Coupled nitrate N and O stable isotope fractionation by a natural marine plankton consortium

The stable nitrogen (N) and oxygen (O) isotope ratios (15N/14N and 18O/16O, respectively) of nitrate (NO) were measured during incubations of freshly collected seawater to 3 investigate the effect of light intensity on the isotope fractionation associated with nitrate assimilation and possible co-occurring regeneration and nitrification by in situ plankton communities. Surface seawater was collected off the coast of Vancouver, Canada in late fall and in late summer and was incubated under different laboratory light conditions for 10 and 30 days, respectively. In the late summer experiments, parallel incubations were supplemented with 15NH and H18O tracers to monitor co-occurring 4 2 nitrification. Differences in irradiance in the fall incubations resulted in slightly reduced nitrate consumption at low light but had no distinguishable impact on the N isotope effect (15ε) associated with NO assimilation, which ranged between 5 and 8‰. The 3 late-summer community incubations, in contrast, showed significantly reduced growth rates at low light and more elevated 15ε of 11.9 ± 0.4‰, compared to 8.4 ± 0.3‰ at high-light conditions. The seasonal differences could reflect physiological adaptations of the fall plankton community to reduced irradiance, such that their incubation at low light did not elicit the increase in proportional cellular nitrate efflux required to raise the isotope effect. In both the fall and summer incubations, the ratio of the coincident rises in the 15 δ N and 18 δ O of NO was comparable to previous monoculture phytoplankton experiments, 3 with a 18 1δ O: 15 1δ N of ∼1, regardless of light level. A decoupling of 18 1δ O: 15 1δ N is expected if nitrification occurs concomitantly with nitrate assimilation. The lack of such decoupling is best explained by the absence of significant nitrification in any of our study’s treatments, an interpretation supported by our inability to identify any tracer 15N and 18O uptake into the NO pool in the late-summer community incubations. 3