平菇原生质体制备、再生及灭活研究

Studies on Preparation, Regeneration and Ultraviolet-inactivatiou of Pleurotus Protoplasts

紫孢侧耳、糙皮侧耳和佛罗里达侧耳的原生质体制备与再生的最佳条件为：用１％溶壁酶＋０．５％蜗牛酶的混合酶液酶解培养３～４天的菌丝体，１．５小时后，原生质体产量均可达１０￣（１０）个／Ｌ；三种侧耳的最佳再生培养基分别为ＳＭ、ＭＰ和ＲＰ，再生率分别为０．６１％、０．６７％和０．７６％。随着紫外线对原生质体处理时间的加长，再生率逐渐下降，当处理４ｍｉｎ时，再生率接近零。

The optimum conditions of isolation and regeneration of protoplasts for Pleurotus sapidus, P.ostreatus and P. florida were 0.6mol/L MgSO_4 as an osmotic stabi- lizer,1%Lywallzyme+0.5 % Helicase,the mycelia being cultivated for 3～4 days in liquidmedium PSA,enzyme reaction temperature at 30℃,under these conditions,over 10￣(10)/Lprotoplasts could be obtained in one and half hour for three species,the optimum regenera-tion mediums of the three sapidus were separately SP,MP and RP,and their regenerationfrequencies were 0.61 %,0.67 % and 0.76%respectively.The longer the time that theprotoplasts were treated with U. V. ,the lower the frequency of protoplast regeneration. When the protoplasts were irradiated for 4 miniters,the frequency of protoplast regenera-tion was almost zero.