Rice blast and bacterial blight disease resistance related protein derived from rice and encoding gene and application thereof

Abstract

The present application discloses a kind of Magnaporthe grisea and Xanthomonas oryzae resistance related protein derived from rice and its coding gene and application.The present application provides a kind of method for improving the disease resistance of plant: the gene of plant coding OsAspRS protein is replaced by the gene of mutant protein.The present application also protects the application of osasprs mutant in the preparation of plant with improved disease resistance.osasprs mutant is a kind of spot-like mutant plant controlled by single recessive nuclear gene, which is screened by EMS on Huanghuazhan plant.Furthermore, the present application successfully locates and clones AspRS gene, and the mutation of the gene site can cause spot-like phenotype on rice leaf, and further improve the broad-spectrum disease resistance of rice.The present application is helpful to improve the disease resistance of crop, provides gene resources and technical support for cultivating new rice varieties with broad-spectrum resistance, and has important significance and application value for crop disease resistance breeding work.

Description

Technical Field

[0001] This invention belongs to the field of biotechnology, specifically relating to a disease resistance-related protein derived from rice blast fungus and bacterial blight fungus, its encoding gene, and its application. Background Technology

[0002] Plant lesion-like mutants refer to cells that spontaneously develop a hypersensitive reaction-like cell death in plants without any pathogen infection or stress. Most mutants exhibiting this hypersensitive reaction form spots on plant leaves or leaf sheaths. Because these spots resemble plant lesions, these mutants are named lesion-like mutants. Extensive research has found that these mutants are widely present in plants such as Arabidopsis thaliana, wheat, maize, and rice. In lesion-like mutants, the production of reactive oxygen species and the accumulation of cellular secondary metabolites enable plant cells to exhibit disease resistance while simultaneously causing cytotoxicity and cell death, thus limiting further infection and spread of pathogens. Over the past 30 years, numerous mutant genes controlling plant lesions have been cloned and isolated. These genes encode proteins with different functions, participating in various regulatory or metabolic pathways, such as photosynthesis, fatty acid metabolism, and signal transduction.

[0003] A common characteristic of lesion-like mutants is that they all exhibit varying degrees of increased resistance to pathogens, displaying histological and cytological features specific to disease resistance responses. As early as 1995, it was proposed that resistance-related genes might be involved in the formation of plant pseudolesions. The causes of lesion-like traits are complex, and current research on the signaling pathways and regulatory processes involved in programmed cell death and hypersensitivity responses is still very limited. Understanding the intrinsic relationship between lesion-like phenotype-controlling genes and plant immunity, besides utilizing mutant genes, can also involve identifying downstream disease resistance-related genes. Through transgenic or gene-editing technologies, transgenic disease-resistant plants or mutant plants can be obtained, which is of great significance for disease resistance breeding.

[0004] Rice, as one of the most important food crops, provides food for more than half of the world's population. Rice sheath blight, rice blast, sesame leaf spot, and bacterial blight are some of the major diseases causing significant yield losses in rice. These diseases not only cause large-scale yield reductions but also damage the nutritional quality of rice. Therefore, continuously improving rice yield and quality and effectively controlling rice diseases are of great significance for ensuring global food security and sustainable agricultural development. Currently, the main methods for dealing with crop diseases are breeding disease-resistant varieties and using chemical pesticides. However, while traditional chemical pesticides can quickly eliminate pests and diseases, long-term use may lead to resistance, and there are also problems with overuse polluting agricultural products and the ecological environment. The "greening" of the entire process of controlling major agricultural and forestry pests and diseases, and the development of sustainable green agriculture, have become the guiding principles for agricultural development now and in the future. Utilizing plant resistance genes to mediate resistance in breeding is an effective green control strategy for diseases. However, breeding crop varieties with broad-spectrum and durable resistance is challenging. Due to increased species exchange, the specialized resistance of pathogen races is easily overcome by pathogens, and the resistance produced by a single resistance gene is quickly lost in the field. Therefore, further exploration and identification of usable resistance genes and analysis of resistance signal transduction networks are not only of key theoretical guiding significance for disease resistance breeding, but also provide potential new resources for disease resistance breeding. Summary of the Invention

[0005] The purpose of this invention is to provide a disease resistance-related protein derived from rice blast fungus and bacterial blight fungus, its encoding gene, and its application.

[0006] This invention provides a method for improving the disease resistance of plants, comprising the following steps: replacing the gene encoding the OsAspRS protein in the plant with a gene encoding a mutant protein.

[0007] This invention also protects the application of the OsAspRS protein in regulating plant disease resistance.

[0008] The regulation is negative, meaning that an increase in OsAspRS protein reduces disease resistance.

[0009] This invention also protects the use of mutant proteins in regulating plant disease resistance.

[0010] The regulation is positive regulation, meaning that an increase in mutant proteins leads to increased disease resistance.

[0011] This invention also protects the use of the OsAspRS gene in the cultivation of plants with altered disease resistance.

[0012] In this application, overexpression of the OsAspRS gene reduces the plant's disease resistance.

[0013] In this application, overexpression of the OsAspRS gene is specifically achieved by importing the recombinant plasmid shown in sequence 4 of the sequence listing.

[0014] This invention also protects the use of genes encoding mutant proteins in the cultivation of plants with altered disease resistance.

[0015] In this application, overexpression of the gene encoding the mutant protein enhances the plant's disease resistance.

[0016] This invention also protects the OsAspRS protein.

[0017] This invention also protects the OsAspRS gene.

[0018] The OsAspRS gene is the gene that encodes the OsAspRS protein.

[0019] This invention also protects mutant proteins.

[0020] This invention also protects genes encoding mutant proteins.

[0021] The OsAspRS protein mentioned above is (a1) or (a2) or (a3) ​​or (a4) as follows:

[0022] (a1) The protein shown in sequence 1 of the sequence listing;

[0023] (a2) A fusion protein obtained by attaching a tag to the N-terminus and / or C-terminus of the protein described in (a1);

[0024] (a3) Proteins related to plant disease resistance obtained by substituting and / or deleting and / or adding one or more amino acid residues of (a1);

[0025] (a4) is a protein derived from rice that shares more than 98% identity with (a1) and is associated with plant disease resistance.

[0026] The mutant protein mentioned above is a protein obtained by mutating the 276th amino acid residue of the OsAspRS protein from Ser to Asn.

[0027] The OsAspRS gene mentioned above is either (b1) or (b2) or (b3) or (b4) as follows:

[0028] (b1) A DNA molecule with a coding region as shown in Sequence 2 of the sequence listing;

[0029] (b2) The DNA molecule shown in sequence 3 of the sequence listing;

[0030] (b3) A DNA molecule derived from rice and having more than 95% identity with (b1) or (b2) and encoding the protein thereon;

[0031] (b4) A DNA molecule that hybridizes under stringent conditions to a nucleotide sequence defined in (b1) or (b2) and encodes the protein.

[0032] The above stringent conditions can be achieved by hybridization at 65°C using a solution of 6×SSC and 0.5% SDS, followed by washing the membrane once each with 2×SSC and 0.1% SDS and 1×SSC and 0.1% SDS.

[0033] The gene encoding the mutant protein described above is a DNA molecule obtained by mutating the codon encoding the 276th amino acid residue in the OsAspRS gene from the codon encoding Ser to the codon encoding Asn.

[0034] The specific codon for encoding Ser can be AGC.

[0035] The specific codon encoding Asn can be AAC.

[0036] The gene encoding the mutant protein described above can be a DNA molecule obtained by mutating nucleotide 827 of the DNA molecule shown in Sequence 2 of the sequence listing from G to A.

[0037] The gene encoding the mutant protein described above can specifically be a DNA molecule obtained by mutating the 2051st nucleotide of the DNA molecule shown in Sequence 3 of the sequence listing from G to A.

[0038] This invention also protects the use of the osasprs mutant in the preparation of plants with enhanced disease resistance.

[0039] In this application, the osasprs mutant serves as the disease-resistant parent.

[0040] The present invention also provides a method for preparing plants with enhanced disease resistance, comprising the following steps: hybridizing the osasprs mutant with other plants, and obtaining plants with enhanced disease resistance from the hybrid offspring.

[0041] In the method described, the osasprs mutant serves as the resistant parent.

[0042] In the method described, other plants serve as susceptible parents.

