高大环柄菇原生质体制备的研究

Study on Protoplast Preparation of Macrolepiota procera

以高大环柄菇为出发菌株,对其原生质体制备条件进行研究,具体分析了酶浓度、菌龄、渗透压稳定剂以及酶解时间和温度等因素对高大环柄菇原生质体产出量的影响。结果表明:采用液体发酵培养第5 d的菌丝体,以0.5 mol/L NaC l作渗透压稳定剂,加入1.5 g/L混合酶(即纤维素酶与蜗牛酶按1∶1混合)在30℃下酶解3.5 h,制备原生质体效果最佳,原生质体产出量可达6.1×107个/mL。

The protoplast preparation of Macrolepiota procera was studied. The results indicated that the best condition for pmtoplast preparation was as follows ： using the mycelium raised for 5 days on the liquid fermentation medium as material, using 0.5 mol/ L NaCl as stabilizing agent of osmotic pressure, adding 1.5 g/L mixed enzyme （cellulose enzyme： snail enzyme = 1 ： 1 ）, conducting enzymolysis for 3.5 hours at 30 ℃. Under the above best condition, the protoplast yield could reach to 6.1 × 10^7/mL.