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以冬虫夏草无性型中国被毛孢为原材料,通过液体培养获得菌丝体,经酶解处理后在不同条件下获得原生质体,以研究酶的种类及浓度、渗透压稳定剂及浓度、酶液pH值以及酶解时间等不同因素对中国被毛孢原生质体制备的影响。结果显示,中国被毛孢菌株原生质体的最佳制备条件为:使用浓度配比分别为4和3 mg·mL-1的Yatalase和Glucanex酶,在1.0 mol·L-1KCl作为渗透压稳定剂,酶液pH值为6,酶解3 h,所得原生质体量最多。
The mycelia of Cordyceps sinensis anopuriensis were used as raw materials to obtain mycelium by liquid culture. After enzymatic treatment, protoplasts were obtained under different conditions to study the type and concentration of enzyme, osmotic pressure stabilizer and concentration, enzyme solution pH Value and enzymolysis time and other factors on the Chinese fir spore protoplast preparation. The results showed that the optimal preparation conditions for Protoplast from Tricholoma matsutake were as follows: Yatalase and Glucanex at concentration of 4 and 3 mg · mL -1, 1.0 mol·L -1 KCl as osmotic pressure stabilizer, Enzyme solution pH 6, enzymatic hydrolysis 3 h, the resulting protoplast volume up.