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Method for establishing mutant strain by utilizing plasma induced lucid ganoderma protoplast

A protoplast and plasma technology, which is applied in the field of crop mutation breeding, can solve the problems of harvesting season limitation, difficult to achieve mutagenesis effect, etc.

Active Publication Date: 2017-08-18
HEFEI INSTITUTES OF PHYSICAL SCIENCE - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] As a multicellular organism, Ganoderma lucidum has a thicker cell wall, and it is difficult to achieve good mutagenesis effects by conventional methods; although spore mutagenesis can obtain single-cell mutagenesis effects, its harvesting season is limited

Method used

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  • Method for establishing mutant strain by utilizing plasma induced lucid ganoderma protoplast
  • Method for establishing mutant strain by utilizing plasma induced lucid ganoderma protoplast
  • Method for establishing mutant strain by utilizing plasma induced lucid ganoderma protoplast

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Embodiment 1

[0041] A method of utilizing plasma mutagenesis ganoderma lucidum strain cgmcc 5.0026 protoplasts of the present embodiment to construct mutant strains comprises the following steps:

[0042] (1) Construction of Ganoderma lucidum protoplasts

[0043] ①Under sterile conditions, inoculate small pieces of mycelium of solid fermented Ganoderma lucidum seeds into sterilized PD liquid medium (1L sterile water, 200g potatoes, 20g glucose), and cultivate them on a shaking table at 27°C and 150rpm , after 5 days, the mycelium and the bacterial solution were taken out, centrifuged in an ultra-clean bench and washed with ddH 2 O cleaning mycelium;

[0044] ②Take 5mL Ganoderma lucidum mycelium and place it in a sterilized homogenizer, add an appropriate amount of mannitol at a concentration of 0.6mol / L; grind it in the homogenizer, and prepare mycelium homogenate in an ultra-clean bench; Observe that the length of the mycelia is appropriate, centrifuge at 2000rpm for 6 minutes to obtain...

Embodiment 2

[0061] A method of using plasma mutagenesis of Chibaihuang Ganoderma lucidum protoplasts to construct mutant strains in this embodiment comprises the following steps:

[0062] (1) Construction of Ganoderma lucidum protoplasts

[0063] ①Under sterile conditions, inoculate small pieces of solid fermented Ganoderma lucidum seed mycelium into sterilized PD liquid medium (1L sterile water, 200g potatoes, 20g glucose), and culture at 29°C and 200rpm on a shaking table After 7 days, the mycelium and the bacterial solution were taken out, centrifuged in the ultra-clean bench and washed with ddH 2 O cleaning mycelium;

[0064] ②Take 5mL Ganoderma lucidum mycelium and place it in a sterilized homogenizer, add an appropriate amount of mannitol at a concentration of 0.6mol / L; grind it in the homogenizer, and prepare mycelium homogenate in an ultra-clean bench; Observe that the length of the mycelia is appropriate, centrifuge at 5000rpm for 4 minutes to obtain a homogenate of the mycelia...

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Abstract

The invention discloses a method for establishing a mutant strain by utilizing a plasma induced lucid ganoderma protoplast. The method comprises the following steps that 1, a lucid ganoderma protoplast is established, wherein a liquid lucid ganoderma culture mycelium is prepared, a mycelium homogenate is prepared, a cell wall resolvase solution is prepared and cell walls are dissolved, the protoplast is separated and collected, and the protoplast concentration is corrected and saved; 2, protoplast mutation is induced by low-temperature plasma, wherein preparation is performed before protoplast mutation, a plasma mutagenesis device is ventilated for balance, a reaction system is electrified for protoplast mutation, a flat protoplast plate is evenly coated and standing regeneration and culture are performed, a mutant strain is selected, the colonial morphology and growing rate are observed and recorded, and hypha morphology is observed under a light microscope, active substances and stability of the mutant strain are detected, and the strain is saved. The method has the advantages of stably and efficiently utilizing plasma to induce the lucid ganoderma protoplast so as to establish the mutant strain.

Description

technical field [0001] The invention relates to the technical field of crop mutation breeding, in particular to a method for using plasma to induce ganoderma protoplasts to construct mutant strains. Background technique [0002] Ganoderma lucidum (Ganoderma Lucidum) is the fruiting body of Basidiomycotina, Laminomycetes, Polyporaceae, Aphphylloidaceae, and Ganoderma lucidum, and is a relatively valuable traditional Chinese herbal medicine. Among them, the active ingredients of Ganoderma lucidum, such as Ganoderma lucidum polysaccharides and triterpenes, have the functions of anti-tumor, anti-oxidation, immune regulation, liver protection, and blood sugar regulation. [0003] As a multicellular organism, Ganoderma lucidum has a thicker cell wall, so it is difficult to achieve a good mutagenic effect by conventional methods; although spore mutagenesis can obtain a single-cell mutagenic effect, its harvesting season is limited. Because the cell wall of protoplasts is artificia...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/01C12N13/00C12N1/14C12R1/645
CPCC12N1/14C12N13/00C12N15/01
Inventor 黄青马玉涵何华奇姚国华
Owner HEFEI INSTITUTES OF PHYSICAL SCIENCE - CHINESE ACAD OF SCI
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