CN110452293A - A kind of rice insect pest modulin and its encoding gene and application - Google Patents

Abstract

A kind of rice insect pest modulin Os04g05560 and its encoding gene and application are disclosed in the present invention.The expression for reducing or knocking out the protein coding gene can make to feel worm plant with high pest-resistant performance, so as to cultivate highly resistance plant hopper plant.The albumen and its encoding gene can be applied to genetic modification of plants.

Description

A kind of rice insect pest modulin and its encoding gene and application

Technical field

The invention belongs to molecular biology fields, and in particular to a kind of rice insect pest modulin and its encoding gene with Using.

Background technique

As the mankind enter the genome times afterwards comprehensively, before development functional genome research has become life science comprehensively Along field.There is synteny since the transgenic technology of rice is relatively easy, and with other gramineous crop genomes, thus By depending on doing model plant.Currently, rice genome deliberate genetic figure and physical map are completed, further to rice functional gene It is studied, is of great significance to socio-economic development and biological study.

The people of half is had more than in the world using rice as staple food at present.Food Security is that the people of the world faces Challenge.The hybrid rice in 20th century 50, the breeding wheat for semidwarfness of the sixties and the seventies, which cultivates scientific and technological revolution twice, keeps rice yield big Amplitude improves.But rice is endangered by the disease pest of large area in recent decades, and Rice Production is made to be on the hazard.Brown paddy plant hopper is China Primary pest in Rice Production, adult and nymph cause yellow leaf or withered with lancet piercing and sucking rice juice, cause the underproduction or Total crop failure.It is recorded according to Chinese agriculture yearbook, brown paddy plant hopper insect pest is national big in 1966,1969,1973,1977,1983 and 2003 Occur, national especially big generation in 1987,1991,2005,2006 and 2007, hazard area reach the 50% of the rice gross area with On, serious loss is caused to China's Rice Production.And the harm of brown paddy plant hopper mostly occurs in the rice mature pustulation period, at this time greatly Amount uses insecticide, and the pollution to paddy is also problem very serious.

Cultivating insect-proof rice kind using brown planthopper resistant gene is method the most cost-effective in brown paddy plant hopper integrated control. International Rice Research Institute (IRRI) even result of study and Southeast Asia Rice Production it was verified that only medium resistance water Flat rice varieties, are also enough the collective control of brown paddy plant hopper is below horizontal what is caused damages, are unlikely to cause reality to rice The harm and production loss on border.Therefore, excavating brown planthopper resistant gene in rice and applying in rice breeding project is prevention and treatment rice The essential measure of brown paddy plant hopper.

To begin from the 1960s, the genetics of resistance of brown paddy plant hopper and breeding research just have begun progress, but with new The generation of bion (or new cause evil type), the risk that pest-resistant cultivar is faced with reduced service life, resistance is lost.Such as international water Rice research institute released the kind IR26 with Bph1 gene in 1973, occurred that the bion 2 of evil can be caused after 2-3； 1977-1978 releases the kind IR36 and IR42 of tool Bph2 resistant gene, and as a result nineteen eighty-two occurs in some countries in succession The new bion of brown paddy plant hopper, it has to cultivate corresponding new resistant variety IR56 and IR64 again in nineteen eighty-three.In Seo in 2006 The rice varieties for carrying different anti-plant hopper genes several to South Korea are tested, wherein Cheongcheongbyeo (base containing Bph1 Cause), the insect resistace of the rice of ASD7 and M63 (gene containing Bph2) is all declined, but Gayabyeo (simultaneously containing Bph1 with Bph2 gene) rice still there is good insect resistace (Seo et al., 2009).Ptb33 (contains Bph2 and Bph3 simultaneously Gene) cause evil it is horizontal lower, aggrieved grade is 2.5, and Performance of cultivar is pest-resistant (making widely known, 2011).

Therefore, it continues deeper into screening and studies anti-source material, finds new resistant gene, and its related gene is determined Position and clone, research and develop the new rice varieties with resistance genetic material, tool has very great significance.

