Search

N2 fixation rates were measured daily in large (∼ 50 m3) mesocosms deployed in the tropical southwest Pacific coastal ocean (New Caledonia) to investigate the temporal variability in N2 fixation rates in relation with environmental parameters and study the fate of diazotroph-derived nitrogen (DDN) in a low-nutrient, low-chlorophyll ecosystem. The mesocosms were fertilized with ∼ 0.8 µM dissolved inorganic phosphorus (DIP) to stimulate diazotrophy. Bulk N2 fixation rates were replicable between the three mesocosms, averaged 18.5 ± 1.1 nmol N L−1 d−1 over the 23 days, and increased by a factor of 2 during the second half of the experiment (days 15 to 23) to reach 27.3 ± 1.0 nmol N L−1 d−1. These later rates measured after the DIP fertilization are higher than the upper range reported for the global ocean. During the 23 days of the experiment, N2 fixation rates were positively correlated with seawater temperature, primary production, bacterial production, standing stocks of particulate organic carbon (POC), nitrogen (PON) and phosphorus (POP), and alkaline phosphatase activity, and negatively correlated with DIP concentrations, DIP turnover time, nitrate, and dissolved organic nitrogen and phosphorus concentrations. The fate of DDN was investigated during a bloom of the unicellular diazotroph UCYN-C that occurred during the second half of the experiment. Quantification of diazotrophs in the sediment traps indicates that ∼ 10 % of UCYN-C from the water column was exported daily to the traps, representing as much as 22.4 ± 5.5 % of the total POC exported at the height of the UCYN-C bloom. This export was mainly due to the aggregation of small (5.7 ± 0.8 µm) UCYN-C cells into large (100–500 µm) aggregates. During the same time period, a DDN transfer experiment based on high-resolution nanometer-scale secondary ion mass spectrometry (nanoSIMS) coupled with 15N2 isotopic labeling revealed that 16 ± 6 % of the DDN was released to the dissolved pool and 21 ± 4 % was transferred to non-diazotrophic plankton, mainly picoplankton (18 ± 4 %) followed by diatoms (3 ± 2 %). This is consistent with the observed dramatic increase in picoplankton and diatom abundances, primary production, bacterial production, and standing stocks of POC, PON, and POP in the mesocosms during the second half of the experiment. These results offer insights into the fate of DDN during a bloom of UCYN-C in low-nutrient, low-chlorophyll ecosystems.

In the oligotrophic ocean characterized by nitrate (NO3−) depletion in surface waters, dinitrogen (N2) fixation and dissolved organic nitrogen (DON) can represent significant nitrogen (N) sources for the ecosystem. In this study, we deployed large in situ mesocosms in New Caledonia in order to investigate (1) the contribution of N2 fixation and DON use to primary production (PP) and particle export and (2) the fate of the freshly produced particulate organic N (PON), i.e., whether it is preferentially accumulated and recycled in the water column or exported out of the system. The mesocosms were fertilized with phosphate (PO43-) in order to prevent phosphorus (P) limitation and promote N2 fixation. The diazotrophic community was dominated by diatom–diazotroph associations (DDAs) during the first part of the experiment for 10 days (P1) followed by the unicellular N2-fixing cyanobacteria UCYN-C for the last 9 days (P2) of the experiment. N2 fixation rates averaged 9.8 ± 4.0 and 27.7 ± 8.6 nmol L−1 d−1 during P1 and P2, respectively. NO3− concentrations (< 0.04 μmol L−1) in the mesocosms were a negligible source of N, indicating that N2 fixation was the main driver of new production throughout the experiment. The contribution of N2 fixation to PP was not significantly different (p > 0.05) during P1 (9.0 ± 3.3 %) and P2 (12.6 ± 6.1 %). However, the e ratio that quantifies the efficiency of a system to export particulate organic carbon (POCexport) compared to PP (e ratio = POCexport/PP) was significantly higher (p < 0.05) during P2 (39.7 ± 24.9 %) than during P1 (23.9 ± 20.2 %), indicating that the production sustained by UCYN-C was more efficient at promoting C export than the production sustained by DDAs. During P1, PON was stable and the total amount of N provided by N2 fixation (0.10 ± 0.02 μmol L−1) was not significantly different (p > 0.05) from the total amount of PON exported (0.10 ± 0.04 μmol L−1), suggesting a rapid and probably direct export of the recently fixed N2 by the DDAs. During P2, both PON concentrations and PON export increased in the mesocosms by a factor 1.5–2. Unlike in P1, this PON production was not totally explained by the new N provided by N2 fixation. The use of DON, whose concentrations decreased significantly (p < 0.05) from 5.3 ± 0.5 μmol L−1 to 4.4 ± 0.5 μmol L−1, appeared to be the missing N source. The DON consumption (~ 0.9 μmol L−1) during P2 is higher than the total amount of new N brought by N2 fixation (~ 0.25 μmol L−1) during the same period. These results suggest that while DDAs mainly rely on N2 fixation for their N requirements, both N2 fixation and DON can be significant N sources for primary production and particulate export following UCYN-C blooms in the New Caledonia lagoon and by extension in the N-limited oceans where similar events are likely to occur.

