Oryza officinalis anti-Xanthomonas oryzae major gene Xa3/Xa26-2 and application for improving disease resistance of rice thereof

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

Inactive Publication Date: 2010-11-24
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 officinalis anti-Xanthomonas oryzae major gene Xa3/Xa26-2 and application for improving disease resistance of rice thereof
  • Oryza officinalis anti-Xanthomonas oryzae major gene Xa3/Xa26-2 and application for improving disease resistance of rice thereof
  • Oryza officinalis anti-Xanthomonas oryzae major gene Xa3/Xa26-2 and application for improving disease resistance of rice thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0023] Example 1: Isolation and clone Xa3 / Xa26-2 gene from medicinal wild rice and analysis of gene structure

[0024] 1. Identification of large fragments of DNA carrying homologous sequences of Xa3 / Xa26 genes

[0025]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 The specific PCR primers RKa-11L (5'-TTGGCTTGAACGGCTTAACT-3') and RKa-1R (5'-AAGATGAAATATGCTCGGTGGT-3') amplified the DNA fragments of Xa3 / Xa26 gene and MRKa from Minghui 63, and amplified Each about 1kb in length ( figure 2 ). The two PCR amplification products were mixed as probes to screen the genomic BAC (bacterial artificial chromosome) library of medicinal wild rice (Oryza officinalis) (Ammiraju et al., 2006), and 9 positive BAC clones were identified. The nine positive BAC clones (Ammiraju et al., 2006) were donated by Professor Rod Win...

Embodiment 2

[0036] Example 2: Functional verification of Xa3 / Xa26-2 gene

[0037] 1. Construction of genetic transformation vector

[0038] 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 11.9kb fragment that comprises the coding region of Xa3 / Xa26-2 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. A ligation reaction was performed with the recovered fragment containing the Xa3 / Xa26-2 gene and the purified vector. The positive clone was verified by enzyme digestion, and the obtained recombinant plasmid was named D101O.

[0039] 2. Genetic Transformation and T 0 Genetic Transformation Plant Ana...

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Abstract

The invention relates to the technical field of the plant genetic engineering, in particular to the isolation and cloning and functional verification of the DNA fragment of oryza officinalis anti-Xanthomonas oryzae major gene Xa3/Xa26-2. The Xa3/Xa26-2 gene is used to code the leucine-rich protein kinase protein. The Xa3/Xa26-2 gene ensures that rice can resist diseases caused by bacterial pathogen-Xanthomonas oryzae pv. oryzae. The DNA fragment and the regulatory sequence thereof are directly transferred in rice, and the resistance capability to Xanthomonas oryzae of the transgenic rice carrying Xa3/Xa26-2 is significantly enhanced.

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 bacterial blight-resistant main gene Xa3 / Xa26-2. 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 resources to ...

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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