平菇种内原生质体分离与融合杂交育种技术的研究

The techniques of intraspecific protoplast isolation and fusion crossbreeding in Pleurotus ostreatus

将16个平菇资源品种的菌丝体,在25℃、pH5.8～6.0环境下,分别用1.5%溶菌酶加0.6mol/LKCl稳渗剂溶解其细胞壁2h,原生质体数得量最高。将获得的原生质体两两配对成39个组合,在30℃、pH9、0.6mol/LKCl稳渗剂、不加融合促进剂CaCl2和PEG6000的融合条件下,得到种内杂交融合子5株。其中4株融合子再生出菌丝体和子实体;同皿接种实验显示,新菌株与其双亲具明显的拮抗线。经栽培试验与生产示范,新育的“优生1号”品系表现适应性强,优质、丰产、抗杂、耐热。

More protoplasts were obtained when 1.5% lysozyme and 0.6 mol/L KCl were used to dissolve the cell walls of 16 species mycelia in Pleurotus ostreatus for 2 h, at 25 ℃, pH 5.8～6.0. Intraspecific protoplast 5 fusion bodies were gained in 39 combinations of partnership wih 30 ℃, pH 9, 0.6 mol/L KCl, no the fusogen CaCl2 and PEG6000 on improving PDY medium. 4 fusants could regenerate mycelia and fruit bodies; inoculating their mycelia at same media revealed that the fusants manifested different resist lines from their parents each other. The cultivation and demonstration production showed that the new strain of Pleurotus ostreatus Yousheng No.1 was resistant to infection, diseases, high temperature, and has good adaptability and quality, steady high yield, delicious taste.