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A method for inducing nuclear fission and normal formation of fruiting bodies in protoplast fusogens of Flammulina velutipes

A technology of protoplast fusion and Flammulina velutipes, which is applied in the fields of botanical equipment and methods, mushroom cultivation, horticulture, etc., can solve problems such as the inability of fusogens to produce fruit, and achieves the effects of low cost, high fruiting rate and easy operation.

Active Publication Date: 2020-03-20
INST OF SOIL FERTILIZER SICHUAN ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Using protoplast fusion technology to screen excellent strains of Flammulina velutipes is the focus of research by workers in this field, but the traditional method has the problem that the fusion sons cannot produce mushrooms after fusion of Flammulina velutipes protoplasts

Method used

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  • A method for inducing nuclear fission and normal formation of fruiting bodies in protoplast fusogens of Flammulina velutipes
  • A method for inducing nuclear fission and normal formation of fruiting bodies in protoplast fusogens of Flammulina velutipes
  • A method for inducing nuclear fission and normal formation of fruiting bodies in protoplast fusogens of Flammulina velutipes

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Effect test

Embodiment 1

[0015] Embodiment 1: a kind of method that protoplast fusion subnuclei divides and normally forms fruiting body in induction Flammulina velutipes species, it comprises the following steps:

[0016] S1. Preparation of induction medium: take bran and cottonseed, mix them evenly, add water, put them in a glass bottle and seal with a cotton plug, and sterilize at 105°C and 0.08MPa for 2.5h; the bran, cottonseed and The weight ratio of water is 1:3:2;

[0017] S2. Inoculation and cultivation: select Flammulina velutipes fusion sub-bacteria, transfer to PDA medium, and cultivate at 18°C ​​for 8 days, which is the mother species, and inoculate the mother species in the induction medium prepared in step S1, at a temperature of 20°C After cultured for 30 days, the protoplasts fused with daughter nuclei and split;

[0018] S3. Induce fruiting: put the fused Flammulina velutipes after inoculation and culture under the condition that the temperature is 10°C, the relative humidity is 90%,...

Embodiment 2

[0019] Embodiment 2: a kind of method that protoplast fusion subnuclei divides and normally forms fruiting body in induction mushroom species, it may further comprise the steps:

[0020] S1. Preparation of induction medium: take bran and cottonseed, mix them evenly, add water, put them in glass bottles and seal them with cotton plugs, and sterilize them at 120°C and 0.12MPa for 3.5h; the bran, cottonseeds and The weight ratio of water is 1:6:4;

[0021] S2. Inoculation and cultivation: Select the excellent fusion sub-bacteria of Flammulina velutipes, transfer to the PDA medium, and cultivate it at 25°C for 10 days, which is the mother species, inoculate the mother species in the induction medium prepared in step S1, and incubate at 25°C Cultivate for 40 days at a temperature above 100°C, and the protoplasts fuse with daughter nuclei to divide;

[0022] S3. Inducing fruiting: Place the inoculated and cultured Flammulina velutipes fused female and female species under the condi...

Embodiment 3

[0023] Embodiment 3: a kind of method that protoplast fusion subnuclei divides and normally forms fruiting body in induction Flammulina velutipes species, it comprises the following steps:

[0024] S1. Preparation of induction medium: take bran and cottonseed, mix them evenly, add water, put them into glass bottles and seal them with cotton plugs, and sterilize them for 3 hours at 110°C and 0.09MPa; the bran, cottonseeds and water The weight ratio is 1:4:3;

[0025] S2. Inoculation and cultivation: select Flammulina velutipes fusion sub-bacteria, transfer to PDA medium, cultivate at 20°C for 8.5 days, which is the mother species, inoculate the mother species in the induction medium prepared in step S1, and incubate at 22°C Cultivate at high temperature for 34 days, and the protoplasts fuse with daughter nuclei to divide;

[0026] S3. Induce fruiting: place the fused Flammulina velutipes after inoculation and culture under the conditions that the temperature is 13°C, the relat...

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Abstract

The invention discloses a method for inducing enoki mushroom intraspecific protoplast fusion daughter nucleus fission and fruiting body normal formation. The method comprises the following steps: S1, preparing an induction culture medium, including uniformly mixing bran and cottonseeds, and adding water, putting into a glass bottle, performing sterilization treatment in high temperature and high pressure; S2, inoculating and culturing, including inoculating cultivated enoki mushroom fusant mother strain on the induction culture medium to culture, protoplast fusion daughter nucleus splitting; S3, inducing to fruiting, including inducing the inoculated and cultured enoki mushroom fusant mother strain to fruiting, to form fruiting bodies. Through preparing the induction culture medium, inoculating and culturing, and inducing to fruiting, one enoki mushroom protoplast fusion daughter nucleus is split into two cell nucleuses, and a fruiting body can be formed normally. The method solves technical problems that fusant after fusion of enoki mushroom protoplast cannot fruit. The method is simple in operation, high in fruiting rate, low in cost, and is suitable for industrialized large scale production.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method for inducing protoplast fusion in the species of Flammulina velutipes to divide and normally form fruiting bodies. Background technique [0002] Commonly used microbial breeding techniques mainly include natural isolation, mutagenesis screening, cell combination, protoplast fusion, gene introduction, and gene destruction. Among them, protoplast fusion refers to artificial methods that make two cells with different genetic traits Fusion of protoplasts to obtain stable recombinants with both parental traits. This breeding method can break the species boundary of microorganisms, realize the gene recombination of distant strains, make the transfer of genetic material more complete, obtain more opportunities for gene recombination, and thus combine more excellent traits into a single strain. This technology has been paid attention to by the majority of edible fungus ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01G18/00
CPCA01G18/00
Inventor 王波
Owner INST OF SOIL FERTILIZER SICHUAN ACAD OF AGRI SCI
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