Application of rice disease resistance related gene Os04g55720 in bacterial leaf blight breeding

Abstract

The invention discloses application of a rice disease resistance related gene Os04g55720 in bacterial leaf blight breeding, and relates to the technical field of modern biotechnology gene engineering. The rice disease resistance related gene Os04g55720 disclosed by the invention has a nucleotide sequence as shown in SEQ ID NO.1, and consists of 2821 nucleotides; the sequence of the coding region is shown in SEQ ID NO.2, and the length of the sequence is 2148 nucleotides; a protein with 613 amino acids is coded, and the amino acid sequence is shown in SEQ ID NO.3. According to the invention, molecular biology and biochemical technologies are utilized to prove that overexpression of the Os04g55720 enhances the resistance of rice to the bacterial leaf blight, which indicates that the rice protein OsPGDH-3 can effectively control the incidence rate of the rice bacterial leaf blight at a quite low level. The invention provides a new thought, a new strategy and a new gene resource for green prevention and control of rice bacterial blight.

Description

Technical Field

[0001] The invention relates to the technical field of molecular biology, and in particular to application of gene Os04g55720 in rice bacterial blight resistance breeding. Background Art

[0002] Rice is one of the most important food crops for human beings and has a long history of cultivation and consumption in my country. According to statistics, the rice planting area in my country accounts for about 35% of the total planting area of ​​food crops, and the output accounts for 30% of the total domestic grain output. Rice bacterial blight caused by Xanthomonas oryzae pv.oryzae (Xoo) is a common and important disease in rice cultivation worldwide. Due to the use of disease-resistant varieties, rice bacterial blight disappeared in my country for a period of time. However, in recent years, the disease has shown a trend of worsening year by year, especially in some local areas. At present, the prevention and control of rice bacterial blight mainly relies on traditional chemical agents, but its environmental protection and drug resistance problems are becoming increasingly serious. The most economical and effective means to prevent and control rice bacterial blight is to use the disease resistance of rice itself.

[0003] In the early stage of the present invention, the laboratory analyzed a large number of gene chip data of rice infected by Xanthoceras oryzae (Xoo) and Magnaporthe oryzae (Mor), and found that the rice Os04g55720 gene strongly responded to Xoo infection and up-regulated expression. Based on this result, the laboratory further studied and found that the rice Os04g55720 gene responded strongly to Xoo induction and up-regulated expression. Therefore, in order to achieve disease-resistant and high-yield rice breeding, obtaining genes related to resistance to rice bacterial blight is of great significance for improving resistance to rice bacterial blight and cultivating new disease-resistant and high-yield rice varieties. Summary of the invention

[0004] In order to solve the problems existing in the prior art and provide new gene resources for disease-resistant and high-yield rice breeding, the present invention aims to provide the application of rice disease-resistant related gene Os04g55720 and its encoded protein in rice bacterial blight resistance breeding.

[0005] To achieve the above purpose, the technical solution adopted by the present invention is as follows:

[0006] The rice disease resistance-related gene Os04g55720, whose nucleotide sequence is shown in SEQ ID NO.1; the rice disease resistance-related gene Os04g55720, whose coding region sequence is shown in SEQ ID NO.2; and the encoded protein of the rice disease resistance-related gene Os04g55720, whose amino acid sequence is shown in SEQ ID NO.3.

[0007] Furthermore, the present invention comprises a recombinant expression vector, an expression cassette, a transgenic cell line or a genetically engineered bacterium of the rice disease resistance-related gene Os04g55720.

[0008] Furthermore, the rice Os04g55720 gene, the protein and recombinant expression vector, expression cassette, transgenic cell line or genetically engineered bacteria described above are used in the following (A1)-(A5):

[0009] (A1) Application in regulating rice resistance to bacterial blight;

[0010] (A2) Use in the preparation of a product for regulating resistance of rice to bacterial blight;

[0011] (A3) Application in breeding rice with high resistance to bacterial blight;

[0012] (A4) Use in the preparation of a product having high resistance to bacterial blight;

[0013] (A5) Application in breeding of rice with high resistance to bacterial blight;

[0014] Wherein, the regulation comprises the following (B1) or (B2):

[0015] (B1) Improving the resistance of rice to bacterial blight by up-regulating the expression level of the Os04g55720 gene, or up-regulating the content or activity of the protein encoded by the Os04g55720 gene.

