New isolation methods and phylogenetic diversity of Actinobacteria from hypersaline beach in Aksu

Abstract:

Abstract:［Objective］We explored 4 new methods to improve the isolation of actinobacterial resources from high salt areas.［Methods］Optimized media based on 4 new strategies were used for isolating actinobacteria from hypersaline beaches． Glycerin-arginine，trehalose-creatine，glycerol-asparticacid，mannitol-casein，casein-mannitol，mannitol-alanine，chitosan-asparagineand GAUZE' No.1 were used as basic media． New isolation strategy includes 4 methods: ten-fold dilution culture，simulation of the original environment，actinobacterial culture guided by uncultured molecular technology detected，and reference of actinobacterial media for brackish marine environment．The 16S rRNA genes of the isolates were amplified with bacterial universal primers． The results of 16S rRNA gene sequences were compared with sequences obtained from GenBank databases． We constructed phylogenetic tree with the neighbor-joining method．[Results] No actinobacterial strains were isolated by 8 media of control group，while 403 strains were isolated by new strategies．The isolates by new methods were members of 14 genera (Streptomyces， Streptomonospora，Saccharomonospora，Plantactinospora，Nocardia， Amycolatopsis，Glycomyces，Micromonospora， Nocardiopsis， Isoptericola，Nonomuraea，Thermobifida，Actinopolyspora，Actinomadura) of 10 families in 8 suborders．The most abundant and diverse isolates were the two suborders of Streptomycineae (69.96%) and Streptosporangineaesuborder (9.68% ) within the phylum Actinobacteria，including 9 potential novel species．［Conclusion］New isolation methods significantly improved the actinobacterial culturability of hypersaline areas，and obtained many potential novel species，which provided a new and more effective way to isolate actinobacteria resources in hypersaline environments.