A kind of long-term preservation method of Trichosporum sinensis

A technology of Chinese hairy spores and mycelium, applied in the field of microorganisms, can solve the problems of easy variation of strains, heavy workload, frequent species transfer, etc., and achieve the effect of reducing the variation and pollution rate

Active Publication Date: 2016-05-04
YUNNAN UNIV
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  • Description
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Problems solved by technology

The separation of Cordyceps sinensis strains is relatively difficult, and the traditional slant 4 preservation can only be preserved for a short period of time (6 months), and its species is changed frequently, which has caused problems such as a large workload and easy mutation of the strains.

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  • A kind of long-term preservation method of Trichosporum sinensis
  • A kind of long-term preservation method of Trichosporum sinensis

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Embodiment Construction

[0013] The present invention will be described in detail below in conjunction with specific embodiments.

[0014] 1. Materials and methods

[0015] 1.1 Bacteria to be tested

[0016] The mycelium was isolated from Cordyceps sinensis Baima Xueshan in Zhongdian, Yunnan Province, and identified as Trichosporum sinensis by ITS sequence.

[0017] 1.2 Mycelium separation method: take fresh Cordyceps sinensis, take the sub-seat part, disinfect the surface with disinfectant alcohol, use tweezers to take a 1*2mm tissue piece and place it on a PDA plate for cultivation at 18°C, and observe the growth under a dissecting microscope every day Condition. Move the germinated mycelia, slow-growing, and dense mycelium of the tissue block to the PDA slope at 18°C ​​to continue culturing, and observe whether there is pollution during the period. Take the non-polluted slant mycelium, extract DNA, PCR amplification with ITS primers, sequence, and identify whether it is Chaelopedia sinensis by s...

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Abstract

The invention discloses a method for the long-term preservation of Hirsutella sinensis. The method comprises the steps of: separating a mycelia from Cordyceps sinensis, and then culturing the mycelia on a PDA slant, then inoculating the mycelia cultured on slant onto PDA plates, culturing at 18 DEG C for 30 days. Cutting into agar blocks with mycelia by using a scalpel, transferring the agar blocks to sterile 10-20% glycerol cryovials, sealing the cryovials with a sealing film, and storing in a refrigerator with temperature of -70 DEG C. The problems of the variation of the strain and the contamination rate due to the multiple times of inoculation are reduced by the method disclosed by the invention.

Description

technical field [0001] The invention relates to the technical field of microbes, in particular to a long-term preservation method of Trichosporum sinensis. Background technique [0002] Cordyceps sinensis (Ophiocordyceps sinensis) is a precious traditional Chinese medicine and a rare tonic. It is a fungus complex formed by its asexual Hirsutellasinensis infecting the larvae of Hepialus spp.. It is distributed in my country's Qinghai, Tibet, Sichuan, Alpine shrub meadows at an altitude of 3500-5000m in Yunnan and Gansu. [0003] Since Cordyceps sinensis has two completely different forms, the complete form (also known as the sexual generation) and the incomplete form (also known as the asexual generation), 10 years ago, the means of strain identification were backward, and molecular identification had not yet been carried out. For a long period of time, most of them mistakenly cultivated Paecilomyces bat moth as the strain of Cordyceps sinensis only by the morphology of isola...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/04C12R1/645
Inventor 沙涛普春龙张亚平田国樑
Owner YUNNAN UNIV
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