槐耳原生质体制备与再生研究

Study on protoplast preparation and regeneration of Trametesrobiniophila

对槐耳原生质体制备与再生进行了研究,结果表明:槐耳原生质体制备最佳条件是以液体静置培养9d的菌丝体,0.6mol/L蔗糖作为渗透压稳定剂,在体积质量比2%溶壁酶、1%纤维素酶和1%蜗牛酶作用下,25℃酶解3h,制备率达2.75×106个/mL;再生最佳条件是液体静置培养12d的菌丝体,0.6mol/L甘露醇为渗透压稳定剂,在2%溶壁酶作用下,35℃酶解2h,再生率为12.7%.

The conditions of preparing and regenerating Trametes robiniophila protoplasts were studied.The results showed that the optimal preparing conditions of Trametes robiniophila protoplast were 9-day-old mycelium,apply-ing 0.6mol/L sucrose for osmotic pressure stabilizer,using 2%Lywallzyme,1%Cellulase,1%Snailase,digesting at 25℃for 3hours.Protoplast achieved 2.75×106/mL.Protoplasts produced in the conditions that 12-day-old mycelium,at 35℃by 0.6mol/L mannitol,2% Lywallzyme digesting 2hours were optimal to regeneration.The regeneration rate was 12.7%.