白灵菇原生质体制备与再生研究

Study on protoplast preparation and regeneration of Pleurtus nebrodensis Lnzenga

借助正交设计法对白灵菇原生质体制备与再生进行研究.结果表明,原生质体最佳制备体系是:液体静置培养7d的菌丝体,在20g/L的溶壁酶作用下,以0.6mol/L蔗糖为渗透压稳定剂,28℃酶解2h,最高制备率为1.34×107个/mL;其最佳再生体系是:以液体静置培养7d的菌丝体,0.6mol/L MgSO4.7H2O为渗透压稳定剂,在15g/L溶壁酶与10g/L蜗牛酶作用下,34℃酶解3h,最高再生率为3.7%.

The conditions of preparing and regenerating of Pleurtus nebrodensis Lnzenga protoplasts were studied through orthogonal design experiments. The results showed that the optimal preparing conditions of Pleurtus nebrodensis Lnzenga protoplast were 7 - day-old mycelium cultured in static liquid, using 20 g/L Lywallzyme, applying 0.6 mol/L sucrose for osmotic pressure stabilizer, digesting at 28 ℃ for 2 hours. And protoplast achieved 1.34× 10^7/ml. Protoplasts produced in the conditions of 7 -day-old mycelium cultured in static liquid, applying 0.6 mol/L MgSO4 ·7H2O for osmotic pressure stabilizer, using 15 g/L Lywallzyme and 10 g/L Snailase, digesting at 34 ℃ for 3 hours were optimal to regeneration. And the regeneration rate was 3.7℃.