Isotopomer labeling and oxygen dependence of hybrid nitrous oxide production

Abstract. Nitrous oxide (N2O) is a potent greenhouse gas and ozone depletion agent, with a significant natural source from marine oxygen-deficient zones (ODZs). Open questions remain, however, about the microbial processes responsible for this N2O production, especially hybrid N2O production when ammonia-oxidizing archaea are present. Using 15N-labeled tracer incubations, we measured the rates of N2O production from ammonium (NH4+), nitrite (NO2-), and nitrate (NO3-) in the eastern tropical North Pacific ODZ and the isotopic labeling of the central (α) and terminal (β) nitrogen (N) atoms of the N2O molecule. We observed production of both doubly and singly labeled N2O from each tracer, with the highest rates of labeled N2O production at the same depths as the near-surface N2O concentration maximum. At most stations and depths, the production of 45N2Oα and 45N2Oβ were statistically indistinguishable, but at a few depths there were significant differences in the labeling of the two nitrogen atoms in the N2O molecule. Implementing the rates of labeled N2O production in a time-dependent numerical model, we found that N2O production from NO3- dominated at most stations and depths, with rates as high as 1600 ± 200 pM N2O d−1. Hybrid N2O production, one of the mechanisms by which ammonia-oxidizing archaea produce N2O, had rates as high as 230 ± 80 pM N2O d−1 that peaked in both the near-surface and deep N2O concentration maxima. Based on the equal production of 45N2Oα and 45N2Oβ in the majority of our experiments, we infer that hybrid N2O production likely has a consistent site preference, despite drawing from two distinct substrate pools. We also found that the rates and yields of hybrid N2O production were enhanced at low dissolved oxygen concentrations ([O2]), with hybrid N2O yields as high as 20 % at depths where [O2] was below detection (880 nM) but nitrification was still active. Finally, we identified a few incubations with [O2] up to 20 µM where N2O production from NO3- was still active. A relatively high O2 tolerance for N2O production via denitrification has implications for the feedbacks between marine deoxygenation and greenhouse gas cycling.