Summary of the invention

The purpose of the present invention is to provide a kind of rice insect pest modulin and its encoding gene and applications.

The technical solution used in the present invention is:

Application of the albumen Os04g05560 as rice insect pest modulin, the amino acid sequence of albumen Os04g05560 Are as follows:

A) amino acid sequence shown in SEQ ID NO:1；

Or b) amino acid sequence shown in SEQ ID NO:1 by one or several amino acid substitution and/or missing and/ Or the amino acid sequence that addition obtains,

Wherein, SEQ ID NO:1 are as follows:

MACLQLQVLLLLACLLLDAPHLSSAAATVPTPPFSFNFDFSNMSTYKPDDLRFEGNATVHGSFVDLTC NAYGLDISQCTAGRMSYNHPVPFYDQTTKEVASFSTQFTFKIIVPRFNNDKEKGDGMAFFLARYPSRMPPDSGGGS LGLITNNNYSSFGPDQFVSVEFDTYNNTWEQPKQTGDHMGININTVTFSTNTTSVSSFSPNESMMKASITFDSKTS MLVASLQYTGNYSNYAPVNVSAKLPDPTTLLPSEVAVGFSAATGAAFELHQIHSWSFNSTIAAPVQKDHKKAIAVG VSIGGGLILVLLVWSILSWWKWRKTNREFDKGTRGACRFNYHRLAAATNHFSMDNRIGAGTFGEVHKGFLTQLGRE VAVKKILRESRAGNKDFFDEVQTISRAKQKNLVELLGWGMKGSSIIDFVMCWRRQKNTDLFLVYEFVDNGNLHMHL YEKEALLSWRIRYKIVKGIISALVYLHHDRHPYILHRDIKPSNILLDKNFNARLADFGLSRTADNGTIQSSMVVGT ENYLDPECRKTGKFNRSSDVFSFGLVLLEIACKKDENSYAQVWERYIDKTLMQAADDRLQGAFDKRQMERVIVLGL WCCQPNIEMRPTMEKAMDFLESDGPLPKLAKPEITSSSAPSN。

Further, the insect pest refers to planthopper；Preferably, the planthopper refers to that brown paddy plant hopper, white backed planthopper, ash fly At least one of lice.

Encode application of the encoding gene of above-mentioned albumen Os04g05560 as Rice Resistance plant hopper controlling gene.

Further, the nucleotide sequence of encoding gene is as shown in SEQ ID NO:2,

Further, the nucleotides sequence of the cDNA of encoding gene is classified as shown in SEQIDNO:3.

A method of improving rice insect pest performance, comprising the following steps: reduce albumen Os04g05560 in rice Expression quantity, the amino acid sequence of albumen Os04g05560 are as follows: a) amino acid sequence shown in SEQ ID NO:1；Or b) SEQ ID The amino acid that amino acid sequence shown in NO:1 is obtained by the substitution and/or deletion and/or addition of one or several amino acid Sequence.

Further, reduce albumen Os04g05560 expression quantity by knock out albumen Os04g05560 encoding gene or The realization of its expression quantity is reduced, the nucleotide sequence of encoding gene is as shown in SEQ ID NO:2.

Further, insect pest refers to planthopper；Preferably, the planthopper refers to brown paddy plant hopper, white backed planthopper, in small brown rice planthopper At least one.

A kind of breeding method of insect pest rice, comprising the following steps: reduce the expression of albumen Os04g05560 in rice Amount, to improve the insect pest performance of rice, wherein the amino acid sequence of albumen Os04g05560 are as follows: a) shown in SEQ ID NO:1 Amino acid sequence；Or b) amino acid sequence shown in SEQ ID NO:1 passes through the substitution of one or several amino acid and/or lacks Lose and/or add obtained amino acid sequence.

Further, reduce albumen Os04g05560 expression quantity by knock out albumen Os04g05560 encoding gene or The realization of its expression quantity is reduced, the nucleotide sequence of encoding gene is as shown in SEQ ID NO:2.

