SCRE10 gene for improving disease resistance of rice and application thereof

Abstract

This invention belongs to the field of plant genetic engineering technology, specifically relating to a method for improving the disease resistance of rice. SCRE10 Genes and their applications, the aforementioned SCRE10 The base sequence of the gene is shown in SEQ ID NO.1, and the amino acid sequence is shown in SEQ ID NO.2. This invention utilizes a dexamethasone-induced expression mechanism. SCRE10 The transgenic rice plants were found to significantly induce... PR Gene expression and reactive oxygen species surges, enhancing rice resistance to rice false smut and bacterial blight. This invention demonstrates heterologous induction of rice false smut pathogen expression. SCRE10 Genes have the function of positively regulating the disease resistance of rice. SCRE10 Genes can be used to improve the disease resistance of rice, which is of great significance for the creation of disease-resistant rice germplasm.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of plant genetic engineering, and particularly relates to a kind of SCRE10 Genes and applications thereof for improving disease resistance of rice. BACKGROUND

[0002] Rice is one of the most important food crops in the world, and bears a huge pressure of food supply. Among rice diseases, rice blast, rice bacterial leaf blight, sheath blight and smut of rice, etc. not only affect the yield of rice, but also seriously threaten the quality of rice.

[0003] In recent years, with the increase of warm and humid environment caused by global warming, and the overuse of nitrogen fertilizer in agricultural production, the occurrence range of these diseases in the main producing areas of rice continues to expand, and the degree of harm is showing a trend of aggravating year by year. Therefore, it has become an urgent need in the field of current rice research and production to explore and effectively use key genes, improve rice germplasm resources, and enhance the disease resistance of rice to various diseases. SUMMARY

[0004] The purpose of the present application is to provide a kind of SCRE10 Genes for improving disease resistance of rice, which solves the problem of limited disease resistance genes in the prior art.

[0005] The technical solution adopted by the present application is as follows:

[0006] The present application provides a kind of SCRE10 Genes for improving disease resistance of rice, the base sequence of the SCRE10 Gene is shown as SEQ ID NO. 1, and the amino acid sequence is shown as SEQ ID NO. 2.

[0007] The second aspect of the present application provides an application of the SCRE10 Gene, the SCRE10 Gene is used to improve the disease resistance of rice.

[0008] The application refers to transforming the SCRE10 Gene into rice, expressing the SCRE10 Gene in rice, preparing a transgenic plant of rice, and then improving the disease resistance of rice.

[0009] Preferably, the SCRE10 Gene is used to improve the resistance of rice to smut and / or bacterial leaf blight.

[0010] Preferably, the preparation method of the transgenic plant is as follows:

[0011] The cDNA of the smut fungus is used as a template for amplification to obtain the SCRE10 Gene;

[0012] The SCRE10 gene is ligated into the expression vector after enzyme digestion to obtain a recombinant overexpression vector;

[0013] The recombinant overexpression vector is transformed into Agrobacterium to obtain a recombinant positive Agrobacterium;

[0014] The recombinant positive Agrobacterium is used to infect rice to obtain the transgenic plant.

[0015] Preferably, the expression vector is pTA7001.

[0016] Preferably, the smut fungus is P1FZ.

[0017] Preferably, the restriction enzyme used for enzyme digestion of the expression vector is Xho I and Spe I .

[0018] Preferably, the Agrobacterium is EHA105.

[0019] Compared with the prior art, the present application has the following beneficial effects:

[0020] The present application provides a kind of to improve the disease resistance of rice SCRE10 Gene, the base sequence of the SCRE10 Gene is as shown in SEQ ID NO.1, and the amino acid sequence is as shown in SEQ ID NO.2.The present application is by constructing the transgenic rice plant of dexamethasone induced expression SCRE10 It is found that transgenic rice can significantly induce PR Gene expression and burst of reactive oxygen species, and can improve the resistance of rice to smut and bacterial leaf blight.The present application proves that heterologous induced expression smut fungus SCRE10 Gene has the function of positively regulating the disease resistance of rice, SCRE10 Gene can be used to improve the disease resistance of rice, which has important significance for the creation of rice disease-resistant germplasm. BRIEF DESCRIPTION OF DRAWINGS

[0021] Figure 1 is heterologous expression SCRE10 The detection results of transgenic rice on PR Gene.A: transgenic rice SCRE10 protein expression detection."+" represents using 10µM DEX treatment,"-" represents Mock treatment;B: SCRE10 The expression of the gene OsPR5 The expression level of flg22 triggered defense gene SCRE10 The expression of the gene OsPR10a The expression level of flg22 triggered defense gene SCRE10 The expression of the gene OsPR5expression level of E: SCRE10 expression of E elevates chitin-triggered defense genes OsPR10a expression level of E.

