CN113308472A - RNA interference sequence of targeted cucumber mosaic virus, expression vector and application thereof - Google Patents

Abstract

The RNA interference sequence of the targeted cucumber mosaic virus, an expression vector and application thereof are double-stranded RNA, consist of a sense strand and an antisense strand, and are the interference sequence GT1 of the targeted CMV-1a protein GT1 gene specifically, or the interference sequence GT2 of the targeted CMV-1a protein GT2 gene specifically, or the interference sequence GT3 of the targeted CMV-2a protein GT3 gene specifically, or the interference sequence GT4 of the targeted CMV-2a protein GT4 gene specifically, or the interference sequence GT5 of the targeted CMV-CP protein GT5 gene specifically, or the interference sequence GT16 of the targeted CMV-MP protein GT6 gene specifically. The double-stranded RNA interference sequence is used for degrading target messenger RNA, reducing the expression of target protein, effectively silencing CMV gene, controlling the infection of CMV and has environment-friendly characteristic and higher commercial application value.

Description

RNA interference sequence of targeted cucumber mosaic virus, expression vector and application thereof

Technical Field

The invention belongs to the field of agricultural pest control, and particularly relates to an RNA interference sequence of a targeted cucumber mosaic virus, an expression vector and application thereof.

Background

Cucumber Mosaic Virus (CMV) is distributed worldwide, has a very wide host range and a plurality of strains, and a large amount of aphid vectors exist, so vegetables and flowers such as solanaceae, cucurbitaceae, compositae, leguminosae, umbelliferae and the like are frequently attacked in agricultural production, and serious loss of yield, quality and ornamental value is caused.

CMV belongs to the genus Bromoviridae virus, whose genome contains three single-stranded RNA molecules, RNA1 encoding protein 1a responsible for replication of the viral genome; RNA2 encodes protein 2a involved in genome replication and protein 2b involved in RNA silencing inhibition; RNA3 encodes protein 3a, a Movement Protein (MP) and a Capsid Protein (CP) belonging to the virus.

Horizontal transmission of the virus is mainly carried out in a non-persistent manner by aphid mediation, and more than 80 aphids have been reported to transmit CMV, with a seed transmission pathway on some plants. At present, the agricultural treatment of the CMV is mainly realized by cultivating resistant crops and controlling the quantity of the spread aphids through pesticides, however, the CMV has strong variation capacity, the resistance of most crops is difficult to resist various CMV virus subspecies, and the non-continuous spread characteristic of the aphids also brings great difficulty to the method for controlling the aphids through pesticides, so that no obvious and effective method for preventing and controlling the CMV in the agriculture is available at present, and the prevention is mainly realized.

In recent years, the RNA interference (RNAi) technology has been increasingly emphasized in the field of plant protection, and is also considered as one of the most promising methods for controlling pests. RNAi is a natural defense mechanism of a plant against viruses, and is induced by specific double-stranded RNA, so that target messenger RNA is degraded, and the expression of target protein is reduced; both exogenous and endogenous dsrnas can induce the RNAi machinery in a plant, thereby improving the antiviral properties of the plant.

The method of exogenous application is more acceptable to consumers and regulatory policies than the means of induction of endogenous dsRNA (i.e. transgenic plants). The method has achieved remarkable effects in the control of some single-stranded RNA viruses. For example, in the control study of Sesbania mosaic virus (Sesbania mosaic virus), dsRNA targeting CP and MP genes by foliar spray could provide 53% and 64% resistance, respectively. In the prevention and control study of Papaya ringspot virus (Papaya ringspot virus), 100% resistance can be provided by using dsRNA targeting CP and HP-Pro genes simultaneously.

At present, the target in CMV RNAi prevention and control mainly aims at the 2b gene on the genome RNA2 of the CMV, and researches show that spraying of dsRNA targeting the CMV 2b gene on leaf surfaces can provide 40% of resistance for tobacco and 60% of resistance for hot pepper, but whether antiviral effects exist in the exogenous application of the dsRNA aiming at other target genes of the CMV, including 1a, 2a, MP and CP, is not reported.

