We report nitrogen isotope data for nitrate from transects of hydrocast and surface samples collected in the eastern Indian and Pacific sectors of the Southern Ocean, focusing here on the data from the upper water column to study the effect of nitrate consumption by phytoplankton. The δ¹⁵N of nitrate increases by 1–2‰ from deep water into the Antarctic summertime surface layer, due to kinetic isotopic fractionation during nitrate uptake. Estimation of the nitrate uptake isotope effect from Antarctic depth profiles yields values in the range of 5–6‰ in east Indian sector and 4–5‰ in the east Pacific sector. Surface transect data from the Pacific sector also yield values of 4–5‰. The major uncertainty in the profile‐based estimation of the isotope effect involves the δ¹⁵N of nitrate from the temperature minimum layer below the summertime Antarctic surface layer, which deviates significantly from the predictions of simple models of isotope fractionation. For the Subantarctic surface, it is possible to distinguish between nitrate supplied laterally from the surface Antarctic and nitrate supplied vertically from the Subantarctic thermocline because of the distinctive relationships between the δ¹⁵N and concentration of nitrate in these two potential sources. Our Subantarctic samples, collected during the summer and fall, indicate that nitrate is supplied to the Subantarctic surface largely by northward transport of Antarctic surface water. Isotopic data from the Pacific sector of the Subantarctic suggest an isotope effect of 4.5‰, indistinguishable from the Antarctic estimates in this sector.