冬虫夏草无性型菌丝体的原生质体制备条件研究-安徽农业大学学报.PDF

安徽农业大学学报, 2012, 39(5): 793-797 Journal of Anhui Agricultural University 网络出版时间：2012-8-27 12:00:50 [URL] /kcms/detail/34.1162.S1200.017.html 冬虫夏草无性型菌丝体的原生质体制备条件研究 潘丹丹，肖岩岩，陈 超，张 磊，李春如* (安徽农业大学微生物防治省重点实验室, 合肥 230036) 摘 要：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 -1 Yatalase 和 Glucanex 酶，在 1.0 mol·L KCl 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。 关键词：冬虫夏草；中国被毛孢；原生质体；制备条件 中图分类号：S476.12; Q933 文献标识码：A 文章编号：1672−352X (2012)05−0793−05 Conditions for protoplast preparation of Ophiocordyceps sinensis PAN Dan-dan, XIAO Yan-yan, CHEN Chao, ZHANG Lei, LI Chun-ru (Anhui Provincial Key Laboratory of Microbial Control, Anhui Agricultural University, Hefei 230036) Abstract: The mycelia of Hirsutella sinensis, the anamorph of O. sinensis, were cultured in liquid flasks and the different parameters for protoplast preparation of H. sinensis were investigated including the enzyme type and its concentration, stabilizer of osmotic pressure and its concentration, pH value of the enzyme and the enzymolysis time. The results showed that protoplasts were obtained with the highest yield of protoplast under the conditions -1 -1 as follows: digested by Yatalase enzyme of 4 mg/ml and Glucanex enzyme of 3 mg·mL , 1.0 mol·mL KCl as osmotic stabilizer, pH value of enzyme of 6.0 and enzymatic hydrolysis for 3 h.s Key words: Ophiocordyceps sinensis; Hirsutella sinensis ; protoplast; preparation condition 冬虫夏草 （Ophiocordyceps sinensis (Berk.) Sung 如：菌龄的选择；制备酶解液时，酶体系的选择、 et al. ）别名虫草、冬虫草、夏草冬虫，其无性型为 渗透压稳定剂的选择、pH 值、酶解液的浓度等；酶 中国被毛孢(Hirsu