Further, the insect pest refers to planthopper；Preferably, the planthopper refers to that brown paddy plant hopper, white backed planthopper, ash fly At least one of lice.

The beneficial effects of the present invention are:

The present invention provides a kind of new pest-resistant modulin Os04g05560 of rice and its encoding gene.It reduces or knocks out The expression of the protein coding gene can make to feel worm plant with high pest-resistant performance, so as to cultivate highly resistance plant hopper plant.It is described Albumen and its encoding gene can be applied to genetic modification of plants.

Specific embodiment

Enumerate embodiment further below with the present invention will be described in detail.It will similarly be understood that following embodiment is served only for this Invention is further described, and should not be understood as limiting the scope of the invention, those skilled in the art are according to the present invention Some nonessential modifications and adaptations that the principle of elaboration is made all belong to the scope of protection of the present invention.Following specific works of example Skill parameter etc. is also only an example in OK range, i.e. those skilled in the art can do suitable model by the explanation of this paper Interior selection is enclosed, and does not really want to be defined in hereafter exemplary specific data.

Embodiment 1

Using gene editing technology, Os04g05560 gene is knocked out in gene knockout experiment (knockout), makes to feel worm Rice varieties OryzasativaLcv.Nipponbare obtains highly resistance insect resistace.

The building of one, paddy gene knockout carrier

According to Os04g05560 gene cDNA sequence, selects the sequence of its front end as target sequence, design and synthesize (sequence is as follows, but target sequence and corresponding gRNA sequence sequence without being limited thereto for gRNA (guide RNA) sequence.) GRNA sequence fragment recombination to include hygromycin resistance label pBWA (V) H carrier in.Utilize this CRISPR/Cas9 gene Group editor's carrier system, by target sequence one of those or do not fall with 3 base mutation for multiple (to delete or one alkali of addition On base to target sequence).Make Os04g05560 gene cDNA sequence that frameshift mutation occur, the product of expression is not original Amino acid product, to achieve the purpose that knock out Os04g05560 gene.

GRNA sequence 1:5'-GACCTCACCTGCAACGCATA-3'(SEQ ID NO:4)；

Target sequence 1:5'-GACCTCACCTGCAACGCATATGG-3'(SEQ ID NO:5).

GRNA sequence 2:5'-GCGGATGTCGTACAATCACC-3'(SEQ ID NO:6)；

Target sequence 2:5'-GCGGATGTCGTACAATCACCCGG-3'(SEQ ID NO:7).

Two, obtain paddy gene by genetic transformation and knock out seedling

1) to feel the mature embryo of worm rice OryzasativaLcv.Nipponbare as material evoked callus: taking cultured Agrobacterium (EHA105) Bacterium solution is placed in centrifuge tube, centrifuging and taking supernatant, agrobacterium suspension processed；The callus for growing to a certain size is chosen, agriculture is placed in Bacillus suspension infects；Callus is placed in and is co-cultured on base.

2) it screens: callus is taken out；The callus dried is transferred to progress first time screening on screening and culturing medium；It will grow The new culture medium of initial callus transduction of resistant callus, carries out programmed screening.

3) induction of kanamycin-resistant callus tissue is broken up and takes root: picking kanamycin-resistant callus tissue, moves into the culture dish that differential medium is housed, envelope Membrana oralis is sealed, and is put into constant temperature incubation room and is waited seedling differentiation.1cm or so is grown to seedling, moves to root media strong sprout.

4) hygromycin gene special primer, the amplification of Standard PCR method the PCR detection of hygromycin (Hyg) resistant gene: are used Whether detection rice seedlings contain the gene.If containing being to convert positive seedling.

Resistant gene special primer:

Hyg-f:5'-ACGGTGTCGTCCATCACAGTTTGCC-3'(SEQ ID NO:8)；

Hyg-r:5'-TTCCGGAAGTGCTTGACATTGGGA-3'(SEQ ID NO:9).

