【摘要】 采用单因素实验研究酶解温度、酶解时间、渗稳剂种类、培养方式对金针菇（Flammulina filiformis）双核菌株W8-Y33原生质体数量的影响；通过单单杂交与显微镜镜检，筛选出与单核菌株W8亲和、与单核菌株Y33不亲和的单核菌株H33，并对其菌落形态和线粒体类型进行分析。结果表明：在实验范围内，摇床培养7 d、酶解温度28℃、酶解时间2 h、甘露醇为渗稳剂时，菌株W8-Y33的原生质体数量(每毫升1.5×106)最多；筛选出的具有菌株W8细胞质的单核菌株H33菌落形态与Y33和W8差异明显；菌株H33的线粒体类型与单核菌株Y33不同，但与单核菌株W8和双核菌株W8-Y33相同。研究结果为金针菇细胞质遗传、品种改良等研究深入开展提供参考。更多还原

【Abstract】 Using single-factor experiments, the effect of enzymolysis temperature, enzymolysis time, type of osmotic stabilizer, and incubation mode on protoplast yield of dikaryotic Flammulina filiformis strain W8-Y33was studied. Through monokaryotic hybridization and microscopic examination, a monokaryotic strain named H33 was selected. It was compatible with the monokaryotic strain W8 but incompatible with the monokaryotic strain Y33. Then H33 was observed for colony morphology and analyzed for mitochondrial type. The results showed that the protoplast yield of W8-Y33 reached the highest(1.5×10~6 cells per milliliter) when mycelia cultured in shaking flasks for 7 d were enzymolyzed at 28 ℃ for 2 h with mannitol as the osmotic stabilizer.The selected monokaryotic strain H33 containing the cytoplasm of W8 exhibited significant differences in colony morphology compared with monokaryotic strains Y33 and W8. H33, W8 and W8-Y33 contained the same type of mitochondria, that was different from that of Y33. These results provided a reference for further research on cytoplasmic inheritance and variety improvement of F. filiformis.更多还原