Cycling of organic nitrogen in marine plankton communities studied in enclosed water columns

Concentrations of fluorescamine-positive substances (primary amines) and turnover rates of L-leucine pools were measured concurrently in seawater samples taken from 1300 m³ plastic enclosures moored in Saanich Inlet, British Columbia, Canada. Concentration and turnover rates of dissolved free amino acids were calculated and then used to determine the instantaneous flux of dissolved free amino acids, which ranged from 0.09 to 2.42 M d^-1 (i.e.,5 to 145 gC l^-1 d^-1). This flux was highest in the euphotic zone, and was related to net primary production but not to the type of dominant primary producer. Comparison of the flux to changes in the concentration of ammonia in deep water suggested that amino acid degradation accounted for 60% of the flux into the ammonia pool. For a given sample, the amino acid carbon flux ranged from 17 to 210% (mean=78%) of the primary production. Such fluxes of amino acid carbon, if used exclusively by the bacterioplankton, would give growth rates ranging from 0.3 to 3.0 (mean=1.7) bacterial doublings d^-1. These calculations indicate that a large fraction of the community carbon and nitrogen flux passes through the bacterioplankton.     

Effects of host feeding and dissolved ammonium on cell division and nitrogen status of zooxanthellae in the hydroid Myrionema amboinense

There is a relationship between host feeding, nitrogen status and mitotic activity of zooxanthellae symbiotic with the marine hydroid Myrionema amboinense. Decreases in the mitotic index of zooxanthellae in starved M. amboinense, and in internal pool sizes of glutamine and glutamate, amino acids involved in ammonium assimilation via the glutamine synthetase-glutamate synthase (GS/GOGAT) pathway, were partially restored by addition of ammonium chloride to seawater in which hydroids were incubated. Levels of glutamine were more sensitive to host starvation than levels of glutamate, resulting in a decrease in the glutamine: glutamate molar ratio to that found in zooxanthellae cultured on nitrate. Hydroids starved for 5 d and then incubated in different concentrations of ammonium chloride showed a positive correlation between ammonium concentration and mitotic index of their symbiotic zooxanthellae. Host starvation caused a decrease in perturbation of levels of glutamine and glutamate during ammonium assimilation, as well as decreases in rates of assimilation of [¹⁴C]-leucine into TCA-insoluble protein, and in photosynthetic incorporation of [¹⁴C]-bicarbonate. These observations suggest that host starvation reduces nitrogen supply to the zooxanthellae, causing nitrogen stress to the symbionts and reduction in metabolic processes associated with nitrogen assimilation and photosynthesis as well as with cell division.     

Relationship between the zooplankton,phytoplankton, particulate matter and dissolved free amino acids in the Celtic Sea

Estimates of the biomass of zooplankton, phytoplankton and particulate matter collected in the Celtic Sea during mixed-water conditions (on 8 and 9 April 1983) were compared to the concentration and diversity of sixteen dissolved free amino acids (DFAA) measured in seawater and in particles. During a day profile, variations of dissolved amino acids with depth reflected the feeding activity of copepods. The relationship was not apparent in a night profile and other processes, such as heterotrophic utilization of dissolved nitrogen by microorganisms, were thought to be involved. The ratios of total DFAA concentration (nM litre^-1) in the particulate phase over the concentration in seawater ranged from 1 to 200 within the water column. Of the sixteen amino acids measured, ornithine, a decomposition product of arginine, was responsible for more than 70% of the total concentration of DFAA in seawater. In the particles, phenylalanine ranged from 30 to 88% of total DFAA. In seawater this amino acid occurred in the 20 to 40 m depths (1.3 to 9.9% of total DFAA) in the day profile and at 5 m (12.4%) and 80 m (6.4%) in the night profile. Previously it has only been found in very low concentrations (<5%) in seawater, and its presence is considered to be the result of zooplankton feeding.     

