Different patterns of growth and nitrogen uptake in two clones of marineSynechococcus spp.

We grew marineSynechococcus Clones WH7803 and WH8018 at 150µE m^–2 s^–1 in dilute batch cultures with NH ₄ ⁺ as the limiting nutrient. The maximal uptake capacities for NH ₄ ⁺ and NO ₃ ^- were measured in frequent experiments during log and stationary phases of growth. Clone WH7803, originally isolated from oceanic waters, had a specific uptake rate of NH ₄ ⁺ that approximated the maximum (log phase) specific growth rate (ca ~ 0.025 h^–1). NO ₃ ^- uptake was observed only after nitrogen in the media was depleted; the NO ₃ ^– uptake capacity was ca 12% the capacity for NH ₄ ⁺ uptake throughout the nitrogen depleted period. Growth was arrested upon nitrogen depletion, but resumed soon after reinoculation into fresh media, even after 5 d of starvation. Clone WH8018, originally isolated from coastal waters, revealed a five-fold enhancement in the NH ₄ ⁺ uptake rate relative to growth rate at the time of nitrogen depletion. As nitrogen starvation proceeded, this enhancement was reduced. This clone, too, was able to take up NO ₃ ^- once nitrogen in the media was depleted, but only after ca 20 h. Growth continued for a limited period during nitrogen depletion, but nitrogen-starved cells were slow to recover upon reinoculation into fresh media. We speculate that clonal differences may reflect differences in the molecular regulation of nitrogen assimilation.     

Phosphorus- and nitrogen-limited photosynthesis and growth of Gracilaria tikvahiae (Rhodophyceae) in the Florida Keys: an experimental field study

The relative effects of NH ₄ ⁺ (N) and PO ₄ ^3- (P) on growth rate, photosynthetic capacity (P_max), and levels of chemical constituents of the red macroalga Gracilaria tikvahiae McLachlan were assayed during winter and summer, 1983 in inshore waters of the Florida Keys by using in-situ cage cultures. During winter, both N and P enrichment enhanced growth over that of ambient seawater; however, P rather than N accounted for more (60%) of the increased winter growth. During summer, P, but not N, enhanced growth over ambient seawater and accounted for 80% of increased growth. Similarly, P_max was enhanced by both P and N during winter (but mostly by P) and only by P during summer. Elevated C:P, C:N and N:P ratios of G. tikvahiae tissue during winter, but only C:P and N:P ratios during summer, support the pattern of winter N and P limitation and summer P-limitation. This seasonal pattern of N vs P limited growth of G. tikvahiae appears to be a response to seasonally variable dissolved inorganic N (twofold greater concentrations of NH ₄ ⁺ and NO ₃ ^- during summer compared to winter) and constantly low to undetectable concentrations of PO ₄ ^3- . Mean C:P and N:P ratios of G. tikvahiae tissue during the study were 1 818 and 124, respectively, values among the highest reported for macroalgae.     

Nitrogen uptake characteristics of Chordaria flagelliformis (Phaeophyta) in batch mode and continuous mode experiments

The uptake of NH ₄ ⁺ and urea by Chordaria flagelliformis was measured in a perturbed system (batch mode) and in an apparent steady-state system (continuous mode). The maximum uptake rates (V_max) measured in short-term depletion experiments greatly exceeded those determined under a steady nutrient supply. C. flagelliformis briefly exposed to high nutrient concentrations has an impressive capacity for rapid uptake (high V_max)_which is uncoupled from growth. In contrast, under continous, homogenous low N concentrations the alga appears well equipped (low half-saturation constant, K_m) to take up the available N. The results show that previous work based on batch mode uptake experiments probably overestimated the N requirements for the growth of many seaweeds.     