The response of the phytoplanktonic community (primary production and algal biomass) to contrasted Saharan dust events (wet and dry deposition) was studied in the framework of the DUNE ("a DUst experiment in a low-Nutrient, low-chlorophyll Ecosystem") project. We simulated realistic dust deposition events (10 g m−2) into large mesocosms (52 m3). Three distinct dust addition experiments were conducted in June 2008 (DUNE-1-P: simulation of a wet deposition; DUNE-1-Q: simulation of a dry deposition) and 2010 (DUNE-2-R1 and DUNE-2-R2: simulation of two successive wet depositions) in the northwestern oligotrophic Mediterranean Sea. No changes in primary production (PP) and chlorophyll a concentrations (Chl a) were observed after a dry deposition event, while a wet deposition event resulted in a rapid (24 h after dust addition), strong (up to 2.4-fold) and long (at least a week in duration) increase in PP and Chl a. We show that, in addition to being a source of dissolved inorganic phosphorus (DIP), simulated wet deposition events were also a significant source of nitrate (NO3−) (net increases up to +9.8 μM NO3− at 0.1 m in depth) to the nutrient-depleted surface waters, due to cloud processes and mixing with anthropogenic species such as HNO3. The dry deposition event was shown to be a negligible source of NO3−. By transiently increasing DIP and NO3- concentrations in N–P starved surface waters, wet deposition of Saharan dust was able to relieve the potential N or NP co-limitation of the phytoplanktonic activity. Due to the higher input of NO3− relative to DIP, and taking into account the stimulation of the biological activity, a wet deposition event resulted in a strong increase in the NO3−/DIP ratio, from initially less than 6, to over 150 at the end of the DUNE-2-R1 experiment, suggesting a switch from an initial N or NP co-limitation towards a severe P limitation. We also show that the contribution of new production to PP strongly increased after wet dust deposition events, from initially 15% to 60–70% 24 h after seeding, indicating a switch from a regenerated-production based system to a new-production based system. DUNE experiments show that wet and dry dust deposition events induce contrasting responses of the phytoplanktonic community due to differences in the atmospheric supply of bioavailable new nutrients. Our results from original mesocosm experiments demonstrate that atmospheric dust wet deposition greatly influences primary productivity and algal biomass in LNLC environments through changes in the nutrient stocks, and alters the NO3−/DIP ratio, leading to a switch in the nutrient limitation of the phytoplanktonic activity.