Nitrogen regeneration during the degradation of several amino acids by plankton communities collected near Halifax,Nova Scotia,Canada

Nitrogen regeneration accompanying the bacterial degradation of a variety of amino acids supplied at 10.0 M to samples of coastal plankton communities collected near Halifax, Nova Scotia, Canada was examined. A lag period characterized by a low rate of amino acid uptake and ammonia release was typically followed by a dramatic increase in the rates of uptake and ammonia release. The duration of the lag period varied with the amino acid tested. The ratio of the final ammonia concentration to the nitrogen supplied as amino acid was taken as the regeneration ratio. This value varied from 0.58 to 0.86 for L-arginine and 0.38 to 1.17 for the other amino acids tested, with an average value of 0.74. The presence of inorganic fixed nitrogen at 10.0 M had no effect on the degradation of L-arginine. Other organic compounds supplied at 10.0 M decreased the lag period for L-arginine uptake and degradation. Glucose supplied at 50.0 M decreased the nitrogen regeneration ratio, but did not further decrease the lag for L-arginine degradation. Carbon respiration ratios for L-arginine, L-glutamate, and L-lysine were 0.70, 0.68, and 0.65 when the nitrogen regeneration ratios were 0.86, 0.38, and 0.77, respectively.     

Effect of nitrogen supply on nitrogen uptake,accumulation and assimilation in Porphyra perforata (Rhodophyta)

Porphyra perforata J. Ag. was collected from a rocky land-fill site near Kitsilano Beach, Vancouver, British Columbia, Canada and was grown for 4 d in media with one of the following forms of inorganic nitrogen: NO ₃ ^- , NH ₄ ⁺ and NO ₃ ^- plus NH ₄ ⁺ and for 10 d in nitrogen-free media. Internal nitrogen accumulation (nitrate, ammonium, amino acids and soluble protein), nitrate and ammonium uptake rates, and nitrate reductase activity were measured daily. Short initial periods (10 to 20 min) of rapid ammonium uptake were common in nitrogen-deficient plants. In the case of nitrate uptake, initial uptake rates were low, increasing after 10 to 20 min. Ammonium inhibited nitrate uptake for only the first 10 to 20 min and then nitrate uptake rates were independent of ammonium concentration. Nitrogen starvation for 8 d overcame this initial suppression of nitrate uptake by ammonium. Nitrogen starvation also resulted in a decrease in soluble internal nitrate content and a transient increase in nitrate reductase activity. Little or no decrease was observed in internal ammonium, total amino acids and soluble protein. The cultures grown on nitrate only, maintained high ammonium uptake rates also. The rate of nitrate reduction may have limited the supply of nitrogen available for further assimilation. Internal nitrate concentrations were inversely correlated with nitrate uptake rates. Except for ammonium-grown cultures, internal total amino acids and soluble protein showed no correlation with uptake rates. Both internal pool concentrations and enzyme activities are required to interpret changes in uptake rate during growth.     

Amino acid uptake by the toxic dinoflagellate Alexandrium fundyense

A toxic axenic strain of Alexandrium fundyense is shown to be capable of removing dissolved free amino acids (DFAAs) until concentrations are similar (low nM) to those found in natural waters. Uptake is greatest during exponential growth, rather than during C and/or N-stress as is usual in diatoms and other flagellates. A wide range of amino acids can be taken up, their concentration being decreased within a few hours to the levels observed prior to DFAA addition. The maximum rate of DFAA-N uptake, during early exponential phase, was 0.8 pmol-N cell⁻¹ h⁻¹, equivalent to ≃20% of the total N requirement. More typically, the contribution of DFAA-N was only ≃5%. However, these uptake rates are not sustainable. It is apparent that this organism cannot use amino-N to support significant growth, even though it can take up DFAAs. This, and the fact that the composition of the internal amino acid pool differed from that externally, is further evidence that the N-physiology of this genus is abnormal (differences to other dinoflagellates include an abnormally high concentration of glutamine and arginine, an effective absence of amine X, and release of nitrite during the␣concurrent assimilation of nitrate and ammonium in␣darkness). There is no evidence that the use of DFAAs enhance toxin content, except when cells are supplied with very high (unnatural) concentrations of arginine. Received: 8 May 1989 / Accepted: 14 September 1998     

Nitrogen regeneration by two surf-zone mysids,Mesopodopsis slabberi andGastrosaccus psammodytes

