A method for cloning and sequence analysis of an active product gene resistant to melon wilt

Abstract

The invention belongs to the technical field of melon wilt prevention and control, and discloses a method for cloning and sequence analysis of an active product gene resistant to melon wilt, including: extraction of soil microbial genome DNA by ectopic lysis; construction of a metagenomic Fosmid library and its quality Detection; screening the metagenomic Fosmid expression library to obtain Fosmid active clones; purification and identification of active substances, determining the structure of active substances, and verifying the activity of compounds against melon wilt; screening and transforming plant endophytic strains, and transforming genes into transformed By selecting different promoters, different media and fermentation conditions, a high-efficiency expression system for drug genes resistant to melon wilt was established. The invention uses the isolation, screening and transformation of plant endophytes as expression strains to establish a heterologous high-efficiency expression system for biologically active target genes screened from a library. Lay a solid foundation for the commercialization of anti-fusarium wilt drugs.

Description

technical field [0001] The invention belongs to the technical field of melon wilt prevention and control, and in particular relates to a method for cloning and sequence analysis of an active product gene resistant to melon wilt. Background technique [0002] Fusarium wilt of melon, also known as vine cutting disease and wilt disease, is one of the main soil-borne diseases in melon production, especially the most serious damage to watermelon, cucumber and muskmelon. The disease has spread to dozens of countries and regions such as China, the United States, Italy, Israel, Japan, and India, and occurs to varying degrees in the watermelon growing areas in the north and south of my country. Especially in the southern tropical and subtropical regions, the rainfall is abundant, the climate is mild, and the pathogenic bacteria do not overwinter, resulting in a large accumulation of spores of the pathogenic bacteria in the soil, and the incidence of diseases is extremely high. %, ev...

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Problems solved by technology

[0003] In summary, the existing problems in the prior art are: at present, most of them are controlled by chemical agents. Since Fusarium wilt is a soil-borne disease, its pathogenic bacteria have a strong masking ability, and it is difficult for chemical fungicides to directly contact with it, so it cannot be fundamentally killed. The cost of application is high to kill pathogenic bacteria; at the same time, a series of side effects such as polluting the environment and destroying ecological balance caused by the widespread use of chemical agents have become increasingly prominent, and production and use are gradually being restricted; in addition, long-term use of chemical drugs, pathogenic bacteria are prone to drug resistance , over time, chemical drugs lose their original effect

The probability of success in discovering new antibiotic drugs is getting smaller and smaller, and there is often a phenomenon of repeated screening

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