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Preparation and regeneration method for hirsutella sinensis protoplast

A technology of Chinese hairy sporophytes and protoplasts, applied in the direction of microorganism-based methods, biochemical equipment and methods, microorganisms, etc., can solve problems that are difficult to meet the needs of research, and achieve easy promotion and implementation, shortened regeneration time, and simple operation Effect

Active Publication Date: 2020-06-26
ZHEJIANG UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] There is no complete and efficient protoplast preparation and regeneration system in the existing technology, which is difficult to meet the needs of follow-up research

Method used

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  • Preparation and regeneration method for hirsutella sinensis protoplast
  • Preparation and regeneration method for hirsutella sinensis protoplast
  • Preparation and regeneration method for hirsutella sinensis protoplast

Examples

Experimental program
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Effect test

Embodiment 1

[0033] (1) Culture and purification of mycelia

[0034] Get Mortierella chinensis mycelium ZJB18002 (from the Bioengineering Research Institute of Zhejiang University of Technology, disclosed in CN109082382A), inoculate it in the seed medium, and cultivate it at 11-26° C. and 60-200 rpm for 15-30 days. Transplanted in the fermentation medium, cultured at 11-26°C, 60-200rpm for 3-12 days, washed mycelium with 0.4-1.0mol / L potassium chloride solution for 2-3 times in a centrifuge at 6000-10000rpm, The purified mycelium is collected; the seed medium is composed of 10-30g of silkworm chrysalis powder, 10-25g of bran, 10-25g of corn flour, 10-30g of glucose, and 3-10g of yeast powder per liter of medium , dextrin 3-10g, peptone 5-20g, potassium dihydrogen phosphate 0.05-2g, anhydrous magnesium sulfate 0.05-2g. The composition of the fermentation medium is that each liter of medium contains 10-30g of glucose, 3-10g of yeast powder, 3-10g of dextrin, 5-20g of peptone, 0.05-2g of pot...

Embodiment 2

[0071] Embodiment 2: adopt the inventive method to prepare and regenerate Mortierella chinensis protoplast

[0072] The specific operation steps are as follows:

[0073] a. Mycelia cultivation and purification

[0074] Take the mycelium of Mortierella sinensis, inoculate it in the seed medium, and cultivate it at 16°C and 150rpm for 21 days, then transfer it to the fermentation medium, cultivate it at 16°C and 150rpm for 7 days, and incubate it in a centrifuge at 8000rpm The mycelia were washed three times with 0.6 mol / L potassium chloride solution, and the purified mycelium was collected. The seed medium consists of: every liter of medium contains silkworm chrysalis powder 15g, bran 15g, corn flour 10g, glucose 15g, yeast powder 5g, dextrin 5g, peptone 10g, potassium dihydrogen phosphate 0.2g, anhydrous magnesium sulfate 0.1g, the balance is water. The composition of the fermentation medium is as follows: every liter medium contains 15g of glucose, 5g of yeast powder, 5g o...

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Abstract

The invention relates to a preparation and regeneration method for hirsutella sinensis protoplast. The method comprises the following steps of: taking a potassium chloride solution to serve as an osmotic pressure stabilizing agent, taking commodity enzymes Driselase from Basidiomycetes sp, Lysing Enzyme from Trichoderma harizianum and Yatalase as compound enzymes to carry out enzymolysis so as toprepare the hirsutella sinensis protoplast with a high content, and successfully regenerating. Meanwhile, a regeneration medium is optimized to shorten protoplast regeneration time, and in addition, the preparation and regeneration method is simple to operate, can be conveniently prompted and implemented and has a good application prospect.

Description

[0001] (1) Technical field [0002] The invention relates to a preparation and regeneration method of Mortierella chinensis protoplast. [0003] (2) Background technology [0004] Cordyceps sinensis (Berk.) Sacc.) referred to as Cordyceps, is Ascomycota (Ascomycota), Sclerotinia (Pyrenomycetes), Ergot (Clavicipitales), Ergotaceae (Clavicipitaceae), Cordyceps (Cordyceps) a species of fungus. Cordyceps fungus infects the larvae of Lepidoptera bat moths, and uses the organic matter in the body as a source of nutrition and energy for parasitic life. After continuous growth, development and differentiation, the mycelium finally kinks and forms a shell that protrudes from the host The sub-seats form complexes with the larval corpses, thus constituting Cordyceps sinensis. Modern pharmacology has confirmed that Cordyceps sinensis has a wide range of biological activities such as immune regulation, antibacterial, anti-tumor, anti-oxidation, anti-aging, hypoglycemic and blood fat, and ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/14C12N1/00C12R1/645
CPCC12N1/14C12N1/005
Inventor 金利群柳志强徐哲文张博郑裕国
Owner ZHEJIANG UNIV OF TECH
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