Disclosure of Invention

The invention aims to provide an RNA interference sequence of a targeted cucumber mosaic virus, an expression strain and application thereof, wherein through two specific targeted CMV-1a proteins, two specific targeted CMV-2a, one specific targeted CMV-3a and one specific targeted RNA interference sequence of CMV-CP, target messenger RNA is degraded, so that the expression of target proteins is reduced, CMV genes are effectively silenced, the infection of CMV is controlled, the foliar symptom phenotype of cucumber plants can be effectively controlled in cucumber CMV prevention and control, and the titer of CMV is controlled at a lower level, wherein when the target CMV-1a protein and the target CMV-2a protein are used independently, the prevention and control efficiency can reach 67 percent, the environmental-friendly characteristic and the higher commercial application value are realized, and the defects of the existing prevention and control method are overcome.

In order to achieve the purpose, the invention provides the following technical scheme:

the RNA interference sequence of the targeted cucumber mosaic virus is double-stranded RNA and consists of a sense strand and an antisense strand, and specifically comprises the following steps:

an interference sequence GT1 of a specificity target CMV-1a protein GT1 gene, wherein the specific nucleotide sequence of a sense chain is shown as SEQ ID NO.1, and the specific nucleotide sequence of an antisense chain is shown as SEQ ID NO. 2;

or an interference sequence GT2 of a specificity targeted CMV-1a protein GT2 gene, wherein the specific nucleotide sequence of a sense chain is shown as SEQ ID NO.3, and the specific nucleotide sequence of an antisense chain is shown as SEQ ID NO. 4;

or an interference sequence GT3 of a specificity target CMV-2a protein GT3 gene, the specific nucleotide sequence of the sense strand is shown as SEQ ID NO.5, and the specific nucleotide sequence of the antisense strand is shown as SEQ ID NO. 6;

or an interference sequence GT4 of a specificity target CMV-2a protein GT4 gene, the specific nucleotide sequence of the sense strand is shown as SEQ ID NO.7, and the specific nucleotide sequence of the antisense strand is shown as SEQ ID NO. 8;

or an interference sequence GT5 of a specificity target CMV-CP protein GT5 gene, the specific nucleotide sequence of the sense strand is shown as SEQ ID NO.9, and the specific nucleotide sequence of the antisense strand is shown as SEQ ID NO. 10;

or an interference sequence GT16 of a specificity target CMV-MP protein GT6 gene, the specific nucleotide sequence of the sense strand is shown as SEQ ID NO.11, and the specific nucleotide sequence of the antisense strand is shown as SEQ ID NO. 12.

Further, the invention provides an RNAi inverted repeat sequence of the RNA interference sequence, which comprises a sense strand sequence and an antisense strand sequence of the double-stranded RNA, wherein the inverted repeat sequence of the interference sequence GT1 is GT1-3, and the nucleotide sequence of the inverted repeat sequence is shown as SEQ ID NO. 13; the inverted repeat sequence of the interference sequence GT2 is GT2-3, and the nucleotide sequence is shown as SEQ ID NO. 14; the inverted repeat sequence of the interference sequence GT3 is GT3-3, and the nucleotide sequence is shown as SEQ ID NO. 15; the inverted repeat sequence of the interference sequence GT4 is GT4-3, and the nucleotide sequence is shown as SEQ ID NO. 16; the inverted repeat sequence of the interference sequence GT5 is GT5-3, and the nucleotide sequence is shown as SEQ ID NO. 17; the inverted repeat sequence of the interference sequence GT6 is GT6-3, and the nucleotide sequence is shown in SEQ ID NO. 18.

The invention provides an expression vector targeting cucumber mosaic virus, which contains an original vector and at least one RNA interference sequence or contains the original vector and at least one RNAi inverted repeat sequence.

Further, the original vector is a vector commonly used in the field of gene recombination, such as virus and plasmid.

