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.     

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.     

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     

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.     

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.     

Sediment ammonium availability and eelgrass (Zostera marina) growth

The interaction of sediment ammonium (NH ₄ ⁺ ) availability and eelgrass (Zostera marina L.) growth, biomass and photosynthesis was investigated using controlled environment and in-situ manipulations of pore water ammonium concentrations. Sediment diffusers were used to create pore water diffusion gradients to fertilize and deplete ammonium levels in sediments with intact eelgrass rhizospheres. Between October, 1982 and September, 1983 controlled environment experiments using plants from shallow (1.3 m) and deep (5.5 m) stations in a Great Harbor, Woods Hole, Massachusetts, USA eelgrass meadow along with in-situ experiments at these stations provided a range of sediment ammonium concentrations between 0.1 and 10 mM (adsorbed+interstitial NH ₄ ⁺ ). The results of the in-situ experiments indicate that nitrogen limitation of eelgrass growth does not occur in the Great Harbor eelgrass meadow. A comparison of NH ₄ ⁺ regeneration rates and eelgrass nitrogen requirements indicates an excess of nitrogen supply over demand and provides an explanation for the lack of response to the manipulations. Results of controlled environment experiments combined with in-situ results suggest that sediment ammonium pool concentrations above approximately 100 mol NH ₄ ⁺ per liter of sediment (interstitial only) saturate the growth response of Zostera marina.     

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.     

Characterization of two glutamate dehydrogenases from the symbiotic microalga Symbiodinium microadriaticum isolated from the coral Acropora formosa

In view of their possible involvement in ammonium assimilation in the coral/algal symbiosis, we have purified two distinct glutamate dehydrogenase isoenzymes from the symbiotic dinoflagellate Symbiodinium microadriaticum (Freudenthal) extracted from the staghorn coral Acropora formosa collected from Magnetic Island, North Queensland, Australia, in 1986–1987. An NADPH-specific glutamate dehydrogenase (GDH) displayed biphasic kinetics with respect to ammonium as the variable substrate; at low substrate concentrations the apparent K _m was below 1 mM, whereas at high substrate concentrations the corresponding value was approximately 200 mM. The NADPH-GDH displayed extremely low activity in the direction of glutamate oxidation; together with the kinetic data this suggests a probable role in ammonium assimilation. A second (NADH-specific) GDH was found to have both amination and deamination activities, and presumably functions in vivo in glutamate oxidation. Kinetic constants are reported for both GDH isoenzymes.     

Characteristics of the uptake system for L-lysine and L-arginine inPhaeodactylum tricornutum

Cells ofPhaeodactylum tricornutum Bohlin develop the ability to take up L-lysine when they are deprived of nitrogen (illuminated in nitrogen-free medium), carbon (incubated in darkness) or both. Cells with a developed uptake system take up and accumulate lysine in an unchanged form. Uptake occurs under either aerobic or anaerobic conditions and is dependent on the presence of sodium⁺ ions (K _s ^{Na +}=,ca. 10 mM). Some potassium⁺ ions are necessary for uptake, presumably within the cells, but with potassium⁺-replete cells, increasing K⁺ concentration depresses lysine uptake. The lysine-uptake porter also transports L-arginine.K _s values are about 1.5 M for lysine and 0.5 M for arginine. It is, however, possible that the uptake system developed by incubating cells in darkness differs from that produced in light; it shows a pronounced pH optimum at pH 8.5, whereas the activity of the light-developed system declines from pH 6.5 to pH 9.0 and correlates well with the concentration of lysine⁺. The uptake system developed in darkness may also have a higher affinity for lysine. Lysine uptake is not inhibited by 1 mM concentrations of nitrate, nitrate, ammonium, or urea nor by similar concentrations of amphoteric or acidic amino acids.     

