Leaves, epilithon, macrophytes, and fine benthic organic material are central ecosystem compartments to food webs and mediate nutrient fluxes in streams. Most estimates of gross primary production (GPP) and ecosystem respiration (ER) are made at a reach scale, averaging across compartments. Thus, there is little information on how individual compartments contribute to and scale up to whole‐stream estimates across watersheds. We compared estimates of GPP, ER, and nitrogen (N) uptake of individual ecosystem compartments (dm) and stream reaches (~100 m) in three sizes of streams in a preserved Atlantic Rainforest watershed. The smallest stream had dense forest canopy cover, whereas the largest was more open. We measured substratum‐specific rates of GPP and ER, as well as ammonium and nitrate 15N uptake in recirculating chambers. We compared these decimeter‐scale measurements to whole‐stream estimates, using single‐station dissolved oxygen (GPP and ER) and pulsed N uptake methods. Epilithon and macrophytes (when present) were the dominant GPP and N uptake compartments in open‐canopy sites, and leaves contributed strongly to ER at all sites, even though they covered <3 percent of the stream bottom. Ammonium and nitrate uptake per unit N content varied significantly among substrata and streams. Upscaled inorganic N uptake per unit area was greater when macrophytes were present. Chamber measurements overestimated metabolic rates in the larger streams, but not in the smallest one. The smallest transient storage zone streams were more active than the biggest one, and this influenced the mismatch between whole‐stream and chamber nutrient uptake estimates. We conclude that scaling to the whole watershed requires information on location in the watershed (e.g., where canopy cover is dense), rates of individual compartments, and reach‐specific hydrodynamic information as influenced by large‐scale geomorphic details (i.e., the size and activity of the transient storage zones).