[0043] In the method, plants with leaf spot-like phenotypes are screened from the hybrid offspring.

[0044] The improved disease resistance refers to the relative improvement in disease resistance compared to the susceptible parent.

[0045] The disease resistance mentioned above can specifically refer to resistance to rice blast and / or resistance to bacterial blight.

[0046] The disease resistance mentioned above refers to resistance to plant pathogens.

[0047] The plant pathogens mentioned above may specifically be rice blast fungus and / or bacterial blight fungus.

[0048] For example, the rice blast fungus is rice blast fungus strain Guy11.

[0049] For example, the bacterial blight pathogen is rice bacterial blight pathogen.

[0050] For example, the bacterial blight pathogen is the pathogenic race PXO99 of rice bacterial blight pathogen.

[0051] Any of the plants mentioned above may be monocotyledonous or dicotyledonous.

[0052] Any of the plants mentioned above can be plants of the Poaceae family.

[0053] Any of the plants mentioned above may be plants of the genus *Rice*.

[0054] Any of the plants mentioned above can be indica rice.

[0055] Any of the plants mentioned above could be *Huang Huazhan*.

[0056] Any of the plants mentioned above can be osasprs, a lesion-like mutant of Huanghuazhan indica rice.

[0057] The lesion-like mutant *osasprs* of the indica rice variety *Huanghuazhan* was deposited on August 17, 2021, at the China General Microbiological Culture Collection Center (CGMCC, address: No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing, Institute of Microbiology, Chinese Academy of Sciences), with accession number CGMCC No. 23084. The lesion-like mutant *osasprs* of *Huanghuazhan* is abbreviated as the *osasprs* mutant.

[0058] The inventors of this invention used EMS mutagenesis on Huang Huazhan rice plants to screen for a lesion-like mutant controlled by a single recessive nuclear gene. Further, the inventors performed phenotypic identification, genetic analysis, and genetic background identification on this mutant plant. Using the SIMM method, PCR technology, and gene information analysis, the inventors successfully located and cloned the AspRS gene, located on chromosome 2 at locus LOC_Os02g46130. Mutations at this gene locus can lead to a lesion-like phenotype in rice leaves, thereby improving the broad-spectrum disease resistance of rice. This invention helps improve crop disease resistance, provides genetic resources and technical support for breeding new rice varieties with broad-spectrum resistance, and has significant meaning and application value for crop disease resistance breeding. Attached Figure Description

[0059] Figure 1 This is a phenotypic comparison of the leaves of the Huang Huazhan plant and the osasprs mutant plant in Example 1.

[0060] Figure 2 This is a result diagram of Example 3.

[0061] Figure 3 This is a result diagram of Example 4.

[0062] Figure 4 This is a result diagram of Example 5.

[0063] Figure 5 The result diagram is shown in Example 6.

[0064] Figure 6 The images show leaf photographs of Huang Huazhan's plant, the osasprs mutant plant, and the AspRS gene-reinforced plant in Example 7, as well as sequencing data near the mutation site in the OsAspRS gene.

[0065] Figure 7 This is a graph showing the disease resistance results of Example 7. Detailed Implementation

[0066] The present invention will now be described in further detail with reference to specific embodiments. The given embodiments are merely illustrative of the invention and not intended to limit its scope. The embodiments provided below can serve as a guide for further improvements by those skilled in the art and do not constitute a limitation on the invention in any way.

[0067] Unless otherwise specified, the experimental methods used in the following examples are conventional methods, performed according to the techniques or conditions described in the literature in this field or according to the product instructions. Unless otherwise specified, the materials and reagents used in the following examples are commercially available. Huang Huazhan, an existing indica rice variety, is denoted as HHZ.

[0068] Unless otherwise specified, the quantitative experiments in the following examples are all repeated three times, and the results are averaged.

[0069] The following examples are provided to better understand the present invention, but do not limit the invention. Unless otherwise specified, the experimental methods in the following examples are conventional methods. Unless otherwise specified, the experimental materials used in the following examples were purchased from conventional biochemical reagent stores. Unless otherwise specified, the quantitative experiments in the following examples were performed in triplicate, and the results were averaged.

[0070] Example 1: Obtaining rice lesion mutants

[0071] Huanghuazhan seeds (F0 generation seeds) were soaked in a 0.7% (w / v) EMS aqueous solution and subjected to mutagenesis treatment for 12 hours. The seeds were then multiplied and harvested to obtain F1 generation seeds. The F1 generation seeds were then cultivated into plants, which are the F1 generation plants. The leaf phenotype of the mature F1 generation plants was observed, and a plant with a lesion-like phenotype was obtained from the F1 generation plants (named the lesion-like mutant osasprs of Huanghuazhan rice, denoted as osasprs).

[0072] The lesion-like mutant *osasprs* of the indica rice variety *Huanghuazhan* was deposited on August 17, 2021, at the China General Microbiological Culture Collection Center (CGMCC, address: No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing, Institute of Microbiology, Chinese Academy of Sciences), with accession number CGMCC No. 23084. The lesion-like mutant *osasprs* of *Huanghuazhan* is abbreviated as the *osasprs* mutant.

[0073] A phenotypic comparison of the leaves of Huang Huazhan plants and the leaves of the osasprs mutant plants is shown in [the original text]. Figure 1 .

[0074] Example 2: Genetic analysis of the osasprs mutant

[0075] The osasprs mutant plants were backcrossed with Huang Huazhan plants to obtain the F1 generation backcross. The leaf phenotype of the F1 plants was observed. Seeds from the F1 plants were planted to obtain the F2 generation segregating population. The segregation ratio of normal-leaved plants to disease-spotted-leaved plants in the F2 generation population was 3:1 (χ²). 2 3:1 <χ 2 0.05 =3.84). The results indicate that the lesion-like phenotype of the osasprs mutant is caused by a recessive nuclear gene mutation, and the osasprs mutant is a homozygous mutant of the mutant gene.

[0076] Example 3: Cloning the mutant gene from the osasprs mutant

[0077] The mutant gene was cloned using the Simultaneous Identification of Multiple Mutations (SIMM) pipeline method. The specific steps were as follows: the osasprs mutant was backcrossed with Huanghuazhan plants. Thirty plants with a lesion-like phenotype were selected from the F2 population. Genomic DNA was extracted from the leaves of each plant and mixed in equal amounts, followed by library construction and resequencing. The sequencing data were analyzed using the SIMM method to locate the mutation site, ultimately pinpointing it to rice chromosome 2. In the F2 segregating population, the homozygous mutation of the LOC_Os02g46130 gene was linked to the lesion-like phenotype. In the osasprs mutant, the protein mutation occurred at amino acid position 276, with the amino acid Ser (corresponding codon AGC) changing to Asn (corresponding codon AAC).

[0078] In Huang Huazhan's genome, the LOC_Os02g46130 gene encodes an aspartic-tRNA synthetase (OsAspRS protein), as shown in Sequence 1 of the sequence listing. The gene encoding the OsAspRS protein is the OsAspRS gene. In CDS, the OsAspRS gene is shown in Sequence 2 of the sequence listing. In genomic DNA, the OsAspRS gene is shown in Sequence 3 of the sequence listing.

[0079] OsAspRS gene location (A), schematic diagram of OsAspRS gene structure (B), schematic diagram of OsAspRS protein structure (C), sequencing data near the mutation site in the OsAspRS gene of osasprs mutant plants and Huang Huazhan plants (D) Figure 2 .

[0080] Example 4: Trypan blue staining, DAB staining, and nitroblue tetrazolium (NBT) staining

[0081] The test subjects were the leaves of the tested plants. The tested plants were either osasprs mutant plants or Huanghuazhan plants.

[0082] I. Trypan Blue Staining

[0083] Trypan blue is a cell-active dye that can penetrate the cell membrane of dead cells, thus staining them blue, while live cells are not stained blue. Trypan blue staining solution contains 0.25% trypan blue, 23% water-soluble phenol, 25% (w / v) lactic acid, and 25% glycerol, diluted to volume with ddH2O.

[0084] Specific procedures: 1. Use a trypan blue staining solution in a 70°C water bath; 2. Immerse the leaf in the trypan blue staining solution, vacuum for 15 minutes, then boil in a water bath for 2 minutes, and remove and cool; 3. Place the leaf in a test tube containing 2.5 g / ml chloral hydrate for decolorization; 4. After decolorization, the leaf is basically transparent and can be photographed.

