The Fate of Nitrogren Fixed by Diazotrophs in the Ocean

While we now know that N2 fixation is a significant source of new nitrogen (N) in the marine environment, little is known about the fate of this N (and associated C), despite the importance of diazotrophs to global carbon and nutrient cycles. Specifically, does N fixed during N2 fixation fuel autotrophic or heterotrophic growth and thus facilitate carbon (C) export from the euphotic zone, or does it contribute primarily to bacterial productivity and respiration in the euphotic zone? For Trichodesmium, the diazotroph we know the most about, the transfer of recently fixed N2 (and C) appears to be primarily through dissolved pools. The release of N varies among and within populations and as a result of the changing physiological state of cells and populations. The net result of trophic transfers appears to depend on the co-occurring organisms and the complexity of the colonizing community. In order to understand the impact of diazotrophy on carbon flow and export in marine systems, we need a better understanding of the trophic flow of elements in Trichodesmium-dominated communities and other diazotrophic communities under various defined physiological states. Nitrogen and carbon fixation rates themselves vary by orders of magnitude within and among studies of Trichodesmium, highlighting the difficulty in extrapolating global rates of N2 fixation from direct measurements. Because the stoichiometry of N2 and C fixation does not appear to be in balance with that of particles, and the relationship between C and N2 fixation rates is also variable, it is equally difficult to derive global rates of one from the other. This paper seeks to synthesize what is known about the fate of diazotrophic production in the environment. A better understanding of the physiology and physiological ecology of Trichodesmium and other marine diazotrophs is necessary to quantify and predict the effects of increased or decreased diazotrophy in the context of the carbon cycle and global change

Peptide Hydrolysis and the Uptake of Dipeptides by Phytoplankton

Rates of peptide hydrolysis (using the fluorescent substrate, lucifer yellow anhydride-labeled tetra-alanine) and dipeptide uptake (using dually labeled, 15N and 13C, dialanine) were measured in phytoplankton cultures and in natural populations during algal blooms dominated by one or two taxa. During most sampling events, both peptide hydrolysis and dipeptide uptake were greatest in the size fraction containing the dominant phytoplankter, suggesting that phytoplankton contribute substantially to or may even dominate observed extracellular peptide hydrolysis and dipeptide uptake in the environment. These are the first data suggesting that dipeptides may be taken up directly by phytoplankton and this may represent a previously unaccounted-for nitrogen source in aquatic systems. Like many other processes in phytoplankton, peptide hydrolysis appears sensitive to the diel light cycle and the nutrient environment, with rates varying depending on the dominant N source, but with no clear pattern. Uptake of dialanine, the dominant product of the hydrolysis of the peptide tetra-alanine, also varied depending on the dominant taxa and the nutrient regime. Most of the time, it appeared that low production of dialanine by tetra-alanine hydrolysis limited the uptake of the dipeptide. Close coupling between peptide hydrolysis and dipeptide uptake may also help explain the absence of correlations between rates of peptide hydrolysis and the concentration and composition of the free amino acid pool

The Effect of Growth Rate, Phosphorus Concentration, and Temperature on N-2 Fixation, Carbon Fixation, and Nitrogen Release in Continuous Cultures of Trichodesmium IMS101

With the use of continuous culture systems, rates of dinitrogen (N2) and carbon (C) fixation and nitrogen (N)- and C-based doubling times were assessed in Trichodesmium IMS101 growing exponentially at steady state dilution rates of 0.10, 0.20, and 0.33 d-1 (doubling times of 10, 5, and 3 d - within the range reported for natural populations). Rates of C fixation, N2 fixation, and N release were examined in replicate culture systems with several techniques. Biomass-specific C uptake varied little with population doubling time, but N2 fixation and N release varied markedly among treatments. Total daily gross N2 fixation rates and estimated N release rates were higher in cultures with higher dilution rates. Cultures grown at lower dilution rates had higher daily C:N2 fixation ratios and lower N release rates. Consistent with other studies, it was estimated that Trichodesmium released about 80-90% of their recently fixed N2 during growth. Turnover of cellular C estimated from carbon fixation was a good estimator of population growth rates in steady state cultures, whereas turnover of cellular N estimated from gross or net N2 fixation was not. Small changes in temperature (24°C vs. 28°C) did not appear to affect gross N2 fixation, whereas inorganic phosphorus (1 vs. 5 μmol L-1) supply had a large effect on N2 fixation. These results suggest that continuous culture systems are excellent for elucidating physiological responses of Trichodesmium under ecologically relevant growth conditions and provide a framework for assessing highly variable field estimates of N2 and C fixation

