Preparation method of tricholoma matsutake culture genome

Abstract

The invention relates to a preparation method of a tricholoma matsutake culture genome, which is characterized by comprising the following steps: 1) separating pure mycelium culture from tricholoma matsutake sporocarp so as to obtain the mycelium culture of the tricholoma matsutake; 2) preparing protection buffer liquid which comprises 50mmol / L of Tris-HCL buffer liquid, 50mmol / L of sodium ethylene diamine tetracetate, 1.4mol / L of NaCl and 4wt% of cetyltrimethylammonium bromide; 3) immersing the mycelium culture of the tricholoma matsutake in the protection buffer liquid, oscillating, freezing with liquid nitrogen, water bathing, and repeating the oscillating, freezing with liquid nitrogen and water bathing for 1-3 times; 4) extracting, centrifuging, and collecting supernate; and 5) precipitating and centrifuging the supernate with isopropanol the volume of which is the same as that of the supernate, and then obtaining precipitate, dissolving the precipitate in TE (Tris+EDTA) buffer liquid, storing, and obtaining the tricholoma matsutake culture genome. The preparation method has the advantages of trace use materials, less steps, short consumed time and high yield, and the prepared tricholoma matsutake culture genome is complete in quality and high in purity.

Description

technical field [0001] The invention belongs to the technical fields of applied microbiology and molecular biology, and in particular relates to a preparation method of a matsutake culture genome. Background technique [0002] Tricholoma matsutake, also known as matsutake, is a symbiotic fungus with plants. Its fruiting body is an expensive mushroom with edible and medicinal value. At present, it cannot be cultivated internationally. The wild matsutake mushrooms in the Changbai Mountains in Northeast my country and the Hengduan Mountains in Southwest China are mainly for export, earning nearly 10 billion U.S. dollars for the country every year. In order to protect and utilize this rare natural resource, it is urgent to decipher the genome of the matsutake mushroom and develop the preparation method of the genome of the matsutake culture. [0003] Due to the rare and expensive fruiting body of matsutake mushroom, the research material is purely cultured mycelium of matsutake...

AI Technical Summary

Problems solved by technology

[0005] 1. Mechanical methods, mainly various methods such as liquid nitrogen grinding, room temperature grinding, glass bead grinding, etc. The disadvantages are cumbersome operation, loss of biomass during operation, low yield, unsafe, genome degradation, and unstable product quality

[0006] 2. Chemical method, using benzyl chloride and various lysozymes for digestion, the disadvantages are poor specificity, high cost, and narrow scope of application

[0007] 3. Physical method, reported the application of ultrasonic, microwave, electric drill, boiling and other methods, the disadvantages are cumbersome operation, unsafe, genome degradation, unstable product quality, loss of biological activity