Comparisons of nutrient uptake rates for Baltic macroalgae with different thallus morphologies

In-situ experiments were performed during different seasons to determine uptake rates of PO _3- ⁴ , NH ₊ ⁴ and NO _- ³ within ecologically representative ranges of nutrient concentrations, of dominant macroalgae in the Baltic Sea. Uptake rates were governed by nutrient concentrations, water temperature and thallus morphology, but not by the phylogenetic affinity of the species. Nitrogen uptake rates were always higher than those of phosphorus at the same concentrations, and NH ₊ ⁴ –N uptake rates exceeded those of NO _- ³ –N. The lowest uptake rates occurred among the late successional, long-lived, coarse species with low surface: volume ratios (Fucus vesiculosus, Furcellaria lumbricalis andPhyllophora truncata). The highest uptake rates were measured for short-lived, opportunistic algae, filamentous or with numerous hairs, (Cladophora glomerata, Enteromorpha ahlneriana, Scytosiphon lomentaria, Dictyosiphon foeniculaceus andCeramium tenuicorne). The latter group also had the highest V_max:k_max ratios, which indicates a more competitive advantage for nutrient uptake at low concentrations.     

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.     

Photosynthetic utilization of CO2(aq) and HCO 3 - in Thalassia testudinum (Hydrocharitaceae)

The effects of total dissolved inorganic carbon (DIC), free carbon dioxide [CO_2(aq)], and bicarbonate (HCO ₃ ^- ) concentrations on net photosynthetic oxygen evolution of the marine angiosperm Thalassia testudinum Banks ex König collected from Biscayne Bay (1988) and from Tampa Bay (1990), Florida, USA, were examined. Rates of photosynthesis declined by 85% from pH 7.25 to 8.75 in buffered seawater media with constant DIC concentration (2.20 mM), suggesting a strong influence of CO_2(aq) concentration. A plateau in the pH-response curve between pH 7.75 and 8.50 indicated possible utilization of HCO ₃ ^- . Responses of photosynthesis measured in buffered seawater media of varying DIC concentrations (0.75 to 13.17 mM) and pH (7.8 to 8.61) demonstrated that photosynthesis is rate-limited at ambient DIC levels. Photosynthesis increased in media with increasing HCO ₃ ^- concentrations but near-constant CO_2(aq) levels, confirming HCO ₃ ^- assimilation. Calculated half-saturation constants (K _s )for CO_2(aq) and HCO ₃ ^- indicated a high affinity for the former [K _s (CO₂)=3 to 18 M] and a much lower affinity for the latter [K _s (HCO ₃ ^- )=1.22 to 8.88 mM]. Calculated V _max values for HCO ₃ ^- were generally higher than those for CO_2(aq), suggesting relatively efficient HCO ₃ ^- utilization, despite the apparent low affinity for this carbon form.     

Stimulation of phytoplankton production in coastal waters by natural rainfall inputs: Nutritional and trophic implications

Recent evaluations of estuarine and coastal nutrient budgets implicate atmospheric deposition as a potentially significant (20 to 30%) source of biologically available nitrogen. We examined the potential growth stimulating impact of atmospheric nitrogen loading (ANL), as local rainfall, in representative shallow, nitrogen limited North Carolina mesohaline estuarine and euhaline coastal Atlantic Ocean habitats. From July 1988 to December 1989, using in situ bioassays, we examined natural phytoplankton growth responses, as¹⁴CO₂ assimilation and chlorophylla production, to rain additions over a range of dilutions mimicking actual input levels. Rainfall at naturally occurring dilutions (0.5 to 5%) stimulated both¹⁴CO₂ assimilation and chlorophylla production, in most cases in a highly significant manner. Parallel nutrient enrichments consistently pointed to nitrogen as the growth stimulating nutrient source. Generally, more acidic rainfall led to greater magnitudes of growth stimulation, especially at lower dilutions. Nutrient analyses of local rainfall from May 1988 to January 1990 indicated an inverse relationship between pH and NO ₃ ^- content. There have been growing concerns regarding increasing coastal and estuarine eutrophication, including ecologically and economically devastating phytoplankton blooms bordering urban and industrial regions of North America, Europe, Japan, and Korea. It appears timely, if not essential, to consider atmospheric nutrient loading in the formulation and implementation of nutrient management strategies aimed at mitigating coastal eutrophication.     