5) the gene knockout detection of positive seedling: detection primer is designed near target and carries out PCR, is sequenced later, base is detected Because knocking out situation (the homozygous seedling for the knockout that whether succeeds).Obtain the Os04g05560 gene success to sense worm rice OryzasativaLcv.Nipponbare The homozygous seedling of knockout.

Three, paddy genes knock out the identification of seedling insect resistace

The homozygous seedling for feeling the Os04g05560 gene knockout of worm rice OryzasativaLcv.Nipponbare is identified it by the method for seedling stage assay To the insect resistace of brown paddy plant hopper.

The survival rate for feeling worm receptor kind OryzasativaLcv.Nipponbare as the result is shown is depositing for 0%, Os04g05560 gene knockout homozygosis seedling Motility rate is 100%, and its resistance rank is 0 grade -1 grade (resistance rank).

Embodiment 2

Os04g05560 is compared with existing brown planthopper resistant gene effect

After the Os04g05560 gene knockout of worm rice varieties OryzasativaLcv.Nipponbare (9 grades of resistance rank) being felt, resistance rank is aobvious Work is increased to 0 grade -1 grade (resistance rank).

Applicant carries out Miao Jian discovery to pervious resistant variety, their insect resistace is lost severe.Currently, Mudgo (contains Have Bph1) 5.4 grades of average resistance rank, 8.89 grades of average resistance rank of ASD7 (containing Bph2), Rathu Heenati 4.61 grades of average resistance rank of (containing Bph3), 8.14 grades of average resistance rank of Babawee (containing Bph4).

By comparing it can be found that being used for rice breeding, tool using Rice Resistance plant hopper gene Os04g05560 of the invention There is good prospect.By molecular breeding method or the method for genetic engineering, the expression of the protein coding gene is reduced or knocked out Can lead to sense worm plant has the pest-resistant function of highly resistance, so as to cultivate highly resistance plant hopper rice.

Embodiment 3

The effect of the anti-white backed planthopper of Os04g05560

After the Os04g05560 gene knockout of worm rice varieties OryzasativaLcv.Nipponbare (9 grades of resistance rank) being felt, pass through seedling stage assay Method identify the resistance rank of its Whitebacked Planthopper.

The survival rate for feeling worm receptor kind OryzasativaLcv.Nipponbare as the result is shown is depositing for 0%, Os04g05560 gene knockout homozygosis seedling Motility rate is 100%, and its resistance rank is 0 grade -1 grade (resistance rank).

Therefore, the breeding that the gene Os04g05560 of the anti-plant hopper of adjusting and controlling rice of the invention is applied to anti-white backed planthopper also has Good effect.

