Special low-temperature cryoprotectant for cordyceps sinensis strain preservation and use method thereof

A technology of low-temperature freezing and Cordyceps strains, applied in the direction of preservation of microorganisms, methods based on microorganisms, biochemical equipment and methods, etc., can solve the problems of preservation reports, no Cordyceps fungal strains, etc.

Pending Publication Date: 2020-07-10
NANJING INST FOR THE COMPREHENSIVE UTILIZATION OF WILD PLANTS CHINA COOP +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are also some reports on the preservation methods for the Cordyceps militaris strains, but there is no report for the preservation of the Cordyceps fungus strains

Method used

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  • Special low-temperature cryoprotectant for cordyceps sinensis strain preservation and use method thereof

Examples

Experimental program
Comparison scheme
Effect test

example 1

[0020] 1. Culture of Cordyceps militaris

[0021] The Cordyceps militaris strains were inoculated on PDA slant medium and cultured in the dark at 25 °C. When the diameter of the colony grew to 4 cm, the strains were cultured under light for 4 days to induce the color change of the strains, reduce the formation of aerial hyphae, and promote the formation of spores.

[0022] 2. Preparation of protective agent

[0023] The protective agent consists of two parts, that is, Cordyceps liquid medium, which consists of: glucose 10 g, peptone 4 g, yeast extract 1 g, water to 1000 mL, natural pH. The other part is glycerin. The Cordyceps nutrient solution was sterilized at 115 °C for 30 min, and the glycerin was sterilized at 121 °C for 20 min. Glycerin was prepared with Cordyceps nutrient solution under aseptic conditions before use, and the mass concentration of glycerin was 30%.

[0024] 3. Preparation of strain mycelium and spore suspension

[0025] Gently scrape the mycelium of...

example 2

[0029] 1. Culture of cicadae strains

[0030] Cicadae strains were inoculated on PDA plates with a diameter of 9 cm, and cultured in the dark at 25 °C. When the diameter of the colony grows to 3 cm, the bacteria are cultured under light for 3 days to promote the formation of spores.

[0031] 2. Preparation of protective agent

[0032] The protective agent is composed of two parts, that is, Cordyceps nutrient solution, which is composed of: 20 g of glucose, 8 g of peptone, 2 g of yeast extract, dilute water to 1000 mL, and natural pH. The other part is glycerin. The two parts were sterilized separately, the Cordyceps nutrient solution was sterilized at 115 °C for 30 min, and the glycerin was sterilized at 121 °C for 20 min. Glycerin was prepared with Cordyceps nutrient solution under aseptic conditions before use, and the mass concentration of glycerin was 40%.

[0033] 3. Preparation of strain mycelium and spore suspension

[0034] Gently scrape the mycelia of the culture...

example 3

[0038] 1. Cultivation of Cordyceps gunis

[0039]Cordyceps guni strains were inoculated on PDA plates with a diameter of 6 cm, and cultured in the dark at 16 °C. When the diameter of the colony grew to 3 cm, the strain was cultured under light to induce the mycelia to form spores.

[0040] 2. Preparation of protective agent

[0041] The protective agent is composed of two parts, that is, Cordyceps liquid medium, which is composed of 18 g of glucose, 5 g of peptone, 1 g of yeast extract, water to 1000 mL, and natural pH. The other part is glycerin. The two parts were sterilized separately, the Cordyceps nutrient solution was sterilized at 115 °C for 30 min, and the glycerin was sterilized at 121 °C for 20 min. Glycerin was prepared with Cordyceps nutrient solution under aseptic conditions before use, and the mass concentration of glycerin was 35%.

[0042] 3. Preparation of strain mycelium and spore suspension

[0043] Gently scrape the mycelia of the cultured plate strain...

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Abstract

The invention belongs to the technical field of fungus strain preservation, and relates to a special low-temperature cryoprotectant for cordyceps sinensis strain preservation and a use method. The method comprises the following steps: carrying out strain culture, and inducing strains to form spores by utilizing illumination; collecting hyphae into a sterilized mortar, adding a protective agent, performing grinding, and preparing a suspension, wherein the protective agent is composed of two parts, one part is a cordyceps sinensis nutrient solution which is composed of 10-20 g of glucose, 4-8 gof peptone, 1-2 g of yeast extract and water performing constant volume to 1000 mL with natural pH value, and the other part is glycerin; respectively sterilizing the two parts, and preparing a 10-50%glycerol solution by using a cordyceps sinensis nutrient solution before use; and performing filtration by adopting a cell sieve, collecting filtrate, sub-packaging the filtrate into sterile freezingtubes, sealing the tubes, and carrying out low-temperature freezing preservation. The validity period of strains preserved by the protective agent provided by the invention is more than 3 years, andthe protective agent can be directly used for producing liquid strains, so that the subculture frequency is reduced.

Description

technical field [0001] The invention belongs to the technical field of fungal strain preservation, and relates to a special low-temperature cryoprotectant for preservation of Cordyceps strains and a use method thereof. Background technique [0002] China is the world's largest producer and exporter of edible fungi. The edible fungus industry has become the fifth largest planting industry after grain, oilseeds, fruits and vegetables. In the past two years, my country's total output of edible fungi has reached nearly 40 million tons. Although the scale of edible fungi continues to expand, there are still many problems in edible fungi in my country, the most prominent of which is the problem of strains. Insufficient provenance, serious strain degradation, backward seed production technology and other problems seriously restrict the development of the industry. Since the traditional seed production technology is carried out by expanding the cultivation of solid strains, the cul...

Claims

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Application Information

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IPC IPC(8): C12N1/14C12N1/04C12R1/645
CPCC12N1/14C12N1/04
Inventor 林群英邓春英杨彝华孙力军侯北伟
Owner NANJING INST FOR THE COMPREHENSIVE UTILIZATION OF WILD PLANTS CHINA COOP
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