尖孢镰刀菌辣椒专化型原生质体制备条件优化

Optimizing Method of Preparation of Protoplast in Fusedum oxysporum f.sp.capsicum

以尖孢镰刀菌辣椒专化型(Fusedum oxysporum f.sp.capsicum)为供试菌株,研究了其原生质体制备的最佳条件,明确了菌龄、酶的种类、酶解时间、酶解温度及转速等因素对原生质体制备的影响。结果表明,当利用孢子培养的14 h的菌丝体以15 mg/mL崩溃酶、0.7 mol/L的NaCl作为渗透压稳定剂,140 r/min,32℃酶解4 h时,可最大限度的收集原生质体,其在再生固体培养基SR上再生率为32.3%。

Several conditions in preparing the protoplast of Fusedum oxysporum f. sp. capsicum were optimized. Effects of the incubation time, enzyme system, the enzymolysis time, temperature and rotate speed were discussed. It was found that when keeping other parameters as ： spore cultured for 14 hours, 15 mg/mL of Drislase, 0.7 mol/L NaCl and 140 r/min, 32% for 4 h, the minimum consummation of protoplasts can be obtained and the regeneration rate of the protoplast was 32.3% on regeneration medium SR.