蛹虫草无性型原生质体制备条件的研究

Preparation Conditions of Protoplast of Anamorphic Strains of Cordyceps militaris

以蛹虫草无性型为材料，通过液体培养获得菌丝体，经不同浓度的蜗牛酶、纤维素酶在不同条件下酶解处理菌丝体获得原生质体．实验结果表明：以培养6d的蛹虫草菌丝体为材料，0．5mol／L甘露醇溶液为稳渗剂，蜗牛酶浓度为1％，纤维素酶浓度为0．5％，两种酶混合的体积比为1：2，pH6．5，35℃条件下酶解3h，可得原生质体产量最高为1．36×10^8个／mL．这一研究结果为蛹虫草通过原生质体技术进行菌株的遗传改良提供了重要的理论依据．

Anamorphic Strains of Cordyceps militaris are used to produce mycelia through liquid culture, which are further processed through the enzymolysis treatment with snailase and cellulose to obtain protoplasts. The experimental statistics are as follows： the optimum age of mycelia is 6 d; 0. 5 mol/L mannitol is the best osmotic stabilizer; concentration of snailase and celluase are 1% and 0.5 % respectively; volume ratio of the two enzymes is 1.2; the optimum enzymolysis temperature, pH value and time are 35℃, pH 6.5, 3h, respectively; the highest protoplast yield is 1.36 × 108/mL. These results and data would provide important parameters for the strain improvement of Cordyceps m. through the protoplasttechnique.