High N2 Fixation Rates Research Articles

Carbon Transfer from the Host Diatom Enables Fast Growth and High Rate of N2 Fixation by Symbiotic Heterocystous Cyanobacteria.

Diatom–diazotroph associations (DDAs) are symbioses where trichome-forming cyanobacteria support the host diatom with fixed nitrogen through dinitrogen (N2) fixation. It is inferred that the growth of the trichomes is also supported by the host, but the support mechanism has not been fully quantified. Here, we develop a coarse-grained, cellular model of the symbiosis between Hemiaulus and Richelia (one of the major DDAs), which shows that carbon (C) transfer from the diatom enables a faster growth and N2 fixation rate by the trichomes. The model predicts that the rate of N2 fixation is 5.5 times that of the hypothetical case without nitrogen (N) transfer to the host diatom. The model estimates that 25% of fixed C from the host diatom is transferred to the symbiotic trichomes to support the high rate of N2 fixation. In turn, 82% of N fixed by the trichomes ends up in the host. Modeled C fixation from the vegetative cells in the trichomes supports only one-third of their total C needs. Even if we ignore the C cost for N2 fixation and for N transfer to the host, the total C cost of the trichomes is higher than the C supply by their own photosynthesis. Having more trichomes in a single host diatom decreases the demand for N2 fixation per trichome and thus decreases their cost of C. However, even with five trichomes, which is about the highest observed for Hemiaulus and Richelia symbiosis, the model still predicts a significant C transfer from the diatom host. These results help quantitatively explain the observed high rates of growth and N2 fixation in symbiotic trichomes relative to other aquatic diazotrophs.

Effect of mesoscale eddies on diazotroph community structure and nitrogen fixation rates in the South China Sea

Mesoscale eddies play a key role in ocean biogeochemistry, yet their effects on dinitrogen (N2) fixation are not well understood. Using quantitative polymerase chain reaction (qPCR) and dinitrogen-15 (15N2) tracer methods, we examined the abundance of nifH genes in N2-fixing microorganisms and N2 fixation rates in the upper water column (0–100 m) of mesoscale eddies in the northeastern South China Sea (SCS). High N2 fixation rates affected by an anticyclonic eddy were associated with high abundances of unicellular nitrogen-fixing cyanobacteria in groups A and B (UCYN-A, UCYN-B), while low nitrogen fixation was found in both cyclonic eddy and reference stations. The estimated N2 fixation rates met 9% ± 5% of the nitrogen demand for primary production affected by the anticyclonic eddy. Our results indicate that mesoscale eddies can affect nitrogen fixation through affecting nutrient availability and eddy transport, and play a crucial role in regulating N2 fixation in the oligotrophic SCS.

N2 fixation in urbanization area rivers: spatial-temporal variations and influencing factors.

While nitrogen (N2) fixation is an important process in nitrogen (N) biogeochemical cycling, supplying a significant portion of the N in natural ecosystems, few quantitative constraints exist concerning its contribution to the N enrichment and export from river ecosystems. This study estimates the N2 fixation rates of urban rivers in the Yangtze Estuary area using acetylene reduction. The results demonstrate that the prominent spatiotemporal variability of river N2 fixation rates is driven by various environmental factors. River N2 fixation rates are significantly higher in the summer (90.57 ± 14.60 ngN·L-1·h-1) than in the winter (57.98 ± 15.73 ngN·L-1·h-1). Spatially, rivers draining urban and suburban areas have higher N2 fixation rates than those draining rural areas. The N2 fixation rates are positively correlated with the N2 fixing cyanobacteria density, water temperature, light, and the water phosphorus (P) concentration, but they are negatively correlated with the dissolved N concentration (NH4+-N and NO3--N). The N2 fixation rates annually range from 53.20 to 89.24 ngN·L-1·h-1 for all of the sampling rivers, which is equivalent to a depth integrated (0-0.6m) N input of 0.163-0.274 gN·m-2·a-1. The determined annual N input via N2 fixation is generally higher than that of marine systems, but it is lower than that of eutrophic lakes. This study provides robust evidence that N2 fixation can supply a substantial portion of the N input to human-impacted river ecosystems, which has not been sufficiently accounted for when determining the N mass balance of riverine ecosystems. A high N2 fixation rate may increase the ratio of N to P input to river systems, and therefore render P the limiting factor in aquatic eutrophication.

