铆钉菇D447黑色素高产菌株诱变育种及遗传稳定性研究

为提高铆钉菇（Gomphidius sp.）D447的诱变育种的效率，首先对该菌的孢子最佳活化时间及定向筛选剂曲酸的最优添加量进行了研究，结果表明孢子最佳活化时间为6.0 h，曲酸的最优添加量为0.4%。在此基础上，以紫外线、硫酸二乙酯、氯化锂、5-溴尿嘧啶等诱变剂对D447的活化孢子进行了单一及复合诱变，通过曲酸平板初筛，摇瓶复筛方法，最终获得一株黑色素产量为（2.45±0.16） g/L的诱变株DB-3，较出发菌株D447提高了1.57倍。对突变株进行传代稳定性试验，结果表明诱变株DB-3经斜面传代9代以后，黑色素产量仍较稳定，无明显的回复突变。

Mutation breeding of Gomphidius sp. D447 with high yield melanin and its genetic stability

To improve the efficiency of mutation breeding of Gomphidius sp. D447, the optimum spores culture time and kojic acid (directional filtering reagent) addition were studied first, and the results showed that the optimal results were 6.0 h and 0.4%, respectively. On that basis, a mutant strain Gomphidius sp. DB-3 with high yield melanin was obtained by treating protoplasts with ultraviolet radiation (UV), diethyl sulfate (DES), LiCl, 5-bromouracil (BU) through first screening with kojic acid plate and second screening with shake flask fermentation. The melanin yield of Gomphidius sp. DB-3 increased as much as 1.57 times than that of the original strain, reached (2.45±0.16) g/L. The genetic stability tests showed that the melanin yield of the mutant strain Gomphidius sp. DB-3 was stable and there was no obvious back mutation after nine generations.