灵芝原生质体诱变选育高产漆酶突变株的研究

Breeding of Ganoderma Strains Producing High Laccase Activity by Mutagenesis of Protoplasts

对一株产漆酶活性较高的灵芝(Ganoderma.lucidumKarstNo.1)原生质体的制备、再生及紫外诱变育种进行了研究,获得了1株漆酶活性比对照菌株提高2.83倍的突变株G301。实验所确定的原生质体最佳制备条件为:以蔗糖为稳渗剂,采用4%(W/V)溶壁酶与2%(W/V)蜗牛酶等体积混合酶液,在30℃、pH6.0下,对液体培养48h的灵芝菌丝体酶解1h;同时实验所确定的原生质体诱变的适宜条件为:30W紫外灯距离40cm照射30s。

The output, regeneration and mutagenesis of the protoplasts of Ganoderma. lucidum Karst No. 1 that would produce high laccase activity were studied. The mutant G301 of enzyme activity as 2.83 fold higher than the original strain was obtained under 30W UV lamp, at a distance 40cm from the protoplasts, for 30s. The optimal conditions of protoplast output and regeneration are as follows： using 0.6mol/L sugar as osmotic stabilizer, the mycelium being incubated for 48 hours, the lyric enzyme solution containing a mixture of equal volume of 4%（W/V） Lywallzyme and 2%（W/V） snailase, pH6.0, and enzymatic time for 1 hour at 30℃.