Biogeography of N2 Fixation Influenced by the Western Boundary Current Intrusion in the South China Sea

AbstractThe N2 fixation and primary production rates were measured simultaneously using 15N2 and 13C incubation assays in the northern South China Sea influenced by the Kuroshio intrusion (KI) seasonally. The degree of KI (KI index, range from 0 to 1) was assessed by applying an isopycnal mixing model. The water column integrated N2 fixation and primary production for stations with KI index larger than 0.5 were 463 ± 260 μmol N·m−2·day−1 and 62 ± 19 mmol C·m−2·day−1, respectively, significantly higher than those for stations with KI index lower than 0.5 (50 ± 10 μmol N·m−2·day−1 and 28 ± 10 mmol C·m−2·day−1, respectively). Trichodesmium was the dominant diazotroph at stations with KI index larger than 0.5, with 2 orders of magnitude higher nifH gene abundance than that at stations with KI index lower than 0.5. However, the highest N2 fixation rates were found in waters with moderate KI index around 0.6, suggesting that frontal zone mixing might stimulate N2 fixation. Our results demonstrated that diazotrophs (mainly Trichodesmium) were tightly associated with the KI, which modulated the biogeographic distribution of N2 fixers. In summary, we found the transportation of Trichodesmium by KI, then, we quantified the fraction of KI and N2 fixation rates in the northern South China Sea. The results suggested that KI generated a new biogeographic regime which could significantly influence the carbon and nitrogen cycles far away from the main stream.

High Rates of N2 Fixation in Temperate, Western North Atlantic Coastal Waters Expand the Realm of Marine Diazotrophy

AbstractDinitrogen (N2) fixation can alleviate N limitation of primary productivity by introducing fixed nitrogen (N) to the world's oceans. Although measurements of pelagic marine N2 fixation are predominantly from oligotrophic oceanic regions, where N limitation is thought to favor growth of diazotrophic microbes, here we report high rates of N2 fixation from seven cruises spanning four seasons in temperate, western North Atlantic coastal waters along the North American continental shelf between Cape Hatteras and Nova Scotia, an area representing 6.4% of the North Atlantic continental shelf area. Integrating average areal rates of N2 fixation during each season and for each domain in the study area, the estimated N input from N2 fixation to this temperate shelf system is 0.02 Tmol N/year, an amount equivalent to that previously estimated for the entire North Atlantic continental shelf. Unicellular group A cyanobacteria (UCYN‐A) were most often the dominant diazotrophic group expressing nifH, a gene encoding the nitrogenase enzyme, throughout the study area during all seasons. This expands the domain of these diazotrophs to include coastal waters where dissolved N concentrations are not always depleted. Further, the high rates of N2 fixation and diazotroph diversity along the western North Atlantic continental shelf underscore the need to reexamine the biogeography and the activity of diazotrophs along continental margins. Accounting for this substantial but previously overlooked source of new N to marine systems necessitates revisions to global marine N budgets.

Surplus supply of bioavailable nitrogen through N2 fixation to primary producers in the eastern Arabian Sea during autumn

Diazotrophs have received recent appreciation as a major source of bioavailable nitrogen in the global oceans. They mostly flourish in warm, stratified, calm and nutrient depleted conditions in the ocean. Such conditions prevail during spring and autumn seasons in the Arabian Sea. Some previous experimental studies conducted during spring have suggested the highest rates of N2 fixation among other oceans in the eastern Arabian Sea, but there are no such records during autumn. In addition, modelling studies have suggested high rates of annual N2 fixation in the Arabian Sea. In this study, we conducted isotope labeling incubation experiments in the eastern Arabian Sea in autumn 2010 to estimate N2 fixation rates and primary production. Unlike previous studies conducted in this region, we did not witness any diazotrophic bloom, but our N2 fixation rates (1300–2500 μmol N m−2 d−1) were still comparable to the rates reported in previous studies conducted in spring, and among the highest rates observed in the global oceans. Our data suggest an important role of excess phosphate to sustain N2 fixation during autumn. Most intriguingly our study shows that N2 fixation supplies a surplus amount of bioavailable nitrogen required for primary producers during autumn in this region.

