秀珍菇原生质体高效制备与再生研究

Study on the Separation and Regeneration of Pleurotus pulmonarius Protoplast

为建立秀珍菇原生质体的高效制备体系,以夏秀作为供试菌株,采用正交试验法研究影响秀珍菇原生质体制备和再生的条件,对菌龄等7个单因素、 4个复合因素条件和接种方式进行系统研究。结果显示,秀珍菇菌丝在改良液体PD培养基中摇瓶培养5 d,以0.6 mol·L^(-1)甘露醇为稳渗剂, 2.5%溶壁酶在30℃条件下水浴4 h,得到的原生质体浓度最大,为4.23×107个·mL^(-1)。在TB3再生培养基中采用单层混菌法接种原生质体,其再生率最大,为2.35%。

In order to establish highly efficient preparation and regenaration system of Pleurotus pulmonarius protoplasts, we use the orthogonal test method to systematically study the conditions for the preparation and regeneration of P. ptdmonarius protoplasts. The results showed that when P. pulmonarius mycelium for shake flask cuhure in improved liquid PD medium for 5 days, and hydrolyzed with lysozymes at a concentration of 2.5% and with 0.6 mol L^-1 mannitol as osmotic stabilizers at 30℃ water bath for 4 h, the maximum concentration of protoplast is 4.23×107 个 mL^-1, the highest regenaration rate inoculationg by mixing the protoplast with unsolidifier medium in TB3 regenaration medium is 2.35%.