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Application of GmPGIP3 protein and coding gene thereof to cultivation of plant for resisting root rot and full rot

A technique for coding genes and transgenic plants, which is applied in the field of application of GmPGIP3 protein and its coding genes in cultivating plants resistant to root rot and take-all disease, and can solve the problems of unreliable selection of single plants, difficulties in identification and selection of resistance, Problems such as the lack of germplasm resources of wheat with high resistance to root rot

Inactive Publication Date: 2012-08-29
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

At present, the large-scale popularized varieties and breeding materials in production generally have poor resistance to root rot and take-all, and the progress of conventional disease-resistant breeding is slow. On the one hand, it is difficult to identify and select resistance to these soil-borne diseases. Single plant selection is very unreliable; on the other hand, there is a lack of germplasm resources of wheat with high resistance to root rot and take-all
Therefore, it is difficult to effectively carry out the resistance breeding of wheat root rot and take-all by conventional breeding methods.

Method used

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  • Application of GmPGIP3 protein and coding gene thereof to cultivation of plant for resisting root rot and full rot
  • Application of GmPGIP3 protein and coding gene thereof to cultivation of plant for resisting root rot and full rot
  • Application of GmPGIP3 protein and coding gene thereof to cultivation of plant for resisting root rot and full rot

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Example 1. Obtaining GmPGIP3 Gene Transformed Plant

[0030] 1. Construction of recombinant expression vector

[0031] 1. Design a pair of primers based on the cDNA sequence of GmPGIP3 gene as follows:

[0032] GmPGIP3-F: 5′-ATGTCAAAGTTAAGCATTCT-3′;

[0033] GmPGIP3-R: 5'-TTAAGTGCATGGTGGAAGAG-3'.

[0034] 2. Extract total RNA from the leaves of Zhonghuang 13 in soybean variety and reverse transcribed into cDNA.

[0035] 3. Using the cDNA of step 2 as a template, a primer pair consisting of GmPGIP3-F and GmPGIP3-R is used for PCR amplification to obtain a PCR amplification product. The sequencing result of the PCR amplified product is shown in sequence 2 of the sequence table.

[0036] 4. Using the PCR amplified product of step 3 as a template, a primer pair consisting of GmPGIP3-SMAF ​​and GmPGIP3-SACR is used for PCR amplification to obtain a PCR amplified product.

[0037] GmPGIP3-SMAF:5′-AT CCCGGG ATGTCAAAGTTAAGCATTCT-3';

[0038] GmPGIP3-SACR:5′-GT GAGCTC TTAAGTGCATGGTGGAAGAG...

Embodiment 2

[0084] Example 2: Identification of Root Rot Resistance of Transgenic Plants with GmPGIP3 Gene

[0085] The T of the GmPGIP3 transgenic wheat obtained in Example 1 2 Generation plants (10 plants per line), T of the GmPGIP3 transgenic wheat obtained in Example 1 1 Generation plants (20 plants per line), the T of the empty carrier wheat obtained in Example 1 2 Generation plants (10 plants per line), the empty carrier wheat obtained in Example 1 T 1 Progeny plants (20 plants per line) and Yangmai 18 (WT) (20 plants) were identified as follows:

[0086] During the wheat tillering period, insert a toothpick full of wheat root rot pathogenic bacteria and a grain of wheat kernel full of wheat root rot pathogenic bacteria into the 1-2 leaf sheaths at the base of the wheat. Try to keep the leaf sheath in a natural state of holding stems; spray water for 5-7 days after inoculation, and investigate the condition during the wax maturity period.

[0087] According to the 0-4 grade standard, the d...

Embodiment 3

[0099] Example 3. Identification of resistance to total erosion of GmPGIP3 transgenic plants

[0100] The T of the GmPGIP3 transgenic wheat obtained in Example 1 2 Seed generation (20 grains per plant), T of the empty carrier wheat obtained in Example 1 2 Generation seeds (20 seeds per plant) and Yangmai 18 (WT) seeds (20 seeds) were identified as follows:

[0101] After sterilizing and accelerating the germination of the seeds, they are moved to the flowerpots that have been put into the wheat rot disease pathogenic bacteria cake (one flowerpot for each seed, and the substrate is a mixture of 1 mass of river sand and 2 mass of nutrient soil), After cultivating for 3 weeks, take pictures and identify the following indicators: measure the total root length and black (diseased) root length of a single plant, calculate the percentage of black root area to the total root area (percentage of diseased roots), and make the total erosion serious The degree is divided into five levels (0-4 ...

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Abstract

The invention discloses application of GmPGIP3 protein and coding gene thereof to cultivation of a plant for resisting root rot and full rot. According to the method provided by the invention, a transgenic plant with the disease resistance higher than that of a target plant is obtained by transferring the coding gene of the GmPGIP3 protein into a target plant; the GmPGIP3 protein is shown as the following (a) or (b): (a) the protein consists of the amino acid sequence shown as the sequence 1 in the sequence table; and (b) the protein related to the plant disease resistance is obtained by replacing and / or deleting and / or adding one or more amino acid residues on the amino acid sequence shown as the sequence 1 and is derived from the sequence 1. The inventor establishes a monocotyledon high expression vector of the coding gene of the GmPGIP3 protein and transfers into wheat to obtain transgenic wheat capable of resisting root rot and full rot. The invention has important value on the breeding of the disease-resistant plants.

Description

Technical field [0001] The invention relates to the application of GmPGIP3 protein and its coding gene in cultivating plants resistant to root rot and total rot. Background technique [0002] Wheat root rot is a worldwide wheat soil-borne disease that is difficult to control in wheat production. In recent years, wheat root rot disease has risen from a minor disease to a major disease in some areas. For example, many wheat fields in North China, Huanghuai and Northeast my country have found dry white ears caused by root rot. Wheat root rot is a disease of multiple and compound infections. The main pathogen is Tripolaris sorokiniana (Bipolaris sorokiniana (Sacc. Ex Sorok.) Shoem. for anamorphism, and Cochliobolus sativus (Ito et Kurib.) Drechsl for anamorphism ]. Wheat root rot is a typical multi-stage disease during the whole growth period, which can occur from the seedling stage to the heading stage. Generally, the output is reduced by 10% to 30%, and the output of serious plo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/63C12N15/82A01H5/00
Inventor 张增艳杜丽璞党良徐慧君
Owner INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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