CN110483631A - Black striped plant bug reproduction GAP-associated protein GAP PG and its encoding gene, dsRNA interference sequence and application - Google Patents
Black striped plant bug reproduction GAP-associated protein GAP PG and its encoding gene, dsRNA interference sequence and application Download PDFInfo
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- CN110483631A CN110483631A CN201910725701.4A CN201910725701A CN110483631A CN 110483631 A CN110483631 A CN 110483631A CN 201910725701 A CN201910725701 A CN 201910725701A CN 110483631 A CN110483631 A CN 110483631A
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
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- A01N57/00—Biocides, pest repellants or attractants, or plant growth regulators containing organic phosphorus compounds
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- C07K14/43504—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from invertebrates
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- C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
- C12N15/113—Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing
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Abstract
The invention belongs to agricultural biological technical fields, and in particular to black striped plant bug reproduction GAP-associated protein GAP PG and its encoding gene, dsRNA interference sequence and application.Black striped plant bug reproduction GAP-associated protein GAP PG of the invention, amino acid sequence is as shown in SEQ ID NO.1.Interference sequence of the present invention to black striped plant bug injection pepsinogen gene, significantly reduce the quantity of ovum in black striped plant bug ovary, the lifelong egg laying amount for inhibiting black striped plant bug has eventually led to the fecundity decline of black striped plant bug, can effectively control the development of its population.Gene and albumen of the invention can be used for the biological control of the exploitation and black striped plant bug of transgenic anti-insect plants.
Description
Technical field
The invention belongs to agricultural biological technical fields, and in particular to black striped plant bug reproduction GAP-associated protein GAP PG and its coding base
Cause, dsRNA interference sequence and application.
Background technique
From after China commercial growth Bt cotton, Pesticide use amount and number are greatly reduced so that fleahopper class pest by
Minor pests rise to primary pest.Black striped plant bug (Adelphocoris suturalis) belongs to Semiptera Miridae, is China
One of the primary pest of Yangtze river basin cotton planting district, can also danger to vegetables, soybean, fruit tree etc..Prior art centering is black blind
The prevention and treatment of stinkbug is mainly controlled by chemical pesticides, but chemical prevention can develop drug resistance, and reduces control efficiency, and can also pollute agriculture
People, animal health and safety and ecological environment are jeopardized in product, air, upper earth and waters.In addition, black striped plant bug activity is quick, easily turn
Shifting increases its difficulty of prevention and cure.Therefore it is badly in need of exploring new and effective black striped plant bug control measure.
Pepsinogen is pepsin precursor, is mainly secreted by stomach lining, can in the case where pH is 2.0 or so acid condition
To be activated into the pepsin for having digestive function.The height of serum pepsinogen level can reflect the area Wei Misuan mucous membrane gland
The health condition of body involvement, antrum and body of stomach mucous membrane pyloric gland and intestinal gland metaplasia.Pepsin is the master for participating in protein digestibility
Enzyme is wanted, it is related to the digestion and metabolism of organism.The pepsin extracted from the gastric mucosa of animal such as pig, ox, sheep is alternatively arranged as moving
Object feed addictive.Pepsin is alternatively arranged as toolenzyme to prepare polypeptide or collagen.
Currently, be mainly the relationship of its content and stomach health condition for the research of pepsinogen, it is right
Effect of pepsinogen during insect generations has not been reported.
Summary of the invention
The purpose of the present invention is to provide a kind of black striped plant bug reproduction related gene PG.
A further object of the present invention is to provide the encoded albumen of said gene.
A further object of the present invention is to provide the dsRNA interference sequences of said gene.
A further object of the present invention is to provide the recombinant expression carriers containing said gene.
A further object of the present invention is to provide the recombinant bacterial strains containing said gene.
A further object of the present invention is to provide the applications of black striped plant bug PG gene.
A further object of the present invention is to provide the applications of the dsRNA interference sequence of black striped plant bug PG gene.
