以疏绵状嗜热丝孢菌(Thermomyces lanuginosus)为供试菌株,研究了菌龄、酶的种类及浓度、酶解时间、酶解温度和稳渗剂对原生质体制备的影响及稳渗剂对原生质体再生的影响。结果表明,制备嗜热丝孢菌原生质体比较适宜的条件为:PDB液体培养基培养28h,以0.7mol/L NaCl为稳渗剂,0.15 mol/L的溶壁酶,30℃酶解4h。原生质体再生以0.7mol/L蔗糖作稳渗剂为最佳。
The effects of some factors on the preparation and regeneration of protoplasts of Thermomyces lanuginosus were studied, including the effect of the hyphal age, enzymes and their concentration, time and temperature of digestion, osmotic stabilizers on the protoplast preparation and the effect of osmotic stabilizers on the protoplast regeneration. The results indicated that the optimum conditions for preparing protoplasts were: the condia cultured in liquid PDB medium for 28 hours, 0.7 mol/L NaC1 used as the osmotic stabilizer, 0. 15 mol/L LywaUzyme used to digest the hyphae for 4 hours at 30℃. And 0. 7 mol/L sucrose was the osmotic stabilizer for the regeneration of protoplasts.