[0016] (B2) Reducing the resistance of rice to bacterial blight pathogen by inhibiting the expression of the Os04g55720 gene, or inhibiting the content or activity of the protein encoded by the Os04g55720 gene.

[0017] Preferably, in the application of breeding rice with high resistance to bacterial blight, rice strains resistant to bacterial blight are obtained by screening rice plants that efficiently express the Os04g55720 gene.

[0018] Furthermore, the present invention provides a method for regulating rice resistance to bacterial blight, wherein knocking out the Os04g55720 gene in the rice will reduce the resistance of the rice to bacterial blight; overexpressing the Os04g55720 gene in the rice will increase the resistance of the rice to bacterial blight;

[0019] Furthermore, the present invention provides a method for creating transgenic rice resistant to bacterial blight, wherein the rice Os04g55720 gene is transferred into rice plants to obtain transgenic rice that efficiently expresses the rice Os04g55720 gene.

[0020] Furthermore, the coding region sequence of the rice Os04g55720 gene was cloned into a plant expression vector, first transferred into Agrobacterium, and then transferred into rice cells through callus transformation to obtain transgenic rice that efficiently expressed the rice Os04g55720 gene.

[0021] Beneficial effects of the present invention:

[0022] The present invention has found that the rice Os04g55720 gene plays an important role in the resistance of rice to bacterial blight. After knocking out the Os04g55720 gene, the rice reduced its disease resistance to bacterial blight; while overexpressing the Os04g55720 gene increased the disease resistance of rice to bacterial blight. Therefore, overexpressing the rice Os04g55720 gene can be used to improve the resistance of rice to bacterial blight. Moreover, there is no significant difference in plant height, tiller number, seed length, seed width, and seed thickness between the rice strains overexpressing the Os04g55720 gene and the wild type. Therefore, the present invention can be used as a rice germplasm resource and intermediate material, and is of great significance for rice breeding for resistance to bacterial blight. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] Figure 1 The figures show the resistance phenotypes of rice Os04g55720 gene knockout mutants (including three strains: KO 1-3, KO 2-6 and KO 3-7) and overexpression plants (including three strains: OE 4-2, OE 4-5 and OE 4-8) to bacterial blight. Figure A shows the lesion of two-week-old transgenic material 14 days after leaf inoculation, with a scale of 1 cm. Figure B shows the statistics of lesion lengths of wild-type and mutant materials.

[0024] Figure 2 These are the rice Os04g55720 gene knockout mutant, overexpression plants and wild-type Nipponbare plants in the mature stage; Figure A: wild-type Nipponbare; Figure B: rice Os04g55720 gene knockout mutant; Figure C: rice Os04g55720 gene overexpression plant.

[0025] Figure 3 Agronomic traits of rice Os04g55720 gene knockout mutants, overexpression plants and wild-type Nipponbare plants; Figure A: seed size; Figure B: seed length, seed width and seed thickness (n=10); Figure C: plant height (cm); D: number of tillers; E: 1000-grain weight (g). DETAILED DESCRIPTION

[0026] In order to illustrate the present invention in detail, the following examples will help those skilled in the art to further understand the present invention, but are not intended to limit the present invention in any form. It should be noted that the embodiments are preferred embodiments of the present invention and are intended to illustrate the implementation conditions that can be used to implement the present invention, rather than to limit the experimental conditions.

