Rice disease resistance relevant gene OsWRKY45-2 and application thereof in improving rice disease resistance

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

Inactive Publication Date: 2009-03-18
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 relevant gene OsWRKY45-2 and application thereof in improving rice disease resistance
  • Rice disease resistance relevant gene OsWRKY45-2 and application thereof in improving rice disease resistance
  • Rice disease resistance relevant gene OsWRKY45-2 and application thereof in improving rice disease resistance

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Example 1: Analysis of the expression pattern of OsWRKY45-2 gene in different rice varieties

[0026] The sequence [TIGR (The Institute for Genomic Research, The Institute for Genomic Research, http: / / rice.tigr.org ) database gene locus number: LOC_Os05g25770] search indica rice expressed sequence tag (expressed sequence tag, EST) database REDB (Rice EST DataBase, http: / / redb.ricefgchina.org , Zhang et al., 2005), found a 1437bp cDNA sequence EI77K16 derived from the indica rice variety Minghui 63 (Oryza sativa ssp. The gene derived from Minghui 63 was named OsWRKY45-2.

[0027] In order to confirm whether the OsWRKY45-2 gene is involved in the regulation of the disease resistance response, the present invention uses quantitative reverse transcription-PCR (quantitative reverse transcription-PCR, qRT-PCR) technology (Qiu et al., 2007) to analyze the OsWRKY45-2 gene in different Expression patterns in rice cultivars after inoculation with bacterial blight strain PXO61...

Embodiment 2

[0029] Example 2: Isolation and clone OsWRKY45-2 gene and gene structure analysis

[0030] 1. Prediction of OsWRKY45-2 gene structure

[0031] Using the cDNA sequence EI77K16 of the OsWRKY45-2 gene as a template, the KOME database (http: / / cdna01.dna.affrc.go.jp / cDNA / ) of the full-length cDNA database of japonica rice was searched by the BLAST method (Altschul et al., 1997) and found that EI77K16 The sequence has a homology of 98% with a 1453bp full-length cDNA sequence (registration number: AK066255) from the japonica rice variety Nipponbare in the database. The AK066255 sequence is the cDNA sequence of the gene numbered LOC_Os05g25770 in the whole genome sequence of rice, that is, the cDNA sequence of the allele of OsWRKY45-2 (OsWRKY45). Alignment analysis of these two sequences confirmed that the cDNA sequence EI77K16 of OsWRKY45-2 gene contained the full-length open reading frame encoding OsWRKY45 protein.

[0032] 2. Isolate and clone the full-length DNA of OsWRKY45-2 ge...

Embodiment 3

[0038] Example 3: Functional verification of the OsWRKY45-2 gene

[0039] 1. Construction of genetic transformation vector

[0040] The carrier used in the present invention is pU1301 ( Figure 4 ). pU1301 is a commonly used rice genetic transformation vector (Cao et al., 2007; Qiu et al., 2007; Ding et al., 2008). It is an Agrobacterium-mediated genetic transformation vector carrying a maize ubiquitin promoter with constitutive and overexpression characteristics.

[0041] The DNA fragment ( image 3 ) were digested with restriction endonucleases KpnI and BamHI, and used as exogenous fragments after inactivating the enzymes. At the same time, the genetic transformation vector pU1301 carrying the maize ubiquitin promoter was digested with restriction endonucleases KpnI and BamHI; after digestion, it was extracted with chloroform:isoamyl alcohol (24:1 by volume) to purify the digested product. Use the digested fragment containing the OsWRKY45-2 gene and the purified vector ...

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Abstract

The invention relates to the technical field of plant genetic engineering, in particular relates to isolating clone and functional verification of a DAN fragment containing a gene OsWRKY45-2 related to rice disease resistance. The gene OsWRKY45-2 codes WRKY proteinoids and can endow rice with resistance to the diseases caused by bacterial pathogenic bacteria, namely the leaf blight bacteria (Xanthomonas oryzae pv.oryzae) and fungal pathogenic bacteria, namely the rice blast bacteria (Magnaporthe grisea). The fragment and the exogenous regulatory sequence thereof are directly transferred into rice, thus the resistance capability of transgenic rice with over-expression OsWRKY45-2 against bacterial blight in and rice blast is improved remarkably.

Description

technical field [0001] The invention relates to the technical field of plant genetic engineering. It specifically relates to the isolation and cloning, functional verification and application of a rice disease resistance-related gene OsWRKY45-2. The OsWRKY45-2 gene isolated and cloned in the invention is a positive regulatory factor in rice disease resistance response. The transgenic plants overexpressing the OsWRKY45-2 gene have significantly improved resistance to bacterial blight and rice blast. 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...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/29C12N15/82A01H1/00A01H5/00
Inventor 王石平陶增
Owner HUAZHONG AGRI UNIV
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