猪苓原生质体制备与再生条件的研究  

Study on Protoplast Preparation and Regeneration of Grifola umbellata (Pers.) Fries

作　　者：张丽霞[1] 郭成金[1] 

机构地区：[1]天津师范大学化学与生命科学学院,天津3000387

出　　处：《中国食用菌》2008年第5期35-37,55,共4页Edible Fungi of China

基　　金：天津市科学技术委员会项目(编号:06ZHCXNC05900)

摘　　要：采用L16(45)正交设计,得出猪苓原生质体制备最佳条件是取菌龄7d的菌丝体,以2%溶壁酶,0.6mol·L-1甘露醇作为渗透压稳定剂,在33℃下酶解3h,原生质体数达到2.62×107个·mL-1。原生质体再生的最佳条件是取菌龄5d的菌丝体,以2%溶壁酶,0.6mo·lL-1蔗糖作为渗透压稳定剂,在27℃的条件下,酶解3h。猪苓原生质体再生的最佳培养基为葡萄糖10g、麦芽糖5g、酵母粉4g、0.6mo·lL-1甘露醇、琼脂5.5g,定容到1000mL。再生率可达到0.65%。Author adopted orthogonal experiment and obtained the results. The optimal preparing condition of Grifola umbellata protoplast is that 7-day-old mycelium, applying 0.6mol·L^-1 mannitol for osmotic pressure stabilizer, using 2% lywallzyme, digesting at 33℃ for 3 hours. The number of protoplast is 2.62×10^7·mL^-1. Protoplasts produced in the condition that 5-day-old mycelium, at 27℃ by 0.6mol·L^-1 cane sugar, 2% lywallzyme shacking 3 hours were optimal to regeneration. The medium with the following composition： glucose 10g, yeast extracts 4g, maltose 5g, agar 5.5g, 0.6mol ·L^-1 mannitol, added distilled water to 1000mL was suitable for regeneration. The regeneration rate was 0.65%.

关 键 词：猪苓 原生质体 正交设计 制备 再生 

分 类 号：S567.3[农业科学—中草药栽培]