Nitrate uptake and assimilation in Thalassiosira weissflogii and Phaeodactylum tricornutum: interactions with photosynthesis and with the uptake of other ions

Interactions of the nitrate, phosphate, and ammonium uptake systems and the interactions of these systems with photosynthesis were investigated for Thalassiosira weissflogii and Phaeodactylum tricornutum preconditioned in continuous culture. The cultures were supplied with NO _- ³ and PO ₌ ⁴ in an N:P atomic ratio of 15:1, and residual concentrations of both nutrients in the growth chamber were very low. The rate of NO _- ³ uptake was reduced by the addition of NH ₊ ⁴ or PO ₌ ⁴ . The rate of PO ₌ ⁴ uptake by T. weissflogii was reduced by the addition of NH ₊ ⁴ . The rate of carbon fixation was reduced by NO _- ³ additions and slightly reduced by the addition of PO ₌ ⁴ . There were two components of NO _- ³ uptake, one light-dependent and one light-independent. Uptake inhibition by added PO ₌ ⁴ acted on the light-independent component. The change in the C fixation rate due to added NO _- ³ was equal to the rate of NO _- ³ uptake by the light-dependent component on a molar basis. Nitrate assimilation (reduction) rates were calculated from the time course of extracellular and intracellular NO _- ³ concentrations. The light-dark change in the assimilation rate was similar to the light-dark change in the uptake rate, suggesting close coupling between the light-dependent components of uptake and assimilation. The assimilation rate dropped upon exhaustion of extracellular NO _- ³ , implying that an uptake-coupled component of assimilation is unavailable for the assimilation of intracellular NO _- ³ . The reduction in the C fixation rate due to NO _- ³ was temporally associated with uptake rather than assimilation, but may reflect interaction with either the light-dependent uptake step or the closely coupled assimilation. Phosphate additions reduced the rate of NO _- ³ uptake, while the rate of assimilation was unaffected.     

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.     

Nitrogen supply,tissue composition and frond growth rates for Macrocystis pyrifera off the coast of southern California

Seasonal changes in ambient NO ₃ ^– and NH ₄ ⁺ , tissue composition (N, C, and C/N ratio), and frond growth rates for Macrocystis pyrifera (L.) Agardh were examined. Ambient NO ₃ ^– showed distinct seasonal variations. Frond growth rates were variable, but showed no clear correlation to ambient NO ₃ ^– . The average N content of plant tissue did, however, show the same seasonal variations as ambient NO ₃ ^– . The longitudinal distribution of total tissue N and various components of tissue N along fronds were also analyzed. Several distinct patterns were found: high levels of protein N at growing tips and elevated levels of soluble N in lower parts of the frond. Free amino acids accounted for a major portion of the soluble N, but neither NO ₃ ^– nor NH ₄ ⁺ accumulated in the plant tissue. The longitudinal distribution of N along the fronds is compared to reported variations in C metabolism, and it is concluded that C and N sourcesink relations do not always coincide and bidirectional translocation may occur.     

Relative growth of different species of marine algae in wasterwater-seawater mixtures

In recent studies, we developed a combined nutrient removal-marine aquaculture process for the tertiary treatment of wastewater and the production of commercially important shellfish. Part of this process consists of an outdoor mass cultivation system for marine algae. During our previous experiments we observed that marine diatoms almost exclusively are the dominant algal species in our outdoor cultures. To better understand this phenomenon of diatom dominance we grew 16 species of marine algae in continuous monoculture under laboratory conditions simulating to some degree the conditions prevailing in our outdoor experiments. Species such as Skeletonema costatum, Monochrysis lutheri and Tetraselmis sp., which were never dominant in our outdoor cultures, grew as well in monoculture, as Phaeodactylum tricornutum, frequently, the prevalent species outdoors. However, when monocultures of Dunaliella tertiolecta and Thalassiosira pseudonana (3H) were purposely contaminated with P. tricornutum, the latter species quickly became dominant. It is suggested that a complex interaction of environmental factors is usually responsible for the dominance of a particular species; one such factor may be the nitrogen source in the growth media. Under conditions of virtually, complete nitrogen assimilation, the carbon: nitrogen ratio in the algae was high (7 to 8) when NO ₃ ^- –N was the source of nitrogen, and low (4 to 6) when NH ₄ ⁺ –N was the prime form of nitrogen. When algal growth was low, resulting in a large inorganic nitrogen residue, the carbon:nitrogen ratio was low regardless of whether NO ₃ ^- –N or NH ₄ ⁺ –N was the main nitrogen source.Contribution No. 3297 from the Woods Hole Oceanographic Institution.     

