Application of rice transcription factor OsLSD5 in rice virus resistance

Abstract

The invention relates to a rice transcription factor OsLSD5 and application of the rice transcription factor OsLSD5 in resisting rice stripe virus, and after an overexpression OsLSD5 vector is transferred into plant rice, an OsLSD5-ox transgenic plant with stable inheritance is obtained.

Description

Technical Field

[0001] The invention relates to the field of plant disease prevention and control and transgenic technology, and mainly to rice transcription factor OsLSD5 and application thereof in rice resistance to viral diseases. Background Art

[0002] Rice stripe virus (RSV) is currently considered to be one of the most important rice viral pathogens in East Asia and a typical representative of the genus Tenuivirus. Rice plants infected with RSV usually show symptoms such as chlorosis, discontinuous yellow stripes, necrosis of new leaves and slow growth. The virus is mainly transmitted by the small brown planthopper (Laodelphax striatellus) and can be transmitted to offspring through the insect ovaries. Rice stripe leaf blight caused by RSV occurs in rice-growing areas in many provinces in my country, seriously affecting rice yield and quality. With the development of biotechnology, researchers have conducted extensive research on the genomics and proteomics of RSV and the interactions between the different viral proteins it encodes and host factors. It was found that the RSV genome consists of four negative-sense single-stranded RNAs, named RNA1, RNA2, RNA3 and RNA4 according to their sizes, encoding a total of 7 proteins.

[0003] Lesion simulating disease proteins (LSDs) are a class of specific zinc finger protein subfamily, belonging to the C2C2 type transcription factor, containing multiple conserved zinc finger domains CxxCRxxLMYxxGASxVxCxxC. This zinc finger sequence has homology with GATA type transcription factors, which exist in plants, algae and protozoa, but not in animals. PAD4 (Enhanced disease susceptibility 1) can encode a pathogen-induced protein that acts upstream of salicylic acid (SA). EDS1 (Enhanced disease susceptibility 1) is an important regulator of innate immunity and participates in the signal transduction of R genes together with PAD4. Studies have shown that the PCD symptoms exhibited by lsd1 Arabidopsis mutants are dependent on EDS1 and PAD4. LSD1, EDS1 and PAD4 form a specific hub responsible for triggering cell death and adaptive responses to adverse environments that depend on salicylic acid, ethylene and reactive oxygen species.

[0004] LSDs play an important role in plant growth and development, and in response to abiotic and biotic stresses. On the one hand, rice LSD1 can interact with the transcription factor OsbZIP58, promote OsbZIP58 to bind to the promoter of the gibberellin synthesis-related gene KO2, and upregulate the expression of OsKO2, thereby activating gibberellin synthesis, which in turn activates amylase expression and promotes seed germination; on the other hand, OsLSD1 can indirectly negatively regulate the expression of OssodCc1, reduce the activity of superoxide dismutase SOD, lead to increased accumulation of reactive oxygen species, accelerate PCD of the aleurone layer, and promote seed germination. Overexpression of the rice gene OsLOL5 significantly upregulated the rice oxidative stress-related genes OsAPX2, OsCAT, OsCu / Zn-SOD, and OsRGRC2, and increased the resistance of rice to salt stress. Soybean rust pathogen (Phakopsora pachyrhizi) is a pathogenic strain of Asian rust. When the rust pathogen infects soybean, the expression level of soybean LSD increases rapidly. Silencing the wheat gene TaLSD1 can promote wheat hypersensitive cell death and improve wheat resistance to stripe rust. Overexpression of the rice gene OsLOL2 can enhance rice resistance to rice bacterial blight. Cassava LSD3 can interact with the deacetylase SRT1, inhibiting its deacetylation modification of histone H3K9Ac, resulting in an increase in the expression level of H3K9Ac, and upregulating the expression of pattern recognition receptor genes SOBIR1 (Suppressor of BIR 1) and FLS2 (Flagellin-sensitive 2), resisting Xanthomonas phaseoli pv.manihotis infection of cassava. These studies have shown that LSD proteins play an important role in plant resistance to pathogen infection. However, there are currently no reports on the relationship between rice viruses and LSD proteins. Summary of the invention

[0005] Based on the defects of the prior art, the present invention relates to a rice LSD gene and its application in resisting rice virus.

