Molecular method for determining virulence of phytophthora sojae on disease-resistant gene Rps3a(Chapman)

Abstract

The invention relates to a molecular method for determining virulence of phytophthora sojae on a disease-resistant gene (i)Rps3a( / i)(Chapman), comprising the steps: extracting genome DNA of phytophthora sojae to be detected, carrying out PCR amplification to obtain a DNA fragment containing avirulence gene(i)Avr3a( / i), carrying out enzyme digestion with restriction enzyme (i)Bpu( / i)10I, carrying out agarose gel electrophoresis on the enzyme digestion product to obtain an agarose gel electrophoretogram of the enzyme digestion product; and observing the agarose gel electrophoretogram of the enzyme digestion product, if 2 DNA bands exist in a 250bp-500bp range, the corresponding phytophthora sojae strain to be detected is an avirulent strain, and if 1 DNA band exists under 750bp in the electrophoretogram after enzyme digestion, the corresponding phytophthora sojae strain to be detected is a virulent strain. The method has the advantages of short determination cycle, no intermediate type, accuracy and reliability, and can determine with a high flux reactor the virulence of phytophthora sojae on the disease-resistant gene (i)Rps3a( / i)(Chapman).

Description

technical field [0001] The invention relates to a molecular determination method, in particular to a molecular method for determining the toxicity of Phytophthora soybean disease resistance gene Rps3a (Chapman). Background technique [0002] Soybean root rot caused by Phytophthora sojae infection is one of the main diseases threatening soybean production worldwide. Soybean root rot was first discovered in Indiana, USA in 1948, and then it was found in major soybean producing countries one after another. In 1989, Shen Chongyao and Su Yanchun first isolated Phytophthora sojae in the main soybean producing areas in Northeast my country in 1989. Later, Phytophthora sojae was also found in Heilongjiang Province, Jilin Province and Beijing. In recent years, Shandong, Anhui, Henan, Jiangsu, Zhejiang Phytophthora sojae has also been isolated and identified in 15 provinces (municipalities) including , Inner Mongolia, Hubei, Fujian, Xinjiang, Sichuan, Tianjin and Guizhou. Among them,...

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Problems solved by technology

Common knowledge (2): For the naming of avirulent genes and disease-resistant genes, the avirulent genes are usually named first in the world, and the genes that can interact with the avirulent genes to identify the host as disease-resistant are named as the corresponding anti-viral genes. disease gene; that is, in theory, there may be multiple disease-resistant genes interacting with the same avirulent gene, but in fact it is true. The disease resistance genes Rps4 and Rps6 in soybean interact, but further research found that Avr4 and Avr6 are actually the same gene, renamed Avr4 / 6, and both disease resistance genes Rps4 and Rps6 can interact with the non-toxic gene Avr4 / 6 , respectively identify different parts of the avirulent gene Avr4 / 6 to trigger the disease resistance response. According to the analysis of common knowledge (2), in theory there may be multiple disease resistance genes interacting with the avirulent gene Avr3a, that is, from different disease resistance genes. The disease resistance gene Rps3a of the material may be a completely different disease resistance gene

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