Molecular method for determining the pathogenicity of the Phytophthora soybean avirulence gene avr3a to the differential host l83-570

Abstract

The invention relates to a molecular method for determining the pathogenicity of phythophthora sojae avirulent genes Avr3a to a differential host L83-570.The genome DNA of phythophthora sojae to be determined is extracted; DNA fragments including complete avirulent genes Avr3a are obtained through PCR amplification; DNA fragments are obtained through enzyme digestion and amplification of restriction endonuclease Bmg BI; the enzyme digestion product is subjected to agarose gel electrophoresis, wherein the corresponding phythophthora sojae strains to be determined have no pathogenicity to the differential host L83-570 if an agarose gel electrophoretogram is provided with two DNA bands within the range of 250 bp to 500 bp, and the corresponding phythophthora sojae strains to be determined have the pathogenicity to the differential host L83-570 if one DNA band exists within the range of below 750 bp after enzyme digestion.The determination period of the method is short, no intermediate form exists, and the pathogenicity of the multiple phythophthora sojae avirulent genes Avr3a to the differential host L83-570 can be simultaneously determined.

Description

technical field [0001] The invention relates to a molecular determination method, in particular to a molecular method for determining the pathogenicity of Phytophthora soybean avirulence gene Avr3a to the identification host L83-570. 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...

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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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