Biogeographic drivers of diazotrophs in the western Pacific Ocean

AbstractThe global budget of marine nitrogen (N) is not balanced, with N removal largely exceeding N fixation. One of the major causes of this imbalance is our inadequate understanding of the diversity and distribution of marine N2 fixers (diazotrophs) as well as their contribution to N2 fixation. Here, we performed a large‐scale cross‐system study spanning the South China Sea, Luzon Strait, Philippine Sea, and western tropical Pacific Ocean to compare the biogeography of seven major diazotrophic groups and N2 fixation rates in these ecosystems. Distinct spatial niche differentiation was observed. Trichodesmium was dominant in the South China Sea and western equatorial Pacific, whereas the unicellular cyanobacterium UCYN‐B dominated in the Philippine Sea. Furthermore, contrasting diel patterns of Trichodesmium nifH genes and UCYN‐B nifH gene transcript activity were observed. The heterotrophic diazotroph Gamma A phylotype was widespread throughout the western Pacific Ocean and occupied an ecological niche that overlapped with that of UCYN‐B. Moreover, Gamma A (or other possible unknown/undetected diazotrophs) rather than Trichodesmium and UCYN‐B may have been responsible for the high N2 fixation rates in some samples. Regional biogeochemistry analyses revealed cross‐system variations in N2‐fixing community composition and activity constrained by sea surface temperature, aerosol optical thickness, current velocity, mixed‐layer depth, and chlorophyll a concentration. These factors except for temperature essentially control/reflected iron supply/bioavailability and thus drive diazotroph biogeography. This study highlights biogeographical controls on marine N2 fixers and increases our understanding of global diazotroph biogeography.

Comparison of dinitrogen fixation rates in two subtropical seagrass communities

Seagrass morphology varies greatly between species and can impact their nitrogen (N) cycling capacity, including dinitrogen (N2) fixation. We used a 15N—N2 stable isotope technique to measure N2 fixation rates in different zones of two morphologically diverse seagrass communities, Zostera muelleri and Halophila ovalis in Moreton Bay, Australia. Isotope label additions were made to both the surface and rhizosphere of sediment cores containing whole plants over an artificial diurnal cycle. In both species the highest rates of N2 fixation were found in the leaves (including epiphytes) in the light with areal leaf related N2 fixation rates higher in the Z. muelleri (46 ± 26 μmol N2 m−2 h−1) community compared to H. ovalis (11 ± 7.2 μmol N2 m−2 h−1). There was a switch in the location of N2 fixation from the leaves to the sediments in the dark in the Z. muelleri community which was not observed in H. ovalis. The change in the active site of N2 fixation in the Z. muelleri community is likely related to the reduction in oxygen supply and associated increase in sulphate reducing bacterial activity at night. Overall the Z. muelleri community fixed N2 at ~3 times (~75 μmol N m−2 h−1) the rate of H. ovalis (~25 μmol N m−2 h−1). The results of this study illustrate how plant morphology, and its associated influence on sediment biogeochemistry, can regulate N2 fixation rates in seagrass communities.

Diazotrophy as the main driver of the oligotrophy gradient in the western tropical South Pacific Ocean: results from a one-dimensional biogeochemical–physical coupled model

Abstract. The Oligotrophy to UlTra-oligotrophy PACific Experiment (OUTPACE) cruise took place in the western tropical South Pacific (WTSP) during the austral summer (March–April 2015). The aim of the OUTPACE project was to investigate a longitudinal gradient of biological and biogeochemical features in the WTSP, and especially the role of N2 fixation in the C, N, and P cycles. Two contrasted regions were considered in this study: the Western Melanesian Archipelago (WMA), characterized by high N2 fixation rates, significant surface production and low dissolved inorganic phosphorus (DIP) concentrations, and the South Pacific Gyre (WGY), characterized by very low N2 fixation rates, surface production and high DIP concentrations. Since physical forcings and mixed layer dynamics in both regions were similar, it was considered that the gradient of oligotrophy observed in situ between the WMA and WGY was not explained by differences in physical processes, but rather by differences in biogeochemical processes. A one-dimensional physical–biogeochemical coupled model was used to investigate the role of N2 fixation in the WTSP by running two identical simulations, only differing by the presence (simWMA) or absence (simWGY) of diazotrophs. We showed that the nitracline and the phosphacline had to be, respectively, deeper and shallower than the mixed layer depth (MLD) to bring N-depleted and P-repleted waters to the surface during winter mixing, thereby creating favorable conditions for the development of diazotrophs. We also concluded that a preferential regeneration of the detrital phosphorus (P) matter was necessary to obtain this gap between the nitracline and phosphacline depths, as the nutricline depths significantly depend on the regeneration of organic matter in the water column. Moreover, the model enabled us to highlight the presence of seasonal variations in primary production and P availability in the upper surface waters in simWMA, where diazotrophs provided a new source of nitrogen (N) to the ecosystem, whereas no seasonal variations were obtained in simWGY, in the absence of diazotrophs. These main results emphasized the fact that surface production dynamics in the WTSP is based on a complex and sensitive system which depends on the one hand on physical processes (vertical mixing, sinking of detrital particles), and on the other hand on biogeochemical processes (N2 fixation, remineralization).

