Processing oil sands to extract bitumen produces large volumes of a tailings slurry comprising water, silt, clays, unrecovered bitumen, and residual solvent used in the extraction process. Tailings are deposited into large settling basins, where the solids settle by gravity to become denser mature fine tailings (MFT). A substantial flux of methane, currently estimated at ~40 million L/day, is being emitted from the Mildred Lake Settling Basin. To better understand the biogenesis of this greenhouse gas, the methanogenic consortia in MFT samples from depth profiles in 2 tailings deposits (Mildred Lake Settling Basin and West In-Pit) were analyzed by constructing clone libraries of amplified archaeal and bacterial 16S rRNA genes. The archaeal sequences, whose closest matches were almost exclusively cultivated methanogens, were com- parable within and between basins and were predominantly (87% of clones) affiliated with acetoclastic Methanosaeta spp. In contrast, bacterial clone libraries were unexpectedly diverse, with the majority (~55%) of sequences related to Proteo- bacteria, including some presumptive nitrate-, iron-, or sulfate-reducing, hydrocarbon-degrading genera (e.g., Thauera, Rhodoferax, and Desulfatibacillum). Thus, MFT harbour a diverse community of prokaryotes presumptively responsible for producing methane from substrates indigenous to the MFT. These findings contribute to our understanding of biogenic methane production and densification of MFT in oil sands tailings deposits.