Agaricus bisporus protoplast and preparation method thereof

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...

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