冬虫夏草原生质体制备与再生条件的研究

采用正交分析方法，对影响冬虫夏草菌株原生质体制备与再生的主要因素进行研究。结果表明，最佳条件组合为培养3d 的菌丝体，在2.0% 溶壁酶+1.0% 蜗牛酶的酶液中，0.6mol/L 的KCl 作为原生质体制备的渗稳剂，0.6mol/L 的甘露醇作为再生培养基的渗稳剂，32℃酶解2h。在此条件下，原生质体得率可达到2.98 × 108/ml，再生率可达到42.09 × 10-2。

Study on Protoplast Preparation and Regeneration of Cordyceps sinensis

The primary factors affecting protoplast preparation and regeneration of Cordyceps sinensis were discussed with orthogonal analysis in this study. The results showed that the optimum preparation and regeneration process is as follows: digesting mycelia, which has been cultured for 3 d with 2.0% lywallzyme and 1.0% snailase at 32 ℃ for 2 h, with 0.6 mol/L KCl and 0.6 mol/L mannitol as osmotic pressure stabilizers for protoplast preparation and regeneration media respectively. Under the above conditions, the protoplast yield and regeneration rate reach 2.98 × 108/ml and 42.09 × 10-2, respectively.