Use of osprmt10 protein or its encoding gene in enhancing rice resistance to pest stress

Abstract

The application discloses use of OsPRMT10 protein or a coding gene thereof in enhancing rice resistance to pest stress, and comprises the following steps: reducing or inhibiting expression amount or expression level of the OsPRMT10 protein coding gene in rice, or reducing activity, losing function or causing defects of the OsPRMT10 protein in rice. The application also provides a method for improving rice resistance to pest stress or a method for creating a rice variety resistant to pest stress, and comprises the following steps: constructing a gene knockout vector, a CRISPR / Cas9 gene editing vector or an RNA interference vector of the OsPRMT10 protein coding gene; knocking out or mutating the OsPRMT10 protein coding gene in rice, or inhibiting expression of the OsPRMT10 protein coding gene in rice, so as to improve the resistance of the rice to pest stress; and the application has application prospects in improving the resistance of the rice to pest stress and creating a rice variety resistant to pest stress.

Description

TECHNICAL FIELD

[0001] The present application relates to a new use of OsPRMT10 protein or its encoding gene, in particular to the use of OsPRMT10 protein or its encoding gene in enhancing the resistance of rice to pest stress, and belongs to the field of new use of OsPRMT10 protein or its encoding gene. BACKGROUND

[0002] Cnaphalocrocis medinalis Guenée, belonging to Lepidoptera Pyralidae, is one of the main pests of rice in China. It mainly harms rice at the booting and heading stages, leading to an increase in the empty grain rate and a decrease in the thousand-grain weight, and seriously affecting the yield. At present, the control of Cnaphalocrocis medinalis Guenée mainly relies on chemical control. However, due to the continuous and unreasonable use of chemical pesticides, the problem of Cnaphalocrocis medinalis Guenée resistance and resurgence is becoming increasingly serious. Breeding rice varieties resistant to Cnaphalocrocis medinalis Guenée can effectively control pests, reduce pesticide use, and improve rice yield. Therefore, the exploration and utilization of Cnaphalocrocis medinalis Guenée resistant germplasm resources, the identification of resistance genes, and the in-depth study of the regulation mechanism have important production application value for pest-resistant rice breeding.

[0003] At present, the genetic control technology of Cnaphalocrocis medinalis Guenée mainly enhances the resistance by expressing exogenous pest-resistant genes in rice.

[0004] Protein arginine methyltransferases (PRMTs) play an important role in plant growth and development and biotic and abiotic stress processes. The loss of function of AtPRMT5 enhances the resistance of Arabidopsis to the pathogenic oomycete Hyaloperonospora arabidopsidis Hyaloperonospora arabidopsidisNoco2 resistance. Analysis showed that AtPRMT5 was co-localized with AtCln and the methylosome component small nuclear ribonucleoprotein D3b in the same protein complex, which was essential for mRNA splicing, suggesting that AtPRMT5 was involved in mRNA splicing and thus regulated plant immune response (Shuai et al. (2016). Identification of methylosome components as negative regulators of plant immunity using chemical genetics). PRMT6 positively regulated tomato resistance to tomato bushy stunt virus, and loss-of-function and overexpression of PRMT6 led to exacerbation and alleviation of infection symptoms, respectively. PRMT6 directly targeted the viral suppressor RNA silencing protein P19 of tomato bushy stunt virus and inhibited its function by methylating its key amino acid residues R43 and R115, thereby weakening the dimerization ability of P19 and its small RNA binding activity. Analysis of natural tomato populations showed that two major allelic genotypes with high and low expression of PRMT6 significantly corresponded to strong and weak viral resistance, respectively (Qiangqianget al. (2024). Protein arginine methyltransferase 6 mediates antiviral immunity in plants). Currently, the research on the function of rice PRMT proteins is very limited. By systematically analyzing the subcellular localization and methyltransferase activity of 8 OsPRMT proteins, it was found that OsPRMT1, OsPRMT5, and OsPRMT10 were distributed in the cytoplasm and nucleus, while OsPRMT6a and OsPRMT6b were mainly expressed in the nucleus; OsPRMT1, OsPRMT4, OsPRMT5, OsPRMT6a, OsPRMT6b, and OsPRMT10 could catalyze the arginine methylation of myelin basic protein and fibrin glycine-arginine-rich domains and bovine thymus core histone (Ayaz et al. (2011). Characterization of the prmt gene family in rice reveals conservation of arginine methylation). Through the analysis of maize ZmPRMT1-8 genes and proteins, it was found that ZmPRMTs genes were expressed in different tissues of maize and were induced by various abiotic stresses, such as heat, drought, and salt stress. Further verification of the function of ZmPRMT1 in Arabidopsis thaliana found that overexpression ZmPRMT1Gene-induced early flowering and improved heat tolerance (Qiqi et al. (2022). Genome-wide identification of maize protein arginine methyltransferase genes and functional analysis of zmprmt1 reveal essential roles in Arabidopsis AtPRMT5, through synergistic action with the E3 ubiquitin ligase COP1, regulates light-induced post-transcriptional splicing events in mesophyll cells (Yane et al. (2023). Light controls mesophyll-specific post-transcriptional alsplicing of photoregulatory genes by AtPRMT5). However, research on the involvement of rice PRMT proteins / encoding genes in the regulation of biological stress and immune responses is limited, and there are no reports of rice PRMT proteins / encoding genes regulating plant resistance to pest stress. Summary of the Invention

