Direct measurements of nanomolar nitrate uptake by the marine diatom Phaeodactylum tricornutum (Bohlin). Implications for studies of oligotrophic ecosystems

A new automated procedure for nanomolar nitrate analysis was applied to the study of uptake of low nitrate concentrations (< 100 ngat l^–1) by phytoplankton. The precision of this analytical method (± 3 ngat l^–1) made it possible to monitor the absorption of very low quantities of nitrate over short term periods by a low cell-density culture of the marine diatom Phaeodactylum tricornutum, where the levels of particulate nitrogen and chlorophyll were equivalent to those found in oligotrophic areas (0.5 µgat N l^–1 and 0.4 µg l^–1 respectively). By continuous monitoring of nitrate disappearance from the culture medium, we are able to describe accurately the transient uptake responses of the diatom after a spike addition of trace quantities of nitrate and thus to provide new information on the still largely unknown small-scale phenomenon of pulsed nitrate supply in the upper layer of stratified oceans and rapid uptake of these nitrate patches by phytoplankton.The results show that a N-limited culture of Phaeodactylum tricornutum is immediately capable of taking up trace quantities of nitrate (< 100 ngat l^–1) at high rates (0.10–0.14 h^–1) . These initial rates are one order of magnitude higher than the theoretical rates calculated from the Michaelis-Menten equation and are close to the level of V _max (0.15 h^–1) obtained when cells are exposed to saturating nitrate concentrations. This rapid initial uptake would be a considerable advantage in oligotrophic areas where nanomolar nitrate supply is thought to be episodic. The present results suggest that phytoplankton evolve adaptations to utilize the available nitrate at the spatial and temporal scales at which it occurs. On the other hand, we can consider this physiological adaptation as evidence of nitrate pulses in the field which would invalidate the steady-state approach to the oligotrophic ecosystems.