Black striped plant bug reproduction GAP-associated protein GAP PG of the invention, amino acid sequence is as shown in SEQ ID NO.1:
MIFVAATALLALWASDCRTATPHGGPNRSNGKRFIVQLTRNTEQSGFHSERNYIEELELNVYKSAYSIN
IFVGNPPQEIKVDFDTGSSDLWVRGNLYRNYCYEYPGNQSEFIAQCSSEFVLLGGRYSGSYGGGNCSGRPGNDLVEV
AGMGQYRLSFGVVSSITDHLLPEHCGGILGLGLQYSQYSFLQHLKRNGVLSRNVFSTKLTNQGEYNFVEFGGWDETI
VSENIHWIPIVHNVFGHWRFHANSMSIGSHEVISHPTDMIADTGCTNIVVLSRDIFNKLMQNMQANESSTGLCVMNE
WNLQLEIDIMGKTYVLDWEDFTSKQADVYCPFKLHISYLDRRDDLRMPEVILGDPFLRKFYSIYDFGSPSQIAFVPY
Q
The protein sequence of black striped plant bug pepsinogen gene coding, encodes 378 amino acid residues.
The nucleotide sequence of black fleahopper reproduction related gene PG of the invention is as shown in SEQ ID NO.2:
ATGATTTTCGTTGCTGCTACAGCGCTTTTGGCACTTTGGGCTTCGGACTGTCGCACAGCGACACCTCAC
GGTGGACCGAACCGTTCAAATGGAAAGAGGTTTATTGTGCAGCTTACCAGGAATACGGAACAATCTGGATTTCATTC
GGAAAGGAATTACATAGAGGAGTTGGAACTCAATGTCTACAAGAGCGCATACAGCATAAATATATTTGTGGGAAATC
CTCCTCAAGAAATCAAGGTGGATTTTGACACTGGTTCGAGCGACCTGTGGGTCCGAGGAAATTTATACAGAAACTAC
TGCTACGAATACCCAGGCAACCAATCCGAATTCATCGCACAATGCTCGTCTGAATTTGTGTTACTCGGAGGCCGATA
CAGCGGTTCTTACGGCGGAGGGAATTGTTCTGGAAGACCGGGGAATGATTTAGTTGAGGTAGCGGGAATGGGTCAGT
ACCGGTTAAGCTTCGGAGTTGTTTCTTCGATTACGGATCATTTGCTCCCGGAACACTGCGGTGGAATTTTGGGGCTT
GGATTGCAATATAGTCAATACTCCTTCCTTCAACATTTAAAACGTAACGGGGTGTTGAGTAGAAACGTATTTTCGAC
TAAGCTAACCAATCAGGGGGAATATAACTTCGTCGAATTCGGTGGTTGGGACGAAACAATTGTAAGTGAAAACATAC
ACTGGATTCCCATTGTTCATAATGTTTTTGGACATTGGAGATTTCACGCTAACAGCATGTCTATAGGCAGTCACGAA
GTAATATCACATCCGACTGACATGATAGCTGATACTGGGTGCACTAACATCGTCGTCTTGAGTCGTGATATTTTCAA
TAAATTAATGCAAAATATGCAAGCCAACGAGAGCTCGACCGGATTATGTGTAATGAATGAATGGAATCTACAACTTG
AAATCGACATCATGGGAAAGACTTACGTTCTAGACTGGGAGGATTTCACCTCAAAACAAGCTGATGTGTATTGTCCC
TTCAAGCTCCACATTTCATATCTTGACAGACGGGATGATCTAAGAATGCCGGAAGTAATACTTGGTGATCCATTCTT
GAGAAAGTTCTACTCCATCTATGATTTTGGATCGCCAAGCCAAATTGCGTTTGTACCTTACCAATGA
Above-mentioned nucleotide sequence, i.e. cDNA sequence, sequence 1137bp.
The present invention also provides recombinant expression carriers and recombinant bacterium containing black striped plant bug PG gene or its dsRNA interference sequence
Strain.
The dsRNA interference sequence of the black stinkbug reproduction related gene PG of specific embodiment according to the present invention, by following
Specific primer sequence expands to obtain:
Upstream primer dsPG-F:5 '-CCCGAACAACTGTGGTGGAA-3 ',
Downstream primer dsPG-R:5 '-TTGGCTTGGCGATCCAAAATC-3 '.
Above-mentioned interference sequence is injected to black striped plant bug newly in emergence female adult body with microinjection, it is fixed by real-time fluorescence
It measures RCR (qPCR) and detects black striped plant bug pepsinogen gene expression amount, after statistics injection 10 days in black striped plant bug female adult ovary
Furthermore the quantity of ovum matches female adult after injection with the male worm newly to sprout wings one by one, the lifelong egg laying amount of statistics female adult, service life, oviposition
Early period.As a result, it has been found that black striped plant bug pepsinogen gene expression amount in entire breeding time by extremely significant inhibition, inhibit its table
The quantity of ovum and female adult fecundity in black striped plant bug ovary are significantly suppressed after reaching, the quantity of ovum is reduced in ovary, is laid eggs throughout one's life
Amount is reduced, and the development of fecundity and population to black striped plant bug produces adverse effect.