[0005] One objective of this invention is to provide the use of the OsPRMT10 protein or its encoding gene in enhancing rice's resistance to pests;

[0006] The second objective of this invention is to provide a method for improving the resistance of rice to pests;

[0007] The third objective of this invention is to provide a method for creating pest-resistant rice varieties.

[0008] The above-mentioned objective of the present invention is achieved through the following technical solution:

[0009] One aspect of the present invention is to provide the use of the OsPRMT10 protein or its encoding gene in enhancing rice's resistance to pests, including: reducing or inhibiting the expression level of the encoding gene of the OsPRMT10 protein in rice, or reducing the activity, losing function, or causing defects in the OsPRMT10 protein in rice.

[0010] The OsPRMT10 protein described in this invention is a protein derived from the insect-susceptible rice material CH1057 or the insect-resistant rice material SE378; wherein, the amino acid sequence of the OsPRMT10 protein derived from the insect-susceptible rice material CH1057 is shown in SEQ ID No. 1, and its encoding gene ( OsPRMT10The nucleotide sequence of the genome of ) is shown in SEQ ID No. 2, which encodes the gene ( OsPRMT10 The nucleotide sequence of the coding sequence (CDS) of ) is shown in SEQ ID No. 3; nucleotides 1-213 of the nucleotide sequence shown in SEQ ID No. 2 are exons 1, 290-482 are exons 2, 717-822 are exons 3, 1068-1257 are exons 4, 1583-1768 are exons 5, 1850-1933 are exons 6, and 2318-2494 are exons 7.

[0011] The amino acid sequence of the OsPRMT10 protein derived from the insect-resistant rice material SE378 is shown in SEQ ID No. 4; the encoding gene in the insect-resistant material SE378 ( OsPRMT10 The nucleotide sequence of the genome of ) is shown in SEQ ID No. 5, which encodes the gene ( OsPRMT10 The nucleotide sequence of the coding sequence (CDS) of ) is shown in SEQ ID No. 6; nucleotides 1-213 of the nucleotide sequence shown in SEQ ID No. 5 are exons 1, 290-482 are exons 2, 717-822 are exons 3, 1068-1257 are exons 4, 1583-1768 are exons 5, 1850-1933 are exons 6, and 2318-2494 are exons 7.

[0012] Another aspect of the present invention is to provide a method for improving the resistance of rice to pest stress, comprising: knocking out or mutating the gene encoding the OsPRMT10 protein, reducing, inhibiting or interfering with the expression level or expression amount of the gene encoding the OsPRMT10 protein in rice, thereby reducing the activity, losing function or producing defects of the OsPRMT10 protein in rice.

[0013] Another aspect of the present invention provides a method for creating rice varieties resistant to pest stress, comprising: constructing a gene knockout vector for the OsPRMT10 protein-coding gene, or constructing a CRISPR / Cas9 gene editing vector for the OsPRMT10 protein-coding gene using gene editing technology, or constructing an RNA interference vector for the OsPRMT10 protein-coding gene; transforming rice with the gene knockout vector, CRISPR / Cas9 gene editing vector, or RNA interference vector to knock out, mutate, or inhibit the expression of the OsPRMT10 protein-coding gene in rice, and selecting transgenic rice varieties with improved resistance to pests.