[0006] In one aspect, a rice transcription factor OsLSD5, the nucleotide and protein sequences of which are shown below:

[0007] The nucleotide sequence of OsLSD5 is shown in SEQ ID NO: 1:

[0008] ATGCAGGACCAGCTGATCTGCAGCGGCTGCAGGCGCGTCGTCCAGTACAGGAGAGGGGTCGCCGGCGTCTGCTGCCCGGGCTGCAACACGCTCACCGCCGTCAACCCGTCAGCGGTGGCCGACATGTCGGAGCTCATCTGCAGCGGCTGCCCCACGCTGCTGTTCTACAACCGCGGCGCCTCCAACATCCGCTGCCCCAGCTGCAACAGGCTCAACTCCACC AGATCAGCCAACCAGATTGCACACCTGACATGCGGGCAGTGCCGGACGACTCTGATGCACCCACCTGGAGCCTCAACTGTGCAGTGTGCAACCTGCAGATATGTTAACCATGTCAGGGATGCTCGGCCTCAAACTGTCCTTGTAGAATCCTAAGACACTGGATGATAAGGGCAAGCTGGTGAGCAATGTGGTTGTTGGTGTCACCTCATGGAAAAGATGA

[0009] The amino acid sequence of OsLSD5 is shown in SEQ ID NO: 2:

[0010] MQDQLICSGCRRVVQYRRGVAGVCCPGCNTLTAVNPSAVADMSELICSGCPTLLFYNRGASNIRCPSCNRL

[0011] NSTRSANQIAHLTCGQCRTTLMHPPGASTVQCATCRYVNHVRDARPQTVLVENPKTLDDKGKLVSNVVVGVTSWKR*

[0012] In some embodiments, the biological material related to the protein encoded by the rice gene OsLSD5 provided by the present invention is any one of the following A1) to A12):

[0013] A1) a nucleic acid molecule encoding an OsLSD5 gene protein;

[0014] A2) an expression cassette containing the nucleic acid molecule described in A1);

[0015] A3) a recombinant vector containing the nucleic acid molecule described in A1);

[0016] A4) a recombinant vector containing the expression cassette described in A2);

[0017] A5) a recombinant microorganism containing the nucleic acid molecule described in A1);

[0018] A6) a recombinant microorganism containing the expression cassette described in A2);

[0019] A7) a recombinant microorganism containing the recombinant vector described in A3);

[0020] A8) a recombinant microorganism containing the recombinant vector described in A4);

[0021] A9) a transgenic plant cell line containing the nucleic acid molecule described in A1);

[0022] A10) a transgenic plant cell line containing the expression cassette described in A2);

[0023] A11) a transgenic plant cell line containing the recombinant vector described in A3);

[0024] A12) A transgenic plant cell line containing the recombinant vector described in A4).

[0025] On the other hand, the present invention relates to the use of the rice transcription factor OsLSD5 in the breeding of Gramineae food crops resistant to Tenuivirus;

[0026] In some embodiments, the Tenuivirus genus includes Ricestripe virus (RSV), Maize stripe virus (MSpV), preferably Ricestripe virus (RSV);

[0027] In some embodiments, the gramineous crops are preferably rice, corn, wheat, oats and barley; more preferably rice, and most preferably Hordeum vulgare 11;

[0028] In another aspect, the present invention relates to a method for preparing a transgenic plant resistant to rice stripe virus and / or southern rice black-streaked dwarf virus, the steps of which include:

[0029] (1) Cloning of the rice OsLSD5 gene;

[0030] (2) Construction of overexpression vector;

[0031] (3) Agrobacterium transformation and callus induction culture;

[0032] (4) Positive identification of transgenic plants;

[0033] In some embodiments, the cloning of the rice OsLSD5 gene comprises the following steps:

[0034] 1) Design primers OsLSD5-F and OsLSD5-R based on the open reading frame (ORF) of OsLSD5

[0035] ATGCAGGACCAGCTGATCTG(SEQ ID NO:3);

[0036] TCATCTTTTCCATGAGGTGACACC(SEQ ID NO:4);

[0037] 2) Carry out PCR reaction, recover the PCR product, connect it to pMD18-T vector, pick a single clone, send it for testing, and obtain the correct pMD18-T-OsLSD5 recombinant plasmid.