Beneficial effects of the present invention:
The present invention provides the albumen of a kind of isolated black striped plant bug pepsinogen gene and its coding, finds for the first time
Effect of the pepsinogen gene in black striped plant bug reproductive process, inhibit the expression of pepsinogen can be obviously reduced in it is black blind
The fecundity of stinkbug makes the development decline of black striped plant bug population.
The present invention synthesizes black striped plant bug pepsinogen gene dsRNA, is then input to dsRNA using microinjection
In black striped plant bug body, with green fluorescent protein (GFP) control group, find black striped plant bug pepsinogen gene expression amount entire
Breeding time finds after interfering the gene by extremely significant inhibition, the quantity of ovum and lifelong egg laying amount in black striped plant bug ovary
It substantially reduces.
Therefore, the present invention provides new approaches to control the development of black striped plant bug population, also black blind in green prevention and treatment to realize
Stinkbug and other hemipterans provide the foundation.
Detailed description of the invention
After the dsRNA of Fig. 1 display injection pepsinogen gene interference sequence, the silence efficiency of pepsinogen gene；
Wherein, " * " represents p < 0.05, and " * * " represents p < 0.01；" dsPG " represents the processing group of injection pepsinogen gene dsRNA；
" dsGFP " represents the control group of injection GFP gene dsRNA；
Influence of the dsRNA of Fig. 2 display injection pepsinogen gene interference sequence to black striped plant bug fecundity,
In, A is influence of the dsRNA of injection pepsinogen gene interference sequence to ovum quantity in black striped plant bug ovary；B is injection
Influence of the dsRNA of pepsinogen gene interference sequence to the black striped plant bug female adult pest service life；C is injection pepsinogen gene
Influence of the dsRNA of interference sequence to the lifelong egg laying amount of black striped plant bug；D is injection pepsinogen gene interference sequence
Influence of the dsRNA to black striped plant bug preoviposition period；" * " represents the significance of difference: " * " represents P < 0.05, and " * * " P represents < 0.01；
" dsPG " represents the processing group of injection pepsinogen gene interference sequence dsRNA；" dsGFP " represents injection GFP gene dsRNA
Control group.
Specific embodiment
The clone of 1 black striped plant bug pepsinogen gene of embodiment and analysis
The black striped plant bug sample for weighing 30mg is placed in 1.5ml without in enzyme pipe, and Liquid nitrogen precooler grinds female adult using grinding rod,
Total serum IgE is extracted using TRIzol method.
Using PrimeScriptTM RT Master Mix (perfect real time) kit by said extracted
Total serum IgE synthesizes cDNA template.
The primer for designing synthesis is as follows:
Upstream primer sequence PG-F:5'-TGATCTAAGTGTGATCAGTAAAGATGATTT-3',
Downstream primer sequence PG-R:5'-GTTCCATAAAGTATTTAATTCGCTAAAGTCG-3'.
Using black striped plant bug cDNA as template, using above-mentioned primer PG-F and PG-R, PCR amplification is carried out.1% agarose electrophoresis
PCR product is detected, ethidium bromide (EB) dyes, and observes electrophoresis result under ultraviolet lamp, detects correct segment and cuts glue and utilize DNA
Gel Extraction kit recycles target fragment.
The PCR product of recycling and pEASY-T1 carrier are attached, it is thin that recombinant vector is transformed into the full formula competence in Beijing
Born of the same parents T1, ammonia benzyl resistance LB culture medium are incubated overnight.After overnight incubation, 10 positive colonies of picking carry out PCR verifying, by bacterium colony
PCR amplification is that positive clone takes fresh bacterium solution to be sequenced.
Pepsinogen gene open reading frame overall length 1137bp of the invention, encodes 378 amino acid residues, ammonia
Base acid sequence is as shown in SEQ ID NO.1, molecular mass 93.72kDa, and theoretical isoelectric point is 5.07.
Embodiment 2 inject pepsinogen gene dsRNA after gene silence efficiency and its to black striped plant bug fecundity
Influence
2.1 preparation dsRNA templates
According to pepsinogen gene order obtained in embodiment 1, designing specificity amplification primer, (end 5'- adds T7
Promoter sequence), the amplification of the dsRNA segment for pepsinogen gene, the specific primer of design is as follows:
Upstream primer sequence dsPG-F:CCCGAACAACTGTGGTGGAA
Downstream primer sequence dsPG-R:TTGGCTTGGCGATCCAAAATC.
