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Rice disease resistance related gene OsWRKY45-1 and application thereof in improvement on rice disease resistance

A rice resistance technology, applied in the field of genetic engineering, can solve problems such as unclearness, decline in yield and quality, and changes in rice disease-resistant phenotypes

Inactive Publication Date: 2009-12-02
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 resistance related gene OsWRKY45-1 and application thereof in improvement on rice disease resistance
  • Rice disease resistance related gene OsWRKY45-1 and application thereof in improvement on rice disease resistance
  • Rice disease resistance related gene OsWRKY45-1 and application thereof in improvement on rice disease resistance

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Example 1: Sequence and structural analysis of the OsWRKY45-1 gene

[0024] 1. Determination of different alleles at the OsWRKY45 locus

[0025] Researchers have reported that OsWRKY45 plays a very important role in rice blast resistance induced by benzothiadiazole (also known as benzo[1,2,3]thiadiazole-7-carboxylate). role of (Shimono et al., 2007). The applicant of the present invention designed the PCR primer w45F4 (5′-AT GGTACC GCCTACGCATCATCTCCTTC-3′) (the underline represents the KpnI restriction endonuclease digestion site) and w45R4 (5′-CA GGATCC TTATGGCACAACATTTAGCA-3′), from the japonica rice varieties Mudanjiang 8 (a rice variety that was publicly reported and applied) and Dongjin (a rice variety that was publicly reported and applied) and the indica rice (Oryza sativassp.indica) variety Minghui 63 (a rice variety that was publicly reported and applied rice variety), Zhenshan 97 (a rice variety that was publicly reported and used) and 93-11 (a rice variety...

Embodiment 2

[0030] Example 2: Analysis of the expression profile of the OsWRKY45-1 gene after pathogen infection

[0031] In order to verify whether the OsWRKY45-1 gene is involved in the regulation of rice resistance to bacterial diseases, the applicant of the present invention firstly used the method of quantitative reverse transcription-PCR (qRT-PCR) (Qiu et al., 2007) to analyze Expression pattern of OsWRKY45-1 gene after inoculation of bacterial blight strain PXO61 (Sun et al., 2004) in different rice lines. Bacterial blight inoculation was carried out on rice at the adult plant stage by cutting leaves (Sun et al., 2004). The bacterial blight strain PXO61 was kindly provided by the International Rice Research Institute of the Philippines (Sun et al., 2004). The cultivation of bacterial blight followed a published method (Sun et al., 2004). Total RNA was extracted from inoculated leaves at different time points after inoculation (Zhou et al., 2002). Take 1-5 μg of total RNA and tre...

Embodiment 3

[0033] Example 3: Overexpression of OsWRKY45-1 gene to verify its function in rice disease resistance response

[0034] Because the allele OsWRKY45-2 overexpressing OsWRKY45-1 enhances the disease resistance of rice (invention patent application number: 200810197309.9, publication number: CN101386856, rice disease resistance-related gene OsWRKY45-2 and its role in improving rice disease resistance Inventors: Wang Shiping, Tao Zeng), the applicant of the present invention overexpressed OsWRKY45-1 gene in susceptible rice, and detected and analyzed the disease resistance of genetically transformed plants. The specific operation analysis is as follows:

[0035] 1. Isolation of the OsWRKY45-1 gene

[0036] According to the position of the OsWRKY45-1 gene in the japonica rice Nipponbare genome (rice whole genome sequence database, http: / / rice.tigr.org ), two PCR primers w45F4 (5′AT GGTACC GCCTACGCATCATCTCCTTC-3′) (the underline represents the KpnI restriction endonuclease dige...

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Abstract

The invention relates to the technical field of plant gene engineering, in particular to the isolation cloning and function verification of a DNA fragment containing a rice disease resistance related gene OsWRKY45-1. A nucleotide sequence of the OsWRKY45-1 gene and an amino acid sequence encoded by the gene are shown in SEQ ID NO: 1. Genetically transformed rice of which the expression level of the OsWRKY45-1 gene is remarkably reduced apparently strengthens the resistance on xanthomonas oryzae pv. oryzae and xanthomonas oryzae pv.oryzicola, which proves that the OsWRKY45-1 gene is a negative regulatory factor in reactions of resisting bacterial blight and bacterial stripe of rice, and the disease resistance of the rice can be improved by inhibiting the expression of the OsWRKY45-1 gene.

Description

technical field [0001] The invention relates to the technical field of genetic engineering. It specifically relates to the isolation, cloning, functional verification and application of a rice disease resistance-related gene OsWRKY45-1. OsWRKY45-1 gene is a negative regulator in rice disease resistance response. After inhibiting the function of OsWRKY45-1 gene, the ability of rice to resist bacterial blight and bacterial leaf spot (thin stripe disease) was significantly improved. Background technique [0002] Plants are attacked by various pathogens during their growth. There are many types of plant pathogens, including viruses, bacteria, fungi, 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 ...

Claims

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Application Information

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IPC IPC(8): C12N15/29C12N15/82
Inventor 王石平陶增
Owner HUAZHONG AGRI UNIV
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