Specific molecular marker primer for identification of phytopthora sojae avirulence gene PsAvr1k and method

A technology of Phytophthora sojae and molecular markers, which is applied in the fields of biochemical equipment and methods, recombinant DNA technology, determination/inspection of microorganisms, etc.

Inactive Publication Date: 2016-10-12
NANJING AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, due to the rapid evolution of pathogenic bacteria mediated by pathogenic effectors in nature, disease-resistant variet

Method used

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  • Specific molecular marker primer for identification of phytopthora sojae avirulence gene PsAvr1k and method
  • Specific molecular marker primer for identification of phytopthora sojae avirulence gene PsAvr1k and method
  • Specific molecular marker primer for identification of phytopthora sojae avirulence gene PsAvr1k and method

Examples

Experimental program
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Effect test

Embodiment 1

[0025] Example 1 Detection of PsAvr1k sequence polymorphism in different strains of Phytophthora sojae

[0026] 1. Tested strains

[0027] Phytophthora sojae strains with different virulence were donated by Professor Brett M. Tyler (U.S. Virginia Bioinformatics Laboratory). This strain is well known to those skilled in the art and is preserved by the Plant-Phytophthora Interaction Laboratory of the Plant Disease Department of Nanjing Agricultural University. . The strains were stored in 10% V8 solid medium at a storage temperature of 10°C.

[0028] 2. Preparation of soybean seedlings for testing

[0029] Identify the host Soybeans are sown in plastic flowerpots (d=10cm) equipped with vermiculite (2-4mm), and 15 seeds are planted in each pot, and they are placed in a greenhouse with 14h light (strong light irradiation) / 10h darkness for growth, 7 10 seedlings were reserved for inoculation after two true leaves grew two days later. The differential host used was Williams82. ...

Embodiment 2

[0052] Example 2 Phytophthora sojae PsAvr1k avirulent strain identified by specific primer markers

[0053] 1. Design of labeled primers for specific amplification of PsAvr1k in avirulent strains

[0054] Primers were designed according to the specific segments of avirulent strains and virulent strains (Table 1) ( figure 2 ). See SEQ ID No.5-12 for sequence differences of PsAvr1k gene in different strains

[0055] Table 1PsAvr1k full-length primers and specific marker primers for avirulent strains

[0056]

[0057] 2. PCR amplification and product verification

[0058] The full-length PCR product of PsAvr1k was used as a template to perform PCR amplification with specific primers (SEQ ID No.3 and SEQ ID No.4) for the avirulent strain PsAvr1k. PCR products were verified with 1% agarose gel. If the fragment of 504bp can be amplified, it indicates that the bacterial strain is a PsAvr1k avirulent strain; if the fragment of corresponding size cannot be amplified, it indica...

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Abstract

Belonging to the field of biotechnology, the invention discloses a specific molecular marker primer for identification of phytopthora sojae avirulence gene PsAvr1k and a method. The specific molecular marker primer pair has an upstream primer nucleotide sequence shown as SEQ ID No.3 and a downstream primer nucleotide sequence shown as SEQ ID No.4. Through the molecular marker PCR amplification by means of the molecular marker can rapidly and efficiently detect whether a field phytopthora sojae strain is a PsAvr1k virulent strain or avirulent strain. The research not only can be used for studying the distribution of PsAvr1k avirulence gene in the field, but also can reveal the pathotype differentiation variation characteristics of the phytopthora sojae PsAvr1k, and provides the basis for making of a resistant variety selection strategy.

Description

technical field [0001] The invention belongs to the field of biotechnology, and discloses a specific molecular marker primer and a method for identifying the avirulent gene PsAvr1k of Phytophthora soybean Virus gene PsAvr1k. Background technique [0002] Soybean root rot caused by Phytophthora soybean is a devastating disease in soybean production worldwide. It is closely related to cyanobacteria and belongs to the kingdom of velvet whip, so general fungicides are often ineffective against Phytophthora sojae and other oomycetes. The application of plant disease-resistant varieties has become an important means of achieving disease resistance in agricultural production due to overcoming the problems of phytotoxicity and environmental pollution. However, the rapid evolution of pathogenic bacteria mediated by pathogenic effectors in nature makes disease-resistant varieties quickly overcome by new pathogens, so the application of disease-resistant varieties is also facing cert...

Claims

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Application Information

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IPC IPC(8): C12N15/11C12Q1/68C12Q1/04
CPCC12Q1/6895C12Q2600/156
Inventor 窦道龙宋天巧
Owner NANJING AGRICULTURAL UNIVERSITY
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