Composite protective agent for preserving Boletus aereus strain for long and preservation method thereof

A technology of black boletus and protective agent, which is applied in the directions of microorganism-based methods, preservation of microorganisms, biochemical equipment and methods, etc., can solve the problems of reduced yield, slow growth of strains, whitish mycelium, etc., and achieves resistance to stress. The effect of strong sex, long storage time and reducing damage

Inactive Publication Date: 2015-06-10
JINGHONG HONGZHEN AGRI TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The commonly used protective agent, dimethyl sulfoxide, after preservation, the strains grow slowly, and the mycelium turns white; the strains do not germinate after the glycerin preservation; There are less degradation phenomena, and the yield is also reduced when the fruit is cultivated

Method used

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  • Composite protective agent for preserving Boletus aereus strain for long and preservation method thereof
  • Composite protective agent for preserving Boletus aereus strain for long and preservation method thereof
  • Composite protective agent for preserving Boletus aereus strain for long and preservation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Inoculate the mycelium block on the stock culture medium, culture at a constant temperature of 30°C, and obtain the stock boletus for preservation in 15 days, wherein the stock culture medium contains the following components:

[0037]

[0038]

[0039] The color of the obtained black boletus parent species is milky yellow to golden, with sterile skin, sterile strands, no water spit, and good ductility;

[0040] Make the mycelium block of the black boletus parent species with a length and width of 0.9cm×0.5cm, and put it into a sterilized low-temperature freezing tube according to aseptic operation. The specification of the low-temperature freezing tube is 2ml, internal rotation, The cryopreservation tube contains 0.5ml of sterile composite protective agent, wherein the composite protective agent includes the following components: calculated by volume of water, sucrose 100g / L, potassium dihydrogen phosphate 1.0g / L, magnesium sulfate 1.0g / L, Ammonium tartrate 1.0g / ...

Embodiment 2

[0045] Inoculate the mycelium block on the stock culture medium, culture at a constant temperature of 29°C, and obtain the stock boletus for preservation in 18 days, wherein the stock culture medium contains the following components:

[0046]

[0047] The color of the obtained black boletus parent species is milky yellow to golden, with sterile skin, sterile strands, no water spit, and good ductility;

[0048] Make the mycelium block of black boletus parent species with a length and width of 1cm×0.3cm, and put it into a sterilized low-temperature freezing tube according to aseptic operation. The specification of the low-temperature freezing tube is 2ml, internal rotation, low temperature The cryopreservation tube contains 1ml of sterile composite protective agent, wherein the composite protective agent includes the following components: calculated by volume of water, sucrose 200g / L, potassium dihydrogen phosphate 1.5g / L, magnesium sulfate 1.5g / L, ammonium tartrate 1.5g / L;

...

Embodiment 3

[0053] Inoculate the mycelium block on the stock culture medium, culture at a constant temperature of 28° C., and obtain the stock boletus for preservation in 20 days, wherein the stock culture medium contains the following components:

[0054]

[0055] The color of the obtained black boletus parent species is milky yellow to golden, with sterile skin, sterile strands, no water spit, and good ductility;

[0056] Make the mycelium block of the black boletus parent species with a length and width of 1.1cm×0.4cm, and put it into a sterilized low-temperature freezing tube according to aseptic operation. The specification of the low-temperature freezing tube is 2ml, internal rotation, The cryopreservation tube contains 1.5ml of sterile composite protective agent, wherein the composite protective agent includes the following components: calculated by volume of water, 350g / L of sucrose, 1.8g / L of potassium dihydrogen phosphate, 1.8g / L of magnesium sulfate, Ammonium tartrate 1.8g / L...

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Abstract

The invention relates to the field of fungus preservation, and specifically relates to a composite protective agent for preserving a Boletus aereus strain for long and a preservation method thereof. The composite protective agent for preserving the Boletus aereus strain for long comprises ingredients as follows: counting by water volume, 100-400g / L of cane sugar, 1.0-2.0g / L of monopotassium phosphate; 1.0-2.0g / L of magnesium sulfate; and 1.0-2.0g / L of ammonium tartrate. The preservation method comprises the steps as follows: soaking the Boletus aereus strain into the composite protective agent, pre-cooling for 30min to 60min in the condition at 110 degrees centigrade below zero to 130 degrees centigrade below zero, and then dipping into a liquid nitrogen or a gas nitrogen for preservation. Using the composite protective agent for the preservation, each ingredient cooperates with each other so as to prevent the Boletus aereus strain from being damaged in the ultralow temperature condition; the specific preservation method makes the preservation time longer, and makes the hereditary stability better; the preservation method is characterized in that the yield does not have an obvious difference from the yield before the preservation, and the like; the composite protective agent for preserving the Boletus aereus strain for long and the preservation method thereof have an excellent application prospect, and synchronously provide a reliable guarantee for an industrial large-scale cultivation of the Boletus aereus strain.

Description

technical field [0001] The invention relates to the field of fungal preservation, in particular to a composite protective agent for long-term preservation of black boletus strains and a preservation method thereof. Background technique [0002] The temperature of liquid nitrogen is -196°C, at which temperature the metabolism of fungal cells is completely stopped, thereby avoiding the degradation and variation of fungal strains. Since HWANG successfully used the liquid nitrogen preservation method to preserve fungal strains in 1960, the method has been continuously improved and the technology has become more and more mature. The type and concentration of the protective agent is one of the key issues in the liquid nitrogen preservation method. Although MATA has successfully preserved the strains without using the protective agent and the programmed cooling device, the strains sensitive to low temperature such as black boletus still need Use a protectant. Commonly used protec...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/04C12R1/645
CPCC12N1/04
Inventor 周茂纪光燕石泉纪光玉杨太茂杨君巧谢飞郭雪玲
Owner JINGHONG HONGZHEN AGRI TECH CO LTD
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