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The biodiversity of bacterial communities along the Yenisei River at section c. 1800 km was studied using next-generation sequencing of 16S rRNA genes and common biodiversity indices. Overall, 3022 unique operational taxonomic units were identified. Actinobacteria and Proteobacteria were the dominant phyla at all sampling sites. The highest alpha-diversity values were found in the middle section of the studied river. The beta-diversity of bacterial assemblages in the river was related to the surrounding landscape (biome): three distinctly different bacterial assemblages occurred in sections of the river, situated in mountain taiga, plain taiga and in a region of permafrost, covered by forest-tundra and tundra. Tributaries arising from these different landscapes likely contributed substantially to the variations of Yenisei bacterial communities. In contrast to a prediction of the river continuum concept, the proportion of photoautotrophic Cyanobacteria in bacterial assemblages did not increase downstream, but peaked at the middle section.