[0022] Figure 2 heterologous expression in rice SCRE10 Detection of active oxygen in transgenic rice. A: SCRE10 flg22-induced ROS production in gene-inducible expression lines; B: SCRE10 chitin-induced ROS production in gene-inducible expression lines.

[0023] Figure 3 inducible expression in rice SCRE10 enhanced resistance to Magnaporthe oryzae. A: typical disease symptoms of wild type Nip and SCRE10 Typical disease symptoms of transgenic lines inoculated with Magnaporthe oryzae strain JS60-2; B: leaf lesion length of wild type Nip and SCRE10 Statistical chart of rice blast pustule number of transgenic lines inoculated with Magnaporthe oryzae strain JS60-2.

[0024] Figure 4 inducible expression in rice SCRE10 enhanced resistance to Xanthomonas oryzae. A: Xanthomonas oryzae PX099 A inoculated wild type Nip and SCRE10 Typical disease symptoms of transgenic lines 2 weeks after inoculation; B: leaf lesion length. DETAILED DESCRIPTION

[0025] The present application will be further described in the following specific examples without limiting the scope of the present application. Details and forms of the technical solutions of the present application can be modified or replaced without departing from the spirit and scope of the present application, and these modifications or replacements fall within the protection scope of the present application.

[0026] The inventive concept of the present application is as follows:

[0027] Plant disease resistance is a complex process of multiple gene synergistic regulation. Current researches are mostly focused on major disease resistance genes and disease resistance related genes involved in rice disease resistance response, and multiple key disease resistance related genes have been successfully cloned. These genes can encode different types of functional proteins such as kinases and receptor proteins, and participate in the resistance process of rice to different pathogenic bacteria by regulating different response pathways such as signal transduction and secondary metabolism in rice. In the functional research of plant pathogen effector proteins, it is found that a class of secreted proteins can enhance the disease resistance of plants by inducing plant immunity. Therefore, developing and utilizing the genes of this class of effector proteins will become a new strategy to improve the disease resistance of rice.

[0028] To enable those skilled in the art to better understand and implement the technical solutions of this invention, the invention will be further described below with reference to specific embodiments. Unless otherwise specified, all reagents used in this invention are commercially available, and all methods used are conventional techniques in the art.

[0029] The list of abbreviations for this invention is shown in Table 1.

[0030] Table 1 List of Abbreviations

[0031]

[0032] The experimental method of this invention is as follows:

[0033] 1. Rice seedlings PR Gene expression detection methods.

[0034] After sterilization, rice seeds were transferred to 1 / 2 MS solid medium for 5-7 days, then transferred to liquid 1 / 2 MS medium and cultured at 28°C for 2-3 days. Subsequently, under sterile conditions, rice seedlings of suitable size were gently transferred to 1 / 2 MS liquid medium containing 10 µM dexamethasone (DEX) using sterile forceps. The seedlings were sealed, removed, and placed in a climate chamber for 16 hours of static treatment. Afterward, the culture dishes were gently transferred to a sterile operating table, and any remaining MS liquid on the seedlings was aspirated with sterile filter paper. The seedlings were then gently transferred to 1 / 2 MS liquid medium containing chitin and 0.01% surfactant using sterile forceps. The culture dishes were then placed in a climate chamber for 6 hours of static treatment. Finally, the treated seedling leaves were taken and gently placed into sterile centrifuge tubes containing steel balls and placed in liquid nitrogen. RNA extraction and reverse transcription were completed within a short time, and RT-qPCR was performed for verification and analysis.

[0035] 2. ROS determination method.