In-depth characterization of diazotroph activity across the western tropical South Pacific hotspot of N<sub>2</sub> fixation (OUTPACE cruise)

Abstract. Here we report N2 fixation rates from a ∼ 4000 km transect in the western and central tropical South Pacific, a particularly undersampled region in the world ocean. Water samples were collected in the euphotic layer along a west to east transect from 160∘ E to 160∘ W that covered contrasting trophic regimes, from oligotrophy in the Melanesian archipelago (MA) waters to ultra-oligotrophy in the South Pacific Gyre (GY) waters. N2 fixation was detected at all 17 sampled stations with an average depth-integrated rate of 631 ± 286 µmolNm-2d-1 (range 196–1153 µmolNm-2d-1) in MA waters and of 85 ± 79 µmolNm-2d-1 (range 18–172 µmolNm-2d-1) in GY waters. Two cyanobacteria, the larger colonial filamentous Trichodesmium and the smaller UCYN-B, dominated the enumerated diazotroph community (> 80 %) and gene expression of the nifH gene (cDNA > 105 nifH copies L−1) in MA waters. Single-cell isotopic analyses performed by nanoscale secondary ion mass spectrometry (nanoSIMS) at selected stations revealed that Trichodesmium was always the major contributor to N2 fixation in MA waters, accounting for 47.1–83.8 % of bulk N2 fixation. The most plausible environmental factors explaining such exceptionally high rates of N2 fixation in MA waters are discussed in detail, emphasizing the role of macro- and micro-nutrient (e.g., iron) availability, seawater temperature and currents.

Distribution and rates of nitrogen fixation in the western tropical South Pacific Ocean constrained by nitrogen isotope budgets

Abstract. Constraining the rates and spatial distribution of dinitrogen (N2) fixation fluxes to the ocean informs our understanding of the environmental sensitivities of N2 fixation as well as the timescale over which the fluxes of nitrogen (N) to and from the ocean may respond to each other. Here we quantify rates of N2 fixation as well as its contribution to export production along a zonal transect in the western tropical South Pacific (WTSP) Ocean using N isotope (“δ15N”) budgets. Comparing measurements of water column nitrate + nitrite δ15N with the δ15N of sinking particulate N at a western, central, and eastern station, these δ15N budgets indicate high, modest, and low rates of N2 fixation at the respective stations. The results also imply that N2 fixation supports exceptionally high, i.e. ≥ 50 %, of export production at the western and central stations, which are also proximal to the largest iron sources. These geochemically based rates of N2 fixation are equal to or greater than those previously reported in the tropical North Atlantic, indicating that the WTSP Ocean has the capacity to support globally significant rates of N2 fixation, which may compensate for N removal in the oxygen-deficient zones of the eastern tropical Pacific.

Nitrogen-fixation activity and the abundance and taxonomy of nifH genes in agricultural, pristine, and urban prairie stream sediments chronically exposed to different levels of nitrogen loading.

