产纤维素酶原生质体形成和再生条件选择

摘要

The optimal conditions for protoplast isolation and regeneration of 4 auxotrophic mutant strains of Tdchodenna were studied.0.2mol · L-1 (pH5.8) Phosphate buffer is the better buffer system for protoplast isolation of all 4 strains. The best osmotic stablizer for T. Iiride UV2-11 and T. sp UV2-2 is 0.7mol· L-1 KCI, and the best mycelia age is 20 and 18～20h, respectly. The best osmotic stablizer for T. Koningii UV2-1 and UV2-15 is 0.6mol · L-1 NaC1 and the best mycelia age is 16～20 hours. The suitable enzymolysis time is 2～3h. Czapek medium containing 0.5% yeast extract and 0.5% Calium pantotthenic is the suitable regeneration medium for all 4 strains and both 0.7mol · L-1 KCl and 0.6mol · L-1 NaC1 is the best osmotic stablizer for protoplast regeneration.%研究了4种营养缺陷型产纤维素酶菌的原生质体形成和再生条件。结果表明，它们原生质体形成条件均以0.2mo1·L-1(pH5.8)磷酸缓冲液为好。绿色木霉(Trichoderma viride)UV2-11和另一未定木霉(T. sp.)UV2-2以0.7mol·L-1KCl的渗透稳定剂为好，菌丝菌龄分别为20，18～20h；康宁木霉(T. Koningii)UV2-1和UV2-15以0.6mol·L-1NaCl的渗透稳定剂为好，菌丝菌龄均为16～20h，酶解时间均为2～3h。原生质体再生培养基以加入0.5％酵母膏和0.5％泛酸钙的改良czapek培养基效果好，用0.7mol·L-1KCl和NaCl作为渗透稳定剂，菌株原生质体的再生率较高。