Preferably, the primary vector is a pGEM vector plasmid.

The invention provides an expression strain of the RNA interference sequence, which is an engineering strain obtained by transforming the expression vector into escherichia coli.

The invention provides an application of the RNA interference sequence or RNAi inverted repeat sequence of the targeted cucumber mosaic virus or the expression vector in preparing a reagent for preventing and controlling the cucumber mosaic virus.

Preferably, in said application, cucumber seedlings are sprayed with a cell lysate containing one or more of said RNA interference sequences.

The invention has the following beneficial effects:

the six double-stranded RNA interference sequences provided by the invention comprise two RNA interference sequences capable of specifically targeting CMV-1a protein, two specific targeting CMV-2a, one specific targeting CMV-CP and one specific targeting CMV-MP, can effectively silence CMV genes, interfere the synthesis of nucleic acid replicase, movement protein and coat protein in the CMV replication process, can be used for the biological control of CMV, and are environment-friendly and have higher commercial application value.

The expression vector containing the double-stranded RNA interference sequence and the RNAi inverted repeat sequence is converted into escherichia coli to obtain an engineering strain, a dsRNA preparation is prepared, and the dsRNA spray solution is sprayed on leaf surfaces, so that cucumber mosaic virus diseases can be effectively prevented and treated.

Drawings

FIG. 1 is a diagram showing the ApaI and EcoRI double digestion identification of expression vectors pGEM-dsGT1, pGEM-dsGT2, pGEM-dsGT3, pGEM-dsGT4, pGEM-dsGT5 and pGEM-dsGT6 in the examples of the present invention.

FIG. 2 is an electrophoresis diagram of the induced expression of dsRNA in the example of the present invention, wherein 1mM IPTG is added to represent the fermentation of recombinant bacteria;

FIG. 3 is an electrophoresis chart of dsRNA for enzyme digestion verification in accordance with the present invention, wherein the chart represents the dsRNA after DNase I and RNase A (containing 0.5M NaCl) digestion of total RNA after extraction;

FIGS. 4-13 show the prevention effect of dsRNA spray solution on cucumber CMV control in the examples of the present invention, wherein FIGS. 4-11 show the growth status of plants in experimental group, control group and control group; FIG. 12 shows the plant heights of the groups; FIG. 13 shows the DAS-ELISA detection results.

Detailed Description

The present invention is further illustrated by the following specific examples.

The experimental methods in the examples of the present invention are all conventional methods unless otherwise specified. The test materials used in the following examples, unless otherwise specified, were purchased from conventional biochemical stores, and the Marker was purchased from Dalibao Bio Inc.; in the examples, the percentages are by mass unless otherwise specified; in the quantitative tests in the examples, three repeated experiments are set, and the results are averaged.

Examples

1. Construction of dsRNA expression vector pGEM-dsGT

Among the six RNA interference sequences GT1, GT2, GT3, GT4, GT5 and GT6 of the present invention, there are two RNA interference sequences capable of specifically targeting CMV-1a protein, two specifically targeting CMV-2a, one specifically targeting CMV-CP and one specifically targeting CMV-MP, which are detailed in Table 1.

TABLE 1 Gene targets of CMV genomic RNA

Note: MP proteins are also sometimes described as 3a proteins.

Respectively synthesizing RNAi inverted repeat sequences of the targeted CMV virus gene fragment interference sequences, wherein the inverted repeat sequence of the interference sequence GT1 is GT1-3, and the nucleotide sequence is shown as SEQ ID NO. 13; the inverted repeat sequence of the interference sequence GT2 is GT2-3, and the nucleotide sequence is shown as SEQ ID NO. 14; the inverted repeat sequence of the interference sequence GT3 is GT3-3, and the nucleotide sequence is shown as SEQ ID NO. 15; the inverted repeat sequence of the interference sequence GT4 is GT4-3, and the nucleotide sequence is shown as SEQ ID NO. 16; the inverted repeat sequence of the interference sequence GT5 is GT5-3, and the nucleotide sequence is shown as SEQ ID NO. 17; the inverted repeat sequence of the interference sequence GT6 is GT6-3, the nucleotide sequence is shown in SEQ ID NO.18, 5'(Apa I) and 3' (EcoR I) are added at two ends of five inverted repeat sequences except GT4, and 5'(Apa I) and 3' (Nde I) are added at two ends of GT4-3 of the interference sequence GT 4.