Small streams exert great influences on the retention and attenuation of nitrogen (N) within stream networks. Human land use can lead to increased transport of dissolved inorganic N compounds and downstream eutrophication. Microbial activity in streams is important for maintaining an actively functioning N cycle. Chronically high N loading in streams affects the rates of the central processes of the N cycle by increasing rates of nitrification and denitrification, with biota exhibiting decreased efficiency of N use. The LINXII project measured N-cycle parameters in small streams using 15NO3- tracer release experiments. We concurrently measured N2 fixation rates in six streams of three types (agricultural, pristine, and urban prairie streams) as part of this broader study of major N-cycle processes. Nitrogen fixation in streams was significantly negatively correlated with nitrate levels, dissolved inorganic N levels, and denitrification rates. Algal mat and leaf litter samples generally exhibited the highest rates of N2 fixation. The abundance of nifH genes, as measured by real-time PCR, was marginally correlated with N2-fixation rates, but not to other N-cycle processes or stream characteristics. The nifH sequences observed were assigned to cyanobacteria, Deltaproteobacteria, Methylococcus, and Rhizobia. Seasonal changes, disturbances, and varying inputs may encourage a diverse, flexible, stable N2-fixing guild. Patchiness in the streams should be considered when assessing the overall impact of N2 fixation, since algal biomass exhibited high rates of N2 fixation.

N2 production and fixation in deep-tier burrows of Squilla empusa in muddy sediments of Great Peconic Bay

Global marine N budgets often show deficits due to dominance of benthic N2 production relative to pelagic N2 fixation. Recent studies have argued that benthic N2 fixation in shallow water environments has been underestimated. In particular, N2 fixation associated with animal burrows may be significant as indicated by high rates of N2 fixation reported in muddy sands populated by the ghost shrimp, Neotrypaea californiensis (Bertics et al., 2010). We investigated whether N2 fixation occurs at higher rates in the burrow-walls of the deep-burrowing (~0.5–4m) mantis shrimp, Squilla empusa, compared to ambient, estuarine muds and measured seasonal in-situ N2 concentrations in burrow-water relative to bottom-water. Acetylene reduction assays showed lower N2 fixation in burrow-walls than in un-populated sediments, likely due to inhibitory effects of O2 on ethylene production. Dissolved N2 was higher in burrow-water than proximate bottom-water at all seasons, demonstrating a consistent balance of net N2 production relative to fixation in deep-tier biogenic structures.

Catchment-scale alder cover controls nitrogen fixation in boreal headwater streams

AbstractAlder (Alnus spp.) is a woody plant with bacterial symbionts that fix atmospheric N2 into bioavailable N. We studied 12 North American boreal headwater streams spanning a steep gradient of catchment alder cover (0–27%) to test the hypothesis that increasing inputs of inorganic N associated with increasing alder cover would reduce or eliminate in-stream benthic N2 fixation. We measured N2 fixation rates, chlorophyll a, and ash-free dry mass (AFDM) of periphyton in early (May) and late (August) summer 2011. Dissolved inorganic N (DIN) concentrations, composed almost entirely of NO3/NO2-N, ranged from below detection limits to nearly 2 mg/L and were strongly predicted by catchment alder cover in both months. Higher N2 fixation rates were observed in August than in May. N2 fixation rates declined sharply when alder cover exceeded ~2% of catchment cover, corresponding to 20 to 40 µg/L DIN. This pattern also was evident among 3 streams with contrasting catchment alder cover sampled approximately every 2...

High‐nitrogen fixation rates in the particulate and dissolved pools in the Western Tropical Pacific (Solomon and Bismarck Seas)

AbstractDinitrogen (N2) fixation rates were investigated in the euphotic layer of the Bismarck and Solomon Seas using 15N2 incubation assays taking into account both the particulate and the dissolved pools. Average depth‐integrated particulate N2 fixation rates were 203 (range 43–399) and 1396 (range 176–3132) μmol N m−2 d−1 in the Bismarck and Solomon Seas, respectively. In both seas, N2 fixation measured in the dissolved pool was similar to particulate N2 fixation, highlighting the potentially substantial underestimation of N2 fixation in oceanic budgets when only particulate N2 fixation is considered. Among the diazotroph phylotypes targeted using quantitative polymerase chain reaction amplification of nifH genes, Trichodesmium was the most abundant. Regression analyses suggest that it accounted for the major proportion of N2 fixation. However, unicellular cyanobacterial and non‐cyanobacterial diazotrophs were also occasionally abundant. This study reports high pelagic N2 fixation rates and confirms that the Western Tropical South Pacific is a hot spot for marine N2 fixation.

