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Ultralow temperature preservation method for pleurotus eryngii original mother strains

A technology of eryngium eryngii and ultra-low temperature is applied in the field of preservation of the original parent species of eryngium eryngii, which can solve problems such as complicated operation, and achieve the effects of simple operation, good genetic stability and high survival rate.

Active Publication Date: 2016-04-20
湖南果秀食品有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The preservation method of stropharia strains (200810017694.4) adopts the vacuum freeze-drying method of mycelium to improve the low-temperature freezing resistance of the strains, and then preserves them in freezing tubes, which is complicated to operate

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] The present embodiment proposes a kind of cryopreservation method of the original parent species of Pleurotus eryngii, comprising the following steps:

[0018] A. Make a PDA inclined test tube, and after the sterility test, evaporate the water droplets and water on the test tube wall;

[0019] B. Transfer the original mother species to the above-mentioned qualified PDA medium, and culture it in the dark at 22±1°C for 16 days in an incubator, so that the mycelium is basically covered with the medium;

[0020] C, press glycerol: Tween-80: pure water=20%: 5%: 75% volume ratio preparation glycerol agent, then it is packed in the plastic test tube of low temperature resistance; Its 1 / 2-4 / 5 of the total capacity is cooled after autoclaving, and aseptically preserved for later use;

[0021] D. In the ultra-clean workbench, under aseptic conditions, cut the medium in the PDA inclined test tube into 3mm × 3mm medium pieces, and then transfer 3-6 medium pieces with hyphae to the...

Embodiment 2

[0025] The present embodiment proposes a kind of cryopreservation method of the original parent species of Pleurotus eryngii, comprising the following steps:

[0026] A. Make a PDA inclined test tube, and after the sterility test, evaporate the water droplets and water on the test tube wall;

[0027] B. Transfer the original mother species to the above-mentioned qualified PDA medium, and culture it in the dark at 22±1°C for 17 days in an incubator, so that the mycelium is basically covered with the medium;

[0028] C, press glycerol: Tween-80: pure water=25%: 4%: 71% volume ratio preparation glycerol agent, then it is packed in the plastic test tube of low temperature resistance; Its 1 / 2-4 / 5 of the total capacity is cooled after autoclaving, and aseptically preserved for later use;

[0029] D. In the ultra-clean workbench, under aseptic conditions, cut the medium in the PDA inclined test tube into 3mm × 3mm medium pieces, and then transfer 3-6 medium pieces with hyphae to the...

Embodiment 3

[0033] The present embodiment proposes a kind of cryopreservation method of the original parent species of Pleurotus eryngii, comprising the following steps:

[0034] A. Make a PDA inclined test tube, and after the sterility test, evaporate the water droplets and water on the test tube wall;

[0035] B. Transfer the original mother species to the above-mentioned qualified PDA medium, and culture it in the dark at 22±1°C for 18 days in an incubator, so that the mycelium is basically covered with the medium;

[0036] C, press glycerol: Tween-80: pure water=30%: 2%: 68% volume ratio preparation glycerol agent, then it is packed in the plastic test tube of low temperature resistance; Its 1 / 2-4 / 5 of the total capacity is cooled after autoclaving, and aseptically preserved for later use;

[0037] D. In the ultra-clean workbench, under aseptic conditions, cut the medium in the PDA inclined test tube into 3mm × 3mm medium pieces, and then transfer 3-6 medium pieces with hyphae to the...

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Abstract

The invention discloses an ultralow temperature preservation method for pleurotus eryngii original mother strains, which comprises: A, PDA (potato dextrose agar) slant tubes are made, and water beads on tube walls and moisture are evaporated; B, original mother strains are transferred into PDA culture mediums and put into a thermostat at 22+ / -1 DEG C for cultivating in darkness for 16-18 days, and the mediums are full of hyphae on the whole; C, glycerite is prepared from glycerinum, tween-80 and purified water at a volumetric proportion of (15%-30%):(0-5%):(80%-65%), separately loaded into high / low-temperature resistant plastic tubes to reach 1 / 2-4 / 5 of total volume, subjected to high-pressure sterilization and then cooled; D, the plurality of culture mediums with hyphae are transferred into the tubes, and the strain culture mediums are submerged in glycerite and sealed; E, the plastic tubes are precooled for 1-2 days at subzero 18+ / -2 DEG C, and then transferred to a refrigerator at subzero 80+ / -5 DEG C for long-term preservation. The ultralow temperature preservation method can be used for preserving pleurotus eryngii original mother strains for 8 years, and can ensure the vitality of the strains and the heredity stability; the possibility of variation is low and the cost is low.

Description

technical field [0001] The invention relates to the field of preservation methods for the original parent species of Pleurotus eryngii, in particular to a cryogenic preservation method for the original parent species of Pleurotus eryngii. Background technique [0002] In order to keep the good characters of edible fungus strains unchanged, it is necessary to preserve the parent species. At present, the preservation of edible fungus strains mainly uses liquid nitrogen cryopreservation method and slope regular transplantation method, etc. The liquid nitrogen cryopreservation method and the preservation cost are very high, and the slope regular transplantation method has a short storage time, and the strains are not stable and easy to occur Variation or degeneration etc., a kind of cryopreservation method (20061003531.7) of straw mushroom strain in the prior art adds mass fraction in the test tube slope of the straw mushroom strain inoculation and is 10-40% trehalose solution, ...

Claims

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Application Information

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IPC IPC(8): C12N1/04C12R1/645
CPCC12N1/04
Inventor 沈凡超阳国秀易恢满夏志兰姬建军
Owner 湖南果秀食品有限公司
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