Introduction: Urmia Lake, located in the northwest of Iran, is the largest permanent lake in Iran and one of the three permanent hypersaline lakes in the world. Microbial sampling from four western different regions of the lake was done and then, biodiversity of the samples were studied by culture dependent and culture independent methods.Materials and Methods: In culture dependent methods, halophilic and halotolerant bacteria were isolated under aerobic condition on four growth media, MH, SWN, MHLN and SWNLN. Differentiation of bacterial isolates was performed based on colony features, Gram staining and primary biochemical tests. In culture independent method, environmental DNA from water and soil samples were extracted. After 16S rRNA PCR amplification 16S rRNA library was constructed and 20% of clones were sequenced.Results: Within 217 purified isolates in culture dependent methods, 52 strains were selected for 16S rRNA gene sequencing and representatives of Halobacillus, Halomonas, Planococcus, Gracilibacillus, Bacillus, Pontibacillus, Paracoccus, Marinobacter, Providencia, Staphylococcus, Alkalibacterium, Sanguibacter, Lysobacter, Kocuria, Pontibacter, Salicola, Micrococcus, Oceanobacillus, Brevundimonas, Thalassobacillus, Microbacterium and Piscibacillus genera were identified. In culture independent method, selected clones belonged to Salinibacter, Adhaeribacter and Cesiribacter genera.Discussion and Conclusion: In culture dependent selected strains 18 strains showed less than 98.7% sequence similarity to the closest known strains and were representatives as new taxa of Urmia lake. In culture independent method selected clones belonged to Bacteroidetes category. Data obtained from this study were similar to other scientific reports from hypersaline lakes.