Method for direct extraction of pathogen DNA from pathogenic tissue of Graminaceous crop

Abstract

The invention provides a method for direct extraction of pathogenic fungus and bacteria DNA from leaf scabs of gramineous crops such as pathogenetic rice and corn. The method of the invention can directly extract pathogen DNA from leaf and neck of rice and corn leaf scabs infected by rice blast, the extraction method is simple, fast and low-cost, and the extracted DNA is applicable to PCR based analysis on pathogenic organism molecular detection and diagnosis, pathogen nontoxic gene detection and colony genetic diversity.

Description

technical field [0001] The invention belongs to the field of microbial molecular detection, in particular to a method for directly extracting DNA of pathogenic bacteria from susceptible tissues of gramineous crops. Background technique [0002] The extraction of DNA from crop pathogenic microorganisms is the basis for the detection and analysis of related pathogenic molecules. The methods commonly used to extract the DNA of traditional pathogenic fungi are CTAB extraction and cell wall lysis. The CTAB method requires the grinding of liquid nitrogen in the early stage, or the use of enzymes to break the cell wall, and then uses toxic reagents such as chloroform to extract to remove proteins, DNA precipitation and elution and other steps. Although high-quality DNA can be obtained, but The whole process is time-consuming and inefficient. Moreover, the isolation and cultivation of pathogenic microorganisms need to be completed in the early stage of DNA isolation, and the exper...

AI Technical Summary

Problems solved by technology

The CTAB method requires the grinding of liquid nitrogen in the early stage, or the use of enzymes to break the cell wall, and then uses toxic reagents such as chloroform to extract to remove proteins, DNA precipitation and elution and other steps. Although high-quality DNA can be obtained, but The whole process is time-consuming and inefficient

Moreover, the isolation and cultivation of pathogenic microorganisms need to be completed in the early stage of DNA isolation, and the experiment cycle is long.

Although the method of kit extraction is much simpler than the traditional method, it has the problem of high cost, which is not conducive to the analysis and detection of a large number of samples

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