[0036] First, Nipponbare and heterologous transgenic rice plants from the same part and with suitable growth conditions were selected and treated with DEX solution and Mock solution for 24 h, respectively. Then, samples were taken with a punch and placed in a petri dish containing sterile water and treated overnight in the dark to remove reactive oxygen species. Finally, PAMP solution, 100 μL of chemiluminescent substrate L-02 and the treated leaves were mixed into a sterile centrifuge tube and placed in a chemiluminescence detector for detection.

[0037] 3. Methods for inoculating rice with rice false smut.

[0038] First, activate the rice false smut fungus strain and culture it in a shaker at 150 rpm and 28℃ for 5-7 days. Second, break up the mycelia in the bacterial culture and adjust the conidia concentration to 1×10⁻⁶. 6The bacterial solution was then injected into the rice panicle using a needle-free syringe, with 10 replicates. Finally, 28 days after inoculation, the number of rice scald balls and the incidence of disease were counted and photographed in the field.

[0039] The rice scald fungus strain JS60-2 used in the present application is disclosed in the literature:

[0040] Xinhang Z, Anfei F, Shanshan Q, et al. Ustilaginoidea virens secretes a family of phosphatases that stabilize the negative immune regulator OsMPK6 and suppress plant immunity. [J]. The Plant cell, 2022, 34 (8): DOI: 10.1093 / PLCELL / KOAC154.

[0041] 4. A method for inoculating rice with white leaf blight bacteria.

[0042] First, the laboratory-preserved rice white leaf blight PXO99 A strain was activated in NA solid medium; then, an appropriate amount of bacterial solution was transferred to NB liquid medium and treated at 200 rpm and 28℃ for 12h; after the medium became turbid, the bacterial cells were collected by centrifugation at room temperature at 9000 rpm for 1 min; then, the collected bacterial cells were treated with 10 mM magnesium chloride solution twice, resuspended with 10 mM magnesium chloride solution, and adjusted to OD 600 0.8; finally, the bacterial solution was taken with a pair of scissors, lightly shaken, and the same length of leaf was cut on the same position of the rice leaf; after 14-15 days, the disease was photographed and counted.

[0043] The rice white leaf blight PXO99 A strain is disclosed in the reference: Zeng Chen. Construction of rice white leaf blight PXO99 A MCP-free mutant and research on chemotaxis receptor gene [D]. Guangxi University, 2019.

[0044] 5. Western blot experiment method.

[0045] (1) Prepare Western blot samples: take plant tissues and crush, add appropriate amount of 1×SDS Loading Buffer, cook the sample in a dry oven at 100℃ for 10 min, centrifuge at 12000 rpm for 2 min, and store at -20℃.

[0046] (2) Gel running: Spot the samples sequentially in a 10% protein gel and electrophoresis at 80V for 1-2 hours. Stop when the marker bands are completely separated.

[0047] (3) Transfer membrane: Transfer membrane from negative to positive in the order of cotton pad-filter paper-protein gel-PVDF membrane-filter paper-cotton pad, place it in the transfer apparatus, and electrophorese at 200mA current for 2.5h per tank.

[0048] (4) Blocking: After the transfer is completed, the membrane is taken out and placed in the blocking solution, and incubated overnight at 4°C or at room temperature for 2-3 hours.

[0049] (5) Primary antibody: After blocking, discard the waste liquid, wash the membrane 3 times with 1×TBST for 10 min each time, and after the membrane is cleaned, add an appropriate volume of primary antibody and incubate for 1 h to 3 h.

[0050] (6) Secondary antibody: recover the primary antibody, wash the membrane 3 times with 1×TBST for 10 min each time, and after the washing is finished, add an appropriate volume of secondary antibody and incubate for 1 h.

[0051] (7) Wash the membrane, recover the secondary antibody, and wash the membrane 3 times with 1×TBST for 10 min each time.

[0052] (8) Development: Blot dry the liquid on the film, wet it in the developing solution, place it in the developing instrument for development, observe and save the image.

[0053] Example 1

[0054] A method to improve the disease resistance of rice SCRE10 Genes and their applications, as detailed below:

[0055] 1. Preparation SCRE10 Transgenic rice lines.