The response of N2 (dinitrogen) fixation to contrasted (wet and dry) Saharan dust deposition was studied in the framework of the DUNE project (a DUst experiment in a low-Nutrient, low-chlorophyll Ecosystem) during which realistic simulations of dust deposition (10 g m−2) into large mesocosms (52 m3) were performed. Three distinct experimental dust additions were conducted in June 2008 (DUNE-1-P: simulation of a wet deposition, DUNE-1-Q: simulation of a dry deposition) and 2010 (DUNE-2-R: simulation of 2 successive wet depositions) in the northwestern oligotrophic Mediterranean Sea. Here we show that wet and dry dust deposition induced a rapid (24 h or 48 h after dust additions), strong (from 2- to 5.3-fold) and long (at least 4–6 days duration) increase in N2 fixation, indicating that both wet and dry Saharan dust deposition was able to relieve efficiently the nutrient limitation(s) of N2 fixation. This means in particular that N2 fixation activity was not inhibited by the significant input of nitrate associated with the simulated wet deposition (~ 9 mmol NO3− m−2). The input of new nitrogen associated with N2 fixation was negligible relative to the atmospheric NO3− input associated with the dust. The contribution of N2 fixation to primary production was negligible (≤ 1%) before and after dust addition in all experiments, indicating that N2 fixation was a poor contributor to the nitrogen demand for primary production. Despite the stimulation of N2 fixation by dust addition, the rates remained low, and did not significantly change the contribution of N2 fixation to new production since only a maximum contribution of 10% was observed. The response of N2 fixation by diazotrophs and CO2 fixation by the whole phytoplankton community suggests that these metabolic processes were limited or co-limited by different nutrients. With this novel approach, which allows us to study processes as a function of time while atmospheric particles are sinking, we show that new atmospheric nutrients associated with Saharan dust pulses do significantly stimulate N2 fixation in the Mediterranean Sea and that N2 fixation is not a key process in the carbon cycle in such oligotrophic environments.

A better understanding of the factors controlling N2 fixation is a pre-requisite for improving our knowledge on the contribution of N2 fixation process in the nitrogen cycling. Trace-metal clean nutrient/dust addition bioassays (+P, +PFe, +dust) were performed at three stations located in the western, central and eastern Mediterranean Sea, in summer 2008 as part of the BOUM cruise. The main goals were (1) to investigate the nutrient factor(s) limiting N2 fixation (uptake of 15N2) and (2) to evaluate the potential impact of a Saharan dust event on this biological process during the stratification period. Initially, surface waters at the three stations were DIP-depleted (2 fixation (from 130 % to 430 %). The highest dust stimulation of N2 fixation was recorded at the station located in the eastern basin. The response of diazotrophic activity to nutrient additions was variable between the sampled stations suggesting a spatial variability of the factor controlling N2 fixation over the whole basin. At all stations, N2 fixation was not limited by Fe nor co-limited by P and Fe. At the western station, N2 fixation was DIP limited while at the eastern one, N2 fixation was first DIP limited, then was limited by one or several chemical element(s) released by dust. Our results demonstrated that a Saharan dust input was able to relieve these successive on going limitations. Very interestingly, at the station located in the central basin, N2 fixation was not limited by the availability of P yet it was strongly stimulated by dust addition (x3.1). A chemical element or a combination of several, released by the added dust may have been responsible for the observed stimulations of N2 fixation. These results indicated that Saharan dust pulses to the surface Mediterranean waters, in addition to P and Fe, could be a source of chemical(s) element(s) that are necessary for metabolic processes and therefore influence rates of N2 fixation.