[0027] The present invention provides a rice disease resistance-related gene Os04g55720, wherein the coding region sequence of the rice disease resistance-related gene Os04g55720 is shown in SEQ ID NO. 2. The sequencing result shows that the CDS sequence contains a stop codon of 1842 bp in length and encodes a protein of 613 amino acids. Studies have found that overexpression of the Os04g55720 gene can improve the ability of rice to resist bacterial blight.

[0028] The invention provides a recombinant expression vector pCAMBIA1300-Os04g55720 containing the rice disease resistance-related gene Os04g55720.

[0029] The present invention provides a recombinant engineering bacterium containing the rice disease resistance-related gene Os04g55720.

[0030] The present invention also proposes the use of the rice disease resistance-related gene Os04g55720 in improving the resistance of rice to bacterial blight.

[0031] In a preferred embodiment of the present invention, the application as described above comprises the following steps:

[0032] The coding region sequence of the rice Os04g55720 gene was cloned into the pCAMBIA1300 vector to obtain a recombinant expression vector;

[0033] After the recombinant expression vector is transformed into Agrobacterium, the transformed Agrobacterium is used to infect and transform rice callus tissue to further obtain transgenic rice plants.

[0034] In a preferred embodiment of the present invention, the rice variety is Nipponbare. The Os04g55720 gene of the present invention can also be used to transform other varieties of rice to obtain transgenic rice plants with resistance to bacterial blight, which is not specifically limited in the present invention.

[0035] The features and performance of the embodiments of the present invention are further described in detail below in conjunction with the accompanying drawings.

[0036] Example 1: Obtaining Rice Os04g55720 Gene Knockout Mutant Material

[0037] The amino acid sequence of rice Os04g55720 protein is shown in SEQ ID NO. 3, and the coding sequence (CDS sequence) of rice Os04g55720 gene is shown in SEQ ID NO. 2. The present invention uses CRISPR / Cas9 technology to knock out the target gene in a targeted manner, thereby obtaining a knockout rice mutant of the Os04g55720 gene.

[0038] The steps for constructing the knockout vector are as follows:

[0039] (1) First, based on the sequence of the Os04g55720 gene, two target sites were selected and primers 55720-Y1-F / R and 55720-B1-F / R were designed. The primer sequences are as follows:

[0040]

[0041] (2) Preparation of gRNA fragments

[0042] The reaction system for preparing the gRNA fragment of the target site Y1 / B1 is as follows:

[0043]

[0044] The PCR reaction conditions were:

[0045] 95℃ 10min

[0046] 55℃ 10min

[0047] 14℃ 5min

[0048] (3) Enzyme digestion to construct a recombinant plasmid containing gRNA

[0049] Using the yl-cas9 vector and restriction endonuclease Bsa I, a recombinant plasmid containing the target site Y1 / B1 was constructed. The enzyme cutting and ligation system is as follows:

[0050]

[0051] The configured system was placed in a 37°C incubator for 2 hours, and two recombinant plasmids were finally obtained: 55720-Y1 and 55720-B1. The two plasmids were transformed into E. coli using the heat shock method to obtain E. coli containing plasmid 55720-Y1 and E. coli containing 55720-B1, and positive clones were selected for detection.

[0052] (4) Construction of dual-target recombinant plasmid

[0053] Plasmids 55720-Y1 and 55720-B1 were extracted from E. coli using a kit, and then digested and ligated using a dual-target system. The system is as follows:

[0054]

[0055] The configured system was placed in a 37°C incubator for 2 hours, and the dual-target recombinant plasmid yl-cas9-55720 was finally obtained. The two plasmids were transformed into E. coli by heat shock method, and positive clones were selected for detection. After the sequencing was correct, the plasmid was used for rice callus transformation, and the rice Os04g55720 gene knockout mutant was obtained with the Nipponbare strain as the background.

[0056] Example 2: Obtaining rice Os04g55720 gene overexpression material.