[0014] The pest described in the present application is preferably a lepidopteran pyralid insect, and further preferably, the lepidopteran pyralid insect includes a Cnaphalocrocis medinalis.

[0015] The mutation described in the present application includes substitution, deletion and / or addition of one or more nucleotides on the nucleotide sequence of the coding gene or the promoter of the OsPRMT10 protein; specifically, the mutation can be obtained by physical mutagenesis, chemical mutagenesis, gene editing. The physical mutagenesis includes but is not limited to radiation mutagenesis, space breeding, etc.; the method of chemical mutagenesis includes mutagenesis caused by treatment with EMS and the like; the method of gene editing includes but is not limited to ZFN, TALEN and / or CRISPR / Cas, etc.

[0016] The skilled person in the art can use conventional gene knockout, gene editing technology or RNA interference technology and the like to knockout or mutate the coding gene of the OsPRMT10 protein in rice; for example, design sgRNA with the coding gene of the OsPRMT10 protein as the target gene and construct a coding gene knockout vector of the OsPRMT10 protein or use gene editing technology to construct a CRISPR / Cas9 gene editing vector of the coding gene of the OsPRMT10 protein, etc., to knockout or mutate the coding gene of the OsPRMT10 protein in rice, and these methods are all familiar to the skilled person in the art; the skilled person in the art knows that the main principle of the CRISPR / Cas gene editing system or gene editing method is to find the position to be edited by a nucleic acid fragment called guide RNA (guide-RNA, gRNA) in the host genome, that is, the target DNA sequence, and then cut the DNA through Cas protein. In the present application, the Cas protein includes but is not limited to Cas9, Cas12, Cas12a, Cas12j, Cas12e, Cas13 and / or Cas14 protein, etc.

[0017] For reference, the present application provides a CRISPR / Cas9 gene editing vector of a OsPRMT10 gene, comprising: a Cas9 protein and a sgRNA targeting OsPRMT10 the gene; wherein the nucleotide sequence of the sgRNA targeting OsPRMT10 the gene is selected from the nucleotide sequence shown in SEQ ID No. 7 or / and SEQ ID No. 8.

[0018] As a preferred specific embodiment, the present application provides a CRISPR / Cas9 gene editing vector of a OsPRMT10The method for constructing the CRISPR / Cas9 gene editing vector of the gene comprises the following steps: connecting the nucleotide sequence shown in SEQ ID No. 7 or / and SEQ ID No. 8 to a pCBSG032 original vector to obtain OsPRMT10 The CRISPR / Cas9 gene editing vector of the gene.

[0019] The RNA interference technology is a routine technology in the art, which specifically binds to the homologous region of the mRNA expressed by the target gene through 21-23 bp small interfering RNA (siRNA) or double-strand RNA (dsRNA; double-strand RNA), degrades the mRNA, and achieves the effect of inhibiting gene expression; the normal expression or normal function of the coding gene or promoter of the OsPRMT10 protein can be interfered by the RNA interference technology (RNAi) to interfere with the normal expression of the coding gene or the promoter of the OsPRMT10 protein or to make the normal function thereof defective.

[0020] The transformation and the scheme for introducing the gene editing vector into the plant can be changed according to the type of the plant or plant cell to be transformed. Suitable methods for introducing the polynucleotide into the plant cell include: transforming the gene editing vector or the RNA interference vector into the plant cell or tissue by using a Ti plasmid, a Ri plasmid, a plant virus vector, direct DNA transformation, microinjection, electric conduction, Agrobacterium-mediated transformation, and other conventional biological methods, and then using conventional methods to regenerate the stably transformed plant from the transformed cell (McCormick et al. Plant Cell Reports. 1986. 5:81-84).

[0021] The present application effectively improves the resistance of rice to pest stress by reducing or inhibiting the expression amount or expression level of the coding gene of the OsPRMT10 protein in rice, or reducing the activity, losing the function or producing defects of the OsPRMT10 protein in rice; the present application has application prospects in improving the resistance of rice to pest stress and creating rice varieties resistant to pest stress.

[0022] Definitions of terms involved in the present invention

[0023] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs.

