 '-CACTGGATCCACTTCCAGAACGAATTCGAGC-3 ';
Forward sequence downstream primer (SacI): 5 '-GACGAGCTCCCTCCACCACATAACCCATG-3 ';
Reverse sequence upstream primer (SalI): 5 '-GACGTCGACCCTCCACCACATAACCCATG-3 ';
Reverse sequence downstream primer (HindIII): 5 '-CACAAGCTTACTTCCAGAACGAATTCGAGC-3 '.
Earlier forward sequence and reverse sequence are connected to upward intron both sides of pSK-int carrier (Chinese plasmid vector strain cell pnca gene preservation center-Biovector Science Lab); Promptly form hairpin structure; Secondly cut the structure that obtains forward sequence+intron+reverse sequence through SacI and SalI enzyme; And be connected in pCanG-HA (Chinese plasmid vector strain cell pnca gene preservation center-Biovector Science Lab) the conversion carrier T-DNA district between SacI and SalI restriction enzyme site; This carrier is promotor with CaMV35S; With NPTII is selection markers, is transformed into rice varieties Japan fine (Inst. of Rice, Guangdong Academy of Agricultural Sciences) through Agrobacterium LBA4404 (Chinese plasmid vector strain cell pnca gene preservation center-Biovector Science Lab) subsequently, obtains to express the transgenic paddy rice of brown planthopper ecdysone receptor dsRNA with this; Observe and three indexs of fertility through survival rate statistics and phenotype, assess the effect of this transgenic paddy rice in the brown planthopper control.
The endonuclease reaction system:
The endonuclease reaction condition: 37 ℃, enzyme is cut and is spent the night.
The coupled reaction system:
The coupled reaction condition: 16 ℃, reaction is spent the night.
Choose 7 transgenic paddy rice strains system and experimentize, insert on every strain transgenic seedling 25 first age the brown planthopper nymph, continued to feed 11 days, statistics obtains survival rate and is about 90% (shown in figure 13), compares variant with non-transgenic Japan fine contrast; In the process of feeding, the brown planthopper Dysecdysis phenotype that causes death; Transgenic paddy rice to the brown planthopper adult that sprouted wings a day of feeding is carried out the female single male pairing of brachypterism type list, and after laying eggs 15 days, the back algebraically that statistics hatches is compared with contrast, and remarkable reduction (shown in Figure 14 and 15) is all arranged.Above result shows, through the dsEcRc transgenic paddy rice of feeding, can effectively control brown planthopper harm, for the brown planthopper control provides new research direction and thinking.
The foregoing description is a preferred implementation of the present invention; But embodiment of the present invention is not restricted to the described embodiments; Other any do not deviate from change, the modification done under spirit of the present invention and the principle, substitutes, combination, simplify; All should be the substitute mode of equivalence, be included within protection scope of the present invention.
Claims (2)
1. type of casting off a skin suppresses insecticide, and it is characterized in that: main active is a double-stranded RNA, and the template of transcribing of this double-stranded RNA is at least a among NlEcR-A, NlEcR-B or the NlEcR-A/B;
The nucleotide sequence of the positive-sense strand of NlEcR-A is shown in SEQ ID:NO.1;
The nucleotide sequence of the antisense strand of NlEcR-A is shown in SEQ ID:NO.2;
The nucleotide sequence of the positive-sense strand of NlEcR-B is shown in SEQ ID:NO.3;
The nucleotide sequence of the antisense strand of NlEcR-A is shown in SEQ ID:NO.4;
The nucleotide sequence of the positive-sense strand of NlEcR-A/B is shown in SEQ ID:NO.5;
The nucleotide sequence of the antisense strand of NlEcR-A/B is shown in SEQ ID:NO.6.