Nitrogen regeneration by two surf zone mysids,Mesopodopsis slabberi andGastrosaccus psammodytes, was determined under laboratory conditions. The mysids were collected from the lower Sundays River estuary, South Africa, from early spring 1984 to late summer 1985. The forms of nitrogen excreted and the effects of mass, temperature and feeding on excretion rate were determined for each species at three experimental temperatures. Comparison of the forms of nitrogen excreted revealed only slight differences between species, with ammonia the major form and urea and amino acids the secondary excretory products in both cases. Mass significantly influenced the rate of ammonia excretion at all experimental temperatures, with no significant difference in slope (common b=0.602) detected between species. During the day sediment deprivation resulted in a 15% and 20% increase in mean ammonia excretion rates of juvenile and adultG. psammodytes respectively, whereas no significant differences were found at night. The mean ammonia excretion rates of fedM. slabberi andG. psammodytes were 2 and 2.5 times higher than starved excretion rates, respectively.G. psammodytes andM. slabberi recycle 139 to 150 g N per running meter of surf zone per year and 1 007 to 1 208 g N m^-1 yr^-1, respectively. Togehter this constitutes 10% of total phytoplankton nitrogen requirements in the surf zone.     

Light-saturated photosynthesis by phytoplankton size fractions in the New York bight,USA

The relationships between netplankton and nanoplankton assimilation numbers, temperature, and major nutrient concentrations were studied and evaluated in the context of seasonal patterns in the biomass of these phytoplankton size fractions. Netplankton and nanoplankton blooms typically occur during late winter (2° to 8°C) and summer (18° to 24°C), respectively. Variations in nanoplankton and netplankton assimilation numbers were not statistically related to the development or collapse of specific blooms based on weekly sampling, but assimilation numbers were higher during the bloom periods than during transition periods of rapid temperature change (8° to 18°C). Differences in the assimilation numbers between size fractions could account for the dominance of the nanoplankton fraction during the summer bloom period but not for the dominance of netplankton during the winter bloom period. Nanoplankton and netplankton assimilation numbers were exponential functions of temperature between 8° and 24°C and 8° and 20°C, respectively. Below 8°C the assimilation numbers of both fractions were higher than expected on the basis of temperature. Above 20°C netplankton assimilation numbers declined with temperature. Netplankton and nanoplankton assimilation numbers were occasionally correlated with dissolved inorganic nitrogen concentrations from less than 1.0 to more than 15 g-at l^-1. Under these conditions, nanoplankton growth rates (calculated from assimilation number and carbon:chlorophyll) were higher and increased more rapidly with dissolved inorganic nitrogen than netplankton growth rates.     

Effect of nitrogen deprivation on amino acid uptake by the chlorophyte Platymonas subcordiformis

Platymonas subcordiformis (UTEX 171) was cultured axenically for 4 d in constant light in a nitrate-containing medium and harvested in the log-phase of cell division. Cells were resuspended in artificial sea water without nutrients and either kept in constant light or placed in constant darkness. High-performance liquid chromatography was used to measure the free amino acid pools of the cells and to determine rates of net entry of each of a mixture of 18 amino acids at daily intervals for 5 d. Free amino acid pools decreased both in light and darkness in the absence of a nutrient sypply. The influx of amino acids in cells maintained in the light increased selectively. Comparison of the rate of entry of ¹⁴C-labeled glycine and net disappearance of glycine from the medium indicated extrusion of non-volatile labeled carbon that did not interact with reagents specific for amine groups. Light was required for synthesis of additional transporter protein which was apparently responsible for increased influx in cells maintained in the light. This response was blocked in the presence of cycloheximide. Cells maintained in the dark for prolonged periods retained the capacity to respond to light by synthesis of new transporter protein. Analysis of incorporation of amino acids into macromolecules indicated that both the overall rate and the pattern of amino acid incorporation were modified in the light. Analysis of the kinetics of glycine entry at a series of temperatures indicated that the concentration of glycine at which entry is half the maximum rate is approximately 2.7 M at the cell surface.     

