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Disease resistance-related protein RCR1 derived from wheat, related biomaterials thereof, and application for same

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 slow progress in the research of root rot resistant wheat varieties

Active Publication Date: 2013-09-04
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, due to the lack of ideal wheat sheath blight and root rot resistant germplasm resources, conventional breeding methods have made slow progress in the selection of wheat varieties resistant to sheath blight and root rot

Method used

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  • Disease resistance-related protein RCR1 derived from wheat, related biomaterials thereof, and application for same
  • Disease resistance-related protein RCR1 derived from wheat, related biomaterials thereof, and application for same
  • Disease resistance-related protein RCR1 derived from wheat, related biomaterials thereof, and application for same

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0078] Embodiment 1, the cloning of wheat resistance protein RCR1 and its coding gene

[0079] The inventors of the present invention analyzed the gene differential expression data of the sheath blight-resistant wheat CI12633 and the sheath blight-susceptible wheat Wenmai 6 by RNA-Sequencing, combined with expression correlation analysis, isolated and cloned the wheat resistant The important gene of sheath blight-RCR1 gene. The specific cloning method is as follows:

[0080] The total RNA of wheat CI12633 leaf sheath was extracted, and the extracted RNA sample was reverse-transcribed to synthesize the first-strand cDNA according to the procedure of the first-strand cDNA synthesis kit of Invitrogen Company, which was used as a template for gene cloning, with U:5'-CCCAGTCAAAGTCCGTCAGT-3 ' and L: 5'-GTGGCTTACAGGCACGCAG-3' as primers for PCR amplification. The amplification program was as follows: pre-denaturation at 94°C for 3 minutes; then 35 cycles at 94°C for 45 seconds, 60°...

Embodiment 2

[0090] Example 2, the acquisition of sheath blight resistant transgenic wheat and the identification of disease resistance

[0091] 1. Construction of recombinant expression vector

[0092] Using the Fast PCR Clone Kit, the complete ORF sequence of the RCR1 gene was constructed on the high-efficiency expression vector pAHC25 of monocotyledonous plants. The specific steps are as follows:

[0093] 1. Preparation of linearized plasmid: Digest the pAHC25 vector plasmid with SmaI and SacI, electrophoresis on 1% agarose gel, and recover the linearized pAHC25 vector framework with an agarose gel DNA purification and recovery kit.

[0094] 2. PCR amplification of the target gene RCR1: design a pair of primers RCR1-O-F according to the ORF sequence of the RCR1 gene: 5'- AACTCGGTATCTAGA ATGGAGTTGGCAGTAGGTGC-3'

[0095] and RCR1-O-R: 5'- CGATCGGGGAAATTC TCAGCTGCTGATAACATGAT-3', the 5' end of the primer introduces a 15 bp sequence complementary to the sequences at both ends of the li...

Embodiment 3

[0145] Embodiment 3, cultivate the transgenic wheat that sheath blight property reduces

[0146] 1. Silencing the RCR1 gene in wheat CI12633 using virus-mediated gene silencing technology

[0147] 1. The two ends of the DNA fragment shown in the 2716-3030 nucleotides of sequence 1 in the sequence listing (sequence 3 in the sequence listing) are respectively provided with NheI recognition sequences. After NheI digestion, insert the DNA fragment (315bp) shown in the 2716-3030th 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 (antiRCR1) that is reverse complementary to the DNA fragment shown in the 2716-3030th nucleotide of Sequence 1 in the sequence listing is driven by the T7 promoter of the γ vector to obtain the recombinant vector BSMV-γ: antiRCR1 .

[0148] 2. At the two-leaf one-heart stage, use the recombinant vector BSMV-γ:antiRCR1 to transfect the second le...

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Abstract

The invention discloses a disease resistance-related protein RCR1 derived from wheat, the related biomaterials thereof, and an application for the same. RCR1 is the protein a) or the protein b) as follows: a) the protein composed of amino acid sequences shown in a sequence 2 in a sequence table; and b) the protein obtained by performing the substitution and / or deletion and / or addition of one or more amino acid residues on the amino acid sequences shown in the sequence 2 in the sequence table, and related to plant disease resistance. Experiments proved that RCR1 and the biomaterials relates to RCR1 can be used for regulating plant disease resistance, and especially used for regulating the resistance of wheat to banded sclerotial blight.

Description

technical field [0001] The invention relates to the disease resistance-related protein RCR1 derived from wheat and related biological materials and applications. Background technique [0002] Wheat (Triticuma aestivum) is one of the four major crops that humans rely on for survival. More than one-third of the world's population uses wheat as a staple food. The yield and quality of wheat directly affect human survival and quality of life. In recent years, with changes in farming system, fertilizer and water conditions, and climate conditions, soil-borne diseases such as sheath blight and root rot have become one of the important factors restricting high and stable yields of wheat. Wheat sheath blight occurs heavily in Jianghan, Jianghuai, Huanghuai, and southern North China wheat regions such as Anhui and Jiangsu along the Huaihe River, Huaibei, most of Hubei, most of Henan, southern Shandong, and central and southern Hebei. Sheath blight can generally reduce wheat productio...

Claims

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Application Information

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IPC IPC(8): C07K14/415C12N15/29C12N15/82C12N1/15C12N1/19C12N1/21C12N5/10C12N15/11A01H5/00
Inventor 张增艳祝秀亮徐惠君魏学宁杜丽璞
Owner INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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