2. described the casting off a skin of claim 1 suppressed the application of insecticide in preparation control brown planthopper medicine.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 201110335416 CN102499254B (en) | 2011-10-28 | 2011-10-28 | Molt-inhibiting insecticide and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 201110335416 CN102499254B (en) | 2011-10-28 | 2011-10-28 | Molt-inhibiting insecticide and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN102499254A true CN102499254A (en) | 2012-06-20 |
| CN102499254B CN102499254B (en) | 2013-11-06 |
Family
ID=46211436
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 201110335416 Expired - Fee Related CN102499254B (en) | 2011-10-28 | 2011-10-28 | Molt-inhibiting insecticide and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102499254B (en) |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102876691A (en) * | 2012-09-21 | 2013-01-16 | 山西大学 | Beta-N-acetylglucosaminidase genes of insects and application thereof |
| CN108707603A (en) * | 2018-04-13 | 2018-10-26 | 中山大学 | A kind of ESCRT-III cores subunit Snf7 class double-stranded RNAs and preparation method and application |
| CN109439685A (en) * | 2018-12-26 | 2019-03-08 | 菏泽学院 | A kind of construction method of the plant expression vector that can express gypsymoth USP gene dsRNA and its application |
| CN109468336A (en) * | 2018-12-07 | 2019-03-15 | 中国水稻研究所 | Brown paddy plant hopper protein phosphatase gene NlPP1, albumen and its dsRNA and application |
| CN111955486A (en) * | 2020-08-27 | 2020-11-20 | 湖南省植物保护研究所 | Method for preventing and controlling plant pests, nucleic acid pesticide, and preparation method and application thereof |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2001034815A1 (en) * | 1999-11-05 | 2001-05-17 | Cambria Biosciences, Llc | Dsrna as insect control agent |
| WO2005049841A1 (en) * | 2003-11-17 | 2005-06-02 | Commonwealth Scientific And Industrial Research Organisation | Insect resistance using inhibition of gene expression |
-
2011
- 2011-10-28 CN CN 201110335416 patent/CN102499254B/en not_active Expired - Fee Related
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2001034815A1 (en) * | 1999-11-05 | 2001-05-17 | Cambria Biosciences, Llc | Dsrna as insect control agent |
| WO2005049841A1 (en) * | 2003-11-17 | 2005-06-02 | Commonwealth Scientific And Industrial Research Organisation | Insect resistance using inhibition of gene expression |
Non-Patent Citations (5)
| Title |
|---|
| GEANETTE LAM 等: "Inducible expression of double-stranded RNA directs specific genetic interference in Drosophila", 《CURRENT BIOLOGY》 * |
| PAN,Z. 等: "FJ263048", 《GENBANK》 * |
| PAN,Z.等: "FJ263049", 《GENBANK》 * |
| QIONG YAO 等: "Identification of 20-Hydroxyecdysone Late-Response Genes in the Chitin Biosynthesis Pathway", 《PLOS ONE》 * |
| 张文庆 等: "昆虫几丁质合成及其调控研究前沿", 《应用昆虫学报》 * |
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102876691A (en) * | 2012-09-21 | 2013-01-16 | 山西大学 | Beta-N-acetylglucosaminidase genes of insects and application thereof |
| CN108707603A (en) * | 2018-04-13 | 2018-10-26 | 中山大学 | A kind of ESCRT-III cores subunit Snf7 class double-stranded RNAs and preparation method and application |
| CN108707603B (en) * | 2018-04-13 | 2021-07-20 | 中山大学 | ESCRT-III core subunit Snf7 double-stranded RNA and preparation method and application thereof |
| CN109468336A (en) * | 2018-12-07 | 2019-03-15 | 中国水稻研究所 | Brown paddy plant hopper protein phosphatase gene NlPP1, albumen and its dsRNA and application |
| CN109468336B (en) * | 2018-12-07 | 2021-06-04 | 中国水稻研究所 | Brown planthopper protein phosphate gene NlPP1, protein, dsRNA (double-stranded ribonucleic acid) thereof and application |
| CN109439685A (en) * | 2018-12-26 | 2019-03-08 | 菏泽学院 | A kind of construction method of the plant expression vector that can express gypsymoth USP gene dsRNA and its application |
| CN111955486A (en) * | 2020-08-27 | 2020-11-20 | 湖南省植物保护研究所 | Method for preventing and controlling plant pests, nucleic acid pesticide, and preparation method and application thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN102499254B (en) | 2013-11-06 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102776189B (en) | Cytochrome P450 dsRNA (double-stranded ribonucleic acid) and application to aphid growth inhibition | |