[0085] See photos Figure 3 The lesion-like areas on the leaves of the A. osasprs mutant were stained dark blue.

[0086] II. DAB staining

[0087] H2O2 can react with diaminobenzidine (DAB) under the catalysis of peroxidase to produce a reddish-brown substance. Therefore, DAB is used to detect the H2O2 content in leaves.

[0088] The leaves were immersed in 1 mg / ml DAB staining solution (pH 3.8) and vacuumed for 15 min. Then, the leaves were completely submerged in the DAB staining solution and placed under light at room temperature for 12 h. After that, the leaves were removed, destained with 95% ethanol, and photographed after complete decolorization.

[0089] See photos Figure 3 The leaves of the B. osasprs mutant showed numerous brown spots at the sites of lesion formation, while those of the Huanghuazhan plant showed almost none. This indicates that the osasprs mutant leaves accumulate more H2O2.

[0090] III. NBT staining

[0091] NBT staining is used to detect superoxide anion free radicals (O2) in cells. - An effective method for measuring O2 content; the deeper the blue color, the more O2 it indicates. - The higher the content, the lower the O2 content. - The lower the concentration, the better. NBT staining solution: Dissolve NBT dry powder in PBS buffer (pH 7.6, 10mM) to a concentration of 0.5mg / ml, and prepare fresh before use.

[0092] Specific operating procedure: Immerse the leaves completely in NBT staining solution, vacuum for 10-15 minutes, and stain in a sealed container at 28°C in the dark for 3 hours; then discard the staining solution, decolorize with 90% ethanol, and take photos.

[0093] See photos Figure 3 O2 in the leaves of the C. osasprs mutant - It has a higher content.

[0094] Example 5: Disease Resistance Test

[0095] The tested plants were either osasprs mutant plants or Huanghuazhan plants.

[0096] I. Detection of rice blast resistance

[0097] 1. Preparation of rice blast fungal inoculum.

[0098] Spores of the rice blast fungus strain Guy11 were suspended in distilled water containing 0.01% (v / v) Tween-20 to obtain a concentration of 2 × 10⁻⁶. 5 A spore suspension with 1 spore per ml is called rice blast fungus spore solution.

[0099] 2. Detection of rice blast resistance

[0100] Test plants were sown and cultured in a greenhouse for 60 days. A potential circular wound was artificially created on the second-to-last leaf from the top using a punch. Then, 10 μl of rice blast fungus spore liquid was dropped onto the wound and sealed with transparent tape. The plants were then transferred to an environment of 28°C and ≥80% humidity for further culture. Fourteen days after inoculation, the inoculated leaves were photographed and the length of the lesions was measured. The relative biomass of rice blast fungus was also determined.

[0101] Method for detecting the relative biomass of *Magnaporum oryzae*: Total DNA was extracted from diseased portions of inoculated leaves. The abundance ratio of the *Magnaporum oryzae* MoPot2 gene to the rice OsUBQ gene was determined using qRT-PCR and used as the relative biomass of *Magnaporum oryzae*. Primers used for detecting the MoPot2 gene: 5'-ACGACCCGTCTTTACTTATTTGG-3'; 5'-AAGTAGCGTTGGTTTTGTTGGAT-3'. Primers used for detecting the OsUBQ gene: 5'-AACCAGCTGAGGCCCAAGA-3'; 5'-ACGATTGATTTAACCAGTCCATGA-3'.

[0102] See photos of the inoculated leaves Figure 4 A.

[0103] lesion length see Figure 4 B (25 plants, data in mean ± standard deviation).

[0104] The results of the relative biomass of rice blast fungus are shown in [the table below]. Figure 4 C (25 plants, data in mean ± standard deviation).

[0105] Compared with Huang Huazhan plants, the osasprs mutant showed significantly reduced leaf lesion length and a significantly lower relative biomass of rice blast fungus at the lesion sites. These results indicate that the osasprs mutant is resistant to rice blast fungus.

[0106] II. Detection of rice bacterial leaf blight resistance

[0107] 1. Preparation of bacterial suspension of white leaf blight pathogen.

[0108] The pathogenic race PXO99 of rice bacterial blight pathogen was collected, suspended in sterile double-distilled water, and the OD was obtained. 600nm A bacterial solution with a value of 0.5 is a bacterial solution of bacterial blight pathogen.

[0109] 2. Detection of resistance to bacterial leaf blight in rice

[0110] Eight weeks after sowing, the tips of the second-to-last leaves from the top of the test plants were cut off using scissors dipped in bacterial blight solution. The plants were then cultured normally. Fourteen days after inoculation, the leaf lesion phenotype was observed and the lesion length was measured.

[0111] See leaf photos Figure 4 D.

[0112] lesion length see Figure 4 E (30 plants, data in mean ± standard deviation).

[0113] Compared with the Huanghuazhan plant, the osasprs mutant showed significantly less leaf wilting, with lesion length approximately 1 / 8 that of the Huanghuazhan plant leaves. The results indicate that the osasprs mutant is resistant to bacterial blight.

[0114] Example 6: Detection of Rice Resistance Gene Expression Levels

[0115] The tested plants were either osasprs mutant plants or Huanghuazhan plants.

[0116] 1. Take leaves from the test plants and extract total RNA.

[0117] 2. Take the total RNA obtained in step 1 and reverse transcribe it to obtain cDNA.

[0118] 3. Using the cDNA obtained in step 2 as a template, perform real-time quantitative PCR to detect the expression level of disease resistance-related genes.

[0119] The disease resistance genes are: osPR1a gene, osPR1b gene, osPR5 gene, osPBZ1 gene, osNH1 gene, and osAOS2 gene.

[0120] See results Figure 5 Compared with Huang Huazhan plants, the expression levels of resistance-related genes in the osasprs mutant were significantly increased, by approximately 40, 20, 4, 600, 5, and 980 times, respectively.

[0121] Example 7: Obtaining and identifying AspRS gene replacement plants

[0122] I. Preparation of recombinant plasmids

[0123] A recombinant plasmid was prepared to complement the OsAspRS gene. The recombinant plasmid was a double-stranded circular DNA molecule as shown in Sequence 4 of the sequence listing. In Sequence 4 of the sequence listing, nucleotides 8923-9757 constitute the promoter, and nucleotides 9774-11420 constitute the OsAspRS gene.

[0124] II. Preparation of AspRS gene replacement plants

[0125] 1. The recombinant plasmid obtained in step 1 was introduced into Agrobacterium EHA105 to obtain recombinant Agrobacterium.

[0126] 2. The recombinant Agrobacterium obtained in step 1 was used to infect the immature embryo callus tissue of the osasprs mutant using the Agrobacterium infection method. Then, hygromycin screening (screening concentration of 25 mg / L), differentiation culture and rooting culture were carried out in sequence to obtain regenerated plants.

[0127] 3. Plant the regenerated plants in the field, observe the leaf phenotype, and select plants with the same leaf phenotype as Huang Huazhan plants.

[0128] 4. The plants obtained in step 3 were sequenced with the OsAspRS gene as the target gene. The sequencing results showed that plants with OsAspRS gene complementation were obtained, referred to as AspRS gene complementation plants, denoted as OsAspRs-COM. The sequencing results showed that the AspRS gene complementation plants simultaneously possessed the OsAspRS gene and the mutant gene (the OsAspRS gene is shown in sequence 2 of the sequence listing, and the mutant gene is a DNA molecule obtained by mutating the codon encoding the 276th amino acid residue in the OsAspRS gene from AGC encoding Ser to AAC encoding Asn).

[0129] Leaf photographs of Huang Huazhan plants, osasprs mutant plants, and AspRS gene-reinforced plants cultured under parallel conditions are shown below. Figure 6 The leaves of the A. supplemented plants partially restored the leaf phenotype of the wild-type Huang Huazhan plant. Sequencing images of the OsAspRS gene near the mutation site in Huang Huazhan plants, osasprs mutant plants, and AspRS gene supplemented plants are shown in [the image]. Figure 6 B.

[0130] III. Detection of Rice Resistance

[0131] Test plants: Huang Huazhan plants, osasprs mutant plants, or AspRS gene-reinforced plants.

[0132] Detection of resistance to bacterial blight in rice

[0133] The method is the same as step two of Example 5.