[0056] This embodiment uses Agrobacterium-mediated transformation to create transgenic rice lines. First, the cDNA of *Strombus haemolyticus* (rice false smut fungus) is used as a template for PCR cloning to obtain... SCRE10 Genes. SCRE10 The PCR product of the gene and the pUC19-35S-FLAG vector were processed simultaneously. Xho I and BstB I Double digestion with enzymes, followed by recovery and overnight ligation to construct the pUC19-35S-SCRE10-FLAG vector. Then, using the pUC19-35S-SCRE10-FLAG vector as a template, [the following text is incomplete and requires further context: "to construct the pUC19-35S-SCRE10-FLAG vector."] SCRE10 Gene insertion through Xho I and Spe I The recombinant overexpression vector was obtained from the double-digested binary vector pTA7001. Then, the vector carrying... SCRE10The pTA7001 vector of the application is transformed into Agrobacterium GV3101, and transgenic plants are obtained by using Agrobacterium-mediated rice transgenic system. The specific method is as follows:

[0057] (1) Amplification SCRE10 The gene is amplified to obtain a PCR product.

[0058] Based on the sequence of the SCRE10 gene shown as SEQ ID NO. 1, the primers used for PCR are designed, and the forward primer and the reverse primer are shown as SEQ ID NO. 3 and SEQ ID NO. 4. The cDNA of Ustilaginoidea virens P1FZ is used as a template, and the reaction system is mixed in a 200 μL centrifuge tube. The reaction system is shown in Table 2.

[0059] Ustilaginoidea virens P1FZ is disclosed in the literature: Ustilaginoidea virens suppresses floral immunity through promoting 2 GA biosynthesis by the effector SCRE9.

[0060] SEQ ID NO. 3: CTCGGTACCCTCGAGATGAAGTTTTCGATCGTCGT.

[0061] SEQ ID NO. 4: TTTGTAGTCTTCGAAAGCTTTGTGCCTGGCAGAAG.

[0062] Table 2 Reaction system

[0063]

[0064] The mixed reaction system is subjected to gene amplification in a PCR instrument to obtain a PCR product, and the condition settings of the PCR reaction are shown in Table 3.

[0065] Table 3 PCR reaction system

[0066]

[0067] (2) Enzymatic digestion of the vector and the PCR product.

[0068] The required pUC19-35S-FLAG vector and the PCR product are subjected to Xho I and BstB I double enzyme digestion, and the reaction system is mixed in a 200 μL centrifuge tube, as shown in Table 4, and 37°C water bath for 4 h.

[0069] Table 4 Enzymatic digestion reaction system

[0070]

[0071] (3) Obtaining of the gel recovery product.

[0072] After step (2) is completed, 10x Loading Buffer is added to the product, 1% agarose gel is used for electrophoresis at 150V voltage, and the fragment size is verified; the target fragment band in the gel is recovered and purified by using a gel recovery kit, and the recovered product is stored in a -20°C refrigerator. The experimental method is referred to the agarose gel DNA recovery kit of Kang Weishijinian Biological Reagent Co., Ltd., and the steps are as follows:

[0073] ①The cut target gel block is placed in a 2.0mL centrifuge tube, weighed, 1 volume of PG Buffer is added, and the gel is dissolved in a 65°C dry oven, and the mixture is constantly inverted and mixed until completely dissolved, and then taken out and cooled to room temperature.

[0074] ②Adsorption column equilibration: 200µL PS Buffer is added to the adsorption column, and centrifuged at 12000rpm for 1min.

[0075] ③Waste liquid removal, 200µL PS Buffer is added to the adsorption column, and centrifuged at 12000rpm for 1min.

[0076] ④Waste liquid removal, 500µL PW Buffer is added to the adsorption column for rinsing, and centrifuged at 12000rpm for 1min, and rinsed again.

[0077] ⑤Waste liquid removal, 12000rpm empty column centrifugation for 3min to remove residual ethanol.

[0078] ⑥The adsorption column is placed in a new 1.5mL centrifuge tube and uncapped, and ethanol is volatilized at room temperature for 5min.

[0079] ⑦50µL EB Buffer eluent is added to the adsorption column, and centrifuged at 12000rpm for 1min, and the DNA solution is obtained, and stored at -20°C for use.

[0080] (4) Obtaining of pUC19-35S-SCRE10-FLAG vector by homologous recombination.