Growth and Nitrogen Uptake Kinetics in Cultured Procentrum donghaiense

We compared growth kinetics of Prorocentrum donghaiense cultures on different nitrogen (N) compounds including nitrate, ammonium urea, glutamic acid, dialanine, and cynate. P. donghaiense exhibited standard Monod-type growth kinetics over a range of nitrogen concentrations for all N. Compounds tested. Cultures grown on glu and urea had the highest maximum growth rates. However, cultures grown on cyanate had lower half saturation constants. Nitrogen uptake kinetics were measured in nitrate-deplete and replete batch cultures of P. donghaiense. In nitrate-deplete batch cultures, P. donghaiense exhibited Michaelis-Menten type uptake kinetics for nitrate, ammonium, urea and algal amino acids; uptake was saturated at or below 50 umol N L-1. In nitrate replete batch cultures, ammonium, urea, and algal amino acid uptake kinetics were similar to those measured in nitrate deplete batch cultures. Together, our results demonstrate that P. donghaiense can grow well on a variety of N sources, and exhibits similar uptake kinetics under both nutrient replete and deplete conditions. This may be an important factor facilitating their growth during bloom initiation and development in N-enriched estuaries where many algae compete for bioavailable N. and nutrient environment changes as a result of algal growth

Peptide Hydrolysis, Amino Acid Oxidation, and Nitrogen Uptake in Communities Seasonally Dominated by Aureococcus Anophagefferens

Elevated levels of dissolved organic nitrogen (DON) and dissolved inorganic nitrogen (DIN) are among the factors implicated in the initiation of algal blooms. However, the degree to which phytoplankton augment their autotrophic metabolism with heterotrophic uptake of organic carbon that is associated with DON is unknown. We evaluated the relative importance of peptide hydrolysis, amino acid oxidation, and amino acid uptake over a seasonal cycle in an embayment on Long Island, New York, that had high concentrations of dissolved organic matter (DOM) and a bloom of the brown tide pelagophyte, Aureococcus anophagefferens. Amino acids were a significant component (up to 50%) of the total N uptake, particularly during the late summer. About half of the associated amino acid C was also taken up. Amino acid oxidation rates were an order of magnitude lower than free amino acid uptake rates, but still supplied up to 32.5% of the NH4+ taken up. Up to 75% of the amino acid oxidation was in the bacterial size fraction (\u3c1.2 μm), and rates were significantly correlated with bacterial densities. Peptide hydrolysis rates were high, and most (up to 72%) occurred in the brown tide size fraction (1.2–5 μm). The high rates of peptide hydrolysis and amino acid uptake measured in cultures of A. anophagefferens confirm that this species can readily hydrolyze peptides and take up N and C from amino acids. Laboratory findings and size-fractionation studies in the field suggest that A. anophagefferens plays a major role in consumption of both C and N from DOM

Nutrient Cycles and Marine Microbes in a CO2-Enriched Ocean

The ocean carbon cycle is tightly linked with the cycles of the major nutrient elements nitrogen, phosphorus, and silicon. It is therefore likely that enrichment of the ocean with anthropogenic CO2 and attendant acidification will have large consequences for marine nutrient biogeochemistry, and for the microbes that mediate many key nutrient transformations. The best available evidence suggests that the nitrogen cycle may respond strongly to higher CO2 through increases in global N2 fixation and possibly denitrification, as well as potential decreases in nitrification. These trends could cause nitrification to become a nitrogen cycle bottleneck, by increasing the flux of N2 fixed into ammonium while decreasing the fraction being oxidized to nitrate and nitrate. The consequences could include reduced supplies of oxidized nitrogen substrates to denitrifiers, lower levels of nitrate-supported new primary production, and expansion of the regenerated production system accompanied by shifts in current phytoplankton communities. The phosphorus and silicon cycles seem less likely to be directly affected by enhanced CO2 conditions, but will undoubtedly respond indirectly to changing carbon and nitrogen biogeochemistry. A review of culture experiments that examined the effects of increased CO2 on elemental ratios of phytoplankton suggests that for most cyanobacteria and eukaryotes, C:N and N:P ratios will either remain at Redfield values or increase substantially. Natural plankton community CO2 manipulation experiments show much more mixed outcomes, with both increases and decreases in C:N and N:P ratios reported at future CO2 levels. We conclude our review with projections of overall trends in the cycles of nitrogen, phosphorus, and silicon over the next century as they respond to the steady accumulation of fossil-fuel derived CO2 in a rapidly changing ocean

Extracellular Enzyme Activity and Uptake of Carbon and Nitrogen Along an Estuarine Salinity and Nutrient Gradient

Amino acid oxidation (AAO) and peptide hydrolysis (PH) are processes affecting the recycling of organic material and nutrients. We compared extracellular AAO and PH rates to C and N uptake rates along estuarine gradients of salinity, nutrients and productivity in the Pocomoke River, a subestuary of the Chesapeake Bay. This estuary is seasonally depleted in inorganic N, and rich in dissolved organic material (DOM) throughout the year. AAO, PH, and N uptake rates measured in 1999 and 2000 were not limited to particular size fractions measured, or to auto- or heterotrophic groups of organisms. At a station near the turbidity maximum, where chlorophyll a biomass was highest, smaller (\u3c1.2 mum) size-fractions contributed \u3c20% of the AAO in May and up to 80% in August when AAO rates were similar to 10 times lower. Most PH was in the larger (\u3e1.2 mum) size-fraction, except at the least saline station in August of both years. Rates of AAO and PH were not linearly correlated with each other seasonally or spatially. Uptake of NH4+ dominated total N uptake (\u3e50%) at all but the freshwater station, although uptake of organic compounds was measurable at all sites. Rates of dissolved free amino acid uptake, measured using dually labeled compounds, were substantial (up to 11% of the total N uptake) and contributed both C and N for growth. Dual labels unambiguously demonstrated that uptake rates of amino acid C and N were uncoupled; amino acid N was taken up preferentially to amino acid C even when rates were corrected for N uptake from AAO. Conceptual models of DOM cycling should include the realization that enzymatic processes and uptake of DOM occur in both \u27microbial\u27 and larger size fractions. Thus, competition between bacteria and phytoplankton mixotrophs may be an important factor determining the relative uptake of C and N from amino acids and other organic substrates