The Mediterranean Sea is a semi-enclosed basin characterized by a strong thermal stratification during summer during which the atmosphere is the main source of new nutrients to the nutrient-depleted surface layer. From aerosol sampling and microcosm experiments performed during the TransMed BOUM cruise (June–July 2008) we showed that: (i) the Mediterranean atmosphere composition (Al, Fe, P) was homogeneous over ~28° of longitude and was a mixture with a constant proportion of anthropogenic contribution and a variable but modest contribution of crustal aerosols. This quite stable composition over a one month period and a long transect (~2500 km) allowed to define the Mediterranean atmospheric "background" that characterizes the summer season in the absence of major Saharan event and forest fires, (ii) primary production significantly increased at all tested stations after aerosols addition collected on-board and after Saharan dust analog addition, indicating that both additions relieved on-going (co)-limitations. Although both additions significantly increased the N2 fixation rates at the western station, diazotrophic activity remained very low (~0.2 nmol N L−1 d−1), (iii) due to the presence of anthropogenic particles, the probable higher solubility of nutrients associated with mixed aerosols (crustal + anthropogenic contribution), conferred a higher fertilizing potential to on-board collected aerosol as compared to Saharan dust analog. Finally, those experiments showed that atmospheric inputs from a mixed atmospheric event ("summer rain" type) or from a high-intensity Saharan event would induce comparable response by the biota in the stratified Mediterranean SML, during summer.

Dinitrogen (N2) fixation represents a key source of reactive nitrogen in marine ecosystems. While the process has been rather well-explored in low latitudes of the Atlantic and Pacific Oceans, other higher latitude regions and particularly the Indian Ocean have been chronically overlooked. Here, we characterize N2 fixation and diazotroph community composition across nutrient and trace metals gradients spanning the multifrontal system separating the oligotrophic waters of the Indian Ocean subtropical gyre from the high nutrient low chlorophyll waters of the Southern Ocean. We found a sharp contrasting distribution of diazotroph groups across the frontal system. Notably, cyanobacterial diazotrophs dominated north of fronts, driving high N2 fixation rates (up to 13.96 nmol N l-1 d-1) with notable peaks near the South African coast. South of the fronts non-cyanobacterial diazotrophs prevailed without significant N2 fixation activity being detected. Our results provide new crucial insights into high latitude diazotrophy in the Indian Ocean, which should contribute to improved climate model parameterization and enhanced constraints on global net primary productivity projections., (© The Author(s) 2024. Published by Oxford University Press on behalf of FEMS.)

Bacteria play an important role in biogeochemical cycles as they transform and remineralize organic matter. Particles are notable hotspots of activity, hosting particle-attached (PA) communities that can differ largely from their free-living (FL) counterparts. However, long-standing questions remain concerning bacterial community assembly processes and driving factors. This study investigated the FL and PA community compositions and determinants within the Aulne estuary and the Bay of Brest coastal waters (France). Our results revealed that the FL and PA community compositions greatly varied with salinity and season, explaining a larger part of the variance than the sampling fraction. Both the FL and PA communities were driven by deterministic assembly processes and impacted by similar factors. The FL-PA dissimilarity varied across space and time. It decreased in the estuarine stations compared to the freshwater and marine ends, and in summer. Interestingly, a significant proportion of the FL and PA communities' β-diversity and dissimilarity was explained by cohesion, measuring the degree of taxa co-occurrence. This suggested the importance of co-occurrence patterns in shaping the FL and PA community compositionss. Our results shed light on the factors influencing estuarine bacterial communities and provide a first step toward understanding their biogeochemical impacts., (© 2022. The Author(s).)

Heterotrophic microbial communities play a central role in biogeochemical cycles in the ocean by degrading organic matter through the synthesis of extracellular hydrolytic enzymes. Their hydrolysis rates result from the community's genomic potential and the differential expression of this genomic potential. Cell-cell communication pathways such as quorum sensing (QS) could impact both aspects and, consequently, structure marine ecosystem functioning. However, the role of QS communications in complex natural assemblages remains largely unknown. In this study, we investigated whether N -acylhomoserine lactones (AHLs), a type of QS signal, could regulate both hydrolytic activities and the bacterial community composition (BCC) of marine planktonic assemblages. To this extent, we carried out two microcosm experiments, adding five different AHLs to bacterial communities sampled in coastal waters (during early and peak bloom) and monitoring their impact on enzymatic activities and diversity over 48 h. Several specific enzymatic activities were impacted during both experiments, as early as 6 h after the AHL amendments. The BCC was also significantly impacted by the treatments after 48 h, and correlated with the expression of the hydrolytic activities, suggesting that changes in hydrolytic intensities may drive changes in BCC. Overall, our results suggest that QS communication could participate in structuring both the function and diversity of marine bacterial communities., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Urvoy, Lami, Dreanno, Delmas, L’Helguen and Labry.)

