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

Abstract

The application discloses application of a rice disease resistance related gene Os04g55720 in bacterial blight breeding and relates to the technical field of modern biotechnology and genetic engineering technology.The rice disease resistance related gene Os04g55720 has a nucleotide sequence as shown in SEQ ID NO.1, is composed of 2821 nucleotides, has a coding region sequence as shown in SEQ ID NO.2, and has a sequence of 2148 nucleotides; the gene encodes a protein with 613 amino acids, and has an amino acid sequence as shown in SEQ ID NO.3.The application proves that overexpression of Os04g55720 enhances the resistance of rice to bacterial blight by means of molecular biology and biochemical technology, and shows that the rice protein OsPGDH-3 can effectively control the incidence of rice bacterial blight at a relatively low level.The application provides a new idea, strategy and gene resource for green prevention and control of rice bacterial blight.

Description

Technical Field

[0001] This invention relates to the field of molecular biology technology, specifically to the application of gene Os04g55720 in rice bacterial blight resistance breeding. Background Technology

[0002] Rice is one of the most important food crops for humankind, with a long history of cultivation and consumption in my country. Statistics show that rice cultivation area accounts for about 35% of the total grain crop cultivation area in my country, and its yield accounts for 30% of the country's total grain output. Rice bacterial blight, caused by the pathogenic strain *Xanthomonas oryzae* pv. oryzae (Xoo), is a common and important disease in rice cultivation worldwide. Due to the use of 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 increasing severity year after year, especially in some localized areas. Currently, the control of rice bacterial blight mainly relies on traditional chemical agents, but the environmental impact and drug resistance issues are becoming increasingly serious. The most economical and effective means of controlling rice bacterial blight is to utilize the rice's own disease resistance.

[0003] In previous studies, our laboratory analyzed a large amount of gene chip data on rice infected by Xoo (bacterial blight pathogen) and Mor (rice blast pathogen), discovering that the rice Os04g55720 gene strongly responds to Xoo infection with upregulated expression. Based on this result, further research revealed that the rice Os04g55720 gene also exhibits a dramatic upregulated expression response to Xoo induction. Therefore, obtaining genes related to rice bacterial blight resistance is of great significance for improving rice resistance to bacterial blight and cultivating new disease-resistant, high-yielding rice varieties, in order to achieve high-yield and disease-resistant rice breeding. Summary of the Invention

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

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

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

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

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

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

[0010] (A2) Application in the preparation of products that regulate rice resistance to bacterial blight;

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

[0012] (A4) Application in the preparation of rice products with high resistance to bacterial blight;

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

[0014] The regulation includes either (B1) or (B2):

[0015] (B1) By upregulating the expression level of the Os04g55720 gene, or upregulating the content or activity of the protein encoded by the Os04g55720 gene, the resistance of rice to bacterial blight can be improved.

[0016] (B2) By inhibiting the expression level of the Os04g55720 gene, or by inhibiting the content or activity of the protein encoded by the Os04g55720 gene, the resistance of rice to bacterial blight can be reduced.

[0017] Preferably, in the application of rice breeding for high resistance to bacterial blight, rice lines 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 the resistance of rice to bacterial blight. Knocking out the Os04g55720 gene in rice will reduce the resistance of rice to bacterial blight; overexpressing the Os04g55720 gene in rice will increase the resistance of 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 expresses the rice Os04g55720 gene.

[0021] The beneficial effects of this invention are:

[0022] This invention reveals that the rice Os04g55720 gene plays a crucial role in rice resistance to bacterial blight. Knocking out the Os04g55720 gene reduces rice's resistance to the bacterial blight pathogen; conversely, overexpression of the Os04g55720 gene increases resistance. Therefore, overexpression of the rice Os04g55720 gene can enhance rice resistance to bacterial blight. Furthermore, rice lines overexpressing the Os04g55720 gene showed no significant differences in plant height, tiller number, seed length, seed width, and seed thickness compared to wild-type lines. Therefore, this invention can serve as a rice germplasm resource and intermediate material, which is of significant importance for breeding rice strains resistant to bacterial blight. Attached Figure Description

[0023] Figure 1 The bacterial blight resistance phenotypes of rice Os04g55720 gene knockout mutants (including three lines: KO 1-3, KO 2-6, and KO 3-7) and overexpression plants (including three lines: OE 4-2, OE 4-5, and OE 4-8) were studied. Figure A shows the lesion pattern of transgenic materials after two weeks of growth, 14 days after leaf cutting and inoculation, with a scale bar of 1 cm. Figure B shows the statistical analysis of lesion length in wild-type and mutant materials.

