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The bacteria inhabiting brackish lake environments are poorly known, and there are few studies on the microbial diversity of these environments. Lake Bosten, a large brackish inland lake, is the largest lake in Xinjiang Province in northwestern China. Because sediments record past limnic changes, the analysis of sedimentary bacteria in Lake Bosten may help elucidate bacterial responses to environmental change. We employed 454 pyrosequencing to investigate the diversity and bacterial community composition in Lake Bosten. A total of 48 230 high-quality sequence reads with 16 314 operational taxonomic units were successfully obtained from the 4 selected samples, and they were numerically dominated by members of the Deltaproteobacteria (17.1%), Chloroflexi (16.1%), Betaproteobacteria (12.6%), Bacteroidetes (6.6%), and Firmicutes (5.7%) groups, accounting for more than 58.1% of the bacterial sequences. The sediment bacterial communities and diversity were consistently different along the 2 geographic environmental gradients: (i) freshwater-brackish water gradient and (ii) oligotrophic-mesotrophic habitat gradient. Deltaproteobacteria, Chloroflexi, and Betaproteobacteria were amplified throughout all of the sampling sites. More Bacteroidetes and Firmicutes were found near the Kaidu River estuary (site 14). Our investigation showed that Proteobacteria did not display any systematic change along the salinity gradient, and numerous 16S rRNA sequences could not be identified at the genus level. Our data will provide a better understanding of the diversity and distribution of bacteria in arid region brackish lakes.