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Key challenges to regionalization of methane fluxes in the Amazon basin are the large seasonal variation in inundated areas and habitats, the wide variety of aquatic ecosystems throughout the Amazon basin, and the variability in methane fluxes in time and space. Based on available measurements of methane emission and areal extent, seven types of aquatic systems are considered: streams and rivers, lakes, seasonally flooded forests, seasonally flooded savannas and other interfluvial wetlands, herbaceous plants on riverine floodplains, peatlands, and hydroelectric reservoirs. We evaluate the adequacy of sampling and of field methods plus atmospheric measurements, as applied to the Amazon basin, summarize published fluxes and regional estimates using bottom-up and top-down approaches, and discuss current understanding of biogeochemical and physical processes in Amazon aquatic environments and their incorporation into mechanistic and statistical models. Recommendations for further study in the Amazon basin and elsewhere include application of new remote sensing techniques, increased sampling frequency and duration, experimental studies to improve understanding of biogeochemical and physical processes, and development of models appropriate for hydrological and ecological conditions.