Occurrence and significance of nitrogen transport in the brown alga Laminaria digitata

Nitrogen transport in a Laminaria digitata (Huds.) Lamour population growing at Arbroath, Scotland has been investigated (during the period 3 June 1981 to 3 July 1981), using ¹⁵N as tracer. NO ₃ ^- was assimilated both by the blade meristem and by the mature blade. NO ₃ ^- uptake by the blade meristem alone was insufficient to supply the nitrogen demand for growth. This additional demand was met by nitrogen transport from the mature blade to the meristem. It was estimated that 70% of the nitrogen demand of the meristematic region was supplied by nitrogen transport from the mature blade. Although transport occurred, the size of the endogenous pool of stored nitrogen in the mature blade did not change, the quantity of nitrogen transported being equivalent to the amount of NO ₃ ^- assimilated during the experimental period. The possible co-transport of nitrogen and carbon is discussed.     

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.     

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.     

Effect of algal bloom deposition on sediment respiration and fluxes

Using sediment cores collected in November 1989 from Aarhus Bight, Denmark, the fluxes of O₂, CO₂ (total CO₂), NH ₄ ⁺ , NO ₃ ^– +NO ₂ ^– and DON (dissolved organic nitrogen) across the sediment-water interface were followed for 20 d in an experimental continous flow system. On day 7, phytoplankton was added to the sediment surface, to see the result of simulated algal bloom sedimentation. Benthic O₂ consumption and CO₂ efflux, 38 to 41 mmol O₂ m^-2 d^-1 and 25 to 30 mmol CO₂ m^-2 d^-1, respectively, immediately increased by 39% and 100% after the algal addition, but gradually declined to the orginal level. Fluxes of NH ₄ ⁺ (1.0 to 1.2 mmol m^-2 d^-1) and DON (0.3 to 0.9 mmol m^-2 d^-1) increased due to the organic substrate addition. NH ₄ ⁺ and NO ₃ ^– flux changed direction, becoming an efflux and influx, respectively, for a few days and a large amount of DON (max. rate 4.0 mmol m^-2 d^-1) was immediately produced either by bacterial hydrolytic activity or from autolysis of the algae. DON was the most significant nitrogen component in pore water and in terms of N-flux from sediment. A temporary stimulation of anaerobic respiration processes (sulfate reduction and denitrification) and a decrease in nitrification were indicated. After the effect of the organic addition had declined, the fluxes gradually reverted to the original rates. The halflife of the added algal material, of which 20 to 30% was very labile, was estimated to be 2 to 3 wk.     

Bioturbation effects of the amphipod Corophium volutator on microbial nitrogen transformations in marine sediments

Microcosms containing different densities of Corophium volutator, ranging from 0 to 6000 ind m^-2, were incubated in a flow-through system. Benthic fluxes of CO₂, O₂, NO₃ ^- and NH₄ ⁺ were measured regularly. Thirteen days after setup the microcosms were sacrificed and sediment characteristics, pore water NO₃ ^-, NH₄ ⁺ and exchangeable NH₄ ⁺ concentrations, and potential nitrification activity were measured. The presence of C. volutator increased overall mineralization processes due to burrow construction and irrigation. The amphipods increased the ratio CO₂/O₂ fluxes from 0.73 to 0.86 in microcosms inhabited by 0 and 6000 ind m^-2, respectively. Burrow ventilation removed NH₄ ⁺ from the sediment, which was nitrified in the oxic layer and transported NO₃ ^- to the burrow sediment, where denitrification potential was enhanced. Nitrification and total denitrification rates (denitrification of NO₃ ^- coming from the overlying water and of NO₃ ^- generated within the sediment) were calculated and discussed. Bioturbation by C. volutator increased both nitrification and denitrification, but denitrification was stimulated more than nitrification. Denitrification of NO₃ ^- coming from the overlying water was stimulated 1.2- and 1.7-fold in microcosms containing 3000 and 6000 ind m^-2 relative to control microcosms. The presence of C. volutator (6000 ind m^-2) stimulated nitrogen removal from the system, as dinitrogen, 1.5-fold relative to non-bioturbated microcosms. C. volutator individuals used in our study were collected from Norsminde Fjord, Denmark, in 1990.     