Using black striped plant bug cDNA as template, PCR amplification, 1% agarose are carried out using above-mentioned primer dsPG-F and dsPG-F
Electrophoresis detection PCR product, ethidium bromide (EB) dye, and observe electrophoresis result under ultraviolet lamp, cut glue and utilize DNA gel recycling examination
Agent box purification and recovery target fragment.PCR product is connected on pEASY-T1 carrier.The list of picking colony PCR test positive
Bacterium colony, sequencing, detects the correctness of gene order, and to correct single colonie is sequenced, 6ml LB+AMP culture medium shakes bacterium training overnight
It supports.
The plasmid containing target fragment is extracted with AxyPrep plasmid extraction kit, using the plasmid as template, use is above-mentioned
Specific primer dsPG-F and dsPG-F carry out second of PCR amplification.
2.2 synthesis dsRNA
By the product utilization phenol chloroform method purifying of second of PCR, dsRNA is synthesized.With phenol chloroform dsRNA, use
1% agarose gel electrophoresis detects cut back quality, is that 10 μ g/ μ l are spare by dsRNA concentration dilution.
2.3 feeding dsRNA
Using green fluorescence protein gene (GFP) double-strand dsRNA as control, pepsinogen gene dsRNA is passed through aobvious
Microinjection injects in new emergence female adult body from the outermost of black striped plant bug metathorax and abdomen coria.
Silence efficiency detection: 5 days, 10 days, 14 days and 18 days after the injection treatment full worms of black striped plant bug are collected respectively, are extracted
After RNA and reverse transcription are cDNA, use Premix ExTaqTMII and Bio-Rad Detection iQ2System
Detect the silencing effect of pepsinogen gene.
As shown in Figure 1, with injection dsGFP control group compared with, inject pepsinogen gene dsRNA after 5d, 10d,
14d, 18d, the extremely significant decline of pepsinogen gene expression amount, expression quantity is in entire breeding time by extremely significant inhibition.
Therefore, inject black striped plant bug pepsinogen gene interference sequence can lead to pepsinogen gene expression quantity it is significant under
Drop.
Ovum quantity statistics in ovary: after injection dsRNA10 days, female adult ovary, each processing are dissected using Stereo microscope
20 female adults that do not mate are dissected, 3 biology repeat, observe ovarian morphology, count and analyze the number of ovum in black striped plant bug ovary
Amount.
Fecundity: it after female adult injects dsRNA, is matched one by one with the male worm newly to sprout wings, is put into disposal plastic cup (5cm
× 7cm) mating, if discovery male worm is dead, horse back supplements new sexually matured male worm, with 40 pairs of insects for a processing group, 3
Biology repeats, statistics preoviposition period, lifelong egg laying amount and female adult pest service life, evaluates injection pepsinogen gene with this
Influence of the dsRNA to black striped plant bug preoviposition period, egg laying amount and female adult pest service life.
Dissection ovary counts the quantity of ovum and observes ovarian morphology after injection pepsinogen gene dsRNA10d.Such as Fig. 2
Shown in middle A, compared with the control group, after silencing pepsinogen gene, the quantity of ovum is substantially reduced in black striped plant bug ovary, is subtracted
Lack 23.5%, it is seen that the dsRNA of injection pepsinogen gene can substantially reduce the quantity of ovum in black striped plant bug female adult ovary.
It mates after new emergence female adult injection pepsinogen gene dsRNA with male worm, counts female adult pest service life, preoviposition period
With lifelong egg laying amount, the influence of pepsinogen gene pairs black striped plant bug fecundity is evaluated with this.It the results are shown in Table 1 and Fig. 2.
Table 1 injects influence of the dsRNA of pepsinogen gene to black striped plant bug fecundity
Table 1 illustrates: influence of the dsRNA of injection pepsinogen gene to black striped plant bug fecundity.Wherein " ns " generation
Table is without significant difference；" * " represents p < 0.05, and " * * " represents p < 0.01.Figure Chinese and English abbreviation meaning: " dsPG " represents injection
The processing group of pepsinogen gene dsRNA；" dsGFP " represents the control group of injection GFP gene dsRNA.