[0024] The terms "polynucleotide" or "nucleotide" mean deoxyribonucleotides, deoxyribonucleosides, ribonucleosides, or ribonucleotides and their polymers, in single-stranded or double-stranded form. Unless specifically limited, the term encompasses nucleic acids containing known analogs of natural nucleotides, which have binding properties similar to a reference nucleic acid and are metabolized in a manner similar to naturally occurring nucleotides. Unless otherwise specifically limited, the term also means oligonucleotide analogs, including PNAs (peptide nucleic acids), DNA analogs (phosphate thioesters, phosphoramidites, etc.) used in antisense techniques. Unless otherwise specified, a specific nucleic acid sequence also implicitly encompasses variants of its conserved modifications (including (but not limited to) degenerate codon substitutions) and complementary sequences, as well as explicitly specified sequences. Specifically, degenerate codon substitution can be achieved by generating a sequence in which the 3rd position of one or more selected (or all) codons is substituted with a mixed base and / or deoxyinosine residue.

[0025] The terms “polypeptide,” “peptide,” and “protein” are used interchangeably herein to refer to polymers of amino acid residues. That is, the description of a polypeptide is equally applicable to the description of a peptide and the description of a protein, and vice versa. The terms apply to naturally occurring amino acid polymers as well as amino acid polymers in which one or more amino acid residues are non-naturally encoded amino acids. As used herein, the terms cover amino acid chains of any length, including full-length proteins (i.e., antigens), wherein the amino acid residues are linked by covalent peptide bonds.

[0026] The term "conversion" refers to the method of introducing a heterologous DNA sequence into a host cell or organism.

[0027] The term "expression" refers to the transcription and / or translation of endogenous genes or transgenes in plant cells. Attached Figure Description

[0028] Figure 1 The image shows the recombinant vector pWMC025.

[0029] Figure 2 for OsPRMT10 Nucleotide sequence alignment results of the insect-sensitive material CH1057 and the knockout mutant.

[0030] Figure 3 The amino acid sequence alignment results of OsPRMT10 in the insect-sensitive material CH1057 and the knockout mutant are shown.

[0031] Figure 4 for OsPRMT10 Nucleotide sequence alignment results of insect-resistant material SE378 and knockout mutant.

[0032] Figure 5OsPRMT10 in insect-resistant material SE378 and knock-out mutant.

[0033] Figure 6 CH1057 and knock-out mutant.

[0034] Figure 7 CH1057 and knock-out mutant. DETAILED DESCRIPTION

[0035] The present application will be further described below in connection with specific experimental examples, and the advantages and features of the present application will become more apparent from the description. However, these experimental examples are only exemplary and do not constitute any limitation to the scope of the present application. It should be understood by those skilled in the art that modifications or substitutions can be made to the details and forms of the present application without departing from the spirit and scope of the present application, and such modifications and substitutions fall within the scope of the present application.

[0036] Biological material and data processing method

[0037] 1. Biological material

[0038] Rice P1790-5-1M-4-5M-1B-3M-B (CH1057) and ADNY 11::C1 (SE378) are disclosed in the document “Genomic variation in 3,010 diverse accessions of Asian cultivated rice”, Nature, 2018, 557: 43-49.”, and the above biological material can be obtained by the public from the applicant, and the above biological material is only used for repeating the experiments of the present application and cannot be used for other purposes.

[0039] 2. Data processing method

[0040] The following experimental examples use EXCEL statistical software to process data, and the experimental results are expressed as mean ± standard deviation, T test is used, n.s. indicates no significant difference, P < 0.05 (*) and P < 0.01 (**) indicate significant difference, and P < 0.001 (*** ) indicates extremely significant difference.

[0041] Experimental Example 1 OsPRMT10 Construction of gene editing knock-out vector and obtaining, identification and phenotype observation of transgenic plants

[0042] 1. Experimental method

[0043] 1.1 OsPRMT10Gene editing knockout target primer design and CRISPR / Cas9 vector construction

[0044] According to the principle of gene editing, through online Blast homology analysis, two specific segments of 20 bp in length in the full-length cDNA sequence were selected as target segments for gene editing knockout. OsPRMT10 Two target sites were designed for improving editing efficiency, wherein target site 1 was the DNA shown in SEQ ID No. 7, i.e. 5'- AGATGCTCTGCGACCGCGTCCGG -3' (SEQ ID No. 7), and target site 2 was the DNA shown in SEQ ID No. 8, i.e. 5'- CGCCCACCACTTCCGCGGCAAGG -3' (SEQ ID No. 8).

