2种捕食线虫真菌大量原生质体的快速制备与再生技术

 比较了培养基、菌龄、溶壁酶、配制酶液的缓冲溶液和酶解条件等重要因素对2种捕食线虫真菌原生质体形成的影响.采用优化后的制备条件,用较低质量浓度的酶液(5mg/mL纤维素酶,5mg/mL蜗牛酶),在短时间内(5h)蠕虫节丛孢(Arthrobotrys vermicola)原生质体得率可达0.5×10⁷～1.0×10⁷mL^-1,球状单顶孢(Monacrosporium sphaeroides)原生质体得率为0.8×10⁷～1.5×10⁷mL^-1.原生质体的再生除了受到原生质体制备方法的影响,还与再生培养基的营养成分、渗透压稳定剂及其浓度有很大关系.另外还对原生质体的释放和再生进行了详细的观察和描述.

The effective and rapid technique for protoplast generation and regeneration of two nematode-trapping fungi

The factors that influenced the formation of protoplasts of two nematode-trapping fungi,including culture media,mycelial stages,lysing enzymes and its dilution buffers,were studied and the optimized technique for protoplast preparation was determined.By the optimized protocol,the protoplast yields of the two fungi,Arthrobotrys vermicola and Monacrosporium sphaeroides were 0.5×10⁷—1.0×10⁷mL^-1,and 0.8×10⁷—1.5×10⁷mL^-1 respectively.The regeneration of protoplasts was affected not only by the generation method,but also by the nutrients,the osmetic stablizer and its concentration in the regeneration media.Additionally,the process of protoplast generation and regeneration was observed and described in detail.