[0134] See leaf photos Figure 7 A.

[0135] lesion length see Figure 7 B (data in the form of mean ± standard deviation, n≥8).

[0136] The results showed that, compared with the osasprs mutant, the leaves of the AspRS gene-replenished plants exhibited significantly greater wilting and lesion length, approximately twice that of the osasprs mutant. The susceptibility of the AspRS gene-replenished plants to the pathogen was restored to half the level of the Huanghuazhan plants.

[0137] The present invention has been described in detail above. For those skilled in the art, the invention can be practiced in a wide range of ways with equivalent parameters, concentrations, and conditions without departing from its spirit and scope, and without requiring unnecessary experiments. Although specific embodiments have been given, it should be understood that further modifications can be made to the invention. In summary, according to the principles of the invention, this application is intended to include any changes, uses, or improvements to the invention, including changes made using conventional techniques known in the art that depart from the scope disclosed herein. Some of the essential features can be applied within the scope of the following appended claims. sequence list <110> Institute of Microbiology, Chinese Academy of Sciences <120> Disease resistance-related proteins of rice blast fungus and bacterial blight fungus, their encoding genes, and applications <130> GNCYX212623 <160> 4 <170> SIPOSequenceListing 1.0 <210> 1 <211> 549 <212> PRT <213> Oryza sativa <400> 1 Met Ser Ser Glu Pro Pro Pro Asp Ala Ala Ala Ala Ala Ala Ser Ser 1 5 10 15 Ala Gly Asp Leu Ala Ala Asp Leu Ser Ser Ala Thr Ile Ser Lys Lys 20 25 30 Gln Leu Lys Lys Asp Ala Arg Lys Ala Glu Lys Ala Glu Lys Ala Ser 35 40 45 Gln Arg Gln Gln Gln Gln Gln Pro Gln Ala Asp Ala Asp Asp Pro Phe 50 55 60 Ala Ala Asn Tyr Gly Asp Val Pro Val Glu Glu Ile Gln Ser Lys Thr 65 70 75 80 Ile Ser Gly Arg Val Trp Thr Glu Ile Gly Gly Leu Asp Glu Ala Ala 85 90 95 Ala Gly Arg Ser Val Leu Ile Arg Gly Ala Ala Gln Ala Ile Arg Pro 100 105 110 Val Ser Lys Lys Met Ala Phe Val Val Leu Arg Glu Ser Met Ser Thr 115 120 125 Val Gln Cys Val Leu Val Ala Ser Ala Asp Ala Gly Val Ser Thr Gln 130 135 140 Met Val Arg Phe Ala Thr Ser Leu Ser Lys Glu Ser Ile Val Asp Val 145 150 155 160 Glu Gly Val Val Ser Leu Pro Lys Glu Pro Leu Lys Ala Thr Thr Gln 165 170 175 Gln Val Glu Ile Gln Val Arg Lys Ile Tyr Cys Ile Asn Arg Ala Ile 180 185 190 Pro Thr Leu Pro Ile Asn Leu Glu Asp Ala Ser Arg Ser Glu Ala Glu 195 200 205 Ile Glu Lys Ala Glu Gln Ala Gly Glu Lys Leu Val Arg Val Gly Gln 210 215 220 Asp Thr Arg Leu Asn Tyr Arg Ala Ile Asp Leu Arg Thr Pro Ala Asn 225 230 235 240 Gln Ala Ile Phe Arg Ile Gln Cys Gln Val Glu Asn Lys Phe Arg Glu 245 250 255 Tyr Phe Leu Ser Lys Asn Phe Val Gly Ile His Ser Pro Lys Leu Ile 260 265 270 Ala Gly Ser Ser Glu Gly Gly Ala Ala Val Phe Lys Leu Gln Tyr Asn 275 280 285 Gly Gln Pro Ala Cys Leu Ala Gln Ser Pro Gln Leu Tyr Lys Gln Met 290 295 300 Ala Ile Cys Gly Gly Phe Glu Arg Val Phe Glu Val Gly Pro Val Phe 305 310 315 320 Arg Ala Glu Asn Ser Asn Thr His Arg His Leu Cys Glu Phe Val Gly 325 330 335 Leu Asp Ala Glu Met Glu Ile Lys Glu His Tyr Phe Glu Val Cys Asp 340 345 350 Ile Ile Asp Gly Leu Phe Val Ala Ile Phe Lys His Leu Asn Glu Asn 355 360 365 Cys Lys Lys Glu Leu Glu Thr Ile Asn Arg Gln Tyr Pro Phe Glu Pro 370 375 380 Leu Lys Tyr Leu Glu Lys Thr Leu Lys Leu Thr Tyr Glu Glu Gly Ile 385 390 395 400 Gln Met Leu Lys Glu Ala Gly Thr Glu Ile Glu Pro Met Gly Asp Leu 405 410 415 Asn Thr Glu Ala Glu Lys Lys Leu Gly Arg Leu Val Lys Glu Lys Tyr 420 425 430 Gly Thr Glu Phe Phe Ile Leu Tyr Arg Tyr Pro Leu Ala Val Arg Pro 435 440 445 Phe Tyr Thr Met Pro Cys Tyr Asp Asn Pro Ala Tyr Ser Asn Ser Phe 450 455 460 Asp Val Phe Ile Arg Gly Glu Glu Ile Ile Ser Gly Ala Gln Arg Ile 465 470 475 480 His Leu Pro Glu Leu Leu Thr Lys Arg Ala Thr Glu Cys Gly Ile Asp 485 490 495 Ala Ser Thr Ile Ser Ser Tyr Ile Glu Ser Phe Ser Tyr Gly Ala Pro 500 505 510 Pro His Gly Gly Phe Gly Val Gly Leu Glu Arg Val Val Met Leu Phe 515 520 525 Cys Ala Leu Asn Asn Ile Arg Lys Thr Ser Leu Phe Pro Arg Asp Pro 530 535 540 Gln Arg Leu Val Pro 545 <210> 2 <211> 1650 <212> DNA <213> Oryza sativa <400> 2 atgtcgtcgg agcctccacc cgatgccgcc gccgccgccg cctcctccgc gggggatctc 60 gcggccgatc tctcctccgc caccatcagc aagaagcagc tcaagaagga tgcgaggaag 120 gcggagaagg ccgagaaggc gtcgcagcgc cagcagcagc agcagccgca ggccgacgcc 180 gacgaccccct tcgcggccaa ctacggcgac gtccccgtcg aggagatcca gtccaagacc 240 atctccggcc gcgtgtggac cgagatcggc ggcctcgacg aggccgccgc cggccgctcc 300 gtcctcatcc gcggcgccgc gcaggcgatc cggcccgtca gcaagaagat ggccttcgtt 360 gtgctgcgcg agagcatgag caccgtccag tgcgtgctcg tcgccagcgc cgacgcaggg 420 gtcagcaccc agatggtgcg cttcgccacc tccctcagca aggaatcaat cgtcgacgtc 480 gagggcgtcg tctccctccc caaggagccc ctcaaggcca ccacgcagca ggtggagatt 540 caggtgagga agatctattg catcaatagg gcgatcccca cccttccaat caaccttgag 600 gatgcctcac ggagtgaggc tgaaattgaa aaggctgaac aagctggaga gaagctagtt 660 cgtgtgggcc aagatactcg cttgaactat agagctattg atctccggac acctgcaaat 720 caagcaatat tcagaattca atgtcaagtt gagaacaaat tcagggaata tttctgtcg 780 aaaaattttg ttgggattca cagcccaaag ctaattgctg gatccagcga aggtggtgca 840 gctctattca agctacagta caatggacag ccagcatgtt tagcacagtc tcctcagttg 900 tataagcaga tggctatttg tggtgggttt gaacgtgtat ttgaggttgg acctgttttt 960 agagctgaga attcaaacac tcacaggcat ctgtgtgagt ttgttggcct tgatgctgag 1020 atggagatta aggaacatta ttttgaggtt tgcgacataa tagatggttt gtttgtagca 1080 atatttaaac acttgaatga aaattgcaag aaagaactag agacaataaa caggcaatat 1140 ccatttgaac ctctgaagta tttagagaaa actttgaagc tcacgtatga agaaggaatt 1200 caaatgctga aggaagctgg aacagaaatc gaacccatgg gtgacctcaa cactgaagct 1260 gagaaaaaac taggccggct tgttaaggag aagtatggaa cagaattttt catcctctat 1320 cggtatcctt tggctgtgcg tcccttctac accatgcctt gttatgacaa cccagcttac 1380 agtaactctt ttgatgtctt tattcgagga gaggaaataa tatctggagc acaaagaata 1440 catttaccag agctattgac gaaacgtgca acagagtgtg gaattgatgc gagtactatt 1500 tcatcatata tcgaatcgtt cagctatggt gcacctcctc atggtggttt tggtgtcggc 1560 ctggagaggg tggtaatgct gttctgcgcc ctaaacaaca tcaggaagac atcacttttc 1620 cctcgcgatc cacaaaggct ggtgccataa 1650 <210> 3 <211> 5388 <212> DNA <213> Artificial Sequence <400> 3 tggccccaca tgtcatccac tcgtgctacg cagcagttga gcaatcggtg attaaaaaaa 60 aattccacac ccaaacccta gccccgcacc gatccctctc cctctcccgc cgccaccgcc 120 tccgcctccg cctcccctcg cgccgtcgcc gtcgccaaga gccgccactt ccgcttccgc 180 ctctgcctcg tccaagtcca agccaactac ctaaacctaa accctattct tcttctttcg 240 cctccgccgc gatgtcgtcg gagcctccac