Nitrogen uptake by marine phytoplankton : the effects of irradiance, nitrogen supply and diel periodicity
Creators: name: Cochlan, William Patrick
Publisher: name: University of British Columbia
Year: 2011

description: Diel patterns of nitrogen (NO₃⁻, NH₄⁺, urea) uptake were investigated in natural assemblages of phytoplankton from neritic and oceanic environments off the coast of British Columbia. This is the first study to report nitrogen uptake rates and extensive measurements of ambient NH₄⁺ and urea concentrations in these waters. Calculated rates of N uptake, 15 based on ¹⁵N incorporation into particulate matter during time course experiments, were maximal during the day and minimal at night. Besides the obvious effects of irradiance, the amplitude of the periodicity in uptake rate was influenced by phytoplankton community composition, ambient nitrogen concentration, forms of nitrogen available, and depth of sampling. Uptake of nitrogen during the night and in artificial darkness were measurable proportions of daytime and light uptake rates, with the importance of dark uptake generally increasing with increasing N limitation. This is the first study of diel urea uptake by marine phytoplankton in the field. The ratios of dark to light urea uptake over a diel cycle were more similar to those of NO₃⁻than those of the other reduced N form, NH₄⁺. Rates of NO₆⁻ and urea uptake by phytoplankton in the shallow and deep chlorophyll layers of the Strait of Georgia were measured over a gradient of irradiances and results of these experiments could be fitted with a hyperbolic function similar to the Michaelis-Menten equation. Half-saturation constants (KLT) for light-dependent uptake of urea and NO₃⁻ ranged from 0 to 14% of the surface irradiance and dark uptake was a variable, but often substantial (&gt; 50%) portion of the total (light + dark) uptake. The uptake response of nitrate-replete and -starved populations of the picoflagellate, Micromonas pusilla (Butch.) Manton et Parke, to urea, NH₄⁺ and NO₃⁻ perturbations was determined by both ¹⁵N accumulation and nutrient disappearance from the culture medium. Maximum specific uptake rates (Vmax) of NH₄⁺ were 0.13 h⁻¹, more than 2 times the Vmax of NO₃⁻ or urea (ca. 0.05 h⁻¹). The half-saturation constants (Ks) for urea, NH₄⁺ and NO₃⁻ were within ± 0.1 µg-at N•L⁻¹ of each other; the average value of 0.41 µg-at. N•L⁻¹ is within the range reported for small, oceanic diatoms. NO₃⁻ uptake was completely inhibited following NH₄⁺ addition (1-10 µg-at. N•L⁻¹), whereas urea addition resulted in only a 28% reduction in NO3- uptake. Starved cultures of M. pusilla exhibited variable uptake of NH₄⁺ and urea as a function of time, with an initial "surge" uptake response. This is the first laboratory study of N uptake by an eucaroyotic picoplankter and demonstrates that many of the transient uptake responses reported for diatoms, with which it competes in the field, are common to this picoplankter. Diel periodicity of nitrogen uptake and assimilation were measured in N-replete batch cultures of M. pusilla and also in N-limited cyclostat cultures (14L:10D) at three growth rates corresponding to ca. 75, 50 and 25% of it's maximal growth rate. Nitrate uptake was continuous and independent of the L:D cycle in the cyclostat cultures at the lowest dilution rate, but NO₃⁻ uptake rates exhibited pronounced periodicity in the batch and higher dilution rate cultures, a response similar to that seen in previous studies of cyclostat cultures of some diatoms. Diel patterns in cell division, mean cell volume, potential uptake rates and internal pools of NO₃⁻ were also observed and are discussed with respect to the nutritional status of the cells. The effect of irradiance on the uptake of NH₄⁺ and NO₃⁻ by M. pusilla was also described by Michaelis-Menten kinetics; with increasing N limitation the importance of light for nitrogen uptake decreased and dark uptake increased from 5-20% to 21-39% of NO₃⁻ and NH₄⁺ uptake rates, respectively, at saturating irradiance.