Chloride was injected as a conservative tracer with nitrate to examine nitrate retention (storage plus biotic uptake) and transport in a 327—m reach of a third—order stream draining a forested basin in northwestern California. Prior to injections, diel patterns of nutrient concentrations were measured under background conditions. Nitrate concentration of stream water increased downstream, indicating that the reach was a source of dissolved inorganic nitrogen to downstream communities under background, low—flow conditions, despite uptake by photoautotrophs. At the onset of continuous solute injection over a 10—d period, timing the passage of the solute front indicated that storage dominated nitrate retention. Instantaneous concentration differences at the base of the reach at hour 24 indicated that biotic uptake accounted for 13% of the nitrate amendment while hydrologic storage constituted 29%. Corrected for groundwater dilution (11.7%), saturation of the stream's channel and hyporheic zones was not complete until 6.8 d of continuous injection. By day 3 nitrate retention was dominated by biotic processes. Biotic uptake was greatest during daylight hours indicating retention by photoautotrophs, but also occurred during darkness. After 10 d of continuous injection, mass balance calculations indicated that 29% of N (339 g) was retained from the total injected (1155 g), while the balance of injected nitrate was transported downstream. Storage of NO_3—N was 117 g or 10% while biotic uptake was 222 g or 19%. Periphyton biomass on slides, chlorophyll a both on slides and on natural cobbles, and net community primary production all indicated a lag in periphyton response to nitrate amendment. Earliest indicators of a biotic response to nutrient amendment were decreases in both tissue C/N and epilithic respiration.