[0057] (1) Design a pair of specific primers based on the sequence information of the Os04g55720 gene:

[0058]

[0059] (2) extracting total DNA from rice Nipponbare leaves as a PCR reaction template, performing PCR amplification with the above primers, and cloning and sequencing the PCR products, specifically comprising the following steps:

[0060] The plant genomic DNA was extracted using the CATB (cetyltrimethylammonium bromide) method. PCR was performed using primers 55720-F and 55720-R, and the PCR products were sequenced. The PCR reaction system was as follows:

[0061]

[0062] The PCR reaction conditions were:

[0063]

[0064] The sequencing results showed that the CDS of rice Os04g55720 gene was 2148bp long and encoded a protein of 613 amino acids.

[0065] (3) Connecting the obtained Os04g55720 gene to the pCAMBIA1300 vector comprises the following steps:

[0066] The sequence amplified by primers 55720-F and 55720-R was connected to the pCAMBIA1300 vector backbone after double digestion with XbaⅠ and ScaⅠ through the Novozyme recombination kit, and then screened by kanamycin plate, and then identified by colony PCR and sequenced. The recombinant plasmid with the correct sequence was used for rice callus transformation, and rice Os04g55720 gene overexpression plants were obtained with the Nipponbare strain as the background.

[0067] Example 3: Identification of transgenic plants for resistance to bacterial blight

[0068] (1) Rice seedling cultivation: Soak rice seeds in prochloraz for 24 hours and then rinse with clean water. Soak the seeds in clean water for 2-3 days, changing the clean water every 12 hours. Plant the rice seedlings in the rice seedling cultivation medium after the rice turns white. After two weeks, move the rice to flooded soil that has been fertilized with basal fertilizer. Fertilizers are urea and potassium dihydrogen phosphate, 0.5 grams each per pot.

[0069] (2) Pathogen infection: For seedlings at the three- to four-leaf stage, the leaf clipping inoculation method was used. Xoo in NA medium was suspended in 10 mM sterilized MgCl 2 Solution (pH = 7.0) to OD 600 = 0.5, dipped in Xoo suspension with sterilized scissors, cut the leaf tip (about 2-3 cm) from the leaf. After shading for 24 hours, grow under the conditions of 28-32℃ (light, 12 hours), 28-32℃ (dark, 12 hours), and 90% relative humidity, and measure the lesion size after 12 days.

[0070] (3) Disease resistance phenotype determination: Figure 1 Figure A shows a comparison of leaves of wild-type Nipponbare and transgenic Nipponbare rice 14 days after inoculation with Xoo. Figure 1 It can be seen that 14 days after inoculation with Xoo, the length of the lesions on the leaves of knockout (KO-Os04g55720) rice was significantly greater than that on the leaves of the wild-type Nipponbare, while the length of the lesions on the leaves of overexpression (OE-Os04g55720) rice was significantly smaller than that on the leaves of the wild-type Nipponbare.

[0071] In terms of agronomic traits, there were no significant differences in plant height, tiller number, seed length, seed width, and seed thickness between the rice lines overexpressing the Os04g55720 gene and the wild type, and the thousand-grain weight of the seeds was slightly reduced by about 1 g compared with the wild type ( Figure 3 ).

[0072] The above results show that the rice disease resistance-related gene Os04g55720 can significantly regulate the resistance of rice to bacterial blight. The invention can be used as a rice germplasm resource and intermediate material for rice bacterial blight resistance breeding.

[0073] The above is only the optimal implementation mode of the present invention, but the protection scope of the present invention is not limited thereto. Any changes or substitutions that can be understood and thought of by any person familiar with the technical field within the technical scope disclosed by the present invention should be included in the scope of the present invention. Therefore, the protection scope of the present invention should be based on the protection scope of the claims.