The result shows that compared with the control group, the lifelong egg laying amount of female adult is remarkably decreased after interference pepsinogen gene, under
14.81% is dropped, but female adult pest service life, preoviposition period are not significantly different compared with the control group.Inhibit pepsinogen gene
Expression after significantly suppress the quantity of ovum and female adult fecundity in black striped plant bug ovary, the quantity of ovum is reduced in ovary, throughout one's life
Egg laying amount is reduced, and finally affects the fecundity of black striped plant bug and the development of population.
Therefore, interference sequence provided by the invention can be applied to the exploitation of the anti-black striped plant bug plant of transgenosis, exploitation green
The zoophobous of environmental protection realizes the purpose of green pest control.
Sequence table
<110>Hua Zhong Agriculture University
<120>black striped plant bug reproduction GAP-associated protein GAP PG and its encoding gene, dsRNA interference sequence and application
<160> 4
<170> SIPOSequenceListing 1.0
<210> 1
<211> 378
<212> PRT
<213>black striped plant bug (Adelphocoris suturalis)
<400> 1
Met Ile Phe Val Ala Ala Thr Ala Leu Leu Ala Leu Trp Ala Ser Asp
1 5 10 15
Cys Arg Thr Ala Thr Pro His Gly Gly Pro Asn Arg Ser Asn Gly Lys
20 25 30
Arg Phe Ile Val Gln Leu Thr Arg Asn Thr Glu Gln Ser Gly Phe His
35 40 45
Ser Glu Arg Asn Tyr Ile Glu Glu Leu Glu Leu Asn Val Tyr Lys Ser
50 55 60
Ala Tyr Ser Ile Asn Ile Phe Val Gly Asn Pro Pro Gln Glu Ile Lys
65 70 75 80
Val Asp Phe Asp Thr Gly Ser Ser Asp Leu Trp Val Arg Gly Asn Leu
85 90 95
Tyr Arg Asn Tyr Cys Tyr Glu Tyr Pro Gly Asn Gln Ser Glu Phe Ile
100 105 110
Ala Gln Cys Ser Ser Glu Phe Val Leu Leu Gly Gly Arg Tyr Ser Gly
115 120 125
Ser Tyr Gly Gly Gly Asn Cys Ser Gly Arg Pro Gly Asn Asp Leu Val
130 135 140
Glu Val Ala Gly Met Gly Gln Tyr Arg Leu Ser Phe Gly Val Val Ser
145 150 155 160
Ser Ile Thr Asp His Leu Leu Pro Glu His Cys Gly Gly Ile Leu Gly
165 170 175
Leu Gly Leu Gln Tyr Ser Gln Tyr Ser Phe Leu Gln His Leu Lys Arg
180 185 190
Asn Gly Val Leu Ser Arg Asn Val Phe Ser Thr Lys Leu Thr Asn Gln
195 200 205
Gly Glu Tyr Asn Phe Val Glu Phe Gly Gly Trp Asp Glu Thr Ile Val
210 215 220
Ser Glu Asn Ile His Trp Ile Pro Ile Val His Asn Val Phe Gly His
225 230 235 240
Trp Arg Phe His Ala Asn Ser Met Ser Ile Gly Ser His Glu Val Ile
245 250 255
Ser His Pro Thr Asp Met Ile Ala Asp Thr Gly Cys Thr Asn Ile Val
260 265 270
Val Leu Ser Arg Asp Ile Phe Asn Lys Leu Met Gln Asn Met Gln Ala
275 280 285
Asn Glu Ser Ser Thr Gly Leu Cys Val Met Asn Glu Trp Asn Leu Gln
290 295 300
Leu Glu Ile Asp Ile Met Gly Lys Thr Tyr Val Leu Asp Trp Glu Asp
305 310 315 320
Phe Thr Ser Lys Gln Ala Asp Val Tyr Cys Pro Phe Lys Leu His Ile
325 330 335
Ser Tyr Leu Asp Arg Arg Asp Asp Leu Arg Met Pro Glu Val Ile Leu
340 345 350
Gly Asp Pro Phe Leu Arg Lys Phe Tyr Ser Ile Tyr Asp Phe Gly Ser
355 360 365
Pro Ser Gln Ile Ala Phe Val Pro Tyr Gln
370 375
<210> 2
<211> 1137
<212> DNA
<213>black striped plant bug (Adelphocoris suturalis)
<400> 2
atgattttcg ttgctgctac agcgcttttg gcactttggg cttcggactg tcgcacagcg 60
acacctcacg gtggaccgaa ccgttcaaat ggaaagaggt ttattgtgca gcttaccagg 120
aatacggaac aatctggatt tcattcggaa aggaattaca tagaggagtt ggaactcaat 180
gtctacaaga gcgcatacag cataaatata tttgtgggaa atcctcctca agaaatcaag 240
gtggattttg acactggttc gagcgacctg tgggtccgag gaaatttata cagaaactac 300
tgctacgaat acccaggcaa ccaatccgaa ttcatcgcac aatgctcgtc tgaatttgtg 360
ttactcggag gccgatacag cggttcttac ggcggaggga attgttctgg aagaccgggg 420
aatgatttag ttgaggtagc gggaatgggt cagtaccggt taagcttcgg agttgtttct 480
tcgattacgg atcatttgct cccggaacac tgcggtggaa ttttggggct tggattgcaa 540
tatagtcaat actccttcct tcaacattta aaacgtaacg gggtgttgag tagaaacgta 600
ttttcgacta agctaaccaa tcagggggaa tataacttcg tcgaattcgg tggttgggac 660
