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Rice disease-resistant related gene OsDR9 and its use in improving rice disease resistance

A disease resistance, rice technology, applied in the field of plant genetic engineering, can solve the problems of decreased yield and quality, change of rice disease resistance phenotype, unclear and other problems

Inactive Publication Date: 2008-08-27
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, although many disease resistance-related genes have been identified in rice (Zhou et al., 2002; Chu et al., 2004), the mechanism of action of these genes in the rice disease resistance response, and whether a single disease resistance-related gene can cause rice disease resistance phenotype changes are not clear
[0006] Rice is an important food crop in the world, but the impact of diseases often leads to the decline of its yield and quality

Method used

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  • Rice disease-resistant related gene OsDR9 and its use in improving rice disease resistance
  • Rice disease-resistant related gene OsDR9 and its use in improving rice disease resistance
  • Rice disease-resistant related gene OsDR9 and its use in improving rice disease resistance

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Example 1: Identify rice disease-resistant lesion-like mutants and isolate and identify mutant genes

[0024] 1. Identification of Lesion-Like Mutants

[0025] Pathogens usually cause plant cell death at the site of infection after infecting plants, forming lesions (Agrios, 1988). Lesion mimics are lesions that resemble pathogenic infestation in the absence of pathogenic infestation. Studies have shown that some genes that cause rice plants to form lesion-like spots due to gene mutations may be disease resistance-related genes (Yamanouchi et al., 2002; Zeng et al., 2004; Mori et al., 2007; Yuan et al., 2007). Therefore, detection of plants that form lesion-like spots in the rice mutant library will help to identify new disease resistance-related genes. Huazhong Agricultural University, where the present invention belongs, created a rice T-DNA insertion mutant library containing about 129,000 independently transformed plants (Wu et al., 2003; Zhang et al., 2006), which l...

Embodiment 2

[0035] Example 2: Determining the OsDR9 gene structure and the relationship between lesion-like spots and OsDR9 gene expression

[0036] 1. OsDR9 gene structure analysis

[0037] Using GENSCAN (http: / / genes.mit.edu / GENSCAN.html), GeneFinder (http: / / genomic.sanger.ac.uk / gf / gf.shtml), Gene Feature Searches (http: / / dot.imgen .bem.tmc.edu: 9331 / ), GeneMark (http: / / genemark.biology.gatech.edu / GeneMark / hmmchoice.html) and other gene structure prediction software and BLAST analysis method (Altschul et al., 1997), Analyze and predict the structural features of the sequence homologous to the flanking sequence of the mutant 02Z15AM37 in a genome sequence AL662948 (section 2 in Example 1) derived from the rice variety Nipponbare and the sequences on both sides. Analysis results showed that in mutant 02Z15AM37, T-DNA was inserted into a predicted gene, which was named OsDR9 (Oryza sativa defense responsive 9) in the present invention.

[0038] The researchers of the present invention de...

Embodiment 3

[0042] Example 3: Verification of functional complementation of OsDR9 gene

[0043] The researchers of the present invention introduced the DNA fragment containing the OsDR9 gene and its own promoter into the mutant 02Z15AM37 by genetic transformation method. The genetic transformation vector used was pCAMBIA2301 (Fig. 7A). The pCAMBIA2301 vector is a series vector of the Agrobacterium-mediated genetic transformation vector pCAMBIA1301 (Sun et al., 2004) commonly used in the world, donated by CAMBIA Laboratory (Center for the Application of Molecular Biology to International Agriculture) in Australia.

[0044] Genetic transformation vectors were constructed using conventional recombinant plasmid construction methods (Sambrook and Russell, 2001). The main steps are: using restriction endonuclease digestion method, the BAC clone OSJNBa0060B20 of the rice variety Nipponbare containing the OsDR9 gene is digested with restriction endonucleases EcoRI and XbaI to obtain the OsDR9 ge...

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Abstract

The invention belongs to the technical field of plant genetic engineering and in particular relates to isolation cloning and functional verification of a DNA fragment which contains rice disease resistance related genes OsDR9. The resistance of rice to rice blast and brown spot disease is intensified through inhibiting expression of the OsDR9 genes in the rice, the OsDR9 genes are proved to be negative regulators in reaction that the rice resists fungous diseases, and disease-resistant rice varieties are cultured through inhibiting the expression of the OsDR9 genes.

Description

technical field [0001] The invention relates to the technical field of plant genetic engineering. It specifically involves the isolation, cloning, functional verification and application of a rice disease resistance-related gene OsDR9. OsDR9 gene is a negative regulator in rice disease resistance response. Inhibiting the function of the OsDR9 gene can significantly enhance the ability of rice to resist diseases. Background technique [0002] Plants are attacked by various pathogens during their growth. There are many types of plant pathogens, including viruses, bacteria, molds and nematodes. Pathogen invasion of plants leads to two results: (1) the pathogen successfully reproduces in the host plant, causing related diseases; (2) the host plant produces a disease-resistant response, killing the pathogen or preventing its growth. Using resistance gene resources to improve plant disease resistance is the fundamental way to prevent diseases and protect the environment at the...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/29A01H1/00A01H5/00
Inventor 王石平丁新华
Owner HUAZHONG AGRI UNIV
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