Surface-film microbial populations: diel amino acid metabolism,carbon utilization,and growth rates

Microheterotrophic dissolved free amino acid (DFAA) utilization, and microbial community and bacterial community carbon production and growth were studied using ³H-labeled organics as tracers in marine surface-film and subsurface (10 cm) waters off Baja California in November 1983. DFAA utilization was generally more rapid during the day (0.14 to 0.38 nM h^-1) than at night (0.04 to 0.14 nM h^-1) in surface-film and subsurface waters, but the percent of utilized amino acid which was respired was always greater during the night (22 to 57%) compared to the day (14 to 18%). Utilization of DFAA-carbon was estimated to range from 0.3 to 5.3 g C l^-1 d^-1 for all stations studied. In six of the 8 samples examined, the percentage of microbial carbon accounted for by the bacterial component of the population (1.4 to 5.9%) was strikingly similar to the percentage of microbial carbon production accounted for by bacterial carbon production (1.9 to 5.1%). In all of these six samples, total microbial specific-growth rates and bacterial specific-growth rates were approximately equivalent (0.9 to 2.2 d^-1 for the microbial community; 0.7 to 1.9 d^-1 for bacteria). The two exceptions were samples apparently influenced by transient flagellate populations migrating into the surface or subsurface waters at night. These observations support the conclusion that surface films contain unique and highly active microbial populations.     

Aspects of nitrogen metabolism of the common mussel Mytilus edulis: Adaptation to abrupt and fluctuating changes in salinity

Mytilus edulis L. were exposed to abrupt (3015 and 1530) and fluctuating (sinusoidal 12 h cycles of 301530) changes in salinity, and the changes in the total osmoconcentration of the haemolymph were recorded. The response of nitrogen metabolism to the altered extracellular osmotic concentrations was investigated in terms of the concentrations of the total NPS (ninhydrin-positive substances) pool and the individual amino acids of the tissues, the concentration of the amino acids of the haemolymph, and the rates of excretion of ammonia and amino acids by whole individuals. The haemolymph became isosmotic with the seawater with abrupt changes in salinity, but with fluctuating salinity was slightly hyperosmotic as the salinity decreased and then slightly hypo-osmotic as the salinity increased. This resulted in a reduction in the extent of the extracellular osmotic change compared to the change in fluctuating salinity to which it was exposed. Total NPS of the tissues decreased with an abrupt decrease in salinity and increased with an abrupt increase in salinity, but a seasonal dependence of the response was indicated. The short-term response of tissue NPS to fluctuating salinity was equivocal, but with long-term exposure the concentration declined. Ammonia and amino acid excretion increased with both an abrupt decrease in salinity and fluctuating salinity and decreased with an abrupt increase in salinity. Haemolymph amino acids increased with an abrupt decrease in salinity. The increased rates of nitrogen excretion accounted for the reductions in the NPS concentrations of the tissues except in the early stages of fluctuating salinity. Taurine, aspartate, threonine, serine, glycine and arginine declined with an abrupt decrease in salinity while alanine and glutamate increased slightly. With an abrupt increase in salinity, alanine and ammonia accumulated in the tissues and then declined while the other amino acids increased slowly over a longer time-course. Similar individual amino acid responses were seen with long-term exposure to fluctuating salinity, except for taurine which did not decrease in concentration. On the basis of the changes in tissue amino acids and ammonia, it is suggested that the alanine dehydrogenase reaction is the primary nitrogen-fixing reaction in marine bivalves such as M. edulis.     

Nitrogenous nutrition in the euphotic zone of the Central North Pacific Gyre

The uptake rates of the three nitrogen compounds ammonium, nitrate, and urea were measured in the oligotrophic North Central Pacific Gyre in August–September 1985. The measurements were performed by using ¹⁵N-labelled substrates and incubating for short-time periods (3 to 4 h) under simulated in situ conditions. Ambient concentrations of the nitrogenous nutrients were generally below 0.10 mol l^-1. The average total daily nitrogen uptake rate, integrated over the euphotic zone, was 12.5 mmol N m^-2 d^-1. Diel studies in the upper water mass resulted in a calculated phytoplankton growth rate of 1.3 d^-1. Ammonium was the dominating nutrient, accounting for on the average 54% of the total nitrogen uptake, while urea uptake represented 32% and nitrate 14%. Ammonium uptake rates at a coastal station off the Hawaiian Islands were very close to the rates found at the oceanic station. Organisms <3 m dominated the nitrogen assimilation, being responsible for about 75% of the ammonium uptake. The nitrogen uptake rates in this study seem to be higher than those found by earlier investigations in the area, but correlated well with other productivity measurements performed during the same cruise.     

