Agaricus bisporus protoplast and preparation method thereof

A technology of Agaricus bisporus and protoplasts, applied in the field of bioengineering, can solve the problems of high price and unsatisfactory effect of lysozyme, and achieve the effect of reducing preparation cost and cost

Inactive Publication Date: 2012-06-13
SOUTH CHINA AGRI UNIV
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AI Technical Summary

Problems solved by technology

[0003] At present, most of the enzymes for preparing Agaricus bisporus protoplasts in China use lytic enzymes, but the single use of lytic enzymes is not only expensive but also unsatisfactory, and the release of protoplasts can only reach 10 5 , and the regeneration rate can only reach 10 -5 , are at a lower level

Method used

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Experimental program
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Effect test

Embodiment 1

[0025] The preparation method of Agaricus bisporus (Agaricus bisporus 2796, purchased from Guangdong Institute of Microbiology) mycelium is as follows:

[0026] On the Agaricus bisporus mycelium plate, use a puncher to take 3 vigorously growing mycelium blocks at the front end, connect them to liquid PDA medium (potato dextrose agar medium), and shake them at 26°C and 160r / min. Bed culture for 7 days, then filter the liquid, take 0.4 g of the above-mentioned liquid and transfer the mycelia obtained after filtration to a 250 mL Erlenmeyer flask containing 160 mL of liquid PDA medium, and culture at 26 ° C for 6 days, that is, the mycelium is 6 days bacteria age, and the supernatant was filtered to obtain Agaricus bisporus mycelium.

[0027] The configuration method of the mixed enzyme solution is as follows:

[0028] Weigh cellulase and helicase and add sterile 0.6mol / L KCl and 0.6mol / L mannitol to dissolve in the osmotic pressure stabilizer obtained by mixing 1:1 by volume, s...

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Abstract

The invention belongs to the field of bioengineering, and discloses an agaricus bisporus protoplast and a preparation method thereof. The preparation method comprises the following steps of: pretreating agaricus bisporus hyphae of which the cell ages are 6-9 days with a permeation pressure stabilizing agent prepared by mixing 0.6 mol / L of KCl and 0.6 mol / L of mannitol in the volume ratio of 1:1; performing enzymolysis by using a mixed enzyme solution containing 7.6 percent by mass of cellulase and 4 percent by mass of helicase at the temperature of 28-30 DEG C for 2-3 hours; and centrifugally separating to obtain the agaricus bisporus protoplast, wherein the release quantity of the protoplast is 9.9*106 / mL, and the regeneration rate in a regeneration culture medium is up to 2.11 percent. Cellulase and helicase are taken as a mixed enzyme system of which the price is lower than single lywallzyme by over a half, the preparation cost of the agaricus bisporus protoplast in scientific research is reduced, and a foundation is laid for agaricus bisporus breeding with a protoplast technology.

Description

technical field [0001] The invention belongs to the field of bioengineering and relates to a preparation method of Agaricus bisporus protoplast, in particular to a cheap Agaricus bisporus protoplast and a preparation method thereof. Background technique [0002] Agaricus bisporus, also known as white mushroom, is an edible fungus belonging to the genus Agaricaceae of the Basidiomycetes Agaricaceae. It is currently the mushroom with the largest cultivation area and consumption in the world. The protein and essential amino acids of the human body, as well as a variety of vitamins, are a kind of fungus with the same source of medicine and food. Because the totipotency of protoplasts of edible fungi is easier to manifest, protoplast mutagenesis and fusion breeding technology is one of the most efficient and economical methods in the breeding of high-yielding and new strains of edible fungi. The successful preparation of protoplasts is a necessary condition for protoplast mutage...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/04
Inventor 莫美华梁大伟
Owner SOUTH CHINA AGRI UNIV
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