Accurately accounting for flows of energy through food webs is challenging because of the spatial and temporal variability associated with energy production and consumption. Wet–dry tropical rivers have a highly seasonal discharge regime where wet season flows allow access to energy sources (inundated wetlands) that are not available during the dry season when aquatic consumers are confined to disconnected waterholes. We combined measures of fish community biomass with previously published feeding guild specific stable isotope analyses to explore how opposing wet- and dry-season habitat templates influence spatial and temporal trends in the sources of energy supporting fish biomass throughout a river network in the wet–dry tropics of northern Australia. Accounting for the relative contribution of each feeding guild to fish community biomass was a critical component of our analyses, as a single feeding guild (invertivore/piscivore) influenced spatial and temporal patterns in the sources of energy supporting overall fish biomass. During the early dry season, the reliance of fish communities on autochthonous sources of energy (periphyton) decreased from the upper to lower reaches of the river network, which correlates with increasing floodplain area and wet season inundation times. These patterns disappeared by the late dry season as fish in both upper and lower reaches became increasingly reliant on autochthonous sources produced within waterholes over the course of the dry season, indicating that the large wet-season gains in fish biomass are maintained through the dry season by energy produced within waterhole refuges. Collectively these results indicate that a combination of autochthonous and allochthonous sources of energy work in unison to support fish community biomass throughout the Mitchell River catchment and that access to these sources of energy is dictated by seasonal patterns in discharge interacting with spatial variability in river geomorphology (channel geometry and floodplain area). Many rivers are experiencing decreased flows due to water resource development and more frequent and severe droughts. Thus, we suggest our study provides insight into how changes in discharge regime could influence food web energetics throughout river networks.