Longitudinal gradients in streamwater nutrient concentrations in Walker Branch are generated as a result of instream nutrient uptake and spatially confined groundwater inputs during the period from November to May. The response of the stream periphyton community to these longitudinal nutrient reductions was determined by measuring periphyton biomass, productivity, species composition, and phosphorus (P) cycling indices at four stations along a longitudinal transect in the stream. Phosphorus cycling indices (chlorophyll-specific phosphatase activity, phosphorus content of periphyton) exhibited significant changes along the longitudinal transect during those times of the year when streamwater soluble reactive phosphorus (SRP) concentrations also decreased along the transect. During the period from June to October, however, neither streamwater phosphorus concentrations nor phosphorus cycling characteristics exhibited longitudinal trends. Regressions between phosphatase activity and streamwater SRP concentration and between phosphorus content and streamwater SRP were highly significant for all data combined, with SRP explaining >74% of the variation in phosphatase activity and P content. Measures of periphyton biomass (chlorophyll a, total biovolume), and productivity (areal carbon fixation rate, chlorophyll-specific carbon fixation rate) exhibited no consistent longitudinal patterns, even during the period of longitudinal streamwater phosphorus depletion. Regressions between productivity measures and streamwater SRP concentration for all data combined were significant, but SRP explained <56% of the variation in productivity. Periphyton biomass and productivity at all stations along the longitudinal transect appear to be maintained at low levels by high and longitudinally uniform rates of herbivory throughout the year. Algal species composition exhibited some response to longitudinal nutrient depletion. The biovolume and percentage of the blue-green alga Chamaesiphon investiens increased and the percentage of the chlorophyte Stigeoclonium sp. declined longitudinally when nutrients also declined. Our results demonstrate an upstream-downstream biotic linkage in Walker Branch. We show that instream nutrient uptake can reduce the concentrations of nutrients in stream water and thereby influence the structure and functioning of downstream periphyton communities. However, increases in nutrient cycling in response to lower streamwater concentrations can partially compensate for nutrient depletion by upstream organisms, thereby buffering primary productivity in downstream periphyton communities from changes in nutrient supply.