The synthesized gene fragment is cloned to an expression vector pGEM-T Easy (Ampicillin) through double enzyme digestion to obtain corresponding expression vectors which are named as pGEM-dsGT-1, pGEM-dsGT-2, pGEM-dsGT-3, pGEM-dsGT-4, pGEM-dsGT-5 and pGEM-dsGT-6 respectively.

The obtained expression vector is identified by enzyme digestion with Apa I and EcoR I, the size of the obtained electrophoresis band is consistent with the size of a predicted fragment, the construction success of the CMV dsRNA expression vector is shown (figure 1), the expression vector is transformed into Escherichia coli HT115(DE3) to obtain recombinant bacteria E.coli HT115/pGEM-dsGT-1, E.coli HT115/pGEM-dsGT-2, E.coli HT115/pGEM-dsGT-3, E.coli HT115/pGEM-dsGT-4, E.coli HT115/pGEM-dsGT-5 and E.coli HT115/pGEM-dsGT-6, and glycerol tubes at the temperature of minus 80 ℃ are frozen and stored.

Expression of dsRNA

The bacterial liquid into which the plasmid was successfully introduced was mixed in a ratio of 1: 100, adding the mixture into TB liquid culture medium containing ampicillin and tetracycline resistance, and shaking at 37 ℃ and 200rpm for overnight culture; transferring 2mL of bacterial liquid to 50mL of TB medium the next day, shaking and culturing at 37 ℃ and 220rpm for 2h, adding 1mM IPTG, inducing for 8h, and finishing fermentation.

Extracting total RNA of bacteria by using Trizol reagent, digesting with DNase I and RNase A, wherein DNase I can specifically digest DNA, RNase A can specifically digest single-stranded RNA under high salt concentration (containing 0.5M NaCl), and verifying through 1.5% agarose gel electrophoresis, and referring to FIGS. 2-3, wherein FIG. 2 is an induced expression electrophoretogram of dsRNA; + represents the addition of 1mM IPTG in the fermentation of the recombinant strain; FIG. 3 is an electrophoresis chart of dsRNA for enzyme-catalyzed verification, which represents dsRNA after total RNA extraction and digestion with DNase I and RNase A.

As can be seen from FIG. 3, a clear target band is obtained after IPTG induction and still exists at the target band after enzymolysis, which indicates that the recombinant bacteria successfully induce and express dsRNA.

Preparation of dsRNA spray solution

After the fermentation of the recombinant bacteria is finished, centrifuging at 12000rpm for 1min to collect cells, adding equal volume of TE buffer solution (containing 0.5M NaCl), shaking for dissolution, placing on ice, crushing the cells by using an ultrasonic crusher for 1s and stopping 3s for 20min, and centrifuging at 12000rpm for 20min to obtain a crude enzyme solution. Heating the crude enzyme solution at 60 ℃ for 40min, centrifuging at 12000rpm for 20min, and taking the supernatant as dsRNA spray solution of subsequent experiments.

Obtaining dsRNA by a phenol chloroform extraction method, digesting by endonuclease DNase I and RNase A, and determining the content of the dsRNA by a NanoDrop ONE method.

