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Long-term storage method of cordyceps militaris strains high in cordycepin productivity

A long-term preservation, high cordycepin technology, applied in the field of agricultural microorganisms, can solve problems such as preservation of unfavorable offspring strains, fast strain reproduction, and reduced cordycepin yield, and achieve the effect of maintaining cordycepin-producing activity.

Active Publication Date: 2020-10-09
EASTERN LIAONING UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] In the prior art, the preservation methods of the Cordyceps militaris bacterial classification mainly include 4-15 ℃ test tube slant refrigeration, 26 ℃ air-conditioned room preservation, 4-15 ℃ test tube slant plus paraffin oil refrigeration, 26 ℃ test tube slant plus sterile water air conditioning Preservation and other methods, among them, 26°C air-conditioned room preservation and 26°C test tube slope plus sterile water air-conditioning method are suitable for short-term preservation, which can effectively guarantee the activity of contemporary strains and can be used as they are taken, but the strains reproduce at this temperature Fast, not conducive to the preservation of offspring strains
4-15°C test tube slant storage is an important method for long-term preservation of Cordyceps militaris strains due to the low temperature and the bacteria can survive for a long time. However, the activity of the bacteria will decrease after long-term storage on the slope, and rejuvenation is often required. Some The rejuvenation of the strain after long-term refrigeration is unsuccessful, and there is a risk of reduced production activity of cordycepin, resulting in a decrease in the production of cordycepin

Method used

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  • Long-term storage method of cordyceps militaris strains high in cordycepin productivity
  • Long-term storage method of cordyceps militaris strains high in cordycepin productivity
  • Long-term storage method of cordyceps militaris strains high in cordycepin productivity

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Effect test

Embodiment 1

[0024] A long-term preservation method of the Cordyceps militaris strain with high production capacity of cordycepin, comprising the following steps:

[0025] Inoculate the strain of Cordyceps militaris on the slant medium of the test tube, wrap the outer surface of the test tube with newspaper (that is, cultivate in a dark light environment), and cultivate it in a constant temperature incubator at 20±1°C, until the mycelium grows all over the slant, the slant strain is obtained;

[0026] The slant strains are cut into pieces under a sterile environment to obtain the bacterium blocks containing mycelium, and set aside;

[0027] Add 30% sterile glycerol and mycelia-containing bacterial blocks into the cryopreservation tube, and completely submerge the bacterial blocks with 30% sterile glycerol. Preserve the cryopreservation tubes using variable temperature treatment, and then take out the cryopreservation tube. Tubes were tested, and the surviving strains were selected for long...

Embodiment 2

[0035] A long-term preservation method of the Cordyceps militaris strain with high production capacity of cordycepin, comprising the following steps:

[0036] Inoculate the strain of Cordyceps militaris on the slant medium of the test tube, wrap the outer surface of the test tube with newspaper, and cultivate it in a constant temperature incubator at 20±1°C, until the mycelium grows all over the slant, the slant strain is obtained;

[0037] The slant strains are cut into pieces under a sterile environment to obtain the bacterium blocks containing mycelium, and set aside;

[0038] Add liquid paraffin and bacteria blocks containing mycelium to the cryopreservation tube, and make the liquid paraffin completely immerse the bacteria block, and use the variable temperature treatment method to store and pre-treat the cryopreservation tube, then take the cryopreservation tube for inspection, and select the surviving bacteria Long-term storage at -80°C;

[0039] The variable temperatu...

Embodiment 3

[0042] A long-term preservation method of the Cordyceps militaris strain with high production capacity of cordycepin, comprising the following steps:

[0043] Inoculate the strain of Cordyceps militaris on the slant medium of the test tube, wrap the outer surface of the test tube with newspaper, and cultivate it in a constant temperature incubator at 20±1°C, until the mycelium grows all over the slant, the slant strain is obtained;

[0044] The slant strains are cut into pieces under a sterile environment to obtain the bacterium blocks containing mycelium, and set aside;

[0045] Add 30% sterile glycerol and mycelia-containing bacterial blocks into the cryopreservation tube, and completely submerge the bacterial blocks with 30% sterile glycerol. Preserve the cryopreservation tubes using variable temperature treatment, and then take out the cryopreservation tube. Tubes were tested, and the surviving strains were selected for long-term preservation at -80°C;

[0046] The variab...

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Abstract

The invention belongs to the technical field of agricultural microorganisms, and particularly relates to a long-term storage method of cordyceps militaris strains high in cordycepin productivity. Thelong-term storage method comprises the steps of inoculating test tube slant culture media with the cordyceps militaris strains, performing culturing in a constant-temperature incubator of 18-25 DEG C,and when a slant is covered with mycelia, obtaining slant strains; cutting the slant strains into blocks in sterile environment to obtain bacterium blocks containing mycelia; adding protective liquidand the bacterium blocks containing mycelia into a frozen storage tube, enabling the protective liquid to completely immerse the bacterium blocks, performing preservation pretreatment on the frozen storage tube by a poikilothermic treatment method, then taking the frozen storage tube for inspection, and selecting viable strains for long-term preservation at minus 80 DEG C. According to the long-term storage method disclosed by the invention, culture media containing ginkgo fruit powder are adopted, so that the cordycepin production activity of the cordyceps militaris strains can be maintained. The poikilothermic treatment method including the steps of reducing room temperature to minus 10 DEG C, then raising the temperature to 4 DEG C, and finally reducing the temperature to minus 80 DEGC is also adopted, so that the low temperature resistant properties of the strains can be improved, and the activity of the cordyceps militaris strains can be reserved.

Description

technical field [0001] The invention belongs to the technical field of agricultural microorganisms, and in particular relates to a method for long-term preservation of Cordyceps militaris strains with high production capacity of cordycepin. Background technique [0002] Cordyceps militaris belongs to the Ascomycota subphylum Ergotaceae Cordyceps genus, is an important traditional Chinese medicine. Cordycepin contained in Cordyceps militaris is a nucleoside antibiotic, which has antibacterial, anti-inflammatory, antiviral, and antitumor activities, and has high nutritional and medicinal value. The screening and preservation of Cordycepin militaris strains with high production capacity of cordycepin is an important prerequisite for the separation and extraction of cordycepin. The number of Cordyceps militaris grown in nature is small and the yield is low, so the technology of artificially cultivating high-cordycepin Cordyceps militaris is gradually rising. [0003] In the pr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01G18/40A01G18/20
CPCA01G18/40A01G18/20
Inventor 刘晓红
Owner EASTERN LIAONING UNIV
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