Designing novel isolation methods could enhance the diversification of the available bacterial strains to biotechnology. In this study, the new ex situ and in situ cultivation methods are introduced for the isolation of actinobacteria. In the ex situ experiments, the soil suspension was spread on the isolation media located above some ordinary papers in immediate contact with the slurry of soil substrate and incubated for 16 weeks. The paper was wholly immersed in the cave soil for in situ cultivations, and the containers were buried under layers of soil in Hampoeil cave for 10 weeks. Fruit wrapping kraft coated paper, with 68.8% recovery of isolates, was a better choice in isolation of actinobacteria than other studied filter paper. Based on the molecular identification results, 19% of the isolates obtained from the in situ cultivation method had less than 98.5% similarity to known taxa of actinobacteria and potentially may represent new species. In contrast, in the standard cultivation method, 1.3% of the isolates had less than 98.5% similarity 16Sr RNA gene. This data shows that the introduced cultivation method is a promising technique for isolating less culturable or new actinobacteria.

设计新的分离方法可以促进生物技术中可用细菌菌株的多样化。本研究引入了新的异地和原位培养方法来分离放线菌。在异地实验中，将土壤悬浮液铺在一些普通纸上方的隔离介质上，与土壤基质的浆液直接接触并孵育16周。将纸完全浸入洞穴土壤中进行原位栽培，并将容器埋在Hampoeil洞穴的土层下10周。水果包装牛皮纸涂布纸的分离物回收率为 68.8%，与其他研究的滤纸相比，是分离放线菌的更好选择。根据分子鉴定结果，从原位培养方法获得的分离株中有 19% 与已知放线菌类群的相似性低于 98.5%，并且可能代表新物种。相比之下，在标准培养方法中，1.3% 的分离株具有低于 98.5% 的相似性 16Sr RNA 基因。该数据表明，引入的培养方法是一种分离不易培养或新放线菌的有前途的技术。