In-situ measurements of nitrogenous nutrient uptake by kelp (Ecklonia maxima) and phytoplankton in a nitrate-rich upwelling environment

Nitrogen uptake by the kelp Ecklonia maxima Osbeck and phytoplankton was examined under different conditions of nutrient availability in a kelp bed off the Cape of Good Hope by measuring nutrient depletion in large plastic bags by the kelp and ¹⁵N uptake by phytoplankton. E. maxima took up nitrate and ammonia, but not urea, and showed only a weak preference for reduced nitrogen. Phytoplankton absorbed all three forms of nitrogen available, with a preference for ammonia and urea. Ambient nitrate concentration exhibited a marked and rapid decrease with northerly winds and an increase in response to offshore southerly winds. Nitrogen uptake by E. maxima was linearly related to ambient concentration and did not saturate even at nitrate concentrations >20g-at N l^-1, resulting in a significantly higher tissue nitrogen content under upwelling conditions. Nitrate imported by upwelling was the chief source of nitrogen utilised within the kelp bed. Locally regenerated nitrogen (ammonia and urea) was calculated to contribute only ca 4% of total nitrogen uptake during upwelling and 30% during the relaxation or downwelling phase.     

Nitrogen assimilation by phytoplankton and other microorganisms in the surface waters of the central North Pacific Ocean

Assimilation rates of ¹⁵N-labelled ammonium, urea, and nitrate by plankton in the upper euphotic zone were measured in 2 summer, 2 winter, and 1 spring cruise in the central North Pacific Ocean. Average rates of ammonium plus urea assimilation could not be determined precisely, but were estimated to be 7 to 25 g-at. N m^-3 day^-1. Indirect evidence suggested that non-photosynthetic microorganisms contributed to these rates. Nitrate assimilation was negligible in the upper waters considered in this report (above the chlorophyll maximum and the nutricline). Potential, nitrate-saturated rates were in the range 1 to 8 g-at. N m^-3 day^-1. Seasonal comparison showed lowest rates of both carbon and nitrogen assimilation rates per chlorophyll a in winter.     

Amino acid uptake and respiration by marine heterotrophs

The concentration and turnover of dissolved free amino acids were measured in samples from 25 and 100 m on three occasions at a station 6 miles off the California (USA) coast. Individual amino acid concentrations varied from undetectable (<0.05 g/l) to 3 g/l, the total amino acid concentration from 1.8 to 8.5 g/l. The greater concentration of total amino acids was always found at 25 m. The predominant amino acids were serine, lysine, aspartate, glutamate and alanine; reliable analyses could not be made for glycine because of a high blank. For the 10 individual amino acids studied, the rate of heterotrophic turnover ranged from undetectable to 1.2 g/l day^-1; serine, aspartate, alanine and glutamate showed the highest rates. In samples from 25 m, the rates were 15 to 20 times higher than those taken from 100 m. The total calculated flux of the amino acids studied varied from 0.015 to 3.2 g/l day^-1 and amounted to 1–10% of photosynthetic carbon dioxide fixation.     

Implications of salinity fluctuation for growth and nitrogen metabolism of the marine diatom Ditylum brightwellii in comparison with Skeletonema costatum