4. CMV control test for cucumber

Preparing dsRNA spray liquid respectively containing interference sequences GT1, GT2, GT3, GT4, GT5 and GT6 by using TE buffer solution, wherein the dsRNA content in the dsRNA spray liquid is 0.5 mu g/mL, carrying out foliar spray on 1 day before cucumber plants are inoculated with cucumber mosaic virus CMV, setting six experimental groups, respectively and uniformly spraying the spray liquid of the interference sequences GT1, GT2, GT3, GT4, GT5 and GT6, wherein 6 segments are totally, each group is provided with 3 repeats, the TE buffer solution is used as a control group, and plants which are not infected with the virus and normally grow are used as the control group.

Spraying dsRNA 1d, inoculating CMV, detecting virus by DAS-ELISA 7 days and 14 days after inoculation, measuring plant height, observing plant disease, and referring to FIG. 4-13, wherein three left pots are experimental plants and three right pots are control plants in FIG. 4-9; in fig. 10, three pots on the left are control plants, three pots on the right are control plants, in fig. 11, three pots on the left are experimental plants, and three pots on the right are control plants.

The experimental results show that the experimental group and the control group have no obvious symptoms after being inoculated with CMV for 7 days and 14 days, while the control group sprayed with TE buffer solution generates slight flower and leaf symptoms after being inoculated with CMV virus for 7 days, the whole plant shows the phenomenon of atrophy after 14 days, the leaves show the symptoms of flower and leaf abnormality, the whole plant shows the phenomena of severe atrophy and yellowing of the flower and leaf after 24 days, and partial plants die, as shown in figures 4-11.

The plant height measurement results (see fig. 12) of different treatment groups show that the plant height of the experimental group sprayed with the dsRNA spraying liquid is more than 43% higher than that of the control group after the plant is cultured for 14 days, and is obviously higher than that of the control group, the foliar infection symptom and the plant growth condition of the experimental group are obviously improved in the prevention and treatment of CMV, and the plant height measurement results are not obviously different from those of the control group without viruses.

The ELISA detection results (figure 13) show that in the plants sprayed with the dsRNA spray solution containing the interference sequences GT1, GT2, GT3 and GT4, the CMV content (titer is less than 0.3) is obviously lower than that of the control group, the CMV content of the plants sprayed with the dsRNA spray solution containing the interference sequences GT5 and GT6 has no obvious difference with that of the control group, and the leaf surface infection symptoms are improved. The results show that the dsRNA of the target genes GT1, GT2, GT3, GT4, GT5 and GT6 can effectively silence the CMV gene and weaken the symptom of CMV infection, and the dsRNA of the target genes GT1, GT2, GT3 and GT4 has obvious protective effect on cucumber CMV infection.

Sequence listing

<110> Shanghai city academy of agricultural sciences

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gctgaagcgg tgtctgatga gtttgtaact tatggtgctg aagattacct tgaaaaatct 240