[0045] According to the target sequence, annealing primers were designed, DNA molecules containing the target sequence were synthesized by annealing, and were constructed into the pCBSG032 original vector to obtain the pWMC025 recombinant vector. The pWMC025 recombinant vector expresses Cas9 protein and sgRNA targeting the above-mentioned target sequence. The map of the pWMC025 recombinant vector is shown in Figure 1 The nucleotide sequence of the recombinant vector pWMC025 was obtained by sequentially connecting the two nucleotide sequences shown in SEQ ID No. 7 and SEQ ID No. 8.

[0046] 1.2 OsPRMT10 Obtaining of transgenic rice

[0047] The pWMC025 recombinant vector was transformed into Agrobacterium EHA105 by freeze-thaw method (Holsters et al., Molecular & General Genetics, 1978, 163(2): 181-7). The indica rice variety P1790-5-1M-4-5M-1B-3M-B (also referred to as "CH1057", which is referred to as CH1057 hereinafter) was transformed by Agrobacterium-mediated method. The mature seeds of CH1057 were mechanically dehulled, and the seeds with full and smooth and sterile spots were selected and inoculated on induction medium after disinfection for induction culture. The rice callus with good appearance and strong growth was selected as the receptor material, and the pWMC025 recombinant vector was transformed into the rice callus by Agrobacterium-mediated method. The callus was inoculated on the induction medium containing 100 μM acetosyringone and OD 600For Agrobacterium transformation with 0.3 - 0.5, the callus soaked in the transformation solution was placed on co-cultivation medium for co-cultivation, and co-cultivated at 28°C in the dark for 50-55 h. The callus with no obvious Agrobacterium on the surface was selected and transferred to N6 antibiotic medium containing 2.0 mg / L 2,4-D and 500 mg / L cefotaxime, and cultured at 28°C in the dark for 3-4 d. The callus was transferred to the screening medium and cultured for 30 d, and subcultured every 10 d. The callus with hygromycin resistance was transferred to pre-regeneration medium and cultured at 28°C in the dark for 7 d, then placed in a light culture room (12 h light / 12 h dark) for continuous culture for 7 d, and then transferred to regeneration medium (250 mL tissue culture flask), and continued to be cultured under light until the regenerated plantlets grew.

[0048] The medium formula involved in the present test example is shown in Table 1 below:

[0049] Table 1 Medium used for genetic transformation and its formula

[0050]

[0051] 1.3 OsPRMT10 - KO Identification of transgenic lines

[0052] To determine the T0 generation OsPRMT10 - KO In the positive transgenic lines OsPRMT10 Genomic changes, PCR amplification and sequencing of the sequences near the gene editing target site in the transgenic positive plants.

[0053] The gene editing target site specific primers are:

[0054] OsPRMT10-F: 5'-TGGACTTCGCCAACTACTTCTGC-3' (SEQ ID No. 9);

[0055] OsPRMT10-R: 5'-TCCACGGCGTACACCTTCCT-3' (SEQ ID No. 10).

[0056] 1.4 OsPRMT10 - KO Identification of rice leaf roller resistance of transgenic lines

[0057] The background material CH1057, 2 T2 generations OsPRMT10 - KO Transgenic rice and background material SE378, 1 T2 generation OsPRMT10 - KOThe transgenic rice seeds were sowed in the seedling tray with the soil drenched with fungicide, and then transplanted to the net house of Zhejiang Jinhua Agricultural Academy after about 25 days of cultivation in the greenhouse, with 1 row of each material and 10 plants in each row. At the tillering stage of the plants, the rice leaf roller adults were artificially introduced in excess to ensure that the susceptible control material TN11 was all damaged. After 1-2 months of introduction of the insects, the number of rolled leaves and the total number of leaves of each single plant were investigated, and the rolled leaf rate of each single plant was calculated:

[0058] Single plant rolled leaf rate = single plant rolled leaf number / single plant total leaf number x 100%.