[0081] The experimental method is referred to the cloning kit of Nanjing Nuowezan Biological Technology Co., Ltd., and the steps are as follows:

[0082] ①The cut target gel block is placed in a 2.0mL centrifuge tube, weighed, 1 volume of PG Buffer is added, and the gel is dissolved in a 65°C dry oven, and the mixture is constantly inverted and mixed until completely dissolved, and then taken out and cooled to room temperature.

[0083] Table 5 Connection reaction system

[0084]

[0085] 2. Thaw the competent cells on ice.

[0086] 3. Take 10 μL ligation product and add to 100 μL competent cells, flick to mix, and incubate on ice for 30 min.

[0087] 4. Heat shock at 42 °C for 45 s, then immediately place on ice for 3 min.

[0088] 5. Add 900 μL LB without antibiotics, and shake at 250 rpm and 37 °C for 1 h.

[0089] 6. Pre-warm the LB solid medium with the corresponding antibody in a 37 °C incubator.

[0090] 7. Centrifuge at 50,000 rpm for 5 min, and remove 900 μL supernatant in a clean bench. Resuspend the bacterial cells with the remaining medium, and spread evenly on the plate with the correct resistance using a sterile spreader.

[0091] 8. Incubate in a 37 °C constant temperature incubator for 16 h.

[0092] 9. Pick a single colony of the transformed bacteria on the plate, and add 5 mL LB liquid medium with antibiotics to a test tube, and place it in a 37 °C constant temperature shaker at 180 rpm for 12 h.

[0093] 10. Take part of the bacterial solution to purify the plasmid using a plasmid purification kit, and use PCR and first-generation sequencing techniques to verify whether the target gene is successfully and correctly connected to the plasmid vector. If successful, mix the remaining bacterial solution with 40% glycerol at a ratio of 1:1, and store in a -80 °C refrigerator for long-term preservation.

[0094] (5) Construct the pTA7001-SCRE10-FLAG recombinant overexpression vector.

[0095] Based on SCRE10 Design the primers for PCR, and the forward primer and reverse primer are shown in SEQ ID NO. 5 and SEQ ID NO. 6. Use the pUC19-35S-SCRE10-FLAG obtained in step (4) as a template for amplification.

[0096] SEQ ID NO. 5: GACTCTAGCCTCGAGATGAAGTTTTCGATCGTCGT.

[0097] SEQ ID NO. 6: CCTGGATCGACTAGTTCACTTATCGTCATCGTCCT.

[0098] Connect the amplification product to the pTA7001 vector digested by Xho I andSpe I After double digestion, the recombinant overexpression vector pTA7001-SCRE10-FLAG was obtained, and the successfully constructed recombinant overexpression vector was transformed into Agrobacterium EHA105 to obtain a recombinant positive Agrobacterium.

[0099] (6) A method for constructing a transgenic rice plant.

[0100] ①Induction of rice callus: After shelling, the mature rice seeds were placed in a sterile triangular flask, treated with 75% ethanol for 3 min, the waste liquid was poured out, 50% sodium hypochlorite solution was poured in, and the flask was shaken at 150 rpm at room temperature for 40 min. Then the waste liquid was poured out, and the seeds were dried on filter paper after being washed with sterile water for 12 times. Then the seeds were transferred to the induction medium and cultured at 28°C under light for 4 weeks. Note that all operations should be sterile.

[0101] ②Subculture: The grown rice callus was carefully picked and transferred to the subculture medium and cultured at 28°C under light for 2 weeks. At this time, the rice callus was golden yellow and granular.

[0102] ③Agrobacterium infection:

[0103] The successfully constructed recombinant overexpression vector was transformed into Agrobacterium EHA105 strain. The specific transformation method was as follows: 2 μL of the recombinant overexpression vector was added to 50 μL of EHA105 competent cells, which were placed on ice for 30 min, treated with liquid nitrogen for 1 min, and then placed in a 37°C water bath for 90 s. After being placed on ice for 3 min, 500 μL of LB medium was added. After being shaken at 28°C and 180 rpm for 4 h, the medium was coated on a LA plate resistant to Rif and the vector. The plate was cultured at 28°C for 3 d.