gatgatgagc tccttgtcgc ttttgagacg atggtcaaac ccatgcgtat cggacaacta 300

tggtgccctg cgtttaataa atgttctttt atttccagca ttgctatggc cagagctttg 360

ttgttggcac ctagaacatc ccaccgaacc atgaagtgtt ttgaagacct ggtcgcggct 420

atttacacta aatcaatgct ggaaataaaa gaacatttat taaacgcagg gcaccatagt 480

tgtccgatac gcatgggttt gaccatcgtc tcaaaagcga caaggagctc atcatcagat 540

ttttcaaggt aatcttcagc accataagtt acaaactcat cagacaccgc ttcagcggga 600

gctccgtcag gagttcggag ggaatgcgcg tcctctgtga cactctcgct gacatccaca 660

gcgggtaggg gcctgtaatt acgacaggcc gcagcagcct cttcgcgttg ctcagatcgc 720

aaacgttcca catcctcggg agtgtcgaca ccgtaactgc cgttcaaaag attggc 776

<210> 16

<211> 748

<212> DNA/RNA

<213> cucumber mosaic virus

<400> 16

ccatatattt gttcgaagtt cttactctct gacgagttcg gtaacacatt ttccgttcca 60

gatccattgc gcgaggttca gcggttagga acaaagaaaa ttccctattc tgacaatgat 120

gaattcttgt ttgctcactt catgagcttt gttgatcgat tgaagttttt ggaccgaatg 180

tctcagtcgt gtatcgatca actttcgatt ttcttcgaat tgaaatacaa gaagtctggg 240

gaagaggctg ctttaatgtt aggcgccttt aagaagtata ccgctaattt ccagtcctac 300

aaagaactct attattcaga tcgtcgtcag tgcgaattga tcaattcgtt ttgtagtaca 360

gagttcaggg ttgagcgtgt aaattccaac aaacagcgaa agaattatgg aattgaacgt 420

gacgacgatc tgaataatag agttctttgt aggactggaa attagcggta tacttcttaa 480

aggcgcctaa cattaaagca gcctcttccc cagacttctt gtatttcaat tcgaagaaaa 540

tcgaaagttg atcgatacac gactgagaca ttcggtccaa aaacttcaat cgatcaacaa 600

agctcatgaa gtgagcaaac aagaattcat cattgtcaga atagggaatt ttctttgttc 660

ctaaccgctg aacctcgcgc aatggatctg gaacggaaaa tgtgttaccg aactcgtcag 720

agagtaagaa cttcgaacaa atatatgg 748

<210> 17

<211> 724

<212> DNA/RNA

<213> cucumber mosaic virus

<400> 17

acagtgtgtt agatttcccg aggcatggct ttccaaggta ccagtaggac tttaactcaa 60

cagtcctcag cggctacgtc tgacgatctt caaaagatat tatttagccc tgaagccatt 120

aagaaaatgg ctactgagtg tgacctaggc cggcatcatt ggatgcgcgc tgataatgct 180

atttcagtcc ggcccctcgt tcccgaagta acccacggtc gtattgcttc cttctttaag 240

tctggatatg atgttggtga attatgctca aaaggataca tgagtgtccc tcaagtgtta 300

tgtgctgtta ctcgaacagt ttccactgat gctgaagggt ctttgagaat ttacttagct 360

gatctgggcg acaaggagtt atctcccata gatgggcaat gcgtttcgtt cgagtaacag 420

cacataacac ttgagggaca ctcatgtatc cttttgagca taattcacca acatcatatc 480

cagacttaaa gaaggaagca atacgaccgt gggttacttc gggaacgagg ggccggactg 540

aaatagcatt atcagcgcgc atccaatgat gccggcctag gtcacactca gtagccattt 600

tcttaatggc ttcagggcta aataatatct tttgaagatc gtcagacgta gccgctgagg 660

actgttgagt taaagtccta ctggtacctt ggaaagccat gcctcgggaa atctaacaca 720

ctgt 724

<210> 18

<211> 820

<212> DNA/RNA

<213> cucumber mosaic virus

<400> 18

aaatctgaat caaccagtgc tggtcgtaac cgtcgacgtc gtccgcgtcg tggttcccgc 60

tccgccccct cctccgcgga tgctaacttt agagtcttgt cgcagcagct ttcgcgactt 120

aataagacgt tagcagctgg tcgtccaact attaaccacc caacctttgt agggagtgaa 180

cgctgtagac ctgggtacac gttcacatct attaccctaa agccaccaaa aatagaccgt 240

gggtcttatt acggtaaaag gttgttacta cctgattcag tcacggaata tgataagaag 300

cttgtttcgc gcattcaaat tcgagttaat cctttgccga aatttgattc taccgtgtgg 360

gtgacagtcc gtaaagttcc tgcctcctcg gacttatccg ttgccgccat ctctgctatg 420

ttcgcggacg gagcctcacc ggtactggtt tatcagtcac ccacacggta gaatcaaatt 480

tcggcaaagg attaactcga atttgaatgc gcgaaacaag cttcttatca tattccgtga 540

ctgaatcagg tagtaacaac cttttaccgt aataagaccc acggtctatt tttggtggct 600

ttagggtaat agatgtgaac gtgtacccag gtctacagcg ttcactccct acaaaggttg 660

ggtggttaat agttggacga ccagctgcta acgtcttatt aagtcgcgaa agctgctgcg 720

acaagactct aaagttagca tccgcggagg agggggcgga gcgggaacca cgacgcggac 780

gacgtcgacg gttacgacca gcactggttg attcagattt 820

Claims (8)