[0074] Sequence Listing:

[0075] A rice disease resistance-related gene Os04g55720, consisting of 2833 nucleotides, and its nucleotide sequence is as SEQ ID NO.1:

[0076] ACCCACATCGCCGCCTCCTCCTCCTTCCTCGGTTTCCCCCAAAAACCCCAAACCCCTCCTCCTCGCCAT

[0077] GGCGGCGCCGTCCCAGACCACCGCCACCACCACCCACCACCGCGTCCTCCTCCCGTCCCACCACCACCA

[0078] CCGAGCGGTGCCTCCCTCGCTCCTCCGCCTCCCGCTCCGCGCCGCGCGCCGGGGCCGCCTCTCCGCCGC

[0079] CGCCGCGGCCGCGGCGCCCGCCGCGTCGACCGCGGCGCCGTCGGAGCCCGCCGCGGGTGCCGTACCGGG

[0080] GAAGCCGACGGTGCTCGTGGCCGAGAAGCTCGGCGCCGCGGGGCTCGAGCTGCTCCGGGGGTTCGCCAA

[0081] CGTGGACTGCTCCTACGGCCTCTCCCCCGAGGAGCTCCGCGCCAAGATCCCGCTCTGCGACGCGCTCAT

[0082] CGTGCGGTCGGGGACCAAGGTGGGGCGCGACGTGTTCGAGGCATCCGGGGGAAGGCTCCGCGTCGTGGG

[0083] GCGCGCCGGGGTCGGGATCGACAACGTCGATCTCGCCGCCGCCACCGAGCACGGGTGCCTCGTCGTCAA

[0084] CGCACCCACGGCGAACACCGTCGCCGCGGCCGAGCACGGCATCGCGCTGCTCACCGCTATGTCCAGGAA

[0085] CATCGCCCAGGCCGACGCGTCGCTCAAGGCTGGTGAGTGCGCACACCCTCCTCAAGTCTGGAAAAGACA

[0086] CCTCTTTAAAAAATGATACTGTACTTGTCTCGATCTCTGTTTAAAGCATTAAAAAAATCTTGCATCAAC

[0087] GTGGCACCTGTAACAGTGGATCGTGCTAGAACGTCTGTTTGATGAATTGCTAGCAAATCGAGAAACACG

[0088] AATGGGTGCTTTTTCACTAGAAGCAGTCCTCAGAGATTACTTTCGTTACTTCATTCAATTTGACATAAT

[0089] CAGAAAAAGTTAATATCTGCTTCAGTTGACAAAGCTAGATGCATGAAGATGTGGAGTAAGG

[0090] GTAGTCTTAGATGAAGCATCTGCCATGCAATCTTCGAATGGCATCTTCGGAAGAAGGGCAACATACCTT

[0091] TTCAATTATGAGTTCCCATGTTCTTAACAATGTGTCTTGTATAGGTAAATGGCAGCGCAACAAGTA

[0092] TGTGGGTGTATCTCTCGTTGGTAAAACTATTGCAATTCTTGGATTTGGAAAGGTTGGGTCAGAAGTCGC

[0093] TCGTCGTGCTAAAGGTTTGGGAATGCATGTGATTGCACACGATCCGTATGCTTCTGCTGATCGTGCCCA

[0094] TGCAATTGGAGTTGAGCTAGTGAGCATGGAAGACGCTTTGACAACTGCTGACTTTATTCTCGTTGCATAT

[0095] GCCTCTTACCCTGCAACAAACAAGATGCTCAATGATGAAACTTTCGCTAAGATGAAGAAGGGTGTTAG

[0096] AATTATAAATGTTGCGCGTGGTGGTGTAATTGATGAAGATGCTCTAGTCAGGGCTCTTGATTCAGGAAT