[0024] Figure 2 The images show mature rice plants with the Os04g55720 gene knockout mutant, overexpressing plants, and wild-type Nipponbare plants. Figure A shows wild-type Nipponbare; Figure B shows the Os04g55720 gene knockout mutant; and Figure C shows the Os04g55720 gene overexpressing plants.

[0025] Figure 3 Agronomic traits of rice Os04g55720 gene knockout mutant, overexpressing plants and wild-type Nipponbare plants; where, 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: thousand-grain weight (g). Detailed Implementation

[0026] To illustrate the content of this invention in detail, the following embodiments will help those skilled in the art to further understand the invention, but do not limit the invention in any way. It should be noted that the embodiments are preferred embodiments of the invention and are intended to illustrate the implementation conditions that can be used to carry out the invention, rather than limiting the experimental conditions.

[0027] This invention provides a rice disease resistance-related gene, Os04g55720, the coding sequence of which is shown in SEQ ID NO.2. Sequencing results show that its CDS sequence contains a 1842 bp stop codon encoding a protein of 613 amino acids. Studies have found that overexpression of the Os04g55720 gene can improve the resistance of rice to bacterial blight.

[0028] This invention proposes a recombinant expression vector pCAMBIA1300-Os04g55720 containing the above-mentioned rice disease resistance-related gene Os04g55720.

[0029] This invention proposes a recombinant engineered bacterium containing the aforementioned rice disease resistance-related gene Os04g55720.

[0030] This invention also proposes the application of the above-mentioned rice disease resistance-related gene Os04g55720 in improving rice resistance to bacterial blight.

[0031] In a preferred embodiment of the present invention, the application as described above includes 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 transforming the recombinant expression vector into Agrobacterium, the transformed Agrobacterium was used to infect and transform rice callus tissue to 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 rice varieties to obtain transgenic rice plants with resistance to bacterial blight; however, the present invention does not impose specific limitations on this.

[0035] The features and performance of the embodiments of the present invention will be further described in detail below with reference to the accompanying drawings.

[0036] Example 1: Obtaining rice Os04g55720 gene knockout mutant material

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

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

[0039] (1) 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 targeting site Y1 / B1 is as follows:

[0043]

[0044] The PCR reaction conditions are as follows:

[0045] 95℃ for 10 minutes

[0046] 55℃ for 10 minutes

[0047] 14℃ for 5 minutes

[0048] (3) Enzymatic digestion to construct recombinant plasmids containing gRNA

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

[0050]

[0051] The prepared system was incubated at 37℃ for 2 hours, resulting in two recombinant plasmids: 55720-Y1 and 55720-B1. The two plasmids were then transformed into *E. coli* using a heat shock method, yielding *E. coli* containing plasmid 55720-Y1 and plasmid 55720-B1, respectively. Positive clones were selected for testing.

[0052] (4) Constructing dual-target recombinant plasmids

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

[0054]

[0055] The prepared system was incubated at 37℃ for 2 hours to obtain the dual-target recombinant plasmid yl-cas9-55720. The two plasmids were transformed into *E. coli* using a heat shock method, and positive clones were selected for detection. After successful sequencing, the plasmids were transformed into rice callus tissue using the Nipponbare variety as a background to obtain the rice Os04g55720 gene knockout mutant.

[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) Total DNA was extracted from the leaves of Nipponbare rice and used as a template for PCR reaction. PCR amplification was performed using the primers described above, and the PCR products were cloned and sequenced. The specific steps included:

[0060] 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 is as follows:

[0061]

[0062] The PCR reaction conditions are as follows:

[0063]

[0064] Sequencing results showed that the CDS of the rice Os04g55720 gene is 2148 bp long and encodes a protein of 613 amino acids.

[0065] (3) The obtained Os04g55720 gene was ligated into the pCAMBIA1300 vector, including the following steps:

[0066] The sequences amplified by primers 55720-F and 55720-R were ligated to the pCAMBIA1300 vector backbone digested with XbaI and ScaI using a Novizan recombinant kit. After screening with kanamycin plates, colony PCR was performed for identification and sequencing. The recombinant plasmids with correct sequences were used for rice callus transformation, and plants overexpressing the Os04g55720 gene were obtained using the Nipponbare cultivar as the background.

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

[0068] (1) Rice seedling raising: Soak rice seeds in prochloraz for 24 hours, then rinse the seeds with clean water and soak them in clean water for 2-3 days, changing the water every 12 hours. After the rice seedlings show signs of sprouting, transplant them into the rice seedling substrate. Two weeks later, transplant the rice seedlings into flooded soil that has been fertilized with base fertilizer. Use urea and potassium dihydrogen phosphate, 0.5 grams of each per pot.

