Extraction and purification method of total plant endophyte genome DNA for colony analysis

Abstract

The invention discloses an extraction and purification method of total plant endophyte genome DNA for colony analysis. The method comprises the following steps of: shearing and grinding the surface bacteria removed fresh plant tissues into paste, soaking the plant tissues into sterilized phosphate buffer solution, performing thermostatic shaking for 1 hour in a table concentrator to separate out microbial cells from the plant tissues, standing, transferring the suspension to a centrifuge tube, performing centrifugal collection on microbial bacteria, then adding lysing solution, shaking and uniformly mixing the solution to release DNA from the cells, repeatedly extracting protein by chloroform-isoamylol, performing centrifugal sedimentation with isopropanol and washing with 70 percent ice ethanol, performing purification by using a DNA specific centrifugal adsorption column to obtain total crude extracted genome DNA, and finally, performing polymerase chain reaction (PCR) amplificationby using special primers to obtain high-quality plant endophyte specific DNA fragments for subsequent analysis. The invention has the advantages that: the method is simple and effective, high in quality, low in pollution and the like, and provides high-quality guarantee for comprehensively and completely researching a plant endophyte colony structure.

Description

technical field [0001] The invention belongs to the technical field of molecular biology, and in particular relates to the extraction and purification of plant endophyte genome total DNA applied to community analysis. Background technique [0002] Microorganisms generally exist in healthy plant tissues (including roots, stems, leaves, fruits, etc.), and they not only actively exist in plants, but also produce many biological functions. As a natural biological resource, it has broad theoretical research value and development and application prospects. Through the study of the microbial community in plants, we can not only understand the population distribution of endophytes, but also explore their physiological functions that are beneficial to the growth of host plants, and provide rich microbial resources for the sustainable development of agriculture and environmental pollution bioremediation. The study of plant endophytes has important theoretical and practical significan...

AI Technical Summary

Problems solved by technology

With the development of molecular biology, various methods for extracting total DNA from environmental samples have been gradually established, but for various environmental samples, none of the methods can be applied to all environmental samples. Unique physical, chemical and biological properties require optimization of a unique method for extracting total DNA

At present, there are many studies on the extraction of genomic DNA in other environments such as soil, activated sludge, and water. However, due to the lack of domestic research on microbial communities in plant tissues, the microbial genome in plants is the premise and basis of its research. The extraction method of total DNA is rarely reported, which requires us to find a fast, effective and low-cost method for the extraction and purification of high-quality, high-reliability and complete genome total DNA

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