[0059] 2 Test results

[0060] 2.1 OsPRMT10 - KO Identification of transgenic lines

[0061] Figure 2 For OsPRMT10 The nucleotide sequence alignment results of the susceptible material CH1057 and the knockout mutant OsPRMT10 - KO1 and OsPRMT10 KO2 - Figure 3 The amino acid sequence alignment results of OsPRMT10 in the susceptible material CH1057 and the knockout mutant OsPRMT10 - KO1 and OsPRMT10 - KO2 .

[0062] OsPRMT10 Homozygous mutant OsPRMT10-KO1 Compared with the background material CH1057, the genomic sequences in the two homologous chromosomes were changed as follows: in the genome of rice CH1057, 6 nucleotides from position 149 to position 154 of SEQ ID No. 2 were deleted, and a C was inserted after the 207th nucleotide of SEQ ID No. 2, OsPRMT10 A total of 5 nucleotide deletions caused a frameshift mutation, thereby knocking out the OsPRMT10 gene and causing premature termination of the OsPRMT10 protein. The amino acid sequence of OsPRMT10 in OsPRMT10 is shown in SEQ ID No. 11: OsPRMT10-KO1

[0063] MASLPNGAASASAASSAAGGGPAVVDKEVDFANYFCTYSYLYHQKEMLCVRMDAYHSAVFRNAHHFRRQGGS* (SEQ ID No. 11).

[0064] OsPRMT10 Homozygous mutant OsPRMT10 - KO2 ​Compared with the background material CH1057, the genomic sequence in the two homologous chromosomes has the following changes: in the genome of rice CH1057, 8 nucleotides from 147th to 154th in SEQ ID No. 2 in the sequence table are deleted, T is inserted after the 208th nucleotide shown in SEQ ID No. 2, OsPRMT10 a total of 7 nucleotide deletions generate a frameshift mutation, thereby knocking out the gene and causing the premature termination of the OsPRMT10 protein. OsPRMT10 OsPRMT10

[0065] The amino acid sequence of OsPRMT10 in OsPRMT10-KO2 is shown in SEQ ID No. 12:

[0066] MASLPNGAASASAASSAAGGGPAVVDKEVDFANYFCTYSYLYHQKEMLCPDGRLPLRRLPQRPPLPRARWFLMWVPGVASSPSGARRLAPGRCTPWRPPIWRSTRASSRGLTTSPTSSR* (SEQ ID No. 12).

[0067] Figure 4 The nucleotide sequence alignment results of the anti-insect material SE378 and the knockout mutant OsPRMT10 - OsPRMT10 KO3 The amino acid sequence alignment results of OsPRMT10 in the anti-insect material SE378 and the knockout mutant Figure 5 - OsPRMT10 KO3 According to the sequencing results, it can be seen that, the homozygous mutant

[0068] - OsPRMT10 Compared with the background material SE378, the genomic sequence in the two homologous chromosomes has the following changes: in the genome of rice SE378, 55 nucleotides from 154th to 208th in SEQ ID No. 5 in the sequence table are deleted, OsPRMT10 a total of 7 nucleotide deletions generate a frameshift mutation, thereby knocking out the gene and causing the premature termination of the OsPRMT10 protein. KO3 OsPRMT10 OsPRMT10 The amino acid sequence of OsPRMT10 in OsPRMT10 is shown in SEQ ID No. 13:

[0069] OsPRMT10-KO3

[0070] ​​​​​​​MASLPNGAASASAASSAAGGGPAVVDKEVDFANYFCTYSYLYHQKEMLCDRARWFLMWVPGVASSPSGARRLAPGRCTPWRPPIWRSTRASSRGLTTSPTSSR* (SEQ ID No. 13).

[0071] Sequencing results showed that three homozygous mutants targeting specific targets were obtained in the T0 generation of the transformants, namely mutants with a total deletion of 5 bp between the two target sites in the CH1057 background. OsPRMT10- KO 1 Homozygous mutant, with a total deletion of 7 bp at both target sites. OsPRMT10- KO 2 The homozygous mutant and the SE378 background showed a combined deletion of 55 bp at the two target sites. OsPRMT10- KO3 Homozygous mutants indicate a CH1057 background. OsPRMT10- KO1 , OsPRMT10- KO2 and SE378 background OsPRMT10- KO3 In OsPRMT10 The gene has been knocked out.