Regional studies of daily,seasonal and size fraction variability in ammonium remineralization

Rates of ammonium remineralization were determined using a ¹⁵N isotope dilution technique for two oceanic regions, one coastal region, and one estuarine region, covering a wide range of ambient nutrient, light, and temperature conditions. Results showed that NH ₄ ⁺ assimilative and regenerative fluxes were primarily in balance, even when the ambient nitrogenous pool was completely dominated by NO ₃ ^- . Variations in uptake and remineralization rates relative to time of day and season were also determined. Size fraction studies at several of the sites showed that the smallest size fraction (<10 m) was usually the most important in remineralizing NH ₄ ⁺ , and the importance of the apparent bacterial fraction (<1 m) may increase following blooms. The results support the concept that, over a wide variety of conditions, the fluxes of NH ₄ ⁺ remineralization and uptake are tightly coupled; phytoplankton are able to utilize NH ₄ ⁺ at the rate that it is produced by heterotrophic processes.     

The use of metabolic ratios for determining the catabolic substrates of a solitary ascidian

In a previous study, Hatcher (1989: Mar. Biol. 102: 445–452) found that variations in CO₂ and O₂ respiration rates in individual marine invertebrates led to RQ (respiratory quotient) values which were variable and often outside theoretical limits. The present study was designed to examine the variability in several excretionbased metabolic ratios which are often used as alternatives for the RQ in qualitative predictions of catabolic substrates. The experimental organism was a solitary ascidian,Herdmania momus (Savigny), collected near Perth, Western Australia, between January and July 1984. Respiration and excretion rates ofH. momus were examined as a function of a progressive nutritional stress, and covariation was examined. Nutritional stress accounted for more of the variation over time in respiration rates (40 to 50%) than in NH ₄ ⁺ excretion rates (20%). Significant net exchanges of dissolved organic compounds were measured. Qualitative predictions of catabolic substrates were based on a comparison of metabolic ratios with theoretical limits. The O:PO ₄ ^3- ratios were lower than the theoretical limits. The values of the O:NH ₄ ⁺ and NH ₄ ⁺ :PO ₄ ^3- ratios were not influenced by nutritional conditions but changed as a function of reproductive condition of the ascidians. Based on the results of this study, it was concluded that metabolic ratios measured on individual ascidians cannot be reliable predictors of catabolic substrates.     

Effects of methionine sulfoximine on growth and nitrogen assimilation of the marine microalga Chlorella autotrophica

Chlorella autotrophica Shihira and Krauss (clone 580), a euryhaline microalga from the marine coastal environment is subject to large fluctuations in external salinity and nitrogen supply. The alga exhibits maximum growth at salinities lower than 100% ASW (artificial seawater). Cells divide faster and show higher cell yields when the supply of either NH ₄ ⁺ or NO ₃ ^- is increased above 0.2 mM. Cells growing on NH ₄ ⁺ show high levels of NADPH-glutamate dehydrogenase (GDH) activity, and the levels of glutamine synthetase (GS) are decreased to very low levels under these conditions. Methionine sulfoximine (MSX), an inhibitor of GS, has little effect on cell division and nitrogen assimilation of cells growing on NH ₄ ⁺ . Cells growing on NO ₃ ^- , however, show marked inhibition (65%) in nitrogen assimilation in the presence of 5 mM MSX. This MSX concentration also causes growth retardation and a progressive decrease in cell protein and nitrogen content. GS is almost completely inhibited by 5 mM MSX in both NH ₄ ⁺ and NO ₃ ^- -grown cells. Cells growing on NH ₄ ⁺ maintain high levels of NADPH-GDH activity in the presence of MSX. NADPH-GDH activity in MSX-treated NO ₃ ^- -grown cells increases, and, in the presence of 5 mM MSX, reaches 40% of the level found in NH ₄ ⁺ -grown cells. These results are consistent with NADPH-GDH providing an alternate pathway for NH ₄ ⁺ assimilation by this marine Chlorella species.     

