金针菇与凤尾菇科间原生质体融合研究

Studies on the Fusion of Protoplasts from Flammulina velutipes and Pleurotus sajor-caju

金针菇与凤尾菇皆属四极性异宗结合食用菌,其双核异核体菌株都具锁状联合遗传标记。本研究以金针菇和凤尾菇的双核异核菌株为亲本,将热灭活的凤尾菇原生质体,以PEG为融合剂,在高钙、高pH值条件下,与金针菇原生质体融合,结果从1329个再生菌株中选择出6株双亲细胞质和细胞核都融合的无锁状联合单核菌株,经融合核分裂技术处理后,融合核分裂成为具锁状联合的双核菌株。应用RAPD技术分析,证明融合子菌株均含有与双亲株同源的遗传物质,是真正的融合子菌株。

Flammulina velutipes (Fr. ) Singer and Pleurotus sajor-caju (Fr.) Singer were tetrapolar and heterothallic edible fungi and their hetero-dicaryotic strains, selected as parent strains in this study, were both with clamp connection as genetic marker. F. velutipes protoplast and heat-treated P. sajor-caju protoplast were fused under high concentration of Ca2+and high pH conditions using PEG as fusogen. And 6 uninucleate fusants without clamp connection, with cytoplasm and cell nuclei of parent strains being all fused, were identified from 1329 regenerate strains. Using fused nucleus division technique, the fused nucleus was divided into dikaryotic strain with clamp connection. While fusants containing the same genetic material as the parent strains was proved to be the genuine fusants by RAPD technique.