Biogeochemical fluxes and fate of diazotroph-derived nitrogen in the food web after a phosphate enrichment: modeling of the VAHINE mesocosms experiment

Abstract. The VAHINE mesocosm experiment in the oligotrophic waters of the Nouméa lagoon (New Caledonia), where high N2 fixation rates and abundant diazotroph organisms were observed, aimed to assess the role of the nitrogen input through N2 fixation in carbon production and export and to study the fate of diazotroph-derived nitrogen (DDN) throughout the planktonic food web. A 1-D vertical biogeochemical mechanistic model was used in addition to the in situ experiment to enrich our understanding of the dynamics of the planktonic ecosystem and the main biogeochemical carbon (C), nitrogen (N) and phosphate (P) fluxes. The mesocosms were intentionally enriched with ∼ 0.8 µmol L−1 of inorganic P to trigger the development of diazotrophs and amplify biogeochemical fluxes. Two simulations were run, one with and the other without the phosphate enrichment. In the P-enriched simulation, N2 fixation, primary production (PP) and C export increased by 201, 208 and 87 %, respectively, consistent with the trends observed in the mesocosms (+124, +141 and +261 % for N2 fixation, PP and C export, respectively). In total, 5–10 days were necessary to obtain an increase in primary and export productions after the dissolved inorganic phosphate (DIP) enrichment, thereby suggesting that classical methods (short-term microcosms experiments) used to quantify nutrient limitations of primary production may not be relevant. The model enabled us to monitor the fate of fixed N2 by providing the proportion of DDN in each compartment (inorganic and organic) of the model over time. At the end of the simulation (25 days), 43 % of the DDN was found in the non-diazotroph organisms, 33 % in diazotrophs, 16 % in the dissolved organic nitrogen pool, 3 % in the particulate detrital organic pool and 5 % in traps, indicating that N2 fixation was of benefit to non-diazotrophic organisms and contributed to C export.

High rates of microbial dinitrogen fixation and sulfate reduction associated with the Mediterranean seagrass Posidonia oceanica

Seagrass meadows of Posidonia oceanica represent hotspots of productivity in the oligotrophic Mediterranean Sea. The lack of dissolved inorganic nitrogen (DIN) in the seawater suggests that the N-demand of these meadows might be in part supported by microbial dinitrogen (N2) fixation. However, currently there are no direct N2 fixation measurements available for this productive marine macrophyte. Here we investigated N2 fixation activity associated with P. oceanica leaf, rhizome and root pieces. In 15N2 incubations, the roots exhibited highest rates of N2 fixation. The rates varied considerably between replicates, presumably due to a patchy microbial colonization of the roots. Additions of organic carbon compounds (acetate, glucose, sucrose or algal lysate) did not enhance the N2 fixation rates. Sulfate reduction rates measured alongside were also highest in root incubations. Correspondingly, sequences of the nifH gene (a marker gene for the iron protein of the N2-fixing enzyme nitrogenase) related to known sulfate-reducing bacteria were retrieved from P. oceanica roots. Other nifH sequences clustered with known heterotrophic diazotrophs previously identified in other marine macrophytes. In particular, many sequences obtained from P. oceanica roots were similar (>94%) to a saltmarsh rhizosphere-associated heterotrophic diazotroph, indicating that heterotrophic lifestyle might be common among marine macrophyte-associated diazotrophs.

N<sub>2</sub> fixation in eddies of the eastern tropical South Pacific Ocean

Abstract. Mesoscale eddies play a major role in controlling ocean biogeochemistry. By impacting nutrient availability and water column ventilation, they are of critical importance for oceanic primary production. In the eastern tropical South Pacific Ocean off Peru, where a large and persistent oxygen-deficient zone is present, mesoscale processes have been reported to occur frequently. However, investigations into their biological activity are mostly based on model simulations, and direct measurements of carbon and dinitrogen (N2) fixation are scarce.We examined an open-ocean cyclonic eddy and two anticyclonic mode water eddies: a coastal one and an open-ocean one in the waters off Peru along a section at 16° S in austral summer 2012. Molecular data and bioassay incubations point towards a difference between the active diazotrophic communities present in the cyclonic eddy and the anticyclonic mode water eddies.In the cyclonic eddy, highest rates of N2 fixation were measured in surface waters but no N2 fixation signal was detected at intermediate water depths. In contrast, both anticyclonic mode water eddies showed pronounced maxima in N2 fixation below the euphotic zone as evidenced by rate measurements and geochemical data. N2 fixation and carbon (C) fixation were higher in the young coastal mode water eddy compared to the older offshore mode water eddy. A co-occurrence between N2 fixation and biogenic N2, an indicator for N loss, indicated a link between N loss and N2 fixation in the mode water eddies, which was not observed for the cyclonic eddy. The comparison of two consecutive surveys of the coastal mode water eddy in November 2012 and December 2012 also revealed a reduction in N2 and C fixation at intermediate depths along with a reduction in chlorophyll by half, mirroring an aging effect in this eddy. Our data indicate an important role for anticyclonic mode water eddies in stimulating N2 fixation and thus supplying N offshore.