[0038] In some embodiments, the construction of the overexpression vector comprises the following steps:

[0039] 1) Designing recombination primers with GTCACC and CCGTCG restriction sites, using the correctly identified pMD18-T-OsLSD5 recombinant plasmid as a template, amplifying the OsLSD5 gene, and recovering the PCR product; then double-digesting the pCV1300 vector with ApaI enzyme, and recovering the cut vector;

[0040] 2) The above PCR product and vector were connected, transformed into Escherichia coli DH5α, and positive clones were selected and sequenced to confirm that the pCV1300-OsLSD5 overexpression vector had been successfully constructed;

[0041] The sequences of the recombination primers are as follows:

[0042] pCV-OsLSD5-F CGACGACAAGACCGTCACCATGCAGGACCAGCTGATCTG (SEQ ID NO: 5);

[0043] pCV-OsLSD5-R GAGGAGAAGAGCCGTCGTCTTTTCCATGAGGTGACACC (SEQ ID NO: 6);

[0044] In some embodiments, the Agrobacterium transformation and callus induction culture steps include:

[0045] (1) Take an appropriate amount of the constructed overexpression vector plasmid and add it to the Agrobacterium competent cells, mix well, add it to the electrode cup precooled at 4°C, transform it with 2200V voltage, add 800μL of LB liquid medium without resistance, shake and culture it at 28°C for 2-3h, centrifuge it at 5000rpm for 1min, discard the supernatant, and spread the remaining precipitate on a solid medium containing 50μg / ml Kan and 50μg / ml Rif resistance, and culture it in a 28°C incubator for 3d; dehull the rice seeds to be transformed, soak them in 75% alcohol for 5min, rinse them with sterile water twice, soak them in 30% sodium hypochlorite solution for 30min, and soak them in sterile water for 30min; put the seeds into the induction medium with tweezers, and culture them in a 28°C light incubator for 3-4 weeks; clip the grown callus tissue into the subculture medium with pre-sterilized tweezers, and subculture them in a 28°C light incubator for 1 week;

[0046] (2) Pick a single clone of Agrobacterium GV3101 and inoculate it into LB liquid culture medium. Cultivate the culture medium until the concentration reaches OD 600 = about 0.6; soak the induced callus in Agrobacterium suspension culture medium for 5 minutes; take out the callus, put it in symbiotic culture medium, and culture it in a 26℃ light incubator for 2-3 days; then transfer the callus to a culture medium containing hygromycin and grow it for 30-45 days. Finally, put the selected callus into rooting culture medium and culture it under light until the callus takes root. Continue to culture for 2 weeks to obtain transgenic plants.

[0047] In some embodiments, the positive seedling identification step of the transgenic plant comprises:

[0048] Total RNA of transgenic plants selected by hygromycin was extracted and quantitatively reverse transcribed into cDNA, with the OsUBQ5 gene of rice as an internal reference; the quantitative primers of OsLSD5 are shown in SEQ ID NO: 7-10:

[0049] qRT-OsLSD5-F:GACATGTCGGAGCTCATCTG (SEQ ID NO:7);

[0050] qRT-OsLSD5-R: CAATCTGGTTGGCTGATCTG (SEQ ID NO:8);

[0051] OsUBQ5-F:ACCACTTCGACCGCCACTACT(SEQ ID NO:9);

[0052] OsUBQ5-R: ACGCCTAAGCCTGCTGGTT (SEQ ID NO: 10);

[0053] In some embodiments, the induction medium comprises: N6 medium 24.1 g / L, 2 mg / L 2,4-D, pH=5.8.

[0054] The subculture culture medium comprises: 24.1 g / L N6 culture medium, 2 mg / L 2,4-D, 50 mg / L hygromycin, 300 mg / mL cephalosporin, and pH=5.8.

[0055] The co-culture medium comprises: 24.1 g / L N6 culture medium, 2 mg / L 2,4-D, 200 μmol / L acetosyringone, and pH=5.2.

[0056] The rooting medium comprises: 1 / 2MS 39.45 g / L, 0.5 mg / L NAA, 50 mg / L hygromycin, and pH=5.8.

[0057] In another aspect, the present invention also relates to a method for detecting resistance of rice to rice stripe virus, the steps of which include:

[0058] 1) The total RNA of the tested rice and the control Zhonghua 11 rice was extracted using the TRIzol method, and the RNA was reverse transcribed into cDNA; the relative expression level of the OsLSD5 gene was detected using quantitative primers, and the quantitative primers were as follows:

[0059] qRT-OsLSD5-F:GACATGTCGGAGCTCATCTG (SEQ ID NO:7);

[0060] qRT-OsLSD5-R: CAATCTGGTTGGCTGATCTG (SEQ ID NO:8);

[0061] OsUBQ5-F:ACCACTTCGACCGCCACTACT(SEQ ID NO:9);

[0062] OsUBQ5-R: ACGCCTAAGCCTGCTGGTT (SEQ ID NO: 10);

[0063] According to the above gene expression detection results, it is judged whether the rice plant to be tested is resistant rice. If the relative expression level of the OsLSD5 gene in the rice plant to be tested is significantly higher than that of the control, it is resistant rice.

