Disease resistance associated protein TaCPK7-R derived from wheat and related biological materials and application thereof

A disease resistance and protein technology, applied in the fields of application, biochemical equipment and methods, and the use of vectors to introduce foreign genetic materials, etc., can solve the problems of poor resistance and slow progress in breeding for sheath blight resistance

Inactive Publication Date: 2014-08-27
INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, conventional breeding for sheath blight resistance has progressed slowly. At present, the large-scale

Method used

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  • Disease resistance associated protein TaCPK7-R derived from wheat and related biological materials and application thereof
  • Disease resistance associated protein TaCPK7-R derived from wheat and related biological materials and application thereof
  • Disease resistance associated protein TaCPK7-R derived from wheat and related biological materials and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0072] Embodiment 1, the cloning of wheat resistance protein TaCPK7-R and its coding gene

[0073] The inventors of the present invention respond to sheath blight bacteria through gene chips of the anti-sheath blight wheat CI12633 and the sheath blight-susceptible wheat Wenmai 6, and the sheath blight-resistant wheat Shanhong wheat and the sheath blight-susceptible wheat Wenmai 6 respond to sheath blight Based on the analysis of the differential expression data of the two sets of genes of the strain, combined with the correlation analysis between the degree of resistance and the expression level, the protein kinase gene TaCPK7-R gene, which is resistant to sheath blight in wheat, was isolated and cloned from CI12633. The specific cloning method is as follows:

[0074] Extract the total RNA from the stems of wheat CI12633 inoculated with Rhizoctonia graminearum for 21 days, and reverse-transcribe the extracted RNA samples to synthesize the first-strand cDNA according to the pro...

Embodiment 2

[0082] Example 2, the acquisition of sheath blight resistant transgenic wheat and the identification of disease resistance

[0083] 1. Construction of recombinant expression vector

[0084] The complete ORF sequence of the TaCPK7-R gene was constructed on the high-efficiency expression vector pAHC25 of monocotyledonous plants, and the specific steps were as follows:

[0085] 1. Preparation of linearized plasmid: Digest the pAHC25 vector plasmid (about 9706bp) with Sma I and Sac I, excise the GUS gene (1888bp) between the restriction sites, 1% agarose gel electrophoresis, and agarose gel DNA The purification recovery kit recovered the linearized pAHC25 vector framework (about 7818bp).

[0086] 2. Acquisition of the target gene TaCPK7-R containing vector homology sites: Design a pair of primers TaCPK7-R-P25-F according to the full-length cDNA sequence of the TaCPK7-R gene: 5'- AACTCGGTATCTAGA ATGGGCAACTGCTGCGC-3' and TaCPK7-R-P25-R:5'- CGATCGGGGAAATTC CTATTGCGTTATCATCTA-3',...

Embodiment 3

[0132] Example 3, Breeding Gene-Silenced Wheat with Reduced Sheath Blight Resistance

[0133] 1. Silencing the TaCPK7-R gene in wheat CI12633 using virus-mediated gene silencing technology

[0134] 1. The two ends of the DNA fragment shown in the 1586-1829 nucleotides of sequence 1 in the sequence listing are respectively provided with Nhe I recognition sequences. After NheI digestion, the DNA fragment (244bp) shown in the 1586-1829th nucleotide of sequence 1 in the sequence listing is inserted into the BSMV-γ (BMSV virus's BMSV-γ) after the NheI enzyme linearization by the reverse insertion method γ vector), the DNA molecule (named anti TaCPK7-R) that is reverse complementary to the DNA fragment shown in the 1586-1829th nucleotide of sequence 1 in the sequence listing is driven by the T7 promoter of the γ vector to obtain Recombinant vector BSMV-γ: anti TaCPK7-R.

[0135] 2. At the two-leaf one-heart stage, use the recombinant vector BSMV-γ:anti TaCPK7-R to transfect the se...

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Abstract

The invention discloses disease resistance associated protein TaCPK7-R derived from wheat and related biological materials and application thereof. The TaCPK7-R is the protein of a) or b), wherein the amino acid sequence is the protein of sequence 2; and the amino acid sequence shown by the sequence 2 in the sequence table is substituted by one or more amino acid residues and/or deleted and/or is add with the protein related to the disease resistance of plants. Experiments prove that TaCPK7-R and biological materials related to the TaCPK7-R can be used for regulating and controlling the disease resistance of plants, and particularly can be used for regulating and controlling the resistance of wheat to banded sclerotial blight.

Description

technical field [0001] The invention relates to a disease resistance-related protein TaCPK7-R derived from wheat in the field of biotechnology, as well as related biological materials and applications. Background technique [0002] Wheat is one of the most important food crops in the world and plays a pivotal role in ensuring food security. With the improvement of fertilizer and water conditions, the increase of planting density and the change of farming system, wheat sheath blight has developed into a major disease of wheat production in my country, and has become an important factor limiting the high and stable yield of wheat in my country. [0003] Wheat sheath blight, also known as wheat sharp eyespot, is a worldwide soil-borne disease mainly caused by Rhizoctonia cerealis and Rhizoctonia solani. Saprophytic fungal diseases (Chen Yanxi, Tang Wenhua, Zhang Dunhua, Jian Xiaoying. Preliminary research on the etiology of wheat sheath blight in my country. Acta Plant Protecti...

Claims

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Application Information

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IPC IPC(8): C07K14/415C12N15/29C12N15/82C12N1/15C12N1/19C12N1/21C12N15/11A01H5/00
CPCC07K14/415C12N15/8282
Inventor 张增艳洪彦涛杜丽璞申芳嫡魏学宁徐惠君
Owner INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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