[0097] AGTTGCACAGGTATGTTTGATTCTCTTAGTCTACATCCTTTTTTCATTTCATGCTGACATCCGGTGAAT

[0098] TATTTTTGAACAGCTTGTTTCCCAGAGTCATGCTAATTTTTTTGTTTATCCCTCTGCTTTAGGCTGCTC

[0099] TTGATGTGTTCACTAAAGAGCCACCGGCACCAGACAGCAAATTAGTGCTGCATGAGAATGTTACTGTAA

[0100] CACCACACCTTGGTGCCAGCACAGTTGAAGCACAGGTATGCGAATTAGCTCAGAATGCTATCCCTTTGT

[0101] GTTCTGTTATACTAGATACCAAAATGATAAATATCTGAAAATCCCAGGAAGGAGTGGCTATTGAAATAG

[0102] CTGAAGCTGTTATTGGAGCTCTGAAAGGGGAACTTGCAGCTTCAGCAGTCAACGCACCAATGGTTCCTG

[0103] CTGAGGTATGAATCTATTGTTTTCTGCTAAATCCAACTTATTCCACATGGCATTTGATTTTTATCTTAC

[0104] ATCCTTGTGATTTGTTCAGGTGCTGTCAGAGCTTGCACCTTATGTTGTGCTCGCAGAGAAGCTTGGGCG

[0105] CCTGGCTGTGCAGCTAGTGGCTGGTGGCGGTGGTATCAAGTCCGTGAAAGTGACCTATGCTTCTGCAAG

[0106] GGCTCCTGATGATCTTGACACGAGACTTCTTCGTGCAATGATCACCAAGGGGTTGATCGAACCAATCTC

[0107] CAGTGTTTTCGTCAATCTGGTCAATGCTGACTTCACTGCGAAACAGAGGGGAGTTCGTATCACTGAGGA

[0108] GAGAATCTTGTTGGATGGCTCACCCGAGACACCTATTGACTACATCCAAGTTCATATTGCCCATGTTGA

[0109] GTCCAAATTTCCCAGTGCAATATCGGAGAGTGGAGAGATCACTGTTGAGGGGAAGGTGAAGGATGGCAT

[0110] CCCCCATCTGACAAAGGTTGGATCATTCCAGGTTGATGTGAGCTTGGAAGGAAGCCTGATCCTTTGCAG

[0111] GCAGGTCGATCAACCTGGTATGATCGGCGCAGTAGGAAGTGTCCTTGGTGAGGAGAATGTTAATGTCAG

[0112] TTTCATGAGTGTTGGAAGAATCGCTCCTCGCAAGCATGCCGTCATGGCGATTGGTGTTGATGAGGAACC

[0113] TAAAAAGAGCACACTGACAAAGATTGGGGAGATTCCGGCGATTGAAGAATTCGTTTTCCTCAAGCTCTA

[0114] GATTCATGCGTTGTATGTTTGAAGCTGCAAACATGGTGTGCAATTTTGATGTAGCTAGCTATTTAATCA

[0115] GGATCTAGAGGACTGATGGTTCTCGTTGGAATAAGCCTTTTTTGGGTTATGGTGCTGTTTGTCGAGTTT

[0116] GATGCAGTGTCTTGATTAATCTGGCTTAGGTTTGAACATGGTGCTTATTAATAAGAAGCCCTCTGTTAA

[0117] TCTA

[0118] A rice disease-resistant related gene Os04g55720, its coding region consists of 1842 nucleotides, and its nucleotide sequence is as shown in SEQ ID NO.2:

