Triton X-114促进木霉菌落在固体培养基上相互独立的方法研究

Method of Triton X-114 Improves the Independence of Trichoderma Colonies on Solid Medium

木霉菌丝生长迅速,互相接触交织,在附着物上可形成不分彼此的盘状,对菌落计数和筛选造成了巨大的挑战。本研究提出了一种在固体培养基上促进丝状木霉形成独立菌落的方法。该方法采用将0.5~10 g·L^(-1)浓度的Triton X-114与重融化的培养基混合,并加入新鲜的孢子,待凝固后放入培养箱。促使菌落互相独立的作用的机理可能与其对菌丝体呼吸活性有一定程度的抑制有关。产生的菌落密度可达60~120 CFU/培养皿,可提高木霉菌落的筛选数量。此外,研究还进一步讨论了基于时间回溯的菌落可分离的人工菌落计数时,做菌落分割所遵循的3个规则。该研究为木霉菌独立菌落的培养方法和简化木霉菌人工菌落计数提供了有价值的参考意义。

The mycelium of Trichoderma strains grows rapidly on solid plates and usually creates contacts or other interactions with each other,resulting in one or several larger mycelial discs,which poses a great challenge to colony counting and screening.In this study,we provide a procedure for promoting the colony independence of filamentous Trichoderma on solid medium.The method involved addition of 0.5 g·L^(-1) to 10 g·L^(-1) of Triton X-114 to remelted solid medium,followed by mixing with fresh spores and place in an incubator after solidification.The mechanism of colony independence may be related to the inhibition of mycelium respiratory activity.The number of obtained Trichoderma colonies ranged from about 60 to 120 per Petri dish(φ10 cm),thereby representing an improved efficiency in strain screening.In addition,we further discussed three rules of colony segmentation. This study provides valuable reference for the cultivation of Trichoderma independent colonies and the simplification of artificial colony counting of Trichoderma.