植物内生放线菌的选择性分离

The selective isolation of endophytic actinomycetes

摘要:

【目的】更好地发掘内生菌资源, 建立有效的植物内生放线菌分离方法。【方法】比较不同消毒剂和消毒程序、样品预处理、选择性分离培养基等分离内生放线菌的效果, 通过形态及16S rRNA基因序列分析进行菌种鉴定。【结果】用5%的次氯酸钠处理样品4?7 min消毒效果最好; 100 °C处理样品15 min能较好地减少真菌和细菌的干扰。丙酸钠、琥珀酸钠等培养基分离放线菌出菌率较高且类群多样性丰富。【结论】植物样品表面消毒干燥后, 100 °C处理15 min, 用无菌搅拌杯打碎, 直接撒植物于分离培养基中的分离方法效果较好。

Abstract:

[Objective] In order to obtain more resources of endophytic actinomyctes, the aim of the study is to establish an effective method of isolating endophytic actinomycetes. [Methods] Disinfectant and sterilization procedures, pretreatment method, isolation medium were compared. The isolates were identified based on morphology and 16S rRNA gene sequence analysis. [Results] It is very effective for samples sterilized 4?7 minute with 5% NaClO. The pretreatment method of processing sample 15 minutes at 100 °C have a good effect on reducing the interference of fungi and bacteria. Isolation mediums containing sodium propionate and sodium succinate have good effect on isolation. [Conclusion] The isolation method that plant samples were surface sterilized and dried, processed 15 minutes at 100 °C, broken in a sterile mixing cup, and then were separated on isolation medium is very useful.