1. The RNA interference sequence of the targeted cucumber mosaic virus is double-stranded RNA and consists of a sense strand and an antisense strand, and specifically comprises the following steps:

an interference sequence GT1 of a specificity target CMV-1a protein GT1 gene, wherein the specific nucleotide sequence of a sense chain is shown as SEQ ID NO.1, and the specific nucleotide sequence of an antisense chain is shown as SEQ ID NO. 2;

or an interference sequence GT2 of a specificity target CMV-1a protein GT2 gene, the specific nucleotide sequence of the sense strand is shown as SEQ ID NO.3, and the specific nucleotide sequence of the antisense strand is shown as SEQ ID NO. 4;

or an interference sequence GT3 of a specificity target CMV-2a protein GT3 gene, the specific nucleotide sequence of the sense strand is shown as SEQ ID NO.5, and the specific nucleotide sequence of the antisense strand is shown as SEQ ID NO. 6;

or an interference sequence GT4 of a specificity target CMV-2a protein GT4 gene, the specific nucleotide sequence of the sense strand is shown as SEQ ID NO.7, and the specific nucleotide sequence of the antisense strand is shown as SEQ ID NO. 8;

or an interference sequence GT5 of a specificity target CMV-CP protein GT5 gene, the specific nucleotide sequence of the sense strand is shown as SEQ ID NO.9, and the specific nucleotide sequence of the antisense strand is shown as SEQ ID NO. 10;

or an interference sequence GT16 of a specificity target CMV-MP protein GT6 gene, the specific nucleotide sequence of the sense strand is shown as SEQ ID NO.11, and the specific nucleotide sequence of the antisense strand is shown as SEQ ID NO. 12.

2. The RNAi inverted repeat sequence of the RNA interference sequence of claim 1, comprising a sense strand sequence and an antisense strand sequence of the double-stranded RNA, wherein the inverted repeat sequence of the interference sequence GT1 is GT1-3, and the nucleotide sequence is shown in SEQ ID NO. 13; the inverted repeat sequence of the interference sequence GT2 is GT2-3, and the nucleotide sequence is shown as SEQ ID NO. 14; the inverted repeat sequence of the interference sequence GT3 is GT3-3, and the nucleotide sequence is shown as SEQ ID NO. 15; the inverted repeat sequence of the interference sequence GT4 is GT4-3, and the nucleotide sequence is shown as SEQ ID NO. 16; the inverted repeat sequence of the interference sequence GT5 is GT5-3, and the nucleotide sequence is shown as SEQ ID NO. 17; the inverted repeat sequence of the interference sequence GT6 is GT6-3, and the nucleotide sequence is shown in SEQ ID NO. 18.

3. An expression vector targeting cucumber mosaic virus comprising a primary vector and at least one RNA interference sequence of claim 1 or comprising a primary vector and at least one RNAi inverted repeat of claim 2.

4. Expression vector targeting cucumber mosaic virus according to claim 3, characterized in that the original vector is a commonly used viral or plasmid vector.

5. The cucumber mosaic virus-targeted expression vector according to claim 4, wherein the original vector is pGEM vector plasmid.

6. The RNA interference sequence expression strain of claim 1, which is an engineering strain obtained by transforming the expression vector of any one of claims 3-5 into Escherichia coli.

7. Use of the cucumber mosaic virus-targeted RNA interference sequence of claim 1, or the RNAi inverted repeat sequence of claim 2, or the expression vector of claim 3 for the preparation of an agent for the control of cucumber mosaic virus.

8. The use of claim 7, wherein a dsRNA spray containing one or more of the RNA interference sequences is sprayed to cucumber seedlings.

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