Bacterial Diversity in Zabuye Salt Lake Tibet by Culture-Independent Approaches

Abstract:

Abstract: [Objective] To get knowledge about bacterial diversity in Zabuye salt lake With the culture-independent approaches. [Methods] Total DNA was extracted from sediments sample of Zabuye salt lake, Tibet. The 16S rRNA gene was amplified by PCR with Bacterial primer f530/r1492.Then the plasmid library of 16S rRNA genes was constructed. The positive clones were screened on plates with IPTG/X-gal/Ap. Amplified ribosomal DNA restriction analysis ( ARDRA) was carried out with restriction enzymes Hae III, Hha I. With the help of culture-independent approaches, DNA was directly extracted from the sediment samples of Zabuye salt lake. After construction of 16S rRNA gene library, ARDRA (amplified rDNA restriction analysis) analysis was carried out. Some clones were selected to be sequenced according to the ARDRA patterns. These partial sequences of 16S rRNA gene were used for construction of the phylogenetic tree.[Results] The phylogenetic tree showed that some clones (57.14% of total clones) belonged to 23 genera in γ-proteobacteria, ?-proteobacteria, δ-proteobacteria, bacteroidetes, firmicutes, and verrucomicrobia. The rest clones were quite different and were most related with uncultured sequences, forming special division in the tree. [Conclusion] These finding showed much prolific bacteria diversity in Zabuye salt lake.