| CN103571844B (en) | DsRNA and the application be combined in control aphid damage thereof | |
| CN102499254B (en) | Molt-inhibiting insecticide and application thereof | |
| CN103088024B (en) | Application of two dsRNAs (double-stranded ribonucleic acids) and combination thereof in controlling aphid damage | |
| US10808261B2 (en) | Double strand RNA-mediated RNA interference through feeding detrimental to larval Lymantria dispar (gypsy moth) | |
| CN106754948B (en) | Nilaparvata lugens NlMLP gene, encoding protein and application thereof | |
| Ikeno et al. | Molecular characterization of the circadian clock genes in the bean bug, Riptortus pedestris, and their expression patterns under long-and short-day conditions | |
| CN106676106B (en) | MiRNA for regulating seed shape, insect resistance and salt tolerance of plant and application thereof | |
| Xu et al. | Full functional sex reversal achieved through silencing of MroDmrt11E gene in Macrobrachium rosenbergii: Production of all-male monosex freshwater prawn | |
| Wang et al. | Initial characterization of the vitellogenin receptor from a Psocoptera insect: Function analysis and RNA interference in Liposcelis entomophila (Enderlein) | |
| Russo et al. | Aphidius ervi venom regulates Buchnera contribution to host nutritional suitability | |
| CN113248586B (en) | Application of brown planthopper PIB14 protein and coding gene thereof in regulation and control of plant brown planthopper resistance | |
| Bai et al. | CRISPR/Cas9‐mediated mutagenesis of the white gene in an ectoparasitic wasp, Habrobracon hebetor | |
| CN105685092B (en) | A kind of chitin synthesis inhibits insecticides and preparation method and application | |
| CN104109672B (en) | DsRNA of ecdysone receptor (EcR) gene and application thereof to control of damage caused by aphids | |
| Cagliari et al. | Parental RNA interference as a tool to study genes involved in rostrum development in the Neotropical brown stink bug, Euschistus heros | |
| CN102586259B (en) | Chilo venosatus walker ecdysis regulation transcription factor cDNA and cloning method and recombinant application thereof | |
| Adeyinka et al. | Host-induced silencing of the CpCHI gene resulted in developmental abnormalities and mortality in maize stem borer (Chilo partellus) | |
| Tan et al. | Molecular and functional characterization of the ecdysone receptor isoform-A from the cotton mirid bug, Apolygus lucorum (Meyer-Dür) | |
| Apone et al. | Targeting the diuretic hormone receptor to control the cotton leafworm, Spodoptera littoralis | |
| CN104109673B (en) | DsRNA of ecdysone receptor (EcR) gene USP and application thereof to control of damage caused by aphids | |
| CN108949769B (en) | Cotton bollworm ecdysone regulatory factor E78-C gene cDNA and application thereof | |
| Zhang et al. | Four insulin-like peptides orchestrate reproductive signaling of the green lacewing, Chrysopa pallens (Rambur)(Neuroptera: Chrysopidae) | |
| Zeng et al. | Effect of four chitinase genes on the female fecundity in Sogatella furcifera (Horváth) | |
| CN104109671A (en) | DsRNA of gene of symbiotic bacterium of aphids and application thereof to inhibition of growth of aphids |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| EE01 | Entry into force of recordation of patent licensing contract |
Application publication date: 20120620 Assignee: Guangdong Sui Yuan Agricultural Polytron Technologies Inc Assignor: Sun Yat-sen University Contract record no.: 2015440000512 Denomination of invention: Molt-inhibiting insecticide and application thereof Granted publication date: 20131106 License type: Exclusive License Record date: 20150805 |
|
| LICC | Enforcement, change and cancellation of record of contracts on the licence for exploitation of a patent or utility model | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20131106 Termination date: 20201028 |
|
| CF01 | Termination of patent right due to non-payment of annual fee |