Rice gene osge61 and its application in rice blast resistance

A rice blast fungus and rice technology, applied in the fields of application, genetic engineering, plant gene improvement, etc., to achieve the effect of improving the ability of rice blast resistance

Active Publication Date: 2020-09-04
FUJIAN AGRI & FORESTRY UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Currently, there is no information on the rice receptor-like protein kinase OsGE61 Application of Genes in Rice Blast Resistance

Method used

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  • Rice gene osge61 and its application in rice blast resistance
  • Rice gene osge61 and its application in rice blast resistance
  • Rice gene osge61 and its application in rice blast resistance

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Example 1 OsGE61 Gene acquisition

[0025] Harpin is a class of exocrine proteins secreted by plant bacterial pathogens through the three-type secretion system and mainly located in the plant interstitium. The external application of Harpin protein to plants has three main effects: improving disease resistance, promoting growth, and inducing cell death. Treatment of rice seedlings with Harpin protein spraying, found by transcriptome analysis OsGE61 Transcriptional upregulation induced by Harpin treatment. However, at different time points after the rice was inoculated with the virulent blast strain OsGE61 Compared with the control treatment, the transcript level was significantly decreased. Accordingly, we speculate that this gene plays an important regulatory role between rice growth and immune response activation. According to the predicted cDNA sequence (XM_015770104) of the gene in the rice variety Nipponbare genome on the NBCI website, we successfully cloned t...

Embodiment 2

[0026] Example 2 OsGE61 Construction of knockout vector

[0027] exist OsGE61 Select two target sites (sequences are shown in SEQ ID NO.3 and SEQ ID NO.4) in the coding region of the open reading frame of the gene, construct a gRNA expression cassette, and then connect the two target gRNA expression cassettes to the knockout In the vector pC1300-Cas9, obtain OsGE61 knockout vector. The detailed vector construction method refers to the patent "Construction and Application of Plant Multi-gene Knockout Vector, ZL201510485573.2", as follows:

[0028] (1) Selection of target sequence and design of primers

[0029] exist OsGE61 The coding region of the open reading frame of the gene is found to contain 5'-(N) X -The target sequence of the NGG-3' structure, wherein N represents any one of A, T, C and G, and X is 19. Design two target sequences according to the above principles, namely GE61-T1 and GE61-T2, the sequences are as follows:

[0030] GE61-T1 SEQ ID NO.3: 5'-AACTGGGA...

Embodiment 3

[0074] Example 3 OsGE61 Obtaining knockout mutants

[0075] Transform the correctly sequenced binary expression vector pC1300-Cas9-SK-gRNA-1-gRNA-2 into Agrobacterium tumefaciens by electroporation EHA105 In the process, after the transformant was verified to be correct, the callus of the japonica rice variety Yandao No. 8 was transformed to obtain OsGE61 Knockout mutant rice plants.

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Abstract

The invention provides an oryza sativa gene OsGE61 and an application thereof to resistance to piricularia oryzae. The length of an open reading frame of the gene is 1884bp, and receptoroid protein kinase is coded and consists of 627 amino acids. The oryza sativa gene OsGE61is knocked out by a CRISPR / Cas9 method, wild type plants and plants of which mutants are knocked out are respectively subjected to magnaporthe oryzae inoculation experiment, the result shows that after the OsGE61 is knocked out, the resistance of oryza sativa plants to the magnaporthe oryzae is obviously improved, and the gene can be used for breeding or cultivating relevant disease-resistant varieties.

Description

technical field [0001] The invention belongs to the field of biotechnology, in particular to a rice gene OsGE61 and its application in rice blast resistance. Background technique [0002] Rice ( Oryza sativa ) is a staple food for more than half of the world's population. by Magnaporthe grisea ( Magnaporthe oryzae ) caused by rice blast causes annual rice yield losses of 10% to 30%. [0003] During the process of getting along with pathogenic microorganisms, some complex immune response systems are constantly produced to resist the invasion of pathogenic microorganisms. It is currently found that the plant immune system includes at least two levels: the first layer is Pathogen-associated molecular patterns (Pathogen-associated molecular patterns, PAMPs)-triggered immunity (PAMP-triggered immunity, PTI), that is, plants carry out molecular recognition of PAMPs of pathogenic bacteria through cell surface pattern recognition receptors (PRRs), thereby initiating plant de...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/54C12N9/12C12N15/82A01H5/00A01H6/46
CPCC12N9/1205C12N15/8213C12N15/8282
Inventor 鲁国东王莫孙鹏林娅红朱昆鹏王宗华
Owner FUJIAN AGRI & FORESTRY UNIV
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