ccgatgccgc cgccgccgcc gcctcctccg 300 cggggatct cgcggccgat ctctcctccg ccaccatcag aagaagcag ctcaagaagg 360 420 aggccgacgc cgacgacccc ttcgcggcca actacggcga cgtccccgtc gaggagatcc 480 agtccaagac catctccggc cgcgtgtgga ccgagatcgg cggcctcgac gaggccgccg 540 ccggccgctc cgtcctcatc cgcggcgccg cgcaggcgat ccggcccgtc near you 600 tggccttcgt tgtgctgcgc gagagcatga gcaccgtcca gtgcgtgctc gtcgccagcg 660 ccgacgcagg ggtcagcacc cagatggtgc gcttcgccac ctccctcagc aaagcaatcaa 720 tcgtcgacgt cgagggcgtc gtctccctcc ccaaggagcc cctcaaggcc accacgcagc 780 aggtctgctc tctccaccat tatgttgggt caatatgatc atcaattgca tttgcatcaa 840 cccttggccg cagttgatta gaatatgcat tcggatagcc ctcttgtgat agctctagaa 900 tgctgaatat ttcatgttag atgatggttc tgcaatcgat tggaattcac attgtgatag 960 tcctcttatg atacctcttg gaggctgaat attcatgtat ggtggttttt gttggacttt 1020 ttgtatccat cttaatttac cgtcacaatt agctcgattt aatcccgcaa cataacttat 1080 tatacggtgt gcacatttaa aactgtgctt tgtttcagtt agtactagct aatgaagtgt 1140 cttttgtttt tgctggcagg tggagattca ggtgaggaag atctattgca tcaatagggc 1200 gatccccacc cttccaatca accttgagga tgcctcacgg agtgaggctg aaattgaaaa 1260 ggctgaacaa gtatgtatat gtgcaacttg cttgtacatc cacaatgtat atttacagga 1320 tgcctataaa tattataatc attgtgagac ttaagaaatt aaccactgct gccgtttcag 1380 gctggagaga agctagttcg tgtgggccaa gatactcgct tgaactatag agctattgat 1440 ctccggacac ctgcaaatca agcaatattc agaattcaat gtcaagttga gaacgtgagt 1500 atttatttat ttatttttac tgttatcatt tttgtattag tggattgtat tgtgccaacg 1560 atgctatgtg atctatagct cagttaacac attaaatttt ccattgaata aaaactactc 1620 tggcaaatgt tcatgctttc agcttagtca atcttgcagt aataaacatt tgcctcttgg 1680 gaactaagct caaactttgc acaagttatt gctgtttgga gatatactga tttggtgacc 1740 ccaagatgtt cccaataatc tgtgccttgc aacttatcta attgcacttg tcaagggtta 1800 gattcaatat gtttgtcttg agtgaaggcc atgcgtgtag tatatattca tgtccaagcc 1860 tttgtaggc taattcttaa ttcagtggag gttaaacaac gagcttctat tatgttttct 1920 atcgcgtatg agcgtgttat attcattatg gaaattaaat cgttgtcatg tattttgcag 1980 aaattcaggg aatattttct gtcgaaaaat tttgttggga ttcacagccc aaagctaatt 2040 gctggatcca gcgaaggtgg tgcagctgta ttcaagctac agtacaatgg acagccagca 2100 tgttagcac agtctcctca gttgtataag cagatggcta tttgtggtgg gtttgaacgt 2160 gtatttgagg ttggacctgt ttttagagct gagaattcaa acactcacag gcatctgtgt 2220 gagtttgttg gccttgatgc tgagatggag attaaggaac attattttga ggtaaggaaa 2280 attggatgaa agatgctttg tattagtata agcaaagtga tgctgactaa gtgctgattt 2340 ctcacacag gttgcgaca taatagatgg ttgtttgta gcaattta aacacttgaa 2400 tgaaaattgc aagaaagaac tagacaat aaacaggca tattccattg aacctctgaa 2460 ggtgaacaag ttcttgcctc ttttgttcc ccttttataa tattatctaa ctcatatttt 2520 gatactatat tagctactc atattatgct aattcataca gtatttagag aaaactttga 2580 agctcacgta tgagaagga attcaatgc tgaaggtaa tatctgttt cagtcatcat 2640 ctctatgtta cctcttattt ggtatgggtg aatttctgaa catatgtctg atctattgaa 2700 tgaaccatat tctagctgag ggtaatatat gatttcttg ttggcctta tatgagaat 2760 ctactaatgg cccaggtttt tactgagaat cctaaatatt tcatgagcat atttacatg 2820 ttggtatgca cattgtgtat tttagcaatt taatgtagtt cacatctttg ttgcaatttg 2880 gacctactga cccagatatt gggttggtaaa agctcagtc attgctcat cctgaaatag 2940 haaccttgac atttcagtc atagaatctt accaataga tagattgtt ggacattttg 3000 acagatgttc atggactaag cctagtaagc tacctacaat ggataccaaa gaatcttgtc 3060 tggtaatttg gtttgttaac attgaccaat tcatgacatt aaacggaaat tatttttcaa 3120 aaaaaaaaaa agaacagtga tgaatcttga tcaatcaaat ctaaagaatg ggactgggga 3180 gagattcaac cccacagtat tttttttag tacgagggag gaagtgagca caccaaccgt 3240 gttcgcagct cactcaaatt tgatataatg aaatcctgtt tttgctaaat atctagtatg 3300 ttagaaacat gttattggca actttgaagt agaacaggtg tacaacttac agatatactg 3360 ggttatctgg gtcgtctttt ttagtaaaac cagatttaat tgttgaacta atttttgct 3420 taggtgttgg cctaaaatgc tttcacagaa ttatatccat agcgattaac atcgtggtttt 3480 gtcctggcag gaagctggaa cagaaatcga acccatgggt gacctcaaca ctgaagctga 3540 gaaaaaacta ggccggcttg ttaaggagaa gtatgcaaat tcagcctgta gttacttatt 3600 cgcttcttt caaaagaaaa tgtggtttga caacttgcta cattgtctgt aggtatggaa 3660 cagaattttt catcctctat cggtatcctt tggctgtgcg tccctctac accatgcctt 3720 gttatgacaa cccagcttac agtaactctt ttgatgtctt tattcgaggt acgtaaattt 3780 gtttatttct ttgagatgtt tgaatttatt acagtatatt tcagtgtcat gcatgcatgt 3840 gtgaggataa tgttacctga gcaggaaaac attatttca gtcctgctgc ttataaatcc 3900 tttgttattt agagccatat tgttgtgaat cactatccta tatgaaatta gtaatgactt 3960 ccacttttct tcattgcata gtctgacacg caaaaagata gataaaaatc atgacttgac 4020 agaaagtaaa ccagaatatg ctctatcctg ccaaggactt gaattcgcca cggggattac 4080 ttaggatggc acagaaatga ccaacaccta tttttagtta gtcgcttaaa tgcattccat 4140 ccaatatgat gtgctaattg gtcctgacgg ttagcttgat tgagtgtttg atggtgcatc 4200 ctatagccaa cctaggtctg cacaaccaca ttgaactttg ctatcaattt aaatttagtg 4260 aaaatttatt cctttattat agtcgtgttt gcgtcatctt atctacttcc atcagtagtc 4320 actatttgct atcactgaag aagatattca tcatgtgtag aagcatcctg ttgaccattt 4380 ttgtgatatg ataggagagg aaataatatc tggagcacaa agaatacatt taccagagct 4440 attgacgaaa cgtgcaacag agtgtggaat tgatgcgagt actatttcat catatatcga 4500 atcgttcagg taagtttatt tgttatcat gttgttagtc ctctaatatt gcaacatatg 4560 ttgtaatgct catccaattt tcttcttgtc atcatctaaa tcacttcatt caactcattt 4620 caatatatgg gtcatcattg tagctatggt gcacctcctc atggtggttt tggtgtcggc 4680 ctggagaggg tggtaatgct gttctgcgcc ctaaacaaca tcaggaagac atcactttc 4740 cctcgcgatc cacaaaggct ggtgccataa tttgcttttt ttcccaagag caaggtttgg 4800 actcagtacg gactgggcag ttttcctcgg ctggtttttt tacctggaca ttattttcgt 4860 atttattaat gtgctgtact gcaaaagctg ctcctttcca caacatttgg aatagttgcc 4920 gatacatttg gaatagggct caacgttggc gttgtgattt cgttgatgat tccgctattc 4980 gtaacaatgg cgaatccaga gcagaaatga ccctatgtca gaaacatatc acatactcgt 5040 atgcctatat ttccaaaaga tagggatta tggaaggatg acatattcaa gtcccaaaat 5100 acaaggtgtg gtttgactcg tggtcttcaa catccaactt ttgattgtca ttttctgtca 5160 tggtaccacc ttcatcctaa tttgacattc tctaagtaga aggcaccaat accaagaaaa 5220 gtgtgaatga cttatatttt aaatattgga tgtcattgca ccattagaaa ttaatcttgt 5280 tggttgcatg taacacattc aatgatgcac attactcaca ttactctttg tacaaataaa 5340 caaaatgaca cttgtttaat gatgatcatt ttggaaagac cccagaaa 5388 <210> 4 <211> 11762 <212> DNA <213> Artificial Sequence <400> 4 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca 60 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca 120 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc 180 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattggctag agcagcttgc 240 caacatggtg gagcacgaca ctctcgtcta ctccaagaat atcaaagata cagtctcaga 300 agaccaaagg gctattgaga cttttcaaca aagggtaata tcgggaaacc tcctcggatt 360 ccattgccca gctatctgtc acttcatcaa area gaaaaggaag gtggcaccta 420 caaatgccat cattgcgata aaggaaaggc tatcgttcaa gatgcctctg ccgacagtgg 480 tcccaaagat ggacccccac