[0104] The grown Agrobacterium single colony was picked and cultured in LB medium containing the corresponding antibiotic at 28°C overnight. After centrifugation at 4000 rpm for 10 min, the bacterial cells were collected and resuspended in 1 mL of AAM medium containing 150 μM As. Then, an appropriate amount of bacterial suspension was added to 30 mL of AAM, and the bacterial concentration was adjusted to OD600=0.2.

[0105] The callus subcultured in several dishes was transferred to a sterile triangular flask, and the prepared Agrobacterium bacterial solution was added to completely immerse the callus in the bacterial solution. The flask was shaken at 28°C for 15 min.

[0106] The bacterial solution was poured out, and the callus was dried on sterile filter paper for 30 min and then transferred to the co-culture medium and cultured at 28°C in the dark for 3 d.

[0107] (4) Screening of the resistant callus: the rice callus after 3 days of dark culture was picked up in a sterile empty bottle, and repeatedly washed with sterile water for 12 times. The last time of washing, 400 pg / mL of phosphinothricin was added in the water. Then the callus was dried on sterile filter paper and transferred to the selection medium, and cultured at 28°C for 3 weeks.

[0108] (5) The resistant callus grown on the selection medium was picked up and cultured on the differentiation medium at 28°C for 4 weeks to grow seedlings.

[0109] (6) The grown seedlings were transferred to the rooting tube containing the rooting medium, and cultured for 2 weeks to grow root system.

[0110] (7) The rice seedlings with healthy root system were covered with sterile water to grow vertically, and then transferred to the greenhouse or field.

[0111] (8) The seeds of T1 generation were shelled and sterilized, and cultured on the 1 / 2MS medium containing 50 pg / mL of hygromycin for 1 week. The rooting and germination rate was counted to determine whether it was single copy insertion. The leaf of the rice with rooting and germination was taken to extract protein for western blot detection of protein expression.

[0112] (9) The T1 generation seeds with single copy insertion and protein expression were selected for breeding. The harvested T2 generation seeds were shelled and sterilized, and cultured on the 1 / 2MS medium containing 50 pg / mL of hygromycin for 1 week. The rooting and germination rate was counted, and the protein expression was verified by western blot to obtain the rice transgenic plants.

[0113] SCRE10 The base sequence of the gene is shown in SEQ ID NO. 1, and the amino acid sequence is shown in SEQ ID NO. 2.

[0114] SEQ ID NO. 1:

[0115] ATGAAGTTTTCGATCGTCGTCCCAGCGGCCTTTGCGGCTGTCGCTCAAGCCGCCCCCCGCCCCCCCGGCGACCCCGACACCTCGGTGGCTTACCGCGGCTGCGAGGGGCCTCCGAACGGATTTCGGCCAGAAGACTGCCTCTTCTTTTGCAAGACCAACGTCATCAGGGTCAGGTGCGGTTGCAGAATGAGCATGGGAGTGTACAAGTGCGACGTGGGCACTGGGGAAAAGCTGAAAAAGTGCTGGGATGAGTCCGAGAAATACTGCATGGATCTCATCATGCCTTCTGCCAGGCACAAAGCTTGA.

[0116] SEQ ID NO.2:

[0117] MKFSIVVPAAFAAVAQAAPRPPGDPDTSVAYRGCEGPPNGFRPEDCLFFCKTNVIRVRCGCRMSMGVYKCDVGTGEKLKKCWDESEKYCMDLIMPSARHKA, * is the stop codon.

[0118] 2. Heterologous expression SCRE10 Inducing PTI immune response in rice.

[0119] To explore SCRE10 Regarding its role in rice immunity, this invention utilizes an Agrobacterium-mediated transgenic construction method to heterologously express [the gene] in rice. SCRE10 Successfully constructed a dexamethasone-induced expression... SCRE10 Transgenic rice lines IE-300 and IE-301. For example... Figure 1 As shown in Figure A, SCRE10 protein expression was undetectable in the IE-300 and IE-301 lines without DEX induction, indicating no leakage in these two lines. However, SCRE10 protein expression was detected after DEX induction. In conclusion, the IE-300 and IE-301 transgenic lines can be used for subsequent experiments.