SEQUENCE LISTING

<110>Inst. of Plant Protection, Guangdong Prov. Academy of Agricultural Sciences

<120>a kind of rice insect pest modulin and its encoding gene and application

<130>

<160> 9

<170> PatentIn version 3.5

<210> 1

<211> 642

<212> PRT

<213>rice

<400> 1

Met Ala Cys Leu Gln Leu Gln Val Leu Leu Leu Leu Ala Cys Leu Leu

1 5 10 15

Leu Asp Ala Pro His Leu Ser Ser Ala Ala Ala Thr Val Pro Thr Pro

20 25 30

Pro Phe Ser Phe Asn Phe Asp Phe Ser Asn Met Ser Thr Tyr Lys Pro

35 40 45

Asp Asp Leu Arg Phe Glu Gly Asn Ala Thr Val His Gly Ser Phe Val

50 55 60

Asp Leu Thr Cys Asn Ala Tyr Gly Leu Asp Ile Ser Gln Cys Thr Ala

65 70 75 80

Gly Arg Met Ser Tyr Asn His Pro Val Pro Phe Tyr Asp Gln Thr Thr

85 90 95

Lys Glu Val Ala Ser Phe Ser Thr Gln Phe Thr Phe Lys Ile Ile Val

100 105 110

Pro Arg Phe Asn Asn Asp Lys Glu Lys Gly Asp Gly Met Ala Phe Phe

115 120 125

Leu Ala Arg Tyr Pro Ser Arg Met Pro Pro Asp Ser Gly Gly Gly Ser

130 135 140

Leu Gly Leu Ile Thr Asn Asn Asn Tyr Ser Ser Phe Gly Pro Asp Gln

145 150 155 160

Phe Val Ser Val Glu Phe Asp Thr Tyr Asn Asn Thr Trp Glu Gln Pro

165 170 175

Lys Gln Thr Gly Asp His Met Gly Ile Asn Ile Asn Thr Val Thr Phe

180 185 190

Ser Thr Asn Thr Thr Ser Val Ser Ser Phe Ser Pro Asn Glu Ser Met

195 200 205

Met Lys Ala Ser Ile Thr Phe Asp Ser Lys Thr Ser Met Leu Val Ala

210 215 220

Ser Leu Gln Tyr Thr Gly Asn Tyr Ser Asn Tyr Ala Pro Val Asn Val

225 230 235 240

Ser Ala Lys Leu Pro Asp Pro Thr Thr Leu Leu Pro Ser Glu Val Ala

245 250 255

Val Gly Phe Ser Ala Ala Thr Gly Ala Ala Phe Glu Leu His Gln Ile

260 265 270

His Ser Trp Ser Phe Asn Ser Thr Ile Ala Ala Pro Val Gln Lys Asp

275 280 285

His Lys Lys Ala Ile Ala Val Gly Val Ser Ile Gly Gly Gly Leu Ile

290 295 300

Leu Val Leu Leu Val Trp Ser Ile Leu Ser Trp Trp Lys Trp Arg Lys

305 310 315 320

Thr Asn Arg Glu Phe Asp Lys Gly Thr Arg Gly Ala Cys Arg Phe Asn

325 330 335

Tyr His Arg Leu Ala Ala Ala Thr Asn His Phe Ser Met Asp Asn Arg

340 345 350

Ile Gly Ala Gly Thr Phe Gly Glu Val His Lys Gly Phe Leu Thr Gln

355 360 365

Leu Gly Arg Glu Val Ala Val Lys Lys Ile Leu Arg Glu Ser Arg Ala

370 375 380

Gly Asn Lys Asp Phe Phe Asp Glu Val Gln Thr Ile Ser Arg Ala Lys

385 390 395 400

Gln Lys Asn Leu Val Glu Leu Leu Gly Trp Gly Met Lys Gly Ser Ser

405 410 415

Ile Ile Asp Phe Val Met Cys Trp Arg Arg Gln Lys Asn Thr Asp Leu

420 425 430

Phe Leu Val Tyr Glu Phe Val Asp Asn Gly Asn Leu His Met His Leu

435 440 445

Tyr Glu Lys Glu Ala Leu Leu Ser Trp Arg Ile Arg Tyr Lys Ile Val

450 455 460

Lys Gly Ile Ile Ser Ala Leu Val Tyr Leu His His Asp Arg His Pro

465 470 475 480

Tyr Ile Leu His Arg Asp Ile Lys Pro Ser Asn Ile Leu Leu Asp Lys

485 490 495

Asn Phe Asn Ala Arg Leu Ala Asp Phe Gly Leu Ser Arg Thr Ala Asp

500 505 510

Asn Gly Thr Ile Gln Ser Ser Met Val Val Gly Thr Glu Asn Tyr Leu

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Asp Pro Glu Cys Arg Lys Thr Gly Lys Phe Asn Arg Ser Ser Asp Val

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Phe Ser Phe Gly Leu Val Leu Leu Glu Ile Ala Cys Lys Lys Asp Glu

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Asn Ser Tyr Ala Gln Val Trp Glu Arg Tyr Ile Asp Lys Thr Leu Met

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Gln Ala Ala Asp Asp Arg Leu Gln Gly Ala Phe Asp Lys Arg Gln Met

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Glu Arg Val Ile Val Leu Gly Leu Trp Cys Cys Gln Pro Asn Ile Glu

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Met Arg Pro Thr Met Glu Lys Ala Met Asp Phe Leu Glu Ser Asp Gly