ccacgaggag catcgtggaa aaagaagacg ttccaaccac 540 gtcttcaaag caagtggatt gatgtgataa catggtggag cacgacactc tcgtctactc 600 caagaatatc aaagatacag tctcagaaga ccaaagggct attgagactt ttcaacaaag 660 ggtaatatcg ggaacctcc tcggattcca ttgcccagct atctgtcact tcatcaaaag 720 gacagtagaa aaggaaggtg gcacctacaa atgccatcat tgcgataaag gaaggctat 780 cgttcaagat gcctctgccg acagtggtcc caaagatgga ccccaccca cgaggagcat 840 cgtggaaaaa gaagacgttc caaccacgtc ttcaaagcaa gtggattgat gtgatatctc 900 cactgacgta agggatgacg caaatccca ctatccttcg caagaccttc ctctatataa 960 ggaagttcat ttcatttgga gaggaccgc tgaaatcacc agtctctctc tacaaatcta 1020 tctctctcga gctttcgcag atcccgggg gcaatgagat atgaaaaagc ctgaactcac 1080 cgcgacgtct gtcgagaagt ttctgatcga aaagttcgac agcgtctccg acctgatgca 1140 gctctcggag ggcgaagaat ctcgtgcttt cagcttcgat gtaggagggc gtggatatgt 1200 cctgcgggta aatagctgcg ccgatggttt ctacaaagat cgttatgttt atcggcactt 1260 tgcatcggcc gcgctcccga ttccggaagt gcttgacatt ggggagttta gcgagagcct 1320 gacctattgc atctcccgcc gtgcacaggg tgtcacgttg caagacctgc ctgaaaccga 1380 actgcccgct gttctacaac cggtcgcgga ggctatggat gcgatcgctg cggccgatct 1440 tagccagacg agcgggttcg gcccattcgg accgcaagga atcggtcaat acactacatg 1500 gcgtgatttc atatgcgcga ttgctgatcc ccatgtgtat cactggcaaa ctgtgatgga 1560 cgacaccgtc agtgcgtccg tcgcgcaggc tctcgatgag ctgatgcttt gggccgagga 1620 ctgccccgaa gtccggcacc tcgtgcacgc ggatttcggc tccaacaatg tcctgacgga 1680 caatggccgc ataacagcgg tcattgactg gagcgaggcg atgttcgggg attcccaata 1740 cgaggtcgcc aacatcttct tctggaggcc gtggttggct tgtatggagc agcagacgcg 1800 ctacttcgag cggaggcatc cggagcttgc aggatcgcca cgactccggg cgtatatgct 1860 ccgcattggt cttgaccaac tctatcagag cttggttgac ggcaatttcg atgatgcagc 1920 ttgggcgcag ggtcgatgcg acgcaatcgt ccgatccgga gccgggactg tcgggcgtac 1980 acaaatcgcc cgcagaagcg cggccgtctg gaccgatggc tgtgtagaag tactcgccga 2040 tagtggaaac cgacgcccca gcactcgtcc gagggcaaag aaatagagta gatgccgacc 2100 ggatctgtcg atcgacaagc tcgagtttct ccataataat gtgtgagtag ttcccagata 2160 agggaattag ggttcctata gggtttcgct catgtgttga gcatataaga aacccttagt 2220 atgtatttgt atttgtaaaa tacttctatc aataaaattt ctaattccta aaaccaaaat 2280 ccagtactaa aatccagatc ccccgaatta attcggcgtt aattcagtac attaaaaacg 2340 tccgcaatgt gttattaagt tgtctaagcg tcaatttgtt tacaccacaa tatatcctgc 2400 caccagccag ccaacagctc cccgaccggc agctcggcac aaaatcacca ctcgatacag 2460 gcagcccatc agtccgggac ggcgtcagcg ggagagccgt tgtaaggcgg cagactttgc 2520 tcatgttacc gatgctattc ggaagaacgg caactaagct gccgggtttg aaacacggat 2580 gatctcgcgg aggtagcat gttgattgta acgatgacag agcgttgctg cctgtgatca 2640 ccgcggtttc aaaatcggct ccgtcgatac tatgttatac gccaactttg aaaacaactt 2700 tgaaaaagct gttttctggt atttaaggtt ttagaatgca aggaacagtg aattggagtt 2760 cgtcttgtta taattagctt cttggggtat ctttaaatac tgtagaaaag aggaaggaaa 2820 taataaatgg ctaaaatgag aatatcaccg gaattgaaaa aactgatcga aaaataccgc 2880 tgcgtaaaag atacggaagg aatgtctcct gctaaggtat ataagctggt gggagaaaat 2940 gaaaacctat atttaaaat gacggacagc cggtataaag ggaccaccta tgatgtggaa 3000 cgggaaaagg acatgatgct atggctggaa ggaaagctgc ctgttccaaa ggtcctgcac 3060 tttgaacggc atgatggctg gagcaatctg ctcatgagtg aggccgatgg cgtcctttgc 3120 tcggaagagt atgaagatga acaaagccct gaaaagatta tcgagctgta tgcggagtgc 3180 atcaggctct ttcactccat cgacatatcg gattgtccct atacgaatag cttagacagc 3240 cgcttagccg aattggatta cttactgaat aacgatctgg ccgatgtgga ttgcgaaaac 3300 tgggaagaag acactccatt taaagatccg cgcgagctgt atgatttttt aaagacggaa 3360 aagcccgaag aggaacttgt cttttcccac ggcgacctgg gagacagcaa catctttgtg 3420 aaagatggca aagtaagtgg ctttattgat cttgggagaa gcggcagggc ggacaagtgg 3480 tatgacattg ccttctgcgt ccggtcgatc agggaggata tcggggaaga acagtatgtc 3540 gagctatttt ttgacttact ggggatcaag cctgattggg agaaaataaa atattatatt 3600 ttactggatg aattgtttta gtacctagaa tgcatgacca aaatccctta acgtgagttt 3660 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt 3720 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt 3780 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag 3840 ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta 3900 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat 3960 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg 4020 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg 4080 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac 4140 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga 4200 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt 4260 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta 4320 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat 4380 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg 4440 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg gtattttctc 4500 cttacgcatc tgtgcggtat ttcacaccgc atatggtgca ctctcagtac aatctgctct 4560 gatgccgcat agttaagcca gtatacactc cgctatcgct acgtgactgg gtcatggctg 4620 cgccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat 4680 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt 4740 catcaccgaa acgcgcgagg cagggtgcct tgatgtgggc gccggcggtc gagtggcgac 4800 ggcgcggctt gtccgcgccc tggtagattg cctggccgta ggccagccat ttttgagcgg 4860 ccagcggccg cgataggccg acgcgaagcg gcggggcgta gggagcgcag cgaccgaagg 4920 gtaggcgctt tttgcagctc ttcggctgtg cgctggccag acagttatgc acaggccagg 4980 cgggttttaa gagttttaat aagttttaaa gagttttagg cggaaaaatc gccttttttc 5040 tcttttatat cagtcactta catgtgtgac cggttcccaa tgtacggctt tgggttccca 5100 atgtacgggt tccggttccc aatgtacggc tttgggttcc caatgtacgt gctatccaca 5160 ggaaagagac cttttcgacc tttttcccct gctagggcaa tttgccctag catctgctcc 5220 gtacattagg aaccggcgga tgcttcgccc tcgatcaggt tgcggtagcg catgactagg 5280 atcgggccag cctgccccgc ctcctccttc aaatcgtact ccggcaggtc atttgacccg 5340 atcagcttgc gcacggtgaa acagaacttc ttgaactctc cggcgctgcc actgcgttcg 5400 tagatcgtct tgaacaacca tctggcttct gccttgcctg cggcgcggcg tgccaggcgg 5460 tagagaaaac ggccgatgcc gggatcgatc aaaaagtaat cggggtgaac cgtcagcacg 5520 tccgggttct tgccttctgt gatctcgcgg tacatccaat cagctagctc gatctcgatg 5580 tactccggcc gcccggtttc gctctttacg atcttgtagc ggctaatcaa ggcttcaccc 5640 tcggataccg tcaccaggcg gccgttcttg gccttcttcg tacgctgcat ggcaacgtgc 5700 gtggtgttta accgaatgca ggtttctacc aggtcgtctt tctgctttcc gccatcggct 5760 cgccggcaga acttgagtac gtccgcaacg tgtggacgga acacgcggcc gggcttgtct 5820 cccttccctt cccggtatcg gttcatggat tcggttagat gggaaaccgc catcagtacc 5880 aggtcgtaat cccacacact ggccatgccg gccggccctg cggaaacctc tacgtgcccg 5940 tctggaagct cgtagcggat cacctcgcca gctcgtcggt cacgcttcga cagacggaaa 6000 acggccacgt ccatgatgct gcgactatcg cgggtgccca cgtcatagag catcggaacg 6060 aaaaaatctg gttgctcgtc gcccttgggc ggcttcctaa tcgacggcgc accggctgcc 6120 ggcggttgcc gggattcttt gcggattcga tcagcggccg cttgccacga ttcaccgggg 6180 cgtgcttctg