No abstract

[1] We report nitrate (NO 3 À ) nitrogen isotope ratios for seawater samples collected in the Subantarctic Zone of the Southern Ocean during both winter and summer as part of the Australian Antarctic CRC Subantarctic Zone (SAZ) Project. The concentration and 15 N/ 14 N of the wintertime surface nitrate are very close to those of the subantarctic thermocline. The 15 N/ 14 N of nitrate in the surface increases sharply into the summer even though there is little seasonal change in nitrate concentration. There are two possible end-member explanations for this observation. First, there may be significant equatorward nitrate transport during the summer, including a supply from the Antarctic surface. Second, the isotope effect of algal nitrate assimilation may be higher than has been estimated elsewhere, for example, for the seasonal sea ice zone of the Antarctic. We use a simple geochemical box model of the SAZ surface mixed layer as it evolves over the course of the summer to simulate salinity, nitrate concentration, and the 15 N/ 14 N of nitrate and sinking N. Our results suggest that a significant portion ($30%) of the summertime SAZ nitrate is supplied from south of the Subantarctic Front and that N export is !3.5 mmol N m À2 d À1 . Our approach also identifies the necessity of an isotope effect for nitrate assimilation in the SAZ of !7% and probably 8-9%. Comparison to laboratory results suggests that this relatively high isotope effect may result from light limitation of algal growth in the SAZ.

In order to (i) establish the biological systematics necessary to interpret nitrogen (N) and oxygen (O) isotope ratios of nitrate ((15)N/(14)N and (18)O/(16)O) in the environment and (ii) investigate the potential for isotopes to elucidate the mechanism of a key N cycle enzyme, we measured the nitrate N and O isotope effects ((15)ε and (18)ε) for nitrate reduction by two assimilatory eukaryotic nitrate reductase (eukNR) enzymes. The (15)ε for purified extracts of NADPH eukNR from the fungus Aspergillus niger and the (15)ε for NADH eukNR from cell homogenates of the marine diatom Thalassiosira weissflogii were indistinguishable, yielding a mean (15)ε for the enzyme of 26.6 ± 0.2‰. Both forms of eukNR imparted near equivalent fractionation on N and O isotopes. The increase in (18)O/(16)O versus the increase in (15)N/(14)N (relative to their natural abundances) was 0.96 ± 0.01 for NADPH eukNR and 1.09 ± 0.03 for NADH eukNR. These results are the first reliable measurements of the coupled N and O isotope effects for any form of eukNR. They support the prevailing view that intracellular reduction by eukNR is the dominant step in isotope fractionation during nitrate assimilation and that it drives the (18)ε:(15)ε ≈ 1 observed in phytoplankton cultures, suggesting that this O-to-N isotope signature will apply broadly in the environment. Our measured (15)ε and (18)ε may represent the intrinsic isotope effects for eukNR-mediated N-O bond rupture, a potential constraint on the nature of the enzyme's transition state.