Rice disease resistance related gene GH3-2 and application thereof in breeding of broad spectrum disease-resistant rice

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

Inactive Publication Date: 2011-03-30
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 singl...

Method used

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  • Rice disease resistance related gene GH3-2 and application thereof in breeding of broad spectrum disease-resistant rice
  • Rice disease resistance related gene GH3-2 and application thereof in breeding of broad spectrum disease-resistant rice
  • Rice disease resistance related gene GH3-2 and application thereof in breeding of broad spectrum disease-resistant rice

Examples

Experimental program
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Effect test

Embodiment 1

[0023] Example 1: Analysis of the expression pattern of GH3-2 gene in different rice varieties

[0024] The GH3-2 gene sequence [TIGR (The Institute for Genomic Research, The Institute for Genomic Research, http: / / rice.tigr.org ) database gene locus number: LOC_Os01g55940] Retrieve the expressed sequence tag (expressed sequence tag, EST) database REDB (Rice ESTDataBase, http: / / redb.ricefgchina.org, Zhang et al., 2005), found that a 2248bp cDNA sequence BI101D22 derived from Minghui 63 had 99% sequence homology with the GH3-2 gene in Nipponbare, so this gene derived from Minghui 63 was named GH3-2.

[0025] In order to confirm whether the GH3-2 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 expression of the GH3-2 gene in different rice Expression patterns in cultivars after inoculation with ...

Embodiment 2

[0027] Example 2: Isolation and clone GH3-2 gene and gene structure analysis

[0028] In order to verify the above speculation, the present invention isolated and cloned the GH3-2 gene from the disease-resistant rice variety Minghui 63 for gene function verification analysis.

[0029] 1. Prediction of GH3-2 gene structure

[0030] Using the cDNA sequence BI101D22 of the GH3-2 gene in rice variety Minghui 63 as a template, the full-length cDNA database KOME ( http: / / cdna01.dna.affrc.go.jp / cDNA / ), and found that the BI101D22 sequence had a 99% homology with a 2365bp full-length cDNA sequence (registration number: AK102809) from the japonica rice variety Nipponbare in the database. The sequence of AK102809 is the cDNA sequence of the gene numbered LOC_Os01g55940 in the rice genome sequence. The alignment analysis of the two sequences confirmed that the cDNA sequence BI101D22 of the GH3-2 gene of Minghui 63 contained the complete coding sequence (coding sequence) encoding the ...

Embodiment 3

[0040] Example 3: Functional verification of GH3-2 gene

[0041] 1. Construction of genetic transformation vector

[0042] The carrier used in the present invention is pU1301 ( Figure 5 ). 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 gene promoter with constitutive and overexpression characteristics.

[0043] The cDNA clone BI101D22 ( image 3 ) was digested with restriction endonucleases KpnI and BamHI to reclaim a 2326nt fragment as an exogenous ( image 3 The shown 2248nt cDNA fragments contain 39nt vector DNA fragments on both sides). At the same time, the genetic transformation vector pU1301 carrying the maize ubiquitin gene promoter was digested with restriction endonucleases KpnI and BamHI; after digestion, the digested product was extracted with chloroform:isoamyl alcohol (volume ratio 24:1) and purified...

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Abstract

The invention relates to the technical field of plant gene engineering, in particular to separation, cloning and function verification of a DNA fragment containing rice disease resistance related genes GH3-2. The genes GH3-2 code auxin amide synthetase. The genes GH3-2 can endow the rice with function of resisting the diseases caused by bacterial pathogenic bacteria Xanthomonas oryzae pv. oryzae, bacterial Xanthomonasoryzae pv. oryzicola and fungal pathogenic bacteria Magnaporthe grisea. The fragment and the exogenous adjusting sequence thereof are directly transferred into the rice, and the transgenic rice for excessively expressing the GH3-2 has the remarkably enhanced resistance to the Xanthomonas oryzae pv. oryzae, bacterial Xanthomonasoryzae pv. oryzicola and the Magnaporthe grisea.

Description

technical field [0001] The invention relates to the technical field of genetic engineering. It specifically involves the isolation, cloning, functional verification and application of a rice disease resistance-related gene GH3-2. GH3-2 gene is a positive regulator in rice disease resistance response. The ability of the transgenic plants overexpressing the GH3-2 gene to resist bacterial blight, thin stripe disease and rice blast 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 fundamental way to preven...

Claims

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Application Information

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IPC IPC(8): C12N15/52C12N15/82
Inventor 王石平傅晶
Owner HUAZHONG AGRI UNIV
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