cctcgatgcg ttgccgctgg gcggcctgcg cggccttcaa cttctccacc 6240 aggtcatcac ccagcgccgc gccgatttgt accgggccgg atggtttgcg accgtcacgc 6300 cgattcctcg ggcttggggg ttccagtgcc attgcagggc cggcagacaa cccagccgct 6360 tacgcctggc caaccgcccg ttcctccaca catggggcat tccacggcgt cggtgcctgg 6420 ttgttcttga ttttccatgc cgcctccttt agccgctaaa attcatctac tcatttattc 6480 atttgctcat ttactctggt agctgcgcga tgtattcaga tagcagctcg gtaatggtct 6540 tgccttggcg taccgcgtac atcttcagct tggtgtgatc ctccgccggc aactgaaagt 6600 tgacccgctt catggctggc gtgtctgcca ggctggccaa cgttgcagcc ttgctgctgc 6660 gtgcgctcgg acggccggca cttagcgtgt ttgtgctttt gctcattttc tctttacctc 6720 attaactcaa atgagttttg atttaatttc agcggccagc gcctggacct cgcgggcagc 6780 gtcgccctcg ggttctgatt caagaacggt tgtgccggcg gcggcagtgc ctgggtagct 6840 cacgcgctgc gtgatacggg actcaagaat gggcagctcg tacccggcca gcgcctcggc 6900 aacctcaccg ccgatgcgcg tgcctttgat cgcccgcgac acgacaaagg ccgcttgtag 6960 ccttccatcc gtgacctcaa tgcgctgctt aaccagctcc accaggtcgg cggtggccca 7020 tatgtcgtaa gggcttggct gcaccggaat cagcacgaag tcggctgcct tgatcgcgga 7080 cacagccaag tccgccgcct ggggcgctcc gtcgatcact acgaagtcgc gccggccgat 7140 ggccttcacg tcgcggtcaa tcgtcgggcg gtcgatgccg acaacggtta gcggttgatc 7200 ttcccgcacg gccgcccaat cgcgggcact gccctgggga tcggaatcga ctaacagaac 7260 atcggccccg gcgagttgca gggcgcgggc tagatgggtt gcgatggtcg tcttgcctga 7320 cccgcctttc tggttaagta cagcgataac cttcatgcgt tccccttgcg tatttgttta 7380 tttactcatc gcatcatata cgcagcgacc gcatgacgca agctgtttta ctcaaataca 7440 catcaccttt ttagacggcg gcgctcggtt tcttcagcgg ccaagctggc cggccaggcc 7500 gccagcttgg catcagacaa accggccagg atttcatgca gccgcacggt tgagacgtgc 7560 gcgggcggct cgaacacgta cccggccgcg atcatctccg cctcgatctc ttcggtaatg 7620 aaaaacggtt cgtcctggcc gtcctggtgc ggtttcatgc ttgttcctct tggcgttcat 7680 tctcggcggc cgccagggcg tcggcctcgg tcaatgcgtc ctcacggaag gcaccgcgcc 7740 gcctggcctc ggtgggcgtc acttcctcgc tgcgctcaag tgcgcggtac agggtcgagc 7800 gatgcacgcc aagcagtgca gccgcctctt tcacggtgcg gccttcctgg tcgatcagct 7860 cgcgggcgtg cgcgatctgt gccggggtga gggtagggcg ggggccaaac ttcacgcctc 7920 gggccttggc ggcctcgcgc ccgctccggg tgcggtcgat gattagggaa cgctcgaact 7980 cggcaatgcc ggcgaacacg gtcaacacca tgcggccggc cggcgtggtg gtgtcggccc 8040 acggctctgc caggctacgc aggcccgcgc cggcctcctg gatgcgctcg gcaatgtcca 8100 gtaggtcgcg ggtgctgcgg gccaggcggt ctagcctggt cactgtcaca acgtcgccag 8160 ggcgtaggtg gtcaagcatc ctggccagct ccgggcggtc gcgcctggtg ccggtgatct 8220 tctcggaaaa cagcttggtg cagccggccg cgtgcagttc ggcccgttgg ttggtcaagt 8280 cctggtcgtc ggtgctgacg cgggcatagc ccagcaggcc agcggcggcg ctcttgttca 8340 tggcgtaatg tctccggttc tagtcgcaag tattctactt tatgcgacta aaacacgcga 8400 caagaaaacg ccaggaaaag ggcagggcgg cagcctgtcg cgtaacttag gacttgtgcg 8460 acatgtcgtt ttcagaagac ggctgcactg aacgtcagaa gccgactgca ctatagcagc 8520 ggaggggttg gatcaaagta ctttgatccc gaggggaacc ctgtggttgg catgcacata 8580 caaatggacg aacggataaa ccttttcacg cccttttaaa tatccgttat tctaataaac 8640 gctcttttct cttaggttta cccgccaata tatcctgtca aacactgata gtttaaactg 8700 aaggcgggaa acgacaatct gatccaagct caagctgctc tagcattcgc cattcaggct 8760 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa 8820 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg 8880 ttgtaaaacg acggccagtg ccaagcttgc atgcctgcag gtccccagat tagccttttc 8940 aatttcagaa agaatgctaa cccacagatg gttagagagg cttacgcagc aggtctcatc 9000 aagacgatct accgagcaa taatctccag gaatcaat accttcccaa gaaggttaa 9060 gatgcagtca aaagattcag gactactgc atcagaca cagagaaga tatattctc 9120 aagatcagaa gtactattcc agtatggacg attcaggct tgctcacaa accaggcaa 9180 gtaatagaga ttggagtctc taaaaaggta gttcccactg atcaaggc catggagtca 9240 aagattcaa tagaggacct aagaactc gccgtaaga ctggcgaaca gttcatacag 9300 agtctcttac gactcaatga caagaaaa atcttcgtca acatggtgga gcacgacaca 9360 cttgtctact ccaaaatat caagataca gtctcagaag accaagggc attgagact 9420 tttcacaaa gggtatatatc cggaaacctc ctcggattcc attgcccagc tatctgtcac 9480 tttattgtga agatagtgga aaggaaggt ggctcctaca aatgccatca ttgcgataaa 9540 ggaaaggcca tcgttgaaga tgcctgcc zgagtggtc ccaagatgg accccaccc 9600 acgaggagca tcgtggaaaa agaagacgtt ccaaccacgt cttcaaagca agtggattga 9660 tgtgatatct ccactgacgt aagggac gcacaatccc actaccttc gcaagaccct 9720 tcctctatat aaagttc atttcatttg gagagaacac gggggactct agaatgtcgt 9780 cggagcctcc acccgatgcc gccgccgccg ccgcctcctc cgcgggggat ctcgcggccg 9840 atctctcctc cgccaccatc agcaagaagc agctcaagaa ggatgcgagg aaggcggaga 9900 aggccgagaa ggcgtcgcag cgccagcagc agcagcagcc gcaggccgac gccgacgacc 9960 ccttcggcggc caactacggc gacgtccccg tcgaggagat ccagtccaag accatctccg 10020 gccgcgtgtg gaccgagatc ggcggcctcg acgaggccgc cgccggccgc tccgtcctca 10080 tccgcggcgc cgcgcaggcg atccggcccg tcagcaagaa gatggccttc gttgtgctgc 10140 gcgagagcat gagcaccgtc cagtgcgtgc tcgtcgccag cgccgacgca ggggtcagca 10200 cccagatggt gcgcttcgcc acctccctca gcaaggaatc aatcgtcgac gtcgagggcg 10260 tcgtctccct ccccaaggag cccctcaagg ccaccacgca gcaggtggag attcaggtga 10320 ggaagatcta ttgcatcaat agggcgatcc ccacccttcc aatcaacctt gaggatgcct 10380 cacggagtga ggctgaaatt gaaaaggctg aacaagctgg agagaagcta gttcgtgtgg 10440 gccaagatac tcgcttgaac tatagagcta ttgatctccg gacacctgca aatcaagcaa 10500 tattcagaat tcaatgtcaa gttgagaaca aattcaggga atattttctg tcgaaaaatt 10560 ttgttgggat tcacagcca aagctaattg ctggatccag cgaaggtggt gcagctgtat 10620 tcaagctaca gtacaatgga cagccagcat gtttagcaca gtctcctcag ttgtataagc 10680 agatggctat ttgtggtggg tttgaacgtg tatttgaggt tggacctgtt tttagagctg 10740 agaattcaaa cactcacagg catctgtgtg agtttgttgg ccttgatgct gagatggaga 10800 ttaaggaaca ttattttgag gtttgcgaca taatagatgg tttgtttgta gcaatattta 10860 aacacttgaa tgaaaattgc aagaaagaac tagagacaat aaacaggcaa tatccatttg 10920 aacctctgaa gtatttagag aaaactttga agctcacgta tgaagaagga attcaaatgc 10980 tgaaggaagc tggaacagaa atcgaaccca tgggtgacct caacactgaa gctgagaaaa 11040 aactaggccg gcttgttaag gagaagttag gaacagaatt tttcatcctc tatcggtatc 11100 ctttggctgt gcgtcccttc tacaccatgc cttgttatga caacccagct tacagtaact 11160 cttttgatgt ctttattcga ggagaggaaa tatatctgg agcacaaaga atacatttac 11220 cagagctatt gacgaaacgt gcaacagagt gtggaattga tgcgagtact atttcatcat 11280 atacgaatc gttcagctat ggtgcacctc ctcatggtgg ttttggtgtc ggcctggaga 11340 gggtggtaat gctgttctgc gccctaaaca acatcaggaa gacatcactt ttccctcgcg 11400 atccacaaag gctggtgcca gactacaaag accatgatgg agactataag gatcacgaca 11460 tcgattacaa ggacgatgac gataagtgat aagaatttcc ccgatcgttc aaacatttgg 11520 caataaagtt tcttaagatt gaatcctgtt gccggtcttg cgatgattat catataattt 11580 ctgttgaatt acgttaagca tgtaataatt aacatgtaat gcatgacgtt attatgaga 11640 tgggttttta tgattagagt cccgcaatta tacatttaat acgcgataga aaaaaaata 11700 tagcgcgcaa actaggataa attatcgcgc gcggtgtcat ctatgttact agatcgggaa 11760 tt 11762

