松茸菌丝原生质体形成与再生条件的研究

Studies on the protoplast preparation of Tricheloma matsutake

采用多因素实验正交优选法比较不同酶、渗透压稳定剂、菌丝生理状况、菌丝培养基、预处理液和酶解时间等因素对松茸原生质体形成的影响和再生培养基、明胶、酪蛋白对松茸原生质体再生的影响。以 0 6MMgSO4 作为渗透压稳定剂 ,2 -硫基苏糖醇为预处理液 ,用 2 %蜗牛酶在 30℃酶解 6h ,每克菌丝可以得到 1× 10 6 个原生质体。再生最优条件 :0 6MMgSO4 ,PDY再生培养基 ,2 5%明胶 ,松茸菌丝再生率最高 4 97%。

Effect of lysozyme,osmosis stabilizer,filament age,pretreatment and medium on protoplast preparation of Tricholma matsutake was determined by orthogonal test.After pretreatment of DDT,then treated 4 hours at 30℃ in 0.6 M MgSO 4 buffer containing 2% snailase,one gram of filaments could release 1×10 6 protoplasts.Studies on the protoplast regeneration showed,that the regeneration rate is 4.97% when protoplasts were grown on the PDY agar medium containing 2.5% gelatin and 0.6mol/L MgSO 4.