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Pro Leu Pro Lys Leu Ala Lys Pro Glu Ile Thr Ser Ser Ser Ala Pro

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Ser Asn

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<212> DNA

<213>rice

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atggcctgcc tgcagctgca ggttcttctc ctcctcgcct gcctgctgct tgatgctcct 60

catcttagta gtgctgcagc cacagttcca acaccaccct tctccttcaa cttcgacttc 120

tccaacatgt ccacatacaa gccggatgac ttgagatttg agggcaacgc gactgtgcac 180

ggcagcttcg tcgacctcac ctgcaacgca tatgggctgg acatctctca gtgcacggct 240

gggcggatgt cgtacaatca cccggtgccc ttctacgacc aaaccaccaa ggaggttgcc 300

agcttctcca cacagttcac cttcaagatt attgtaccaa gatttaacaa cgacaaggag 360

aagggagatg gtatggcttt cttccttgca cgttacccat caagaatgcc tccagattca 420

ggcggcggca gccttggtct catcaccaac aacaactata gtagctttgg cccggaccag 480

ttcgtatccg tcgagtttga cacgtacaac aacacctggg agcagcctaa gcagactggt 540

gaccatatgg gcatcaacat caacacagtc actttttcga ctaacacaac aagtgtatcc 600

agcttcagcc cgaatgagag catgatgaag gcgtccatca cctttgacag caagacctcg 660

atgctggtcg cctctctgca gtataccggt aattattcta actatgcccc tgttaacgtc 720

agtgcgaaac tcccagatcc tacgaccttg ctcccgtcgg aggtggcagt gggattttct 780

gcagccaccg gggcagcctt cgagctccat caaatacact catggtcttt caactcaact 840

attgctgccc cggtacaaaa aggtatatat gcagttacta catctcttga attgattgtt 900

ggattaatgc attgtactcg ttggaggtac ttctgacgca gcaattgaca ccttagtagt 960

ttgtattttt tttgaatgaa tttgtaatgc agcaattgac accgatctgc cgtaacatcg 1020

atgcagatca caagaaagcc attgccgtag gggtctcaat tggaggaggt ctcatattgg 1080

tgctcctggt gtggtctatt ctctcatggt ggaagtggag aaaaacaaac cgtgaatttg 1140

acaagggaac tcgtggggct tgtcggttca actaccaccg cttggctgct gcaacaaacc 1200

atttttcaat ggataatagg attggagcag gtacgttcgg tgaagtacac aaaggtttct 1260

tgacacaatt gggccgtgaa gtggctgtta agaagatctt gagggaatcc agagcaggaa 1320

acaaggactt ttttgacgaa gtccagacca tcagtagagc aaagcagaag aatcttgttg 1380

aacttcttgg ttggggaatg aagggcagct cgatcattga tttcgtcatg tgctggcgta 1440

ggcagaagaa caccgacctt ttcctggtat atgaatttgt ggataacggc aacctacata 1500

tgcacctgta tgagaaggag gccctgctgt catggagaat aaggtacatg ctgcgagctg 1560

agcaatataa tttatattca aattttaaga acctagctag gtattattat attactaccg 1620

cacacataac cccaaaaatt attgtctaat aatgttttac aaaactccac catgtccatg 1680

ttgagcataa tttttctaaa aataaaatat agattgtata tgaaaattgt aattactgat 1740

ttcttgacta aggttgatag tttaactttt cactgcttta aaaaaattat tttagatatg 1800

tttaaattaa atgacatttc aataatacct atatatgggt tttctgccac atagacccta 1860

aaacatgttg tttcctggaa gtaactattt tgaatatgga agtttgaaaa aacgagctag 1920

taatgttgac aatcattatg tacgttcctc ggaacaagaa aatgattatg catgcaaatg 1980

ttgtcttttc aggtataaaa tagtgaaggg cattatcagc gctcttgttt accttcacca 2040

tgatagacat ccatacattc ttcacaggga tatcaaaccg agcaacatac tcttggacaa 2100

aaatttcaat gctagattag cagatttcgg gctgtcgaga actgccgaca atggaacaat 2160

acaatcgtca atggttgtag gaacggaaaa ttacttggat ccagagtgca ggaagacggg 2220

aaagttcaac cgtagctcag atgtcttcag cttcgggctc gttctgctag agattgcttg 2280

caagaaagac gagaatagct atgcacaagt ctgggaaagg tacatcgaca aaactctgat 2340