[0069] (2) Pathogen infection: For seedlings at the three- to four-leaf stage, use the leaf-cutting inoculation method, suspending Xoo in NA medium in 10mM sterile MgCl2 solution (pH=7.0) until OD. 600 =0.5, use sterile scissors to dip into Xoo suspension and cut the leaf tip (about 2-3 cm) off the leaf. After shading for 24 hours, grow under conditions of 28-32℃ (light, 12 hours) and 28-32℃ (darkness, 12 hours) and 90% relative humidity. Measure the size of the lesion after 12 days.

[0070] (3) Disease resistance phenotype determination: Figure 1 Figure A in the image shows a comparison of leaves of wild-type Nipponbare and transgenic Nipponbare rice 14 days after inoculation with Xoo1. From... Figure 1 It can be seen that 14 days after Xoo inoculation, the leaf lesion length of knockout (KO-Os04g55720) rice was significantly larger than that of wild-type Nipponbare rice, while the leaf lesion length of overexpressed (OE-Os04g55720) rice was significantly smaller than that of wild-type Nipponbare rice.

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

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

[0073] The above description is only the preferred embodiment of the present invention, but the protection scope of the present invention is not limited thereto. Any transformations or substitutions that can be conceived by those skilled in the art within the technical scope disclosed in the present invention should be included within the scope of the present invention. Therefore, the protection scope of the present invention should be determined by the protection scope of the claims.

[0074] Sequence List:

[0075] A rice disease resistance-related gene Os04g55720, consisting of 2833 nucleotides, and its nucleotide sequence is as shown in 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] CCTCTTTAAAAAATGATACTGTACTTGTCTCGATCTCTGTTAAAGCATTAAAAAAATCTTGCATCAAC

[0087] GTGGCACCTGTAACAGTGGATCGTGCTAGAACGTCTGTTTGATGAATTGCTAGCAAATCGAGAAACACG

[0088] AATGGGTGCTTTTTCACTAGAAGCAGTCCTCAGAGATTACTTTCGTTACTTCATTCAATTTGACATAAT

[0089] CAGAAAAAGTTAATATCTGCTTCAGTTGACAAAGCTAGATGCATGAAGATGTGAAAATGTGGAGTAAGG

[0090] GTAGTCTTAGATGAAGCATCTGCCATGCAATCTTCGAATGGCATCTTCGGAAGAAGGGCAACATACCTT

[0091] TTCAATTATGATGAGTTCCCATGTTCTTAACAATGTGTCTTGTATAGGTAAATGGCAGCGCAACAAGTA

[0092] TGTGGGTGTATCTCTCGTTGGTAAAACTATTGCAATTCTTGGATTTGGAAAGGTTGGGTCAGAAGTCGC

[0093] TCGTCGTGCTAAAGGTTTGGGAATGCATGTGATTGCACACGATCCGTATGCTTCTGCTGATCGTGCCCA

[0094] TGCAATTGGAGTTGAGCTAGTGAGCATGGAAGACGCTTTGACAACTGCTGACTTTATTCTCGTTGCATAT

[0095] GCCCTTACCCCTGCAACAAACAAGATGCTCAATGATGAAACTTTCGCTAAGATGAAGAAGGGTGTTAG

[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 the rice defense gene Os04g55720 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. An application of overexpression of the rice disease resistance-related gene Os04g55720 in improving rice resistance to bacterial blight, characterized in that, The nucleotide sequence of the gene Os04g55720 is shown in SEQ ID NO.1, and its coding region sequence is shown in SEQ ID NO.

2.

2. The use of a recombinant expression vector, expression cassette, transgenic cell line, or genetically engineered bacterium comprising the rice disease resistance-related gene Os04g55720 as described in claim 1 in any one or more of (A1)-(A4): (A1) Application in improving rice resistance to bacterial blight; (A2) Application in the preparation of products that enhance rice resistance to bacterial blight; (A3) Application in the cultivation of rice with high resistance to bacterial blight; (A4) Application in the development of rice products with high resistance to bacterial blight.

3. A method for improving the resistance of rice to bacterial blight, characterized in that, Overexpression of the rice Os04g55720 gene enhances rice resistance to bacterial blight. Its nucleotide sequence is shown in SEQ ID NO.1, and its coding region sequence is shown in SEQ ID NO.

2.

4. 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 that highly expresses the rice Os04g55720 gene. The nucleotide sequence of the rice Os04g55720 gene is shown in SEQ ID NO.1, and its coding region sequence is shown in SEQ ID NO.

2.

5. The method for constructing transgenic rice resistant to bacterial blight as described in claim 4, characterized in that: The coding region sequence of the rice Os04g55720 gene was cloned into a plant expression vector, first transformed into Agrobacterium, and then transformed into rice cells through callus transformation to obtain transgenic rice that highly expresses the rice Os04g55720 gene. The coding region sequence of the rice Os04g55720 gene is shown in SEQ ID NO.2.

Images