gaaacaattg taagtgaaaa catacactgg attcccattg ttcataatgt ttttggacat 720
tggagatttc acgctaacag catgtctata ggcagtcacg aagtaatatc acatccgact 780
gacatgatag ctgatactgg gtgcactaac atcgtcgtct tgagtcgtga tattttcaat 840
aaattaatgc aaaatatgca agccaacgag agctcgaccg gattatgtgt aatgaatgaa 900
tggaatctac aacttgaaat cgacatcatg ggaaagactt acgttctaga ctgggaggat 960
ttcacctcaa aacaagctga tgtgtattgt cccttcaagc tccacatttc atatcttgac 1020
agacgggatg atctaagaat gccggaagta atacttggtg atccattctt gagaaagttc 1080
tactccatct atgattttgg atcgccaagc caaattgcgt ttgtacctta ccaatga 1137
<210> 3
<211> 20
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 3
cccgaacaac tgtggtggaa 20
<210> 4
<211> 21
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 4
ttggcttggc gatccaaaat c 21
Claims (10)
1. black striped plant bug reproduction GAP-associated protein GAP PG, which is characterized in that its amino acid sequence is as shown in SEQ ID NO.1.
2. black striped plant bug reproduction related gene PG, which is characterized in that encode black striped plant bug reproduction correlation egg described in claim 1
White PG.
3. black striped plant bug reproduction related gene PG as claimed in claim 2, which is characterized in that its nucleotide sequence such as SEQ ID
Shown in NO.2.
4. the dsRNA interference sequence of black striped plant bug reproduction related gene PG as claimed in claim 2.
5. the dsRNA interference sequence of black stinkbug reproduction related gene PG according to claim 4, which is characterized in that described
Interference sequence expands to obtain by following specific primer sequence:
Upstream primer dsPG-F:5 '-CCCGAACAACTGTGGTGGAA-3 ',
Downstream primer dsPG-R:5 '-TTGGCTTGGCGATCCAAAATC-3 '.
6. the recombination of the dsRNA interference sequence comprising black striped plant bug reproduction related gene PG as claimed in claim 2 or gene PG
Expression vector.
7. the recombination of the dsRAN interference sequence comprising black striped plant bug reproduction related gene PG as claimed in claim 2 or gene PG
Bacterial strain.
8. the application of black striped plant bug reproduction related gene PG as claimed in claim 2.
9. application of the black striped plant bug reproduction related gene PG as claimed in claim 2 in prevention and treatment black striped plant bug.
10. application according to claim 8, which is characterized in that including by the dsRNA of black striped plant bug reproduction related gene PG
Interference sequence feeds the step of black striped plant bug.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910725701.4A CN110483631B (en) | 2019-08-07 | 2019-08-07 | Apolygus intermedius reproduction related protein PG, coding gene thereof, dsRNA interference sequence and application |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910725701.4A CN110483631B (en) | 2019-08-07 | 2019-08-07 | Apolygus intermedius reproduction related protein PG, coding gene thereof, dsRNA interference sequence and application |
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| Publication Number | Publication Date |
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| CN110483631A true CN110483631A (en) | 2019-11-22 |
| CN110483631B CN110483631B (en) | 2022-07-05 |
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Cited By (3)
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| CN111471098A (en) * | 2020-03-01 | 2020-07-31 | 华中农业大学 | Apolygus lineolarus ecdysone receptor protein and coding gene and application thereof |
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| CN111662369A (en) * | 2020-07-10 | 2020-09-15 | 华中农业大学 | Apolygus linens receptor substrate1 gene and application thereof |
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| CN110483631B (en) | 2022-07-05 |
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