小蓬竹内生真菌培养分离条件的优化

Optimization of Culture and Isolation Conditions of Endophytic Fungi from Drepanostachyum luodianense

【目的】探明小蓬竹内生真菌合适的分离培养方法,为小蓬竹内生真菌多样性的研究提供。【方法】以喀斯特地区特有濒危物种小蓬竹[Drepanostachyum luodianense(Yi et R.S.Wang)Keng f.]为研究对象,使用次氯酸钠(NaClO)溶液作为主要消毒剂对小蓬竹组织表面进行消毒。并针对不同组织所需的NaClO溶液浓度和消毒时间设置梯度进行探索,系统优化确立小蓬竹内生真菌的分离方法。【结果】小蓬竹各组织表面消毒的最佳方法,叶为2.5%NaClO溶液浸泡1 min,茎为2.5%NaClO溶液浸泡2 min,根为5%NaClO溶液浸泡3 min;小蓬竹分离内生真菌的最佳分离条件为25%双抗PDA培养基,培养温度26℃。小蓬竹根、茎、叶在最佳表面消毒处理后的定殖率分别为83.3%、70%、100%。【结论】在最佳分离条件下,根部内生真菌分离到10~12种菌株。

【Objective】The optimum isolation and culture method for endophytic fungi from Drepanostachyum luodianense is explored to supply the solid basis for the research on diversity of D.luodianense endophytic fungi.【Method】D.luodianense(Yi et R.S.Wang)Keng f.,an unique endangered species in karst area,is taken as the research object.NaClO solution is used as main disinfectant to disinfect the tissue surface of D.luodianense.The gradient for different tissues is set up based on NaClO solution concentration and disinfectant time to optimize and determine the isolation method of endophytic fungi from D.luodianense systematically.【Result】The optimum method for tissue surface disinfectant of D.luodianense includes:2.5%NaClO solution soaking 1 min for leaf,2.5%NaClO solution soaking 2 min for stem,and 5%NaClO solution soaking 3 min for root.The optimum condition for isolated endophytic fungi from D.luodianense is 25%double anti-body PDA medium and culturing at 26℃.The colonization rate of root,stem and leaf in D.luodianense is 83.3%,70%and 100%after the optimum surface disinfectant treatment.【Conclusion】Under the optimum isolation condition,10—12 species strain of endophytic fungi can be isolated from root.