A method for long-term preservation of Cordyceps militaris strains

A long-term preservation, Cordyceps militaris technology, applied in the preservation of microorganisms and other directions, can solve problems such as need to be placed upright, paraffin preservation method is not applicable, etc., achieve good physiological activities, promote large-scale and industrial development, and prevent pollution.

Inactive Publication Date: 2017-07-07
东莞市生物技术研究所
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This method is simple to make and does not require special equipment. The bacteria can be stored for about 1 year without frequent transplantation. The disadvantage is that it needs to be placed upright during storage.
[0003] However, for some strains with tenacious vitality, such as Cordyceps militaris strains, the existing paraffin preservation method is not applicable. Therefore, it is necessary to develop a long-term preservation method that can better inhibit the physiological activities of the strains

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] A method for long-term preservation of Cordyceps militaris strains, comprising the steps of:

[0024] A, strain culture: under aseptic conditions, inoculate the healthy strain of Cordyceps militaris on the sterilized PDA slant medium, carry out dark culture, and after the mycelium grows over 40% of the slope, carry out light culture;

[0025] B. Dehydration of the culture medium: Put the test tube containing the culture into a desiccant, then place the desiccant in the refrigerator, and take out the test tube when the slant medium is dehydrated to 20% of its volume;

[0026] C. Strain preservation: slowly inject sterilized paraffin oil into the test tube on the ultra-clean workbench to submerge the culture by 1cm, use a small vortex oscillator to remove air bubbles on the surface of the culture, insert sterilized degreased cotton, and use After the sterilized paraffin seals, cover with the sterilized pp film, and place it vertically in a refrigerator at 0°C for storage....

Embodiment 2

[0032] A method for long-term preservation of Cordyceps militaris strains, comprising the steps of:

[0033] A, strain culture: under aseptic conditions, inoculate the healthy strains of Cordyceps militaris on the sterilized PDA slant medium, carry out dark culture, and after the mycelia cover 45% of the slope, carry out light culture;

[0034] B. Dehydration of the culture medium: put the test tube containing the culture into a desiccant, then place the desiccant in the refrigerator, and take out the test tube when the slant medium is dehydrated to 22% of its volume;

[0035] C. Strain preservation: slowly inject sterilized paraffin oil into the test tube on the ultra-clean workbench to submerge the culture by 1.5cm, remove the air bubbles on the surface of the culture with a small vortex oscillator, and insert sterilized degreasing cotton. Seal with sterilized paraffin, cover with sterilized pp film, and place vertically in a 0°C refrigerator for storage.

[0036] Before sa...

Embodiment 3

[0041] A method for long-term preservation of Cordyceps militaris strains, comprising the steps of:

[0042] A. Strain cultivation: under aseptic conditions, inoculate the healthy strains of Cordyceps militaris on the sterilized PDA slant medium, and carry out dark cultivation. After the mycelia cover 50% of the slant, carry out light cultivation;

[0043] B. Dehydration of the culture medium: put the test tube containing the culture into a desiccant, then place the desiccant in the refrigerator, and take out the test tube when the slant medium is dehydrated to 25% of its volume;

[0044] C. Strain preservation: slowly inject sterilized paraffin oil into the test tube on the ultra-clean workbench to submerge the culture by 2cm, use a small vortex oscillator to remove air bubbles on the surface of the culture, insert sterilized degreased cotton, and use After the sterilized paraffin seals, cover with the sterilized pp film, and place it vertically in a refrigerator at 0°C for s...

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PUM

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Abstract

The invention relates to the technical field of preservation of strains and in particular relates to a method for preserving a cordyceps militaris strain for a long time. The method comprises the following steps: A. culturing the strain, namely inoculating a healthy cordyceps militaris strain to a sterilized PDA inclined medium under aseptic condition, carrying out dark culture, and carrying out light culture after hyphae overgrows 40%-60% of the slope; B. dewatering the culture medium, namely putting a test tube filled with culture into a drying vessel, then putting the drying vessel in a refrigerator, and taking out the test tube after the inclined medium is dewatered until 20%-30% of volume is left; and C. preserving the strain, namely slowly injecting sterilized paraffin oil into the test tube till the paraffin oil immerses the culture by over 1cm, shaking and removing bubbles on the surface of the culture, plugging sterilized and degreased cotton, sealing by using sterilized paraffin, covering a sterilized pp film, vertically arranging the test tube in the refrigerator at 0 DEG C for preserving. By virtue of the method, the physiological activity of the strain can be well inhibited; the strain can be preserved for more than 3 years; the preserved strain also has good activity.

Description

technical field [0001] The invention relates to the technical field of strain preservation, in particular to a method for long-term preservation of strains of Cordyceps militaris. Background technique [0002] The general steps of the existing paraffin preservation method are as follows: sterilize the liquid paraffin, put it in a thermostat at 37° C., and evaporate the water vapor for subsequent use. Then culture the purified bacteria to be stored in the most suitable slant medium to obtain robust bacteria, absorb sterilized liquid paraffin with a sterile pipette, inject it into the grown slant medium, and use an amount of 1cm higher than the inclined surface shall prevail, and the test tube shall be erected, labeled, and stored at 4°C. This method is simple to make and does not require special equipment. The strain can be preserved for about one year without frequent transplantation. The disadvantage is that it needs to be placed upright when preserved. [0003] However, ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/04
CPCC12N1/04
Inventor 郑贵朝叶锦泉喻孟君
Owner 东莞市生物技术研究所
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