香菇原生质体工程菌株的构建研究——I.香菇原生质体的分离与再生行为

A Study on the Construction of Lentinula edodes Protoplast Engineering Strain The Behavior of Protoplast Isolation and Regeneration of Lentinula edodes ( I)

本研究通过电镜观察与酶解分析确定香菇(Lentinula edodes)菌丝细胞壁组成.并进行了双核菌丝原生质体的分离和培养.研究发现:酶解过程中,菌丝原生质体一般只从菌丝顶端等新生部位释放出来.合适的渗透剂、菌龄和酶解温度是原生质体得率的重要保证;原生质体再生菌丝过程是一个间歇性的依赖于原生质团涌动流向的过程,再生菌丝的形成经历了原生质体形成突起-哑铃状-分枝状等不同形态类型的再生阶段;再生菌丝体在形态上没有观察到锁状联合,栽培后未能结实,初步判断为单核菌株.

The composition of Lentinuta edodes hypha cell wall is determined by electron microscope observation and enzyme digestion analysis. And the protoplast isolation from dikaryotic mycelium and culture are also carried out in this study. The results showed that the protoplast was released generally from new growth part such as hypha tips through enzymolysis and the appropriate osmotic stabilizer, the strain age and the temperature during enzymolysis were all the important factors for high protoplast yield, that the procedure of protolast hypha regeneration was a procedure of intermittence depending on the direction of protoplasmatic mass movement and the formation of protoplast regenerated hypha experienced protruding-shape stage, dumbbell shape stage and branchy shape stage, and that no clamp connection was observed during mycelium growth period, the fruitbody could not be formed from the protoplast strain and the strain regenerated from protoplast might be the monokaryotic strain.