Dark CO2 uptake by the diatom Chaetoceros simplex in response to nitrogen pulsing

In a continuing investigation of dark CO₂ uptake by nitrogen-limited cultures of the marine diatom Chaetoceros simplex (Bbsm), we expanded on several of our earlier conclusions regarding the potential application of this physiological response for measuring the degree and type of nitrogen limitation in phytoplankton populations. First, the duration over which the maximal enhancement of dark ¹⁴CO₂ uptake was sustained after NH ₄ ⁺ enrichment was a function both of the concentration of added NH ₄ ⁺ and the standing crop of phytoplankton nitrogen — in effect, the total N demand. Second, pulsing with NH ₄ ⁺ for a given degree of N-limitation always produced the same level of enhanced dark CO₂ uptake regardless of whether the cultures were preconditioned with oxidized or reduced nitrogen. In contrast, urea pulsing led to reduced dark CO₂ uptake, but the effect was most pronounced in cells grown on NO ₃ ^– . And third, the assay could be used to distinguish readily between no, moderate, and severe N limitation. The degree of severe N limitation was quantitatively correlated with the degree of enhanced dark CO₂ uptake, but this relationship was not so clear in the region of moderate N limitation. The main advantage of the assay is that it is a relatively simple and effective alternative to more complicated techniques for gauging the degree and form of N limitation in phytoplankton. Further evaluation will be required, both in the laboratory and field, before the assay can be calibrated for quantitative use.Contribution No. 5982 from the Woods Hole Oceanographic Institution     

Application of a 15N tracer method to the study of dissolved organic nitrogen uptake during spring and summer in Chesapeake Bay

The dissolved organic nitrogen (DON) pool in marine waters contains a diverse mixture of compounds. It is therefore difficult to accurately estimate planktonic uptake of DON using the limited number of radiolabeled compounds commercially available. We describe a method to estimate DON uptake rates using ¹⁵N-labeled DON recently released from phytoplankton. To make ¹⁵N-labeled DON, we incubated surface water with ¹⁵NH ₄ ⁺ and then isolated the DON, including any recently released DO¹⁵N, with ion retardation resin. This DON was then added to a freshly collected water sample from the same environment to quantify the rate of DON uptake. The technique was applied to investigate rates of DON uptake relative to inorganic nitrogen in the mesohaline Chesapeake Bay during May 1990 and August 1991. The May experiment took place after the spring bloom, and rates of DON uptake [ranging from 0.31 to 0.53 g-atom (g-at) Nl^-1 h^-1] often exceeded rates of NH ₄ ⁺ and NO ₃ ^- uptake combined. The rates of DON uptake at this time were higher than estimated bacterial productivity and were not correlated with bacterial abundance or bacterial productivity. They were, however, correlated with rates of NO ₃ ^- uptake. In May, we estimate that only 7 to 32% of DON uptake was a result of urea utilization. In contrast, in August, when regenerated nutrients predominate in Chesapeake Bay, rates of DON uptake (ranging from 0.14 to 0.51 g-atom Nl^-1 h^-1) were an average of 50% of the observed rates of NH ₄ ⁺ uptake. Consistent with the May experiment, rates of DON uptake were not correlated with bacterial production. A sizable fraction of DON uptake, however, appeared to be due to urea utilization; rates of urea uptake, measured independently, were equivalent to an average of 74% of the measured rates of DON uptake. These findings suggest that, during both periods of study, at least a fraction of the measured DON uptake may have been due to utilization by phytoplankton.     

