Oryza minuta bacterial blight-resisting major gene Xa3/Xa26-3 and application thereof in improving disease resistance of paddy rice

A technology for leaf blight resistance and leaf blight, which is applied in the field of plant genetic engineering and can solve problems such as yield and quality decline

Inactive Publication Date: 2010-11-10
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] Rice is an important food crop in the world, but the im...

Method used

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  • Oryza minuta bacterial blight-resisting major gene Xa3/Xa26-3 and application thereof in improving disease resistance of paddy rice
  • Oryza minuta bacterial blight-resisting major gene Xa3/Xa26-3 and application thereof in improving disease resistance of paddy rice
  • Oryza minuta bacterial blight-resisting major gene Xa3/Xa26-3 and application thereof in improving disease resistance of paddy rice

Examples

Experimental program
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Effect test

Embodiment 1

[0024] Example 1: Isolation and clone Xa3 / Xa26-3 gene from Oryza sativa and gene structure analysis

[0025] 1. Identification of large fragments of DNA carrying homologous sequences of Xa3 / Xa26 genes

[0026]The researchers of the present invention first use the specific PCR primers RKb-3'race2 (5'-TGGTCAAATACCGGAAGGAG-3') and RKb-2R (5'-CAGTCCACCACATGGACAAG-3') of the Xa3 / Xa26 gene and the Xa3 / Xa26 family member MRKa gene Specific PCR primers RKa-11L (5'-TTGGCTTGAACGGCTTAACT-3') and RKa-1R (5'-AAGATGAAATATGCTCGGTGGT-3') amplified Xa3 from rice variety Minghui 63 (a rice variety popularized in China) / Xa26 gene and MRKa DNA fragments, the length of each amplification is about 1kb ( figure 2 ). The two PCR amplification products were mixed as probes to screen the genomic BAC (bacterial artificial chromosome) library of Oryzaminuta (Ammiraju et al., 2006), and 7 positive BAC clones were identified. These 7 positive BAC clones (Ammiraju et al., 2006) were donated by Professo...

Embodiment 2

[0037] Example 2: Functional verification of Xa3 / Xa26-3 gene

[0038] 1. Construction of genetic transformation vector

[0039] The carrier used in the present invention is pCAMBIA1301 ( Figure 5 ), which is a commonly used rice genetic transformation vector (Sun et al., 2004). Reclaim the 13.7kb fragment that comprises the coding region of Xa3 / Xa26-3 gene, promoter and tail sequence ( Figure 4 ). At the same time, the genetic transformation vector pCAMBIA1301 was digested with restriction endonuclease SmaI; after digestion, dephosphorylated with SAP (shrimp alkaline phosphatase); extracted and purified with chloroform:isoamyl alcohol (volume ratio 24:1) Digestion product. The ligation reaction was performed with the recovered fragment containing the Xa3 / Xa26-3 gene and the purified vector. The positive clone was verified by enzyme digestion, and the obtained recombinant plasmid was named D103O.

[0040] 2. Genetic Transformation and T 0 Genetic Transformation Plant A...

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Abstract

The invention relates to the technical field of plant gene engineering, more particularly to isolation and cloning as well as functional verification of DNA fragment containing Oryza minuta bacterial blight-resisting major gene Xa3/Xa26-3. Xa3/Xa26-3 codes leucine-rich protein kinase-type protein, which can enable paddy rice to resist the diseases caused by bacillary pathogenic bacteria-bacterial blight bacteria(Xanthomonas oryzae pv. Oryzae). The fragment and self-regulatory sequences thereof are directly transplanted into paddy rice, therefore, Xa3/Xa26-3. Xa3/Xa26-3-carrying transgenic rice is prominently strengthened in resisting bacterial blight.

Description

technical field [0001] The invention relates to the technical field of plant genetic engineering. It specifically relates to the isolation, cloning, functional verification and application of a rice bacterial blight-resistant main gene Xa3 / Xa26-3. The DNA fragment can endow rice with resistance to diseases caused by bacterial blight. The fragment and its endogenous regulatory sequence are directly transferred into the plant body, and the transgenic rice can produce a defense response to bacterial blight mediated by the gene. technical background [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 resourc...

Claims

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Application Information

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IPC IPC(8): C12N15/29C07K14/415A01H5/00
Inventor 王石平李弘婧
Owner HUAZHONG AGRI UNIV
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