[0119] ATGGCGGCGCCGTCCCAGACCACCGCCACCACCACCCACCACCGCGTCCTCCTCCCGTCCCACCACCAC

[0120] CACCGAGCGGTGCCTCCCTCGCTCCTCCGCCTCCCGCTCCGCGCCGCGCGCCGGGGCCGCCTCTCCGCC

[0121] GCCGCCGCGGCCGCGGCGCCCGCCGCGTCGACCGCGGCGCCGTCGGAGCCCGCCGCGGGTGCCGTACCG

[0122] GGGAAGCCGACGGTGCTCGTGGCCGAGAAGCTCGGCGCCGCGGGGCTCGAGCTGCTCCGGGGGTTCGCC

[0123] AACGTGGACTGCTCCTACGGCCTCTCCCCCGAGGAGCTCCGCGCCAAGATCCCGCTCTGCGACGCGCTC

[0124] ATCGTGCGGTCGGGGACCAAGGTGGGGCGCGACGTGTTCGAGGCATCCGGGGGAAGGCTCCGCGTCGTG

[0125] GGGCGCGCCGGGGTCGGGATCGACAACGTCGATCTCGCCGCCGCCACCGAGCACGGGTGCCTCGTCGTC

[0126] AACGCACCCACGGCGAACACCGTCGCCGCGGCCGAGCACGGCATCGCGCTGCTCACCGCTATGTCCAGG

[0127] AACATCGCCCAGGCCGACGCGTCGCTCAAGGCTGGTAAATGGCAGCGCAACAAGTATGTGGGTGTATCT

[0128] CTCGTTGGTAAAACTATTGCAATTCTTGGATTTGGAAAGGTTGGGTCAGAAGTCGCTCGTCGTGCTAAA

[0129] GGTTTGGGAATGCATGTGATTGCACACGATCCGTATGCTTCTGCTGATCGTGCCCATGCAATTGGAGTT

[0130] GAGCTAGTGAGCATGGAAGACGCTTTGACAACTGCTGACTTTATCTCGTTGCATATGCCTCTTACCCCT

[0131] GCAACAAACAAGATGCTCAATGATGAAACTTTCGCTAAGATGAAGAAGGGTGTTAGAATTATAAATGTT

[0132] GCGCGTGGTGGTGTAATTGATGAAGATGCTCTAGTCAGGGCTCTTGATTCAGGAATAGTTGCACAGGCT

[0133] GCTCTTGATGTGTTCACTAAAGAGCCACCGGCACCAGACAGCAAATTAGTGCTGCATGAGAATGTTACT

[0134] GTAACACCACACCTTGGTGCCAGCACAGTTGAAGCACAGGAAGGAGTGGCTATTGAAATAGCTGAAGCT

[0135] GTTATTGGAGCTCTGAAAGGGGAACTTGCAGCTTCAGCAGTCAACGCACCAATGGTTCCTGCTGAGGTG

[0136] CTGTCAGAGCTTGCACCTTATGTTGTGCTCGCAGAGAAGCTTGGGCGCCTGGCTGTGCAGCTAGTGGCT

[0137] GGTGGCGGTGGTATCAAGTCCGTGAAAGTGACCTATGCTTCTGCAAGGGCTCCTGATGATCTTGACACG

[0138] AGACTTCTTCGTGCAATGATCACCAAGGGGTTGATCGAACCAATCTCCAGTGTTTTCGTCAATCTGGTC

[0139] AATGCTGACTTCACTGCGAAACAGAGGGGAGTTCGTATCACTGAGGAGAGAATCTTGTTGGATGGCTCA

[0140] CCCGAGACACCTATTGACTACATCCAAGTTCATATTGCCCATGTTGAGTCCAAATTTCCCAGTGCAATA

[0141] TCGGAGAGTGGAGAGATCACTGTTGAGGGGAAGGTGAAGGATGGCATCCCCCATCTGACAAAGGTTGGA

[0142] TCATTCCAGGTTGATGTGAGCTTGGAAGGAAGCCTGATCCTTTGCAGGCAGGTCGATCAACCTGGTATG

[0143] ATCGGCGCAGTAGGAAGTGTCCTTGGTGAGGAGAATGTTAATGTCAGTTTCATGAGTGTTGGAAGAATC

[0144] GCTCCTCGCAAGCATGCCGTCATGGCGATTGGTGTTGATGAGGAACCTAAAAAGAGCACACTGACAAAG

[0145] ATTGGGGAGATTCCGGCGATTGAAGAATTCGTTTTCCTCAAGCTCTAG

[0146] The protein sequence encoded by a rice defense gene Os04g55720, which consists of 613 amino acids, and its amino acid sequence is as shown in SEQ ID NO.3:

[0147] MAAPSQTTATTTHHRVLLPSHHHHRAVPPSLLRLPLRAARRGRLSAAAAAAAPAASTAAPSEPAAGAVP

[0148] GKPTVLVAEKLGAAGLELLRGFANVDCSYGLSPEELRAKIPLCDALIVRSGTKVGRDVFEASGGRLRVV

[0149] GRAGVGIDNVDLAAATEHGCLVVNAPTANTVAAAEHGIALLTAMSRNIAQADASLKAGKWQRNKYVGVS

[0150] LVGKTIAILGFGKVGSEVARRAKGLGMHVIAHDPYASADRAHAIGVELVSMEDALTTADFISLHMPLTP

[0151] ATNKMLNDETFAKMKKGVRIINVARGGVIDEDALVRALDSGIVAQAALDVFTKEPPAPDSKLVLHENVT

[0152] VTPHLGASTVEAQEGVAIEIAEAVIGALKGELAASAVNAPMVPAEVLSELAPYVVLAEKLGRLAVQLVA

[0153] GGGGIKSVKVTYASARAPDDLDTRLLRAMITKGLIEPISSVFVNLVNADFTAKQRGVRITEERILLDGS

[0154] PETPIDYIQVHIAHVESKFPSAISESGEITVEGKVKDGIPHLTKVGSFQVDVSLEGSLILCRQVDQPGM

[0155] IGAVGSVLGEENVNVSFMSVGRIAPRKHAVMAIGVDEEPKKSTLTKIGEIPAIEEFVFLKL。

Claims

1. Rice disease resistance-related gene Os04g55720, characterized in that: Its nucleotide sequence is shown in SEQ ID NO.

1.

2. The rice disease resistance-related gene Os04g55720 according to claim 1, characterized in that: The coding region sequence is shown in SEQ ID NO.

2.

3. The rice disease resistance-related gene Os04g55720 according to claim 1, characterized in that: The amino acid sequence of the encoded protein is shown in SEQ ID NO.

3.

4. Use of the rice disease resistance-related gene Os04g55720 according to any one of claims 1 to 3 in rice disease resistance.

5. The use according to claim 4, characterized in that: The application includes the application of a recombinant expression vector, an expression cassette, a transgenic cell line or a genetically engineered bacterium of the rice disease resistance-related gene Os04g55720 and a protein encoded by the gene in any one or more of (A1) to (A5): (A1) Application in regulating rice resistance to bacterial blight; (A2) Use in the preparation of a product for regulating resistance of rice to bacterial blight; (A3) Application in breeding rice with high resistance to bacterial blight; (A4) Application in breeding products of rice with high resistance to bacterial blight; (A5) Application in breeding of rice with high resistance to bacterial blight; Wherein, the regulation is as follows (B1) or (B2): (B1) improving the resistance of rice to bacterial blight by up-regulating the expression level of the Os04g55720 gene, or up-regulating the content or activity of the protein encoded by the Os04g55720 gene; (B2) Reducing the resistance of rice to bacterial blight by inhibiting the expression of the Os04g55720 gene, or inhibiting the content or activity of the protein encoded by the Os04g55720 gene.

6. The use according to claim 5, characterized in that: In the method (B1), the Os04g55720 gene in rice is overexpressed to improve the resistance of rice to bacterial blight; in the method (B2), the Os04g55720 gene in rice is knocked out to reduce the resistance of rice to bacterial blight.

7. A method for regulating rice resistance to bacterial blight, characterized in that: Knocking out the rice Os04g55720 gene reduces the resistance of rice to bacterial blight; overexpressing the rice Os04g55720 gene increases the resistance of rice to bacterial blight.

8. A method for constructing transgenic rice resistant to bacterial blight, characterized in that: The rice Os04g55720 gene was transferred into rice plants to obtain transgenic rice expressing the rice Os04g55720 gene at a high level.

9. The method for constructing a transgenic rice resistant to bacterial blight according to claim 8, characterized in that: The coding region sequence of rice Os04g55720 gene was cloned into a plant expression vector, firstly transferred into Agrobacterium, and then transferred into rice cells through callus transformation to obtain transgenic rice that efficiently expressed rice Os04g55720 gene.

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