Nitrogen Export from an Agriculture Watershed in the Taihu Lake Area, China

Temporal changes in nitrogen concentrations and stream discharge, as well as sediment and nitrogen losses from erosion plots with different land uses, were studied in an agricultural watershed in the Taihu Lake area in eastern China. The highest overland runoff loads and nitrogen losses were measured under the upland at a convergent footslope. Much higher runoff, sediment and nitrogen losses were observed under upland cropping and vegetable fields than that under chestnut orchard and bamboo forest. Sediment associated nitrogen losses accounted for 8–43.5% of total nitrogen export via overland runoff. N lost in dissolved inorganic nitrogen forms (NO ₃ ^– -N + NH ₄ ⁺ -N) accounted for less than 50% of total water associated nitrogen export. Agricultural practices and weather-driven fluctuation in discharge were main reasons for the temporal variations in nutrient losses via stream discharge. Significant correlation between the total nitrogen concentration and stream discharge load was observed. Simple regression models could give satisfactory results for prediction of the total nitrogen concentrations in stream water and can be used for better quantifying nitrogen losses from arable land. Nitrogen losses from the studied watershed via stream discharge during rice season in the year 2002 were estimated to be 10.5 kg N/ha using these simple models.     

Nitrate-reductase activity and in vivo nitrate-reduction rate in Ulva rigida illuminated by blue light

In the marine green alga Ulva rigida C. Agardh, nitrate reductase (NR) is synergetically induced by blue light and nitrate. The present study examines the effect of blue light and a large NO ₃ ^– pulse (0.3 mM) on relevant variables of NO ₃ ^– -assimilation such as NO ₃ ^– -uptake, intracellular NO ₃ ^– -storage, NR activity, in vivo NO ₃ ^– -reduction rate and NO ₂ ^– and NH ₄ ⁺ -accumulation. Nitrate uptake started immediately upon addition of NO ₃ ^– , suggesting the presence of a constitutive carrier, however in the first 1.5 to 2 h, periods of net NO ₃ ^– efflux were frequent. After this time, NO ₃ ^– -uptake and intracellular NO ₃ ^– -accumulation proceeded linearly with time, suggesting the existence of a different NO ₃ ^– -uptake mechanism, which seems to be inducible. Our results indicate that in vivo NO ₃ ^– -reduction is not exclusively dependent on the potential NR activity. In U. rigida, during the first 2 h after a NO ₃ ^– pulse (300 M) there were clear indications that the induction state of the NO ₃ ^– -carrier limits the reduction rate of NO ₃ ^– . Once the induction of the NO ₃ ^– -transporter had been completed (1.5 to 2 h), the NO ₃ ^– -assimilation pathway reached a steady state, NO ₃ ^– -uptake rate, NO ₃ ^– -reduction rate and NO ₂ ^– and NH ₄ ⁺ -accumulation being linear with time. Since the reduction of NO ₃ ^– leads mainly to the accumulation of NH ₄ ⁺ , we conclude that, after the NO ₃ ^– -reduction itself, NH ₄ ⁺ -fixation into carbon skeletons is the limiting step in the assimilation of NO ₃ ^– by U. rigida under blue light.     

Ingestion-independent rates of ammonium excretion by the copepod Calanus pacificus

The relationship between food ingested and NH ₊ ⁴ excretion rate was investigated for female Calanus pacificus collected in August, 1982, from the San Juan Archipelago, Washington State, USA. The copepods were preconditioned to 6 densities of the diatom Thalassiosira weissflogii (0 to 10⁴ cells ml^–1) for 30 h before the experiment. The experiment was conducted with nutrients added in excess to maintain equal rates of NH ₊ ⁴ uptake by the diatoms at all densities. Although ingestion rates of C. pacificus varied from 0 to over 20% of body N d^–1 at the different food levels, excretion was a constant 6.6 nM NH ₊ ⁴ copepod^–1 h^–1 or about 10% of body N d^–1. This ingestion-excretion relationship, which is consistent with previous respiration and fecundity studies, suggests that the ecological dominance of C. pacificus only under conditions of high food abundance may be due to a dramatic increase in its growth efficiency as ingestion increases above the level supporting a constant metabolic rate. The maintenance of a constant level of metabolism during relatively short periods of low food abundance may be advantageous if it allows the copepod to exploit more effectively short-term variability in its food resulting from environmental heterogeneity or vertical migration.Contribution No. 1360 from the School of Oceanography, University of Washington, Seattle, Washington 98195, USA     

