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探讨了在森林资源综合利用中具重要意义的木腐性食用菌香菇(Lentinus edodes)的原生质体制备及再生条件。对用MYG培养的菌丝,采用以0.6mol甘露醇为稳渗剂的1.5％溶壁酶Lywallzyme进行酶解脱壁,在菌龄7d、酶解液pH值4.5、温度28℃、酶解4h时原生质体产量最高(6×10~6/ml,每0.1g鲜菌丝)。酶解温度>31℃时原生质体产量和再生率都明显降低。用SO培养基代替MYG可使原生质体产量提高1倍,但再生率显著下降(从8.5％下降到1.1％)。从产量和再生率两方面综合考虑,以MYG作为菌丝培养基效果较好。
The protoplast preparation and regeneration conditions of the woody mushroom Lentinus edodes which are of great importance in the comprehensive utilization of forest resources were discussed. The mycelium cultured with MYG was treated with 0.6% mannitol as stabilizing agent for 1.5% of lywallzyme Lywallzyme for enzymolysis. After 7 days of culture, the pH value of the hydrolyzate was 4.5, the temperature was 28 ℃, Protoplast yield the highest (6 × 10 ~ 6 / ml, per 0.1g fresh mycelium). When the temperature of enzymolysis> 31 ℃, the yield of protoplast and the regeneration rate decreased significantly. Protoplast production was double by using SO medium instead of MYG, but the regeneration rate was significantly reduced (from 8.5% to 1.1%). Considering both yield and regeneration rate, it is better to use MYG as mycelial medium.