A strain isolation method for efficiently obtaining effective inoculum of Cordyceps militaris

A technology for strain separation and Cordyceps militaris, applied in botany equipment and methods, horticulture, applications, etc., can solve problems such as unclear boundaries, unstable strains of Cordyceps militaris, and difficulty in distinguishing fertile hyphae from Cordyceps militaris plate colonies , to achieve the effect of simple operation, less variation, and increased number of passages

Inactive Publication Date: 2016-01-20
BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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  • Abstract
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  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, it is difficult to distinguish the fertile mycelium part and the non-fertile mycelium of the Cordyceps militaris plate colony obtained by the conventional tissue separation method, and the boundary is not obvious, which finally shows as the instability of the Cordyceps militaris bacterial classification

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] 1. Experimental time: April 2012:

[0027] 2. Experimental materials and design:

[0028] 1. Strains: The Cordyceps militaris strain was purchased from the Institute of Microbiology, Chinese Academy of Sciences, No. PRB.1235. Strain A was isolated by traditional methods; strain B was isolated by the following method:

[0029] Use tweezers to hold the fruiting bodies used for tissue isolation, stay in 80°C sterile water for 1.5 seconds, and then quickly transfer to 4°C sterile water in the refrigerator for 2 minutes.

[0030] 2. Culture medium for liquid shake flasks: 300 grams of peeled potatoes (boiled juice), 10 grams of glucose, 2 grams of peptone, 0.5 grams of potassium dihydrogen phosphate, 1 gram of magnesium sulfate, 1000 ml of water, the pH is natural.

[0031] 3. Nutrient solution for fruiting body production: 2% soybean, 1% glucose, 5% milk powder, 0.5% potassium dihydrogen phosphate, 0.05% magnesium sulfate, vitamin B110mg∕1000ml to make a nutrient solution...

Embodiment 2

[0040] The basic operation is the same as in Example 1, except that the bacterial strain B is obtained by processing and separating by the following method:

[0041] Clamp the fruiting bodies used for tissue separation with tweezers, stay in 75% alcohol for 60 seconds, then quickly transfer to sterile water and wash three times.

[0042] After harvesting, the average fresh weight of strain A reached 28.5 g / bottle. After harvesting, the average fresh weight of strain B reached 32.3 g / bottle.

Embodiment 3

[0044] The basic operation is the same as in Example 1, except that the bacterial strain B is obtained by processing and separating by the following method:

[0045] Pour a small amount of 95% alcohol into a Petri dish, light it, use tweezers to hold the fruiting bodies used for tissue separation, and go back and forth on the blue flame for 1-3 times, and then quickly transfer to a refrigerator at 4°C in the refrigerator. Stay in sterile water for 4 minutes.

[0046] After harvesting, the average fresh weight of strain A reached 27.8 g / bottle. After harvesting, the average fresh weight of strain B reached 33.6 g / bottle.

[0047] The above examples show that the operation of the present invention is simple and easy, the separation effect is good, and the effective inoculum part of the plate colony is easy to identify. Not only the stability of the isolated strain is strong, but also the effect of increasing production is achieved compared with the traditional isolation method...

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Abstract

The invention relates to a strain isolation method capable of effectively obtaining effective inoculant of cordyceps militaris. The strain isolation method is characterized by adopting a heat shock method to process sporocarps. Compared with a traditional isolation plate, the isolation plate in the strain isolation method is less in aerial hyphae, dense in aerial hyphae and fast in aerial hyphae growth. After the dark cultivation is completed, illumination cultivation is fast in colouring and dark in color. The aerial hyphae twist together easily, and primordia are easily formed on the plate. The cultivation experiment shows that compared with the traditional isolated strains, the primordia of the sporocarpa of the isolated strains in the strain isolation method in a cultivation bottle appear early, the form of the sporocarpa corresponds to the form of a parent, and the sporocarps grow neatly and are low in degree of variation. The strain isolation method is simple and feasible in operation, good in isolation effect and legibile in effective inoculum of plate bacterial colony. The isolated strains are strong in stability and less in variation. Compared with the traditional method, the passage number is increased.

Description

technical field [0001] The invention relates to the technical field of Cordyceps militaris cultivation, in particular to a method for maintaining the strain of Cordyceps militaris and its genetic characteristics. Background technique [0002] Cordyceps militaris (L.Fr) Link, also known as Cordyceps militaris, Cordyceps militaris, belongs to Ascomycota, Pyrenomycetes, Sphaeriales, Clavicipitaceae, Cordyceps The genus (Cordyceps (Fr.) Link) fungi. Cordyceps militaris is the type species of the fungus of the genus Cordyceps, and it parasitizes the larvae and pupae of various lepidopteran sun insects in nature. [0003] In recent years, studies have shown that the chemical composition and pharmacological effects of artificially cultivated Cordyceps militaris are similar to those of Cordyceps sinensis, but the price is far lower than that of Cordyceps sinensis. Therefore, the development and application of Cordyceps militaris has a great potential market. [0004] Artificial cu...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01G1/04
Inventor 邢礼军刘春阁张爱武喻少帆王卫红刘新香闫明明杨海涛芦丽红
Owner BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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