Effects of Temperature, Irradiance and pCO2 on the Growth and Nitrogen Utilization of Prorocentrum Donghaiense

Environmental factors such as temp erature, irradiance, and nitrogen (N) supply affect the growth of Prorocentrum donghaiense, but the interactive effects of these physical factors and the effects of atmospheric CO2 (pCO2) on growth and N uptake have not been examined. We compared growth kinetics of P. donghaiense grown on 4 different N substrates (nitrate [NO3 -], ammonium [NH4 +], urea, and glutamic acid [glu]) with respect to temperature, irradiance, and pCO2. Temperature (15 to 30°C) had a positive effect on growth (max. growth rates: 0.17 to 0.65 d-1; optimal temperature: 25 to 30°C); maximum specific growth rates (μmax) declined when cultures were grown at 30°C. P. donghaiense grew well on all 4 N sources, under irradiances ranging from 10 to 180 μmol quanta m-2 s-1. μmax (2.0 ± 0.1 d-1) was observed in cultures growing with NH4+ as the sole N source in the highest irradiance treatment (180 μmol quanta m-2 s-1). These rates were significantly higher than those measured in cultures grown on NO3 -, urea, and glu (all ∼1.4 d-1). Half-saturation constants (Ks) ranged from 66.5 ± 4.6 to 99.4 ± 6.7 μmol quanta m-2 s-1 for cultures grown with glu or NH4+, respectively, as the sole source of N. Both growth and N uptake rates were higher in cultures grown under elevated pCO2. Our results suggest that P. donghaiense exhibits flexible adaptation for N utilization under broad environmental conditions (temperature, irradiance, pCO2), which may play an important role in the formation and duration of P. donghaiense blooms

Toward Resolving Disparate Accounts of the Extent and Magnitude of Nitrogen Fixation in the Eastern Tropical South Pacific Oxygen Deficient Zone

Examination of dinitrogen (N2) fixation in the Eastern Tropical South Pacific oxygen deficient zone has raised questions about the range of diazotrophs in the deep sea and their quantitative importance as a source of new nitrogen globally. However, technical considerations in the deployment of stable isotopes in quantifying N2 fixation rates have complicated interpretation of this research. Here, we report the findings of a comprehensive survey of N2 fixation within, above and below the Eastern Tropical South Pacific oxygen deficient zone. N2 fixation rates were measured using a robust 15N tracer method (bubble removal) that accounts for the slow dissolution of N2 gas and calculated using a conservative approach. N2 fixation was only detected in a subset of samples (8 of 125 replicated measurements) collected within suboxic waters (\u3c 20 μmol O2 kg−1) or at the oxycline. Most of these detectable rates were measured at nearshore stations, or where surface productivity was high. These findings support the hypothesis that low oxygen/high organic carbon conditions favor non-cyanobacterial diazotrophs. Nevertheless, this study indicates that N2 fixation is neither widespread nor quantitatively important throughout this region

Phosphorus Dynamics in Cultures and Natural Populations of Trichodesmium spp

Trichodesmium spp. fix atmospheric N2 and so an element other than N limits production by these species in the oligotrophic ocean. Because dissolved inorganic phosphorus (DIP) is in short supply in many marine systems, it has been hypothesized that P is a growth-limiting nutrient for these species in nature. However, Trichodesmium is capable of hydrolyzing dissolved organic P (DOP) compounds and the inorganic products from hydrolysis may provide an additional source of P for growth. We investigated P dynamics and alkaline phosphatase activity in cultures and natural populations of Trichodesmium from the Atlantic Ocean and the north coast of Australia to determine whether hydrolysis of DOP could supply enough P to fuel growth. During the Atlantic cruise, concentrations of DIP were lower and chlorophyll (chl a)-specific rates of alkaline phosphatase activity by Trichodesmium were higher than during the Australian transect. However, because Trichodesmium were much more abundant during the Australian transect, where they represented the bulk of the surface chl a biomass, total water column rates of alkaline phosphatase activity were higher along the Australian transect than in the Atlantic. In both systems, DOP could potentially supply a significant portion of the cellular P necessary for growth. In cultures and natural populations, alkaline phosphatase activity was inhibited when DIP was present and increased in the presence of DOP. Cultures of Trichodesmium IMS101 grew equally well on media enriched with DOP or DIP at all but the highest enrichment levels. From these studies, we conclude that if the supply of appropriate DOP substrates is adequate, DOP can represent an important P source for Trichodesmium growth which should be included in estimates of P availability in surface waters