In 1987 effects of salinity fluctuations on growth of Ditylum brightwellii (West) Grunow, isolated from the Eastern Scheldt estuary (SW Netherlands) in 1981, were studied. D. brightwellii was grown in a 12 h light: dark cycle at constant salinity in brackish media. Ammonium-limited cultures were subjected to a salinity fluctuation. By decreasing the salinity to 4.8 photosynthesis and cell division were inhibited; cells were deformed. Protein and carbohydrate contents increased slightly, dark respiration was stimulated and cellular levels of glucose decreased at low salinity; this indicated a possible role of sugars in osmoregulation. Ammonium was accumulated in cultures, amino acids may have been stored; the role of the vacuole as a storage compartment was discussed. Both the ammonium uptake capacity and the affinity for ammonium decreased. Nitrogen limitation was relieved in the transient state. [With the activity of the nitrogen assimilation enzymes glutamine synthetase (GS) and glutamate synthase (GOGAT) being uninhibited by lower salinity.] Recovery from hypo-osmotic stress during a salinity increase was initiated by stimulated photosynthesis; chlorophyll a increased, but persistant contractions of cytoplasm (with chloroplasts) may have delayed cell growth. The glutamate dehydrogenase (GDH) activity decreased further whereas the cellular level of alanine increased in the presence of large ammonium pools; this may indicate a temporary activity of ADH (alanine dehydrogenase). Skeletonema costatum (Greville) Cleve, recovered faster from hypoosmotic stress than did D. brightwellii. Due to an osmotic shock from 13.6 to 7.1 S both species excreted amino acids and glucose; S. costatum accumulated more glucose, D. brightwellii accumulated more amino acids. S. costatum may with the competition for nitrogen in waters with an unstable salinity; it will replace D. brightwellii.Contribution no. 427 Delta Institute for Hydrobiological Research, Yerseke, The Netherlands     

Availability of two forms of dissolved nitrogen to the coral Pocillopora damicornis and its symbiotic zooxanthellae

The relative contribution of dissolved nitrogen (ammonium and dissolved free amino acids DFAAs) to the nitrogen budget of the reef-building coral Pocillopora damicornis was assessed for colonies growing on control and ammonium-enriched reefs at One Tree Island (southern Great Barrier Reef) during the ENCORE (Enrichment of Nutrient on Coral Reef; 1993 to 1996) project. P. damicornis acquired ammonium at rates of between 5.1 and 91.8 nmol N cm⁻² h⁻¹ which were not affected by nutrient treatment except in the case of one morph. In this case, uptake rates decreased from 80.5 to 42.8 nmol cm⁻² h⁻¹ (P < 0.05) on exposure to elevated ammonium over 12 mo. The presence or absence of light during measurement did not influence the uptake of ammonium ions. Nitrogen budgets revealed that the uptake of ammonium from concentrations of 0.11 to 0.13 μM could completely satisfy the demand of growing P. damicornis for new nitrogen. P. damicornis also took up DFAAs at rates ranging from 4.9 to 9.8 nmol N cm⁻² h⁻¹. These rates were higher in the dark than in the light (9.0 vs 5.1 nmol m⁻² h⁻¹, P < 0.001). Uptake rates were highest for the amino acids serine, arginine and alanine, and lowest for tyrosine. DFAA concentrations within the ENCORE microatolls that received ammonium were undetectable, whereas they ranged up to 100 nM within the control microatolls. The contribution of DFAAs to the nitrogen budget of P. damicornis constituted only a small fraction of the nitrogen potentially contributed by ammonium under field conditions. Even at the highest field concentrations measured during this study, DFAAs could contribute only ≃11.3% of the nitrogen demand of P.␣damicornis. This contribution, however, may be an important source of nitrogen when other sources such as ammonium are scarce or during periods when high concentrations of DFAAs become sporadically available (e.g. cell breakage during fish-grazing). Received: 22 April 1998 / Accepted: 3 November 1998     

Physiology and chemical composition of nitrogen-fixing phytoplankton in the central North Pacific Ocean

The magnitude and physiological characteristics of biological nitrogen fixation have been studied in the oligotrophic waters of the North pacific gyre. The filamentous blue-green algae Trichodesmium spp. and Richelia intracellularis were the important nitrogen-fixing phytoplankton. Most of the nitrogen fixation occurs in the upper 40 m of the water column, with detectable fixation as deep as 90 m, which corresponds to about the 1 % light depth. There was no evidence of photoinhibition of nitrogen fixation, although CO₂ reduction was depressed slightly at the highest light levels. The rate of nitrogen fixation in the water column varied throughout the day, being highest in mid-morning and in late afternoon. Relatively high fixation rates were also found during periods of darkness. Elevated oxygen concentrations had a marked inhibitory effect on rates of nitrogen fixation, a pO₂ of 0.4 atm causing a 75% inhibition. Data from studies of nitrogen fixation and assimilation rates of ¹⁵N-labelled nitrate, ammonium, and urea indicate that nitrogen fixation furnished about 3% of the total daily fixed nitrogen requirement for phytoplankton growth. Studies with isolated colonies of Trichodesmium spp. indicated that 100% of their nitrogen requirement was met by nitrogen fixation. Chemical composition of the Trichodesmium colonies showed that the C:N ratio was 4.1 and that their phosphorus content relative to carbon or nitrogen was much lower than that of the total particulate material in the water column. Elevated ratios of carbon: adenosine triphosphate (ATP) also suggest that phosphorus deficiency may be limiting the growth of Trichodesmium. The magnitude of nitrogen fixation in the gyre is seasonally dependent, with high rates in late summer and autumn. At these times the water column is stratified, with phosphate and nitrate barely detectable in the upper 100 m. Our data suggest that during these months of stratification, biological fixation of nitrogen amounts to about 33 g-at N/m²/day.     

