木质层孔菌原生质体的制备与再生条件的研究

Studies on Formation and Regeneration of the Protoplast from Fomes lignosns (Brek) Ccoke

目的:提高木质层孔菌原生质体的产量和再生率,为进一步诱变育种提高菌株产木质纤维素酶活力打下基础。方法:通过单因素实验分别筛选合适的酶浓度、菌丝的菌龄、酶解温度、酶解时间、渗透压稳定剂与pH值。结果:在酶解液浓度为2.0%纤维素酶+2.0%蜗牛酶、菌龄60h、酶解温度32℃、酶解时间3h,以0.8mol/L的NaCl溶液为渗透压稳定剂,pH值为5.0时,原生质体形成量达4.13×105个/mL,再生率可达6.95%。有效地提高了木质层孔菌原生质体的产量和再生率。

Objective： Raising the output and the regeneration rate of protoplast from Fomes lignosns（Brek） Ccok which acts as a base of mutation breeding improving lignocellulose production. Method： Suitable enzyme concentration, filamentous age, enzymolysis temperature and time, stabilizing agent of osmotic pressure and pH value were screened by a mono- factor experiment. Result： When the concentration of cellulose and glusulase was 2.0%, age of the strain up to 60h, enzymolysis temperature was 32℃ and time was 3h, NaGl solution was 0.8mol/L, and pH value was 5.0, the output of protoplasts reached 4.13 ×10^5/mL and the rate of regeneration achieved 6.95%. In a word, the output and the regeneration rate of protoplast from Fomes lignosns （Brek） Ccok increased.