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Disease-resistance gap-associated protein TaPK-R1 derived from wheat as well as related biological material and application thereof

A disease resistance, protein technology, applied in application, biochemical equipment and methods, introduction of foreign genetic material using vectors, etc., can solve the problems of slow progress of wheat resistance to sheath blight, lack of easy utilization, etc.

Active Publication Date: 2014-07-16
INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, conventional breeding methods have been slow in breeding sheath blight-resistant wheat varieties due to the lack of readily available germplasm resources for wheat sheath blight resistance

Method used

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  • Disease-resistance gap-associated protein TaPK-R1 derived from wheat as well as related biological material and application thereof
  • Disease-resistance gap-associated protein TaPK-R1 derived from wheat as well as related biological material and application thereof
  • Disease-resistance gap-associated protein TaPK-R1 derived from wheat as well as related biological material and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0077] Embodiment 1, the cloning of wheat resistance protein TaPK-R1 and its coding gene

[0078] 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 TaPK-R1 gene, which is resistant to sheath blight in wheat, was isolated and cloned from CI12633. The specific cloning method is as follows:

[0079] The total RNA of wheat CI12633 leaf sheaths was extracted, and the extracted RNA samples were reverse-transcribed to synthesize the first-strand cDNA according to the procedure of the first-strand cDNA synthesis ki...

Embodiment 2

[0093] Example 2, the acquisition of sheath blight resistant transgenic wheat and the identification of disease resistance

[0094] 1. Construction of recombinant expression vector

[0095] The complete ORF sequence of the TaPK-R1 gene was constructed on the high-efficiency expression vector pAHC25 of monocotyledonous plants, and the specific steps were as follows:

[0096] 1. Preparation of linearized plasmid: Digest the pAHC25 vector plasmid with Spe I and Sac I, electrophoresis on 1% agarose gel, and recover the linearized pAHC25 vector framework with an agarose gel DNA purification and recovery kit.

[0097]2. Acquisition of the target gene TaPK-R1 containing restriction sites: Design a pair of primers TaPK-R1-P25-F: 5'-GA according to the ORF sequence of the TaPK-R1 gene ACTAGT ATGGATTCCGCGAGAAGT-3' and TaPK-R1-P25-R: 5'-GCGAGCTCCTATTTACGTCGAGGTTG-3' were used to extract total RNA from wheat CI12633 leaf sheaths, and reverse-transcribed the extracted RNA samples to synt...

Embodiment 3

[0144] Example 3, Breeding Gene-Silenced Wheat with Reduced Sheath Blight Resistance

[0145] 1. Silencing the TaPK-R1 gene in wheat CI12633 using virus-mediated gene silencing technology

[0146] 1. The two ends of the DNA fragment shown in the 1634-1936 nucleotides of Sequence 1 in the sequence listing are respectively provided with NheI recognition sequences. After NheI digestion, insert the DNA fragment (303bp) shown in the 1634-1936th nucleotide of Sequence 1 in the sequence table into the BSMV-γ (γ vector of BMSV virus) linearized by NheI enzyme by reverse insertion method ), the DNA molecule (the name is anti TaPK-R1, and the nucleotide sequence is sequence 3) that is reverse complementary to the DNA fragment shown in the 1634-1936 nucleotides of sequence 1 in the sequence listing was injected into the gamma vector Driven by T7 promoter, the recombinant vector BSMV-γ: anti TaPK-R1 was obtained.

[0147] 2. At the two-leaf one-heart stage, use the recombinant vector BS...

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Abstract

The invention discloses a disease-resistance gap-associated protein TaPK-R1 derived from wheat as well as a related biological material and an application thereof. The TaPK-R1 is a protein (a) formed by amino acid sequences as shown in a sequence 2 in a sequence table or protein (b) related to plant disease resistance and formed by the amino acid sequences as shown in the sequence 2 in the sequence table, which are replaced and / or deleted and / or added by one or several amino acid residues. The experiments prove that the TaPK-R1 and the biological material related to the TaPK-R1 can be used for adjusting and controlling plant disease resistance, and particularly can be used for adjusting and controlling the banded sclerotial blight resistance of wheat.

Description

technical field [0001] The invention relates to a disease resistance-related protein TaPK-R1 derived from wheat and 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. Wheat sheath blight, also known as wheat sharp eyespot, is caused by saprophytic trophic pathogenic fungi Rhizoctonia cerealis CAG-1 and Rhizoctonia solani AG4, A worldwide soil-borne fungal disease of wheat caused by the AG5 fusion group. The main pathogen of wheat sheath blight in my country is Rhizoctonia cerealis. Sheath blight can generally reduce wheat production by...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/82C12N1/15C12N1/19C12N1/21C12N5/10C12N15/11A01H5/00
CPCC12N9/1205C12N15/8282C12Y207/01037
Inventor 张增艳杨坤荣玮杜丽璞魏学宁叶兴国
Owner INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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