[0064] The present invention uses plant transgenic technology to transform the full-length OsLSD5 gene into rice, and then screens and obtains transgenic strains with high expression levels. Experiments show that after RSV infects rice, the resistance of the OsLSD5 overexpressing transgenic to virus infection is significantly increased compared with the wild type control. OsLSD5 plays a positive regulatory role in the antiviral process of rice.

[0065] The present invention mainly adopts the following scheme to achieve the above invention objectives:

[0066] The present invention takes rice transcription factor OsLSD5 as the object, constructs OsLSD5 recombinant expression vector pCV1300-OsLSD5, uses Agrobacterium GV3101-mediated genetic transformation method to transfer into rice Zhonghua 11 (ZH11) embryo, and puts into symbiotic culture medium to induce callus formation. After obtaining transgenic rice seedlings, the expression amount of OsLSD5 gene in rice is detected by real-time fluorescence quantitative PCR technology (qRT-PCR), and after several generations of screening and identification, T3 generation stable inherited homozygous transgenic rice is obtained. By artificial inoculation of RSV, resistance of OsLSD5 transgenic rice is evaluated. The results show that the symptoms of transgenic rice overexpressing OsLSD5 infected with RSV are milder than those of wild type, and the virus content in the body is also significantly lower than the virus content in wild type rice. This shows that OsLSD5 overexpressing transgenic rice can significantly enhance the resistance of rice to RSV. The research results not only show that OsLSD5 affects the infection efficiency of RSV, but also lay the foundation for the study of the interaction between rice and viruses.

[0067] Compared with the prior art, the advantages and beneficial effects of the present invention are:

[0068] (1) The present invention will overexpress OsLSD5 in Zhonghua 11 rice to obtain stably inherited transgenic rice overexpressing OsLSD5, and analyze the resistance of the transgenic rice to rice stripe disease, which will help deepen our understanding and knowledge of rice viruses and host factors and enrich the molecular mechanism of the interaction between viruses and rice.

[0069] (2) The present invention studies the mechanism of action of OsLSD5 in viral infection and discovers the interaction between plants and viruses, laying a foundation for further research on rice disease resistance theory. BRIEF DESCRIPTION OF THE DRAWINGS

[0070] Figure 1 :Experimental results of relative expression level of OsLSD5 in OsLSD5 overexpressing transgenic rice.

[0071] Figure 2 : Comparison of symptoms between OsLSD5 overexpressing transgenic and wild type control after RSV infection.

[0072] Figure 3 : Experimental results of virus content of OsLSD5 overexpressing transgenic and wild type control after RSV infection. DETAILED DESCRIPTION

[0073] The rice varieties used in this series of experiments are: Zhonghua 11 (ZH11)

[0074] The relevant culture medium components are as follows:

[0075] Induction medium: N6 medium (manufacturer: Haibo Biotechnology Co., Ltd., catalog number: HBZ0601)

[0076] 24.1g / L, 2.5mg / L 2,4-D, PH=5.8.

[0077] Subculture medium: N6 medium (manufacturer: Haibo Biotechnology Co., Ltd., product number: 15HBZ0601) 24.1 g / L, 2,4-D, 50 mg / L hygromycin, 300 mg / mL cephalosporin, PH = 5.8.

[0078] Co-culture medium: N6 culture medium (manufacturer: Haibo Biotechnology Co., Ltd., product number: HBZ0601) 24.1 g / L, 2.5 mg / L 2,4-D, 200 μmol / L acetosyringone, pH = 5.8.

[0079] Rooting medium: 1 / 2MS (Manufacturer: Haibo Biotechnology Co., Ltd., Product No.: HB8469-6)

[0080] 39.45g / L, 0.5mg / L NAA, 50mg / L hygromycin, PH=5.8.

[0081] Example 1: Construction of rice OsLSD5 overexpression vector

[0082] (1) Cloning of the rice OsLSD5 gene

[0083] Primers OsLSD5-F and OsLSD5-R were designed based on the open reading frame (ORF) of OsLSD5. The primer sequences are as follows:

[0084] Table 1: Primer sequences involved in gene cloning

[0085] OsLSD5-F ATGCAGGACCAGCTGATCTG(SEQ ID NO:3); OsLSD5-R TCATCTTTTCCATGAGGTGACACC(SEQ ID NO:4);

[0086] PCR amplification system: total volume 50 μL, including 2×PCR Buffer for KOD FX Neo 25 μL, upstream and downstream primers (10 μM) 1.5 μL each, dNTP Mix (2.5 mM) 5 μL, cDNA template 1 μL, KOD FX Neo 1 μL, ddH2O 15 μL.