Claims

1. A method for improving disease resistance of a plant, comprising the step of: replacing a gene encoding an OsAspRS protein in the plant with a gene encoding a mutant protein; wherein the OsAspRS protein is as follows (al) or (a2): (al) a protein represented by SEQ ID NO: 1 in the sequence listing; (a2) a fusion protein obtained by linking a tag to the N terminus and / or C terminus of the protein of (al); wherein the mutant protein is obtained by mutating the 276th amino acid residue of the OsAspRS protein from Ser to Asn; wherein the plant is rice; and wherein the disease resistance is blast disease resistance and / or bacterial blight disease resistance.

2. Use of a mutant protein in regulating disease resistance of a plant, wherein the mutant protein increases the disease resistance; wherein the mutant protein is as defined in claim 1; wherein the plant is rice; and wherein the disease resistance is blast disease resistance and / or bacterial blight disease resistance.

3. Use of a gene encoding a mutant protein in breeding a plant with improved disease resistance; wherein the mutant protein is as defined in claim 1; wherein the plant is rice; and wherein the disease resistance is blast disease resistance and / or bacterial blight disease resistance.

4. A mutant protein obtained by mutating the 276th amino acid residue of an OsAspRS protein from Ser to Asn; wherein the OsAspRS protein is as follows (al) or (a2): (al) a protein represented by SEQ ID NO: 1 in the sequence listing; (a2) a fusion protein obtained by linking a tag to the N terminus and / or C terminus of the protein of (al).

5. A gene encoding the mutant protein of claim 4. osasprs osasprs osasprs wherein the plant is rice; and wherein the disease resistance is blast disease resistance and / or bacterial blight disease resistance. osasprs osasprs osasprs wherein the plant is rice; and wherein the disease resistance is blast disease resistance and / or bacterial blight disease resistance. ​ ​ ​ ​ ​ ​ ​ 6. ​ Use of the mutant in the preparation of plants with improved disease resistance; the mutant is a lesion-mimic mutant of Oryza sativa L. cv. Xiehuazhan ​ The mutant is a lesion-mimic mutant of Oryza sativa L. cv. Xiehuazhan ​ with the preservation registration number of CGMCC No. 23084; ​ ​ 7. A method for preparing a plant with improved disease resistance, comprising the step of: crossing a mutant with other plants to obtain a plant with improved disease resistance from the hybrid progeny; wherein the mutant is a lesion-mimic mutant of Oryza sativa L. ssp. indica cv. Huanghuazhan. ​ ​ ​ with a preservation registration number of CGMCC No. 23084.​​ ​ ​

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