Nitrate reductase activity in Zostera marina

Eelgrass (Zostera marina L.) has access to nutrient pools in both the water column and sediments. We investigated the potential for eelgrass to utilize nitrate nitrogen by measuring nitrate reductase (NR) activity with an in vivo tissue assay. Optimal incubation media contained 60 mM nitrate, 100 mM phosphate, and 0.5% 1-propanol at pH 7.0. Leaves had significantly higher NR activity than roots (350 vs 50 nmoles NO ₂ ^– produced g FW^–1 h^–1). The effects of growing depth (0.8 m MLW, 1.2 m, 3.0 m, 5.0 m) and location within the eelgrass meadow (patch edge vs middle) on NR activity were examined using plants collected from three locations in the Woods Hole area, Massachusetts, USA, in July 1987. Neither depth nor position within the meadow appear to affect NR activity. Nitrate enrichment experiments (200 M NO ₃ ^– for 6 d) were conducted in the laboratory to determine if NR activity could be induced. Certain plants from shallow depth (1.2 m) showed a significant response to enrichment, with NR activity increasing from >100 up to 950 nmoles NO ₂ ^– g FW^–1 h^–1 over 6 d. It appears that Z. marina growing in very shallow water (0.8 m) near a shoreline may be affected by ground water or surface run-off enrichments, since plants from this area exhibited rates up to 1 600 nmol NO ₂ ^– g FW^–1 h^–1. Water samples from this location consistently had slightly higher NO ₃ ^– concentrations (1.4 M) than all other collection sites (0.7 M). Thus, it is possible that chronic run-off or localized groundwater inputs can create sufficient NO ₃ ^– enrichment in the water column to induce nitrate reductase activity in Zostera leaves.     

Isotopic characterization of atmospheric nitrogen inputs as sources of enhanced primary production in coastal Atlantic Ocean waters

Current estimates indicate that atmospheric nitrogen deposition is responsible for 26 to over 70% of new nitrogen (N) input to North Carolina estuaries and coastal waters. Concentrations of N in coastal rainfall events in a 2-yr period (August 1990 to 1992) ranged from 0.7 to 144 M for NO ₃ ^- and 0.5 to 164 M for NH ₄ ⁺ . The ¹⁵N values of the NO ₃ ^- and the NH ₄ ⁺ were determined in 15 rain events. NH₄ ⁺ values averaged-3.13 (range:-12.5 to+3.6), while NO ₃ ^- plus dissolved organic N fractions had an average ¹⁵N of+1.0 (range:-2.0 to+4.7). The uptake of this isotopically light N into particulate N, in parallel with primary productivity and biomass (as chlorophyll a) determinetions, was examined in microcosm and mesocosm bioassays. As phytoplankton productivity and biomass increased with added rainwater N, the ¹⁵N of particulate N decreased. To investigate the effects of significant atmospheric N loading with stable isotope tracers, we measured the ¹⁵N of the>1 m fraction from surrounding coastal waters. Owing to the episodic nature of atmospheric deposition and the great variation in N loading with each event, a simple assessment of the atmospheric contribution was not possible. During a period in which rainfall inputs were significant and frequent (August 1992), ¹⁵N values were several more negative than during periods of drought (Fall 1990). These experiments and observations emphasize the contribution of atmospheric nitrogen deposition to new production in coastal waters.