College of Plant Protection of China Agricultural University, Beijing, 100193, China.
Chongqing Academy of Chinese Materia Medica, Chongqing, 400065, China.
Sci Rep. 2018 Oct 23;8(1):15623. doi: 10.1038/s41598-018-33965-z.
Isaria farinosa is a pathogen of alpine Thitarodes larvae that are hosts for the Chinese medicinal fungus, Ophiocordyceps sinensis. A matrix analysis indicated that the optimal culture conditions for the mycelial growth of I. farinosa are a 50-mL liquid broth in a 250-mL flask at more than 100-rpm rotation and 15-25 °C. Illumination does not affect the mycelial growth. The optimal nutrition requirements are D-(+)-galactose and D-(-)-fructose as carbon resources and D-cysteine as well as yeast powder, peptone, and beef extract as nitrogen resources at a carbon-to-nitrogen ratio of 1:1 to 1:7. The mineral component and vitamins also significantly increase the mycelial growth of I. farinosa. Based on the optimal culture conditions and nutrition requirements for the mycelial growth of I. farinosa, the effects of altitude on mycelial growth and its metabolome were evaluated using quadrupole-time-of-flight/mass spectrometry, principal component analysis and partial least squares discriminant analysis. The altitude did not affect the mycelial production but significantly regulated its metabolome. The study presents a new approach to better select a method for producing more useful metabolites and highlights the necessity of establishing standards for culturing methods related to altitude to preserve fungal quality; additionally, the results indicate that the use of a fermenter may meet the demands of large-scale mycelial production.
虫生真菌离褶伞是高寒地区幼虫期蝠蛾属昆虫的病原真菌,其可被用于中国药用真菌——冬虫夏草的人工培养。基质分析表明,离褶伞菌丝生长的最佳培养条件是:在 250ml 摇瓶中,50ml 液体培养基,转速超过 100rpm,温度为 15-25℃。光照不影响菌丝生长。最佳营养需求为 D-(+)-半乳糖和 D-(-)-果糖作为碳源,D-半胱氨酸、酵母粉、蛋白胨和牛肉浸膏作为氮源,碳氮比为 1:1 至 1:7。矿物质成分和维生素也显著增加了离褶伞的菌丝生长。基于离褶伞菌丝生长的最佳培养条件和营养需求,利用四极杆飞行时间/质谱、主成分分析和偏最小二乘判别分析,评估了海拔对菌丝生长及其代谢组的影响。海拔不影响菌丝的产生,但显著调节其代谢组。该研究提出了一种更好地选择生产更有用代谢物的方法,强调了建立与海拔相关的培养方法标准的必要性,以保持真菌质量;此外,结果表明使用发酵罐可能满足大规模菌丝生产的需求。