Nitrogen exchange between a southeastern USA salt marsh ecosystem and the coastal ocean

The salt marsh ecosystem at North Inlet, South Carolina, USA consistently exported dissolved inorganic nitrogen via tidal exchange with the coastal Atlantic Ocean. Concentrations centrations of NH ₄ ⁺ and NO ₃ ^- +NO ₂ ^- displayed distinct tidal patterns with rising values during ebb flow. These patterns suggest the importance of biogeochemical processes in the flux of material from the salt marsh. NH ₄ ⁺ export peaked during the summer (15 to 20 mg m^-2 tide^-1) during a net balance of tidal water exchange. Remineralization of NH ₄ ⁺ within the salt marsh system appears to be contributing to the estimated annual net export of bout 4.7 g NH ₄ ⁺ -N m^-2 yr^-1. NO ₃ ^- +NO ₂ ^- exports were higher in the fall and winter of 1979 (2 to 4 mg N m^-2 tide^-1). The winter export coincided with a considerable net export of water with no distinctive concentration patterns, suggesting a simple advective export. However, the fall peak of NO ₃ ^- +NO ₂ ^- export occurred during a period of net water balance in tidal exchange and an insignificant freshwater input from the western, forested boundary. During the summer and fall, tidal concentration patterns were particularly apparent, suggesting that nitrification within the salt marsh system was contributing to the estimated annual net export of ca 0.6 g NO ₃ ^- +NO ₂ ^- -N m^-2 yr^-1.Contribution No. 637 from the Belle W. Baruch Institute of Marine Biology and Coastal Research     

Urea as a nitrogen source for the phytoplankton in the Oslofjord

Ambient concentrations of urea in the inner Oslofjord, Norway, showed a pronounced yearly cycle in 1980, with values in the range 0.1 to 10.0 μg-at N l^-1; this cycle resemble that of ammonia although urea concentrations were usually lower. The uptake of urea by phytoplankton was investigated using ¹⁵N. Urea was usually a less important N source than NH ₄ ⁺ , and accounted for 0 to 53% (mean 19%) of summed NH ₄ ⁺ +NO ₃ ^- + urea uptake rates from April to October. Absolute as well as relative (specific) uptake rates of urea were higher in the summer (June–August) than at other times. Uptake of urea was inhibited by NH ₄ ⁺ concentrations higher than 1 to 2 μg-at N l^-1. The summed NH ₄ ⁺ +NO ₃ ^- + urea uptake rate was exponentially related to temperature.     

Phosphate uptake by intertidal algae in relation to zonation and season

The removal of phosphate from ambient seawater by whole plants of five species of fucoid algae, collected from the east coast of N. Ireland in 1988 and 1989, was followed over 6-h periods. A transient uptake pattern was observed forPelvetia canaliculata (L.) Dcne. et Thuret,Fucus spiralis L.,F. vesiculosus L. andF. serratus L., consisting of an initial period of high uptake, followed by a phase of zero uptake and then a period at an intermediate rate.Ascophyllum nodosum (L.) Le Jolis had a constant slow rate of uptake over 6 h. The initial uptake rate ofF. spiralis was significantly greater than that of any other species. Phosphate uptake over a 2-h period was measured at concentrations ranging from that of ambient seawater to 25µg-at. l^–1 for whole plants ofF. spiralis andF. serratus, using a large scale batch method. A small scale batch method was used for whole plants ofP. canaliculata and sections of the other four species investigated. Uptake abilities of the algae at low concentrations of phosphate were compared using the parameterV ₁ (the uptake rate at 1µg-at. l^–1) and at high concentrations usingV _max, the maximum uptake rate. These kinetic parameters of uptake were calculated using a method that avoids bias and permits statistical evaluation of the results. The fucoid algae studied could be divided into two distinct groups on the basis of their abilities to take up phosphate from seawater.P. canaliculata andA. nodosum had low values ofV ₁ in winter, which were also correlated with their positions on the shore and did not vary between winter and summer. TheFucus species had higher values ofV ₁ in winter, which were also correlated with their positions on the shore. In summer, however,V ₁-values for these species decreased and no longer correlated with their shore heights. TheV _max-value forF. spiralis was higher in winter than in summer but was signifcantly greater than that of any other species at all times of year. The ecological significance ofV _max is discussed in relation to nutrient limitation and the possible occurrence of patches of high nutrient concentration in the intertidal environment.     

