程东升; 邹莉
东北林业大学; 东北林业大学 哈尔滨 150040; 哈尔滨 150040
【中文摘要】 原生质体制备是通过体细胞杂交或基因转导进行种质改良的基础。探讨了糙皮侧耳(Pleurotus ostreatus)原生质体制备及再生条件。用 MYG 培养菌丝,以甘露醇为稳渗剂,用1.5%溶壁酶 Lywallzyme 进行酶解脱壁,在菌龄7天、酶液 pH 值4.5、温度28℃、酶解4hr 的条件下原生质体产量最高(20×106/ml,0.01g 湿菌丝),其再生率大于8%,菌丝培养前对接种物进行予切碎处理可进一步使产量提高约2倍,而再生率并不显著下降。酶解液 pH 值为5.5和6.5时的原生质体产量低于 pH4.5,但再生率有所提高。
【英文摘要】 Preparation and culture of protoplasts are basic techniques for genetic improvement of plants and fungi through somatic cell fusion or gene transfer.Conditions for preparing the protoplasts from edible fungus Pleurotus ostreahs were examined.The highest production of protoplasts with plating efficiency of more than 8% was abtained with the enzyme mixture of pH 4.5 containing 1.5% Lywallzyme,0.6mol/L mannitol as osmotic stabilizer, and incubated at 28℃,from 7-day-age mycelia cultured in MYG medium.Shredding the inocula before mycelium incubation increased by 2 times the production of protoplasts. with plating efficiency not markedly affected.The protoplast production was lower at pH5.5 or pH6.5 than at pH4.5,but plating efficiency was increased at the former two pH value.
【中文关键词】 糙皮侧耳; 原生质体制备; 细胞工程
【英文关键词】 Pleurotus ostreatus; Protoplast preparation; Cytological engineering
【基金】国家教委资助优秀年轻教师基金项目
【文献出处】 中国食用菌,Edible Fungi of China,编辑部邮箱,1993年05期 【DOI】CNKI:SUN:ZSYJ.0.1993-05-005