Biofilms of heterotrophic bacteria cover organic matter aggregates and constitute hotspots of mineralization, primarily acting through extracellular hydrolytic enzyme production. Nevertheless, regulation of both biofilm and hydrolytic enzyme synthesis remains poorly investigated, especially in estuarine ecosystems. In this study, various bioassays, mass spectrometry and genomics approaches were combined to test the possible involvement of quorum sensing (QS) in these mechanisms. QS is a bacterial cell-cell communication system that relies notably on the emission of N-acylhomoserine lactones (AHLs). In our estuarine bacterial collection, we found that 28 strains (9%), mainly Vibrio, Pseudomonas and Acinetobacter isolates, produced at least 14 different types of AHLs encoded by various luxI genes. We then inhibited the AHL QS circuits of those 28 strains using a broad-spectrum lactonase preparation and tested whether biofilm production as well as β-glucosidase and leucine-aminopeptidase activities were impacted. Interestingly, we recorded contrasted responses, as biofilm production, dissolved and cell-bound β-glucosidase and leucine-aminopeptidase activities significantly increased in 4%-68% of strains but decreased in 0%-21% of strains. These findings highlight the key role of AHL-based QS in estuarine bacterial physiology and ultimately on biogeochemical cycles. They also point out the complexity of QS regulations within natural microbial assemblages., (© 2021 Society for Applied Microbiology and John Wiley & Sons Ltd.)

Phytoplankton is composed of a broad-sized spectrum of phylogenetically diverse microorganisms. Assessing CO 2 -fixation intra- and inter-group variability is crucial in understanding how the carbon pump functions, as each group of phytoplankton may be characterized by diverse efficiencies in carbon fixation and export to the deep ocean. We measured the CO 2 -fixation of different groups of phytoplankton at the single-cell level around the naturally iron-fertilized Kerguelen plateau (Southern Ocean), known for intense diatoms blooms suspected to enhance CO 2 sequestration. After the bloom, small cells (<20 µm) composed of phylogenetically distant taxa (prymnesiophytes, prasinophytes, and small diatoms) were growing faster (0.37 ± 0.13 and 0.22 ± 0.09 division d -1 on- and off-plateau, respectively) than larger diatoms (0.11 ± 0.14 and 0.09 ± 0.11 division d -1 on- and off-plateau, respectively), which showed heterogeneous growth and a large proportion of inactive cells (19 ± 13%). As a result, small phytoplankton contributed to a large proportion of the CO 2 fixation (41-70%). The analysis of pigment vertical distribution indicated that grazing may be an important pathway of small phytoplankton export. Overall, this study highlights the need to further explore the role of small cells in CO 2 -fixation and export in the Southern Ocean., (© 2021. The Author(s).)

Installation of an Ocean Thermal Energy Conversion pilot plant (OTEC) off the Caribbean coast of Martinique is expected to use approximately 100,000 m 3  h -1 of deep seawater for its functioning. This study examined the potential effects of the cold nutrient-rich deep seawater discharge on the phytoplankton community living in the surface warm oligotrophic waters before the installation of the pilot plant. Numerical simulations of deep seawater upwelled by the OTEC, showed that a 3.0 °C temperature change, considered as a critical threshold for temperature impact, was never reached during an annual cycle on the top 150 m of the water column on two considered sections centered on the OTEC. The thermal effect should be limited, <1 km 2 on the area exhibited a temperature difference of 0.3 °C (absolute value), producing a negligible thermic impact on the phytoplankton assemblage. The impact on phytoplankton of the resulting mixed deep and surface seawater was evaluated by in situ microcosm experiments. Two scenarios of water mix ratio