Phosphorus metabolism of oceanic dinoflagellates: phosphate uptake,chemical composition and growth of Pyrocystis noctiluca

The phosphorus metabolism of Pyrocystis noctiluca Murray (Schuett) 1886 has characteristics which may enhance its potential for success in orthophosphate impoverished waters. The steady-state phosphate uptake rates were equal in the light and dark, and were directly proportional to both the phosphorus cell quota and the cell division rate. In contrast, nutrient-saturated uptake rates were multiphasic, faster in the light than the dark, 2 to 4 orders of magnitude greater than steady-state rates, and were inversely proportional to both the phosphorus cell quota and the cell division rate. These uptake characteristics suggest that P. noctiluca may take up phosphate coincidently at their typically low ambient concentrations as well as to exploit episodic nutrient events in nature. Cell division rates were a hyperbolic function of the ambient orthophosphate concentration. The shortest doubling time was 8.7 d, the phosphate concentration at half the maximum division rate was 0.15 M and the threshold, concentration for cell division was ca 0.05 M PO ₄ ^3- . Division rates of P. noctiluca in the ocean are much faster than predicted from the measured ambient orthophosphate concentrations. Since this dinoflagellate has high naturally occurring alkaline phosphatase activities, and can utilize organic-P compounds, we suggest that organic-P can be as important as orthophosphate in supporting the observed division rates of P. noctiluca in the sea.     

Dynamics and physiology of saxitoxin production by the dinoflagellatesAlexandrium spp.

Toxin content (fmol cell^–1) and a suite of elemental and macromolecular variables were measured in batch cultures of the dinoflagellatesAlexandrium fundyense, A. tamarense andAlexandrium sp. from the southern New England region, USA. A different perspective was provided by semicontinuous cultures which revealed sustained, steady-state physiological adaptations by cells to N and P limitation. Two types of variability were investigated. In batch culture, changes in nutrient availability with time caused growth stage variability in toxin content, which often peaked in mid-exponential growth. A second type of variability that could be superimposed on growth stage differences is best exemplified by the high toxin content of cells grown at suboptimal temperatures. Calculations of the net rate of toxin production (R _tox ; fmol cell^–1 d^–1) for these different culture treatments and modes made it possible to separate the dynamics of toxin production from cell division. Over a wide range of growth rates, cells produced toxin at rates approximating those needed to replace losses to daughter cells during division. The exception to this direct proportionality was with P limitation, which was associated with a dramatic increase in the rate of toxin production as cells stopped dividing due to nutrient limitation in batch culture. Growth stage variability in batch culture thus reflects small imbalances (generally within a factor of two) between the specific rates of toxin production and cell division. N limitation and CO₂ depletion both affect pathways involved in toxin synthesis before those needed for cell division; P limitation does the opposite. The patterns of toxin accumulation were the same as for major cellular metabolites or elemental pools. The highest rates of toxin production appear to result from an increased availability of arginine (Arg) within the cell, due to either a lack of competition for this amino acid from pathways involved in cell division or to increased de novo synthesis. There were no significant changes in toxin content with either acclimated growth at elevated salinity, or with short term increases or decreases of salinity. These results demonstrate that toxin production is a complex process which, under some conditions, is closely coupled to growth rate; under other conditions, these processes are completely uncoupled. Explanations for the observed variability probably relate to pool sizes of important metabolites and to the differential response of key biochemical reactions to these pool sizes and to environmental conditions.