Department of Microbial Biotechnology, School of Biology and Center of Excellence in Phylogeny of Living Organisms, College of Science, University of Tehran, Tehran, 14155-6455, Iran.
Microbial Technology and Products Research Center, University of Tehran, Tehran, Iran.
Folia Microbiol (Praha). 2021 Dec;66(6):1047-1054. doi: 10.1007/s12223-021-00907-8. Epub 2021 Sep 6.
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.
设计新型的分离方法可以增强可用于生物技术的细菌菌株的多样性。在这项研究中，介绍了用于放线菌分离的新型离体和原位培养方法。在离体实验中，将土壤悬浮液涂在与土壤基质泥浆直接接触的隔离培养基上，并孵育 16 周。纸完全浸入洞穴土壤中进行原位培养，容器埋在 Hampoeil 洞穴的土层下 10 周。在分离放线菌方面，果包装牛皮纸的回收率为 68.8%，优于其他研究过的滤纸。基于分子鉴定结果，从原位培养方法获得的分离物中，有 19%与已知的放线菌分类群的相似度小于 98.5%，可能代表新的种。相比之下，在标准培养方法中，有 1.3%的分离物的 16SrRNA 基因相似度小于 98.5%。这些数据表明，所介绍的培养方法是一种有前途的分离培养能力较低或新型放线菌的技术。