Succession within the prokaryotic communities during the VAHINE mesocosms experiment in the New Caledonia lagoon

Abstract. N2 fixation fuels ∼ 50 % of new primary production in the oligotrophic South Pacific Ocean. The VAHINE experiment has been designed to track the fate of diazotroph-derived nitrogen (DDN) and carbon within a coastal lagoon ecosystem in a comprehensive way. For this, large-volume ( ∼ 50 m3) mesocosms were deployed in the New Caledonian lagoon and were intentionally fertilized with dissolved inorganic phosphorus (DIP) to stimulate N2 fixation. This study examined the temporal dynamics of the prokaryotic community together with the evolution of biogeochemical parameters for 23 consecutive days in one of these mesocosms (M1) and in the Nouméa lagoon using MiSeq 16S rRNA gene sequencing and flow cytometry. Combining these methods allowed for inference of absolute cell numbers from 16S data. We observed clear successions within M1, some of which were not mirrored in the lagoon. The dominating classes in M1 were Alpha- and Gammaproteobacteria, Cyanobacteria, eukaryotic microalgae, Marine Group II Euryarchaeota, Flavobacteriia, and Acidimicrobia. Enclosure led to significant changes in the M1 microbial community, probably initiated by the early decay of Synechococcus and diatoms. However, we did not detect a pronounced bottle effect with a copiotroph-dominated community. The fertilization with ∼ 0.8 µM DIP on day 4 did not have directly observable effects on the overall community within M1, as the data samples obtained from before and 4 days after fertilization clustered together, but likely influenced the development of individual populations later on, like Defluviicoccus-related bacteria and UCYN-C-type diazotrophic cyanobacteria (Cyanothece). Growth of UCYN-C led to among the highest N2-fixation rates ever measured in this region and enhanced growth of nearly all abundant heterotrophic groups in M1. We further show that different Rhodobacteraceae were the most efficient heterotrophs in the investigated system and we observed niche partitioning within the SAR86 clade. Whereas the location in- or outside the mesocosm had a significant effect on community composition, the temporal effect was significantly stronger and similar in both locations, suggesting that overarching abiotic factors were more influential than the enclosure. While temporal community changes were evident, prokaryotic diversity (Shannon index) only declined slightly from ∼ 6.5 to 5.7 or 6.05 in the lagoon and M1, respectively, throughout the experiment, highlighting the importance of multiple and varying sources of organic matter maintaining competition.

High cell-specific rates of nitrogen and carbon fixation by the cyanobacterium Aphanizomenon sp. at low temperatures in the Baltic Sea.

Aphanizomenon is a widespread genus of nitrogen (N2)-fixing cyanobacteria in lakes and estuaries, accounting for a large fraction of the summer N2-fixation in the Baltic Sea. However, information about its cell-specific carbon (C)- and N2-fixation rates in the early growth season has not previously been reported. We combined various methods to study N2-fixation, photosynthesis and respiration in field-sampled Baltic Sea Aphanizomenon sp. during early summer at 10°C. Stable isotope incubations at in situ light intensities during 24 h combined with cell-specific secondary ion mass spectrometry showed an average net N2-fixation rate of 55 fmol N cell(-1) day(-1). Dark net N2-fixation rates over a course of 12 h were 20% of those measured in light. C-fixation, but not N2-fixation, was inhibited by high ambient light intensities during daytime. Consequently, the C:N fixation ratio varied substantially over the diel cycle. C- and N2-fixation rates were comparable to those reported for Aphanizomenon sp. in August at 19°C, using the same methods. High respiration rates (23% of gross photosynthesis) were measured with (14)C-incubations and O2-microsensors, and presumably reflect the energy needed for high N2-fixation rates. Hence, Aphanizomenon sp. is an important contributor to N2-fixation at low in situ temperatures in the early growth season.