[0087] PCR amplification program: pre-denaturation at 94°C for 3 min; denaturation at 94°C for 30 s, annealing at 58°C for 30 s, extension at 72°C for 3 min, 40 cycles; final extension at 72°C for 10 min.

[0088] The PCR product was recovered, connected to the pMD18-T vector, single clone was picked, and sent for testing to obtain the correct pMD18-T-OsLSD5 recombinant plasmid, which was sent to Hangzhou Youkang Biological Company for sequencing.

[0089] (2) Construction of overexpression vector

[0090] Primers with BamHI and SacI restriction sites were designed to amplify OsLSD5. The pMD18-T-OsLSD5 plasmid with correct sequencing was used as the template. The pCV1300 vector was double-digested with BamHI and SacI enzymes for the construction of the OsLSD5 gene binary expression vector PCV1300. The primer sequences are as follows:

[0091] Table 2: Primer sequences involved in the construction of overexpression vectors

[0092]

[0093] PCR amplification system: The reaction system (50 μL) was as follows: 2×PCR KOD FX Neo Buffer 25 μL, 2 mM dNTPs, KOD FX Neo 1 μL, upstream primer 1.5 μL, downstream primer 1.5 μL, cDNA 1 μL, ddH2O 15 μL, total volume 50 μL.

[0094] PCR program: pre-denaturation at 95°C for 3 min; denaturation at 95°C for 30 s, annealing at 60°C for 30 s, extension at 72°C for 2 min, 35 cycles; final extension at 72°C for 10 min.

[0095] After the pCV1300 vector was digested with enzymes, the PCR products were recovered and connected to the vectors respectively; positive clones were selected and sequenced to confirm that the pCV1300-OsLSD5 expression vector had been successfully constructed.

[0096] Example 2: Rice genetic transformation

[0097] (1) Agrobacterium transformation and callus induction culture: The constructed recombinant vector plasmid was transformed into Agrobacterium GV3101. The specific steps include: taking 2-3 μL of plasmid and adding it to 100 μL of competent medium, mixing it, sucking it into an electrode cup precooled at 4°C, electroporating it with a voltage of 2200V, adding 800 μL of LB liquid medium without resistance, shaking and culturing it at 28°C and 200rpm for 2-3h, centrifuging it at 5000rpm for 1min, discarding part of the supernatant, and coating the remaining precipitate on a solid medium plate containing 50 μg / ml Kan and 50 μg / ml Rif, and culturing it in an incubator at 28°C for 3d. Mature rice seeds were shelled, soaked in 75% alcohol for 5min, rinsed several times in ddH2O, soaked in 30% sodium hypochlorite solution for 30min, and soaked in ddH2O for 30min. Use tweezers to place the seeds into the induction medium and culture them in a 28°C light incubator for 3 weeks. Use pre-sterilized tweezers to clip the grown callus into the subculture medium and culture them in a 28°C light incubator for 1 week.

[0098] (2) Screening and culture of Agrobacterium-transfected callus and resistant callus: A single clone of GV3101 was selected and inoculated into LB liquid medium and cultured until OD 600 = about 0.6. The induced callus was immersed in the Agrobacterium suspension culture solution for 5 minutes, and the immersed callus was transferred to the culture medium. After culturing in a 26°C light incubator for 2-3 days, the callus was placed on a culture medium containing hygromycin for 30-45 days. The selected callus was transferred to a rooting medium and cultured at 26°C light for 2 weeks to produce green plants with roots, and the OsLSD5-ox transgenic lines OsLSD5-6# and OsLSD5-8# overexpressing the OsLSD5 gene were obtained.

[0099] Example 3: Positive identification of transgenic plants

[0100] Total RNA of the transgenic plants to be tested was extracted and reverse transcribed into cDNA, and the OsUBQ5 gene of rice was used as an internal reference. The quantitative primers are shown in SEQ ID No: 7-10. The relative expression levels of the OsLSD5-ox transgenic lines OsLSD5-#6 and OsLSD5-#8 overexpressing the OsLSD5 gene were significantly higher than those of the control ( Figure 1 ).