[0072] Will OsPRMT10 The homozygous mutants were transplanted to a greenhouse for cultivation. Seeds of the T1 generation homozygous mutants were obtained from individual plants, and then propagated to obtain homozygous T2 generation seeds, resulting in three T2 generation plants. OsPRMT10 - KO Genetically modified rice.

[0073] 2.2 OsPRMT10 KO Identification of rice leaf folder resistance in transgenic lines

[0074] After the rice leaf roller infestation, the average leaf rolling rate of 10 plants in the background material CH1057 was 94.3%. OsPRMT10- KO1 The average leaf curling rate of the 9 plants was 53.97%. OsPRMT10-KO2 The average leaf curling rate of the 10 plants was 54.5%, and 2 samples... OsPRMT10 - KO The leaf curling rate of the transgenic lines was significantly lower than that of the background material CH1057. Figure 6 The average leaf curling rate of SE37810 plants was 71.75%. OsPRMT10-KO 3 The average leaf curling rate of the 10 plants was 32.86%. OsPRMT10-KO3 The leaf curling rate of the strain was significantly lower than that of the background material SE378. Figure 7 ).

[0075] Phenotypic survey results indicate that: OsPRMT10 - KO The transgenic lines showed significantly enhanced resistance to rice leaf folder compared to the background materials CH1057 and SE378, demonstrating that transgenic rice lines...OsPRMT10 Knocking out, mutating or suppressing the expression of the genes can effectively increase the resistance of rice to Cnaphalocrocis medinalis.

Claims

1. Use of an OsPRMT10 protein or a coding gene thereof in enhancing the resistance of rice to Cnaphalocrocis medinalis.

2. Use according to claim 1, characterized in that, The expression amount or expression level of the coding gene of the OsPRMT10 protein in rice is reduced or inhibited, or the activity of the OsPRMT10 protein in rice is reduced, lost or defective.

3. Use according to claim 1 or 2, characterized in that, The amino acid sequence of the OsPRMT10 protein is represented by SEQ ID No. 1 or SEQ ID No. 4; the nucleotide sequence of the coding gene of the OsPRMT10 protein is represented by SEQ ID No. 2 or SEQ ID No. 5; wherein the nucleotide sequence of the CDS of the coding gene represented by SEQ ID No. 2 is represented by SEQ ID No. 3, and the nucleotide sequence of the CDS of the coding gene represented by SEQ ID No. 5 is represented by SEQ ID No.

6.

4. A method of increasing resistance to pest stress in rice, comprising: The coding gene of the OsPRMT10 protein is knocked out or mutated, the expression amount or expression level of the coding gene of the OsPRMT10 protein in rice is reduced, inhibited or interfered, the activity of the OsPRMT10 protein in rice is reduced, lost or defective; and the pest is Cnaphalocrocis medinalis.

5. A method of creating a rice variety that is resistant to a pest stress comprising: A gene knockout vector of the coding gene of the OsPRMT10 protein is constructed, or a CRISPR / Cas9 gene editing vector of the coding gene of the OsPRMT10 protein is constructed by using a gene editing technology, or an RNA interference vector of the coding gene of the OsPRMT10 protein is constructed; the gene knockout vector, the CRISPR / Cas9 gene editing vector or the RNA interference vector is transformed into rice, the coding gene of the OsPRMT10 protein in the rice is knocked out, mutated or inhibited, and a transgenic plant rice variety with improved resistance to pests is selected and bred; and the pest is Cnaphalocrocis medinalis.

6. The method according to claim 4 or 5, characterized in that, The amino acid sequence of the OsPRMT10 protein is represented by SEQ ID No. 1 or SEQ ID No. 4; the nucleotide sequence of the coding gene of the OsPRMT10 protein is represented by SEQ ID No. 2 or SEQ ID No. 5; wherein the nucleotide sequence of the CDS of the coding gene represented by SEQ ID No. 2 is represented by SEQ ID No. 3, and the nucleotide sequence of the CDS of the coding gene represented by SEQ ID No. 5 is represented by SEQ ID No.

6.

7. OsPRMT10 Use of CRISPR / Cas9 gene editing vector of a gene in enhancing resistance of rice against a pest; the pest is a rice leaf roller.

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