Method for low-temperature preservation and unfreezing of agrocybe aegirit strains

A technology for cryopreservation and tea tree mushroom, applied in the direction of preservation of microorganisms, methods based on microorganisms, biochemical equipment and methods, etc., can solve the problems of expensive equipment and less equipment, and achieve convenient management, easy degradation, and reduced puncture The effect of cell probability

Inactive Publication Date: 2016-07-06
LUDONG UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Ultra-low temperature refrigerators have better freezing effect, but the equipment is expensive and rarely equipped in enterprises; while household refrigerators are cheap and generally equipped in enterprises

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] (1) Low-temperature preservation of strains:

[0035] ① Cultivation of Agrocybe agrocybe liquid strain:

[0036] Under aseptic conditions, pick a test tube of Agrocybe aegypti which is the size of soybean grains, and inoculate them in GPY liquid medium. The formula components of GPY liquid medium are calculated in g / L as: yeast extract 10, peptone 6, glucose 20, the remainder Water; 25-28°C, 160rpm constant temperature culture for 8-10 days.

[0037] ② Preparation of protection solution:

[0038] 60% concentration of glycerin: accurately measure 60ml glycerin, add water to make the volume up to 100ml;

[0039] 1% xanthan gum: accurately weigh 1g of xanthan gum solid, dissolve it in 100ml of water, 121℃, sterilize for 30min;

[0040] ③In the ultra-clean workbench, using aseptic operation, use a pipette to add 0.5ml of 1% xanthan gum and 1ml of 60% glycerol as a protective solution to a 2ml cryotube, and then add the ready 0.5ml of tea tree mushroom liquid strain.

[0041] ④Put the...

Embodiment 2

[0046] (1) Low-temperature preservation of strains:

[0047] ① Cultivation of Agrocybe agrocybe liquid strain:

[0048] Under aseptic conditions, pick a test tube of Agrocybe aegypti which is the size of soybean grains, and inoculate them in GPY liquid medium. The formula components of GPY liquid medium are calculated in g / L as: yeast extract 10, peptone 6, glucose 20, the remainder Water; 25-28°C, 160rpm constant temperature culture for 8-10 days.

[0049] ② Preparation of protection solution:

[0050] 60% concentration of glycerin: accurately measure 60ml glycerin, add water to make the volume up to 100ml;

[0051] 20% mannitol: accurately weigh 20g of mannitol and dissolve it in 100ml of water;

[0052] ③In the ultra-clean workbench, using aseptic operation, use a pipette to add 0.5ml of 20% mannitol and 1ml of 60% glycerol as a protective solution to a 2ml cryotube, and then add the prepared tea tree Mushroom liquid strain 0.5ml.

[0053] ④Put the prepared cryopreservation tube stra...

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Abstract

The invention discloses a method for low-temperature preservation and unfreezing of agrocybe aegirit strains. The method is characterized by comprising the following steps: (1) low-temperature preservation of the strains: taking 0.5ml of agrocybe aegirit liquid strains, adding 1ml of 60% glycerol and 0.5mlof 1% xanthan gum as a protective solution, or adding 1ml of 60% of glycerol and 0.5ml of 20% mannitol as a protective solution, and performing freezing preservation in a liquid pipe at -20 DEG C; and (2) unfreezing: taking the strains in frozen preservation, putting the strains into water of 35-40 DEG C immediately, shaking the strains quickly for unfreezing, then transferring the strains into a GPY solid medium, and performing constant temperature culture at 25-28 DEG C.The method disclosed by the invention keeps the agrocybe aegirit strains to be high in activity and not to deteriorate, is simple and convenient to operate, and is low in price, low in equipment requirement, small in occupied space, and easy to popularize.

Description

Technical field: [0001] The invention relates to a method for low-temperature preservation and thawing of tea tree mushroom strains, and relates to the technical field of microbial strain preservation. Background technique: [0002] tea tree mushroom (Agrocybeaegirit) It belongs to Basidiomycota, Basidiomycetes, Copromataceae, and Agrocybe genus. It is a medium-temperature edible fungus; it contains 18 kinds of amino acids needed by the human body, rich in nutrients, high in protein, low in lipids and sugars, and healthy The curative effect is obvious; moreover, the nutritional value of tea tree mushroom exceeds that of many other edible fungi. Above; At present, there are certain researches on the function and cultivation methods of tea tree mushroom, but there are few researches on its strain preservation. In the face of such a large-scale cultivation area, strain preservation is particularly important. Efficient and cheap strain preservation methods are conducive to the...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/04C12N1/14C12R1/645
CPCC12N1/04C12N1/14
Inventor 刘静孙磊程显好李维焕
Owner LUDONG UNIVERSITY
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