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Northern post-glacial lakes are significant, increasing sources of atmospheric carbon through ebullition (bubbling) of microbially-produced methane (CH₄) from sediments. Ebullitive CH₄flux correlates strongly with temperature, reflecting that solar radiation drives emissions. However, here we show that the slope of the temperature-CH₄flux relationship differs spatially across two post-glacial lakes in Sweden. We compared these CH₄emission patterns with sediment microbial (metagenomic and amplicon), isotopic, and geochemical data. The temperature-associated increase in CH₄emissions was greater in lake middles—where methanogens were more abundant—than edges, and sediment communities were distinct between edges and middles. Microbial abundances, including those of CH₄-cycling microorganisms and syntrophs, were predictive of porewater CH₄concentrations. Results suggest that deeper lake regions, which currently emit less CH₄than shallower edges, could add substantially to CH₄emissions in a warmer Arctic and that CH₄emission predictions may be improved by accounting for spatial variations in sediment microbiota.