Preparing method of protoplast of edible mushrooms and protoplast

A technology of protoplasts and edible fungi, applied in the direction of fungi, etc., can solve the problems of low efficiency, slow speed, and many steps, and achieve the effect of reducing the total amount of solution, reducing filtration steps, and fast preparation process

Pending Publication Date: 2017-08-22
SHANGHAI ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, there are many steps, slow speed and low efficiency in the existing preparation method of edible fungus protoplasts.

Method used

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  • Preparing method of protoplast of edible mushrooms and protoplast
  • Preparing method of protoplast of edible mushrooms and protoplast
  • Preparing method of protoplast of edible mushrooms and protoplast

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Experimental program
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preparation example Construction

[0027] On the one hand, the preparation method of edible fungus protoplast provided by the invention, this preparation method comprises the following steps:

[0028] S1 mycelium culture steps

[0029] The edible fungus strains are inoculated in a container (such as a petri dish) containing a solid medium for mycelial culture to obtain a colony with mycelium.

[0030] Further, in some embodiments of the present invention, a 7-point inoculation method (such as figure 1 Shown) Inoculate the edible fungus strains on the solid medium for mycelial culture.

[0031] The present invention adopts solid medium for mycelia culture to obtain colonies with mycelium, and avoids reducing the production of mycelium fragments, so that the usage of lysozyme solution can be reduced in the subsequent enzymolysis step, and the enzyme can be simplified. The cultivation conditions of hydrolysis and the shortening of enzymolysis time greatly improved the preparation speed of edible fungus protoplas...

Embodiment 1

[0056] In this embodiment, taking shiitake mushrooms as an example, the preparation method of the edible fungus protoplast provided by the present invention is described, specifically as follows.

[0057] The preparation method of the edible fungus protoplast that present embodiment provides, it comprises the steps:

[0058] (1) The mushroom strain "Shenxiang 215" (purchased from the Institute of Edible Fungi, Shanghai Academy of Agricultural Sciences) was inoculated on a PDA plate covered with cellophane by the 7-point inoculation method, and cultured in the dark at 25°C;

[0059] (2) The shiitake mushroom strain was cultivated for about 4 days, and when the colonies at the 7 inoculation points expanded to the time when the hyphae of each colony were about to be connected, the size of the colony was as follows: figure 1 As shown, the preparation of protoplasts can be carried out;

[0060] (3) Place the cellophane with bacterial colonies in another empty sterilized petri dish...

Embodiment 2

[0073] In this embodiment, taking Flammulina velutipes as an example, the preparation method of the edible fungus protoplast provided by the present invention is described, specifically as follows.

[0074] (1) Inoculate Flammulina velutipes production strain "G1" (purchased from Edible Fungi Research Institute, Shanghai Academy of Agricultural Sciences) on a PDA plate covered with cellophane by 7-point inoculation method, and culture in the dark at 26°C;

[0075] (2) Flammulina velutipes strain was cultivated for about 3 days, and when the bacterium colonies of 7 inoculation points expanded to the time when the mycelium of each bacterium colony will be connected, the preparation of protoplast was carried out;

[0076] (3) Place the cellophane with bacterial colonies in another empty sterilized petri dish after being uncovered from the PDA plate, rinse the bacteria with a small amount of 0.6mol / L mannitol solution, and pour off the excess mannitol solution;

[0077] (3) Use 0....

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Abstract

The invention discloses a preparing method of protoplast of edible mushrooms and the protoplast, and relates to the technical field of preparation of engineering cells of filamentous fungi. The preparing method of the protoplast of the edible mushrooms comprises the steps of inoculating edible mushroom strains into a container containing a solid culture medium to be subjected to mycelium culture so that bacterial colonies in which hyphal is grown can be obtained; conducting cell wall enzymolysis on the bacterial colonies to obtain a protoplast coarse solution; filtering, centrifugating and re-suspending the protoplast coarse solution to obtain a protoplast solution. According to the preparing method, a large amount of protoplast can be quickly obtained. Compared with a conventional method, the preparing method of the protoplast of the edible mushrooms is greatly improved in the aspect of speeds, simple in step, and more convenient to operate.

Description

technical field [0001] The invention relates to the technical field of preparation of engineering cells of filamentous fungi, in particular to a method for preparing edible fungus protoplasts and the protoplasts. Background technique [0002] Protoplasts are cells that have lost their cell walls. During the preparation of protoplasts, the cell walls are artificially degraded. After removing the original cell walls, a spherical osmotic pressure-sensitive protoplast that is only wrapped by the cell membrane is obtained. In the process of protoplast formation in filamentous fungi such as edible fungi, the cell membrane wraps the different parts and sizes of the cells after the cells are broken by the filamentous hyphae, so that the formed protoplasts are single (cell) nucleated and binucleated. Or multinucleated and non-nucleated, monokaryotic and dikaryotic hyphae can be obtained after regeneration. [0003] Therefore, the preparation of edible fungus protoplasts can be used ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/14
CPCC12N1/14
Inventor 章炉军李菲于海龙张美彦宋春艳尚晓冬谭琦
Owner SHANGHAI ACAD OF AGRI SCI
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