Emissions of the potent greenhouse gas methane (CH 4 ) from streams and rivers are a significant component of global freshwater methane emissions. The distribution of CH 4 production and oxidation within stream sections and in vertical sediment profiles is not well understood, and the environmental controls on CH 4 production and emission in such systems create a significant challenge for assessing larger-scale dynamics. Here we investigate factors driving the spatial variability of sediment potential methane production (PMP) and potential methane oxidation (PMO) in a temperate stream network in Germany. PMP was highly variable, ranging from 5 × 10 −4 to 28.58 μg CH 4 gDW −1 d −1 and PMO ranged from 0.43 μg CH 4 gDW −1 d −1 to 14.41 μg CH 4 gDW −1 d −1 . Important drivers of spatial variability of PMP and PMO in the sediments of the stream main-stem were related to fine sediment fraction and organic carbon content. At smaller spatial scale, that is, in a sub-catchment stream section, the drivers were more complex and included sediment nitrogen and organic carbon content, as well as porewater dissolved organic carbon, dissolved organic matter quality, and metal concentrations. As with reservoirs and impounded rivers, fine sediment deposition and organic carbon content were found to be key controls on the spatial variability of CH 4 production and oxidation. These findings enhance our understanding of CH 4 dynamics, improve the potential for identifying CH 4 production hotspots in small streams, and provide a potential means for upscaling emission rates in larger-scale assessments.Plain Language Summary Globally, streams and rivers emit a significant amount of the highly potent greenhouse gas methane. The methane emitted from streams is mainly produced in sediments. The distribution of production and consumption in stream sections and sediment profiles is not well understood, which creates a significant challenge for estimating, for example, regional methane emissions from streams and rivers. We investigated possible factors controlling the variability from site to site and different depths of methane production and consumption in sediments of a stream network in south-west Germany. Sediment properties, the amount of fine sediment and organic carbon content, were key drivers of this variability in the main stream. In a smaller side arm of the main stream, the drivers were more complex, including nitrogen and organic carbon content of the sediment, but also porewater dissolved organic carbon, dissolved organic matter quality, and metals. As for reservoirs and dammed rivers, the accumulation of fine sediments and organic carbon content was found to be key controls of sediment methane production and consumption. These findings enhance our understanding of methane dynamics, improve the potential for identifying methane production hotspots in small streams, and provide a potential means for upscaling emission rates in larger-scale assessments.