Ammonium regeneration by microzooplankton in the Oslofjord

Concentrations of dissolved inorganic nitrogen compounds above the pycnocline in the Oslofjord are very low in the summer, with turnover times of the inorganic N pools of no more than a few hours. To investigate the possibility that continued phytoplankton growth in the summer depends on ammonium excretion by microzooplankton, rates of NH ₄ ⁺ regeneration and assimilation were measured by a ¹⁵N isotope dilution method. Daytime regeneration rates at 0–2 m depth were 0–28% of the calculated assimilation rates at ambient NH ₄ ⁺ concentrations. Regeneration was faster during a dinoflagellate bloom in August than in mixed diatom-dinoflagellate blooms in June and September. Most of the NH ₄ ⁺ appeared to be produced by juvenile copepods, rotifers, tintinnids, and heterotrophic dinoflagellates in the size fraction 45–200 m.     

Marine diatoms grown in chemostats under silicate or ammonium limitation. IV. Transient response of Chaetoceros debilis,Skeletonema costatum,and Thalassiosira gravida to a single addition of the limiting nutrient

The kinetic response of ammonium- or silicate-limited and ammonium- or silicatestarved populations of Chaetoceros debilis, Skeletonema costatum, and Thalassiosira gravida was determined by a single addition of the limiting nutrient to a steady-state culture and subsequent monitoring of the nutrient disappearance of the limiting and non-limiting nutrients at frequent time intervals. The kinetic response of nonlimited (nutrient) populations of these three species was also determined. Three distinct modes of the uptake of the limiting nutrient were observed for ammonium-or silicate-limited populations of these three species, surge uptake (V _s ), internally (cellular) controlled uptake (V _i ), and externally (ambient limiting nutrient concentration) controlled uptake (V _e ). Non-limited populations did not exhibit the three distinct segments of uptake, V _s , V _i and V _e . Estimates of the maximal uptake rate (V _max) and the Michaelis constant (K _s ) were obtained from nutrient-limited populations during the V _e segment of the uptake curve. Pooled values of V _e for the three ammonium-limited populations yielded V _max and K _s estimates of 0.16 h^-1 and 0.5 g-at NH₄–N l^-1. Kinetic data derived from the V _e segment of the uptake curve for silicate-limited populations yielded different values of V _max and K _s for each of the three species. In a number of parameters that were measured, T. gravida was clearly different from C. debilis and S. costatum and its recovery from nutrient starvation was the slowest. Recovery of all species from silicate limitation or starvation was slower than from ammonium limitation or starvation. Ammonium-starved populations maintained a maximal uptake rate at a substrate concentration an order of magnitude lower (0.1 g-at NH₄–N l^-1) than that observed for NH₄-limited populations (1.0 g-at NH₄–N l^-1). Adaptation to the severity of the nutrient limitation occurred as changes in the magnitude of cellular characteristics, such as short-term uptake potential (V _s ) and affinity for the substrate (K _s ). The consequence of these results are discussed in terms of another possible mechanism to explain changes in species composition and succession in nutrient-depleted environments.Contribution No. 944 from the Department of Oceanography, University of Washington, Seattle, Washington 98195, USA.