[0101] Table 3: Primers involved in positive identification of transgenic plants

[0102] qRT-OsLSD5-F GACATGTCGGAGCTCATCTG (SEQ ID NO:7); qRT-OsLSD5-R CAATCTGGTTGGCTGATCTG (SEQ ID NO:8); OsUBQ5-F ACCACTTCGACCGCCACTACT(SEQ ID NO:9); OsUBQ5-R ACGCCTAAGCCTGCTGGTT (SEQ ID NO: 10);

[0103] Example 4: Artificial inoculation of RSV

[0104] OsLSD5 transgenic rice seeds and control rice seeds were placed in a 37°C incubator for 2-3 days. After the seeds turned white, they were sown in 1L glass beakers, with 30-35 seedlings per cup and 3 biological replicates. They were placed in a 25°C artificial climate chamber and cultured under 16h light and 8h dark conditions. 1-2-year-old non-toxic gray planthoppers were transferred to RSV-infected rice seedlings for 3-5 days, and then moved to healthy rice seedlings using a vacuum pump for the transition period (10-12 days).

[0105] The infection rate of insects was detected, and the number of insects inoculated per seedling was calculated based on the infection rate. Rice seedlings at the three- to four-leaf stage (seedlings of about 15 days) were inoculated with virus-carrying / non-virus-carrying Laodelphax striatellus. All insects were swept out after 3 days, and the rice seedlings were placed in a 30℃ greenhouse for growth. After 30 days, the symptoms of rice disease were observed and the incidence rate was calculated to determine the disease status of rice. The two OsLSD5-ox transgenic lines (#6 and #8) showed obvious resistance to RSV, such as Figure 2 As shown. qRT-PCR further detected the virus content, and the results showed that the virus content of the susceptible transgenic rice lines was significantly lower than that of the wild type, such as Figure 3 shown.

[0106] The sequences of the quantitative primers are as follows:

[0107] qRSV-CP-F AGGCAATCAATGACATCTCC; SEQ ID NO:11

[0108] qRSV-CP-R ATCTCTCACAAAGCCAGTGC; SEQ ID NO:12

[0109] It should be understood that the specific examples and schemes described in the present invention are only given as examples for illustrative purposes and are not to be construed as limitations of the present invention. Various further modifications or changes within the spirit and scope of the present invention are to be considered as being included within the scope of the disclosure of the present invention.

Claims

1. An application of rice transcription factor OsLSD5 in breeding of Gramineae food crops resistant to Tenuivirus; the nucleotide sequence of the OsLSD5 gene is shown in SEQ ID NO:

1.

2. The use according to claim 1, wherein the amino acid sequence of the OsLSD5 gene is shown in SEQ ID NO:

2.

3. The use according to any one of claims 1 to 2, wherein the Tenuivirus genus includes Rice stripe virus (RSV) and Maize stripe virus (MSpV); preferably Rice stripe virus.

4. The use according to any one of claims 1 to 3, wherein the gramineous food crop is selected from rice, corn, wheat, oats and barley; preferably rice, more preferably japonica rice varieties such as Huaidao No. 5, Zhonghua 11, Wuyujing No. 3, and Zhonghua 11 is the most preferred.

5. A method for preparing resistant transgenic rice, comprising the following steps: transferring an OsLSD5 overexpression vector into wild-type rice to obtain resistant transgenic rice, wherein the resistant transgenic rice has resistance to rice stripe virus (RSV) compared with wild-type rice, and the nucleotide sequence of the OsLSD5 gene is shown in SEQ ID NO:

1.

6. A method for identifying rice resistant to rice stripe virus, comprising the following steps: extracting total RNA from the rice plant to be tested, reverse transcribing it into cDNA, using the OsUBQ5 gene of rice as an internal reference, using OsLSD5 and OsUBQ5 specific primers for detection, and judging whether the rice plant to be tested is resistant rice based on the gene expression detection results.

7. The method according to claim 6, wherein the specific primer sequence is as follows: qRT-OsLSD5-F GACATGTCGGAGCTCATCTG (SEQ ID NO:7); qRT-OsLSD5-R CAATCTGGTTGGCTGATCTG (SEQ ID NO:8); OsUBQ5-F ACCACTTCGACCGCCACTACT(SEQ ID NO:9); OsUBQ5-R ACGCCTAAGCCTGCTGGTT (SEQ ID NO: 10).

8. The method according to any one of claims 6-7, wherein the determination method is: if the relative expression level of the OsLSD5 gene in the rice plant to be detected is significantly higher than that in the control, the rice plant is resistant rice.

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