[0120] Furthermore, the present invention detected SCRE10 The effect on PTI immunity in rice was investigated. Firstly, [the following was tested]. SCRE10 flg22 and chitin-induced in rice PR Effects on gene expression. Results showed that after treatment with flg22 or chitin, OsPR5 and OsPR10a Significantly upregulated expression. Figure 1 middle, OsActin As an internal reference gene, Mock treatment involves replacing flg22 or chitin with an equal volume of ddH2O. Different letters indicate significant differences in gene expression levels. P <0.01; Duncan's multiple comparison method.

[0121] Next, tests were conducted. SCRE10 The effects of flg22-induced oxygen burst in rice were investigated. Experimental results showed that flg22 or chitin treatment induced a burst of reactive oxygen species, with significant upregulation compared to the wild type. In summary, heterologous expression... SCRE10 It can induce immunity in rice. Results are shown below. Figure 2 .

[0122] 3. Heterologous expression SCRE10Rice lines have increased resistance to rice false smut.

[0123] In order to investigate SCRE10 The effect on disease resistance in rice, in heterologous expression SCRE10 The transgenic lines were inoculated with rice false smut fungus. The results showed that, compared to the wild type, DEX-induced expression significantly improved rice false smut. SCRE10 The transgenic lines produced a significantly reduced number of rice blast balls, indicating induced expression. SCRE10 Transgenic rice showed significantly enhanced resistance to rice false smut, as shown in the results. Figure 3 . Figure 3 In B, different letters indicate that the number of rice koji balls differs significantly after Mock or DEX processing.

[0124] 4. Heterologous expression SCRE10 Rice lines showed increased resistance to Bacillus subtilis.

[0125] In order to investigate SCRE10 The effect of Nip and heterologous expression on resistance to bacterial diseases in rice. SCRE10 Transgenic lines were used to treat rice pathogens caused by *Rhizoctonia solani* (bacterium blight). Xanthomonas oryzae pv . oryzae PXO99 A Inoculation, after DEX-induced expression SCRE10 The transgenic lines showed a significantly shorter lesion length compared to rice lines treated with other methods, as shown in the results below. Figure 4 This indicates induced expression. SCRE10 Genetically modified rice can reduce the infection of Blight Coccidia elegans and enhance the disease resistance of rice.

[0126] The technical features of the above embodiments can be combined in any way. For the sake of brevity, not all possible combinations of the technical features in the above embodiments are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.

[0127] The embodiments described above are merely examples of several implementations of the present invention, and while the descriptions are relatively specific and detailed, they should not be construed as limiting the scope of the invention. It should be noted that those skilled in the art can make various modifications and improvements without departing from the concept of the present invention, and these modifications and improvements all fall within the scope of protection of the present invention.

Claims

1. A gene for improving disease resistance of rice, characterized by, SCRE10 The SCRE10 The base sequence of the gene is shown as SEQ ID NO. 1, and the amino acid sequence is shown as SEQ ID NO.

2. ​ 2. The method of claim 1 SCRE10 Use of a gene, characterized in that, Said SCRE10 genes are used to increase the disease resistance of rice; the disease resistance refers to the resistance to sheath blight and / or white leaf blight bacteria; The application refers to... SCRE10 The gene was transformed into rice and expressed in rice. SCRE10 Genes are used to prepare transgenic rice plants, thereby improving the disease resistance of rice.

3. Use according to claim 2, wherein the compound is ###0002### The preparation method of the transgenic plant is as follows: The cDNA of rice sheath blight fungus was used as a template to amplify and obtain SCRE10 genes; The SCRE10 gene into the expression vector after enzyme digestion, to obtain a recombinant overexpression vector; The recombinant overexpression vector is transformed into Agrobacterium to obtain the recombinant positive Agrobacterium; The transgenic plant is obtained by infecting the rice with the recombinant positive Agrobacterium.

4. Use according to claim 3, wherein the compound is ###0002### The expression vector is pTA7001.

5. The use according to claim 3, wherein the compound is ###0002### The smut fungus is P1FZ.

6. The use according to claim 3, wherein the compound is ###0003### The restriction enzymes used to cut the expression vector are Xho I and Spe I .

7. The use according to claim 3, wherein the compound is ###00003### 3 The Agrobacterium is EHA105.

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