gcaggctgct gacgataggt tacaaggtgc gttcgataag aggcagatgg agcgtgtgat 2400

cgtcttgggg ctgtggtgtt gtcagcctaa tattgagatg cgacctacga tggagaaagc 2460

aatggatttc ctggagagtg atgggccgtt gcctaaactg gccaaacccg aaattacctc 2520

ctcaagtgcg ccaagcaatt ga 2542

<210> 3

<211> 1929

<212> DNA

<213>rice

<400> 3

atggcctgcc tgcagctgca ggttcttctc ctcctcgcct gcctgctgct tgatgctcct 60

catcttagta gtgctgcagc cacagttcca acaccaccct tctccttcaa cttcgacttc 120

tccaacatgt ccacatacaa gccggatgac ttgagatttg agggcaacgc gactgtgcac 180

ggcagcttcg tcgacctcac ctgcaacgca tatgggctgg acatctctca gtgcacggct 240

gggcggatgt cgtacaatca cccggtgccc ttctacgacc aaaccaccaa ggaggttgcc 300

agcttctcca cacagttcac cttcaagatt attgtaccaa gatttaacaa cgacaaggag 360

aagggagatg gtatggcttt cttccttgca cgttacccat caagaatgcc tccagattca 420

ggcggcggca gccttggtct catcaccaac aacaactata gtagctttgg cccggaccag 480

ttcgtatccg tcgagtttga cacgtacaac aacacctggg agcagcctaa gcagactggt 540

gaccatatgg gcatcaacat caacacagtc actttttcga ctaacacaac aagtgtatcc 600

agcttcagcc cgaatgagag catgatgaag gcgtccatca cctttgacag caagacctcg 660

atgctggtcg cctctctgca gtataccggt aattattcta actatgcccc tgttaacgtc 720

agtgcgaaac tcccagatcc tacgaccttg ctcccgtcgg aggtggcagt gggattttct 780

gcagccaccg gggcagcctt cgagctccat caaatacact catggtcttt caactcaact 840

attgctgccc cggtacaaaa agatcacaag aaagccattg ccgtaggggt ctcaattgga 900

ggaggtctca tattggtgct cctggtgtgg tctattctct catggtggaa gtggagaaaa 960

acaaaccgtg aatttgacaa gggaactcgt ggggcttgtc ggttcaacta ccaccgcttg 1020

gctgctgcaa caaaccattt ttcaatggat aataggattg gagcaggtac gttcggtgaa 1080

gtacacaaag gtttcttgac acaattgggc cgtgaagtgg ctgttaagaa gatcttgagg 1140

gaatccagag caggaaacaa ggactttttt gacgaagtcc agaccatcag tagagcaaag 1200

cagaagaatc ttgttgaact tcttggttgg ggaatgaagg gcagctcgat cattgatttc 1260

gtcatgtgct ggcgtaggca gaagaacacc gaccttttcc tggtatatga atttgtggat 1320

aacggcaacc tacatatgca cctgtatgag aaggaggccc tgctgtcatg gagaataagg 1380

tataaaatag tgaagggcat tatcagcgct cttgtttacc ttcaccatga tagacatcca 1440

tacattcttc acagggatat caaaccgagc aacatactct tggacaaaaa tttcaatgct 1500

agattagcag atttcgggct gtcgagaact gccgacaatg gaacaataca atcgtcaatg 1560

gttgtaggaa cggaaaatta cttggatcca gagtgcagga agacgggaaa gttcaaccgt 1620

agctcagatg tcttcagctt cgggctcgtt ctgctagaga ttgcttgcaa gaaagacgag 1680

aatagctatg cacaagtctg ggaaaggtac atcgacaaaa ctctgatgca ggctgctgac 1740

gataggttac aaggtgcgtt cgataagagg cagatggagc gtgtgatcgt cttggggctg 1800

tggtgttgtc agcctaatat tgagatgcga cctacgatgg agaaagcaat ggatttcctg 1860

gagagtgatg ggccgttgcc taaactggcc aaacccgaaa ttacctcctc aagtgcgcca 1920

agcaattga 1929

<210> 4

<211> 20

<212> DNA

<213>artificial synthesized

<400> 4

gacctcacct gcaacgcata 20

<210> 5

<211> 23

<212> DNA

<213>artificial synthesized

<400> 5

gacctcacct gcaacgcata tgg 23

<210> 6

<211> 20

<212> DNA

<213>artificial synthesized

<400> 6

gcggatgtcg tacaatcacc 20

<210> 7

<211> 23

<212> DNA

<213>artificial synthesized

<400> 7

gcggatgtcg tacaatcacc cgg 23

<210> 8

<211> 25

<212> DNA

<213>artificial synthesized

<400> 8

acggtgtcgt ccatcacagt ttgcc 25

<210> 9

<211> 24

<212> DNA

<213>artificial synthesized

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ttccggaagt gcttgacatt ggga 24

Claims (10)

1. application of the albumen Os04g05560 as rice insect pest modulin, the amino acid sequence of albumen Os04g05560 Are as follows:

A) amino acid sequence shown in SEQ ID NO:1；

Or b) amino acid sequence shown in SEQ ID NO:1 by the substitution of one or several amino acid and/or missing and/or adds The amino acid sequence added.

2. application according to claim 1, the insect pest refers to planthopper；Preferably, the planthopper refer to brown paddy plant hopper, At least one of white backed planthopper, small brown rice planthopper.

3. encoding the encoding gene of albumen Os04g05560 of any of claims 1 or 2 as Rice Resistance plant hopper controlling gene Using.

4. application according to claim 3, it is characterised in that: the nucleotide sequence of encoding gene such as SEQ ID NO:2 institute Show.

5. a kind of method for improving rice insect pest performance, comprising the following steps: reduce the table of albumen Os04g05560 in rice Up to amount, the amino acid sequence of albumen Os04g05560 are as follows: a) amino acid sequence shown in SEQ ID NO:1；Or b) SEQ ID The amino acid that amino acid sequence shown in NO:1 is obtained by the substitution and/or deletion and/or addition of one or several amino acid Sequence.

6. according to the method described in claim 5, it is characterized by: reducing the expression quantity of albumen Os04g05560 by knocking out egg The encoding gene of white Os04g05560 reduces the realization of its expression quantity, the nucleotide sequence of encoding gene such as SEQ ID NO:2 institute Show.

7. method according to claim 5 or 6, it is characterised in that: insect pest refers to planthopper；Preferably, the planthopper Refer at least one of brown paddy plant hopper, white backed planthopper, small brown rice planthopper.

8. a kind of breeding method of insect pest rice, comprising the following steps: the expression quantity of albumen Os04g05560 in rice is reduced, To improve the insect pest performance of rice, wherein the amino acid sequence of albumen Os04g05560 are as follows: a) shown in SEQ ID NO:1 Amino acid sequence；Or b) amino acid sequence shown in SEQ ID NO:1 passes through the substitution and/or missing of one or several amino acid And/or the amino acid sequence that addition obtains.

9. breeding method according to claim 8, it is characterised in that: reduce the expression quantity of albumen Os04g05560 by striking The encoding gene of removing protein Os04g05560 reduces the realization of its expression quantity, the nucleotide sequence of encoding gene such as SEQ ID NO: Shown in 2.

10. breeding method according to claim 8 or claim 9, it is characterised in that: the insect pest refers to planthopper；Preferably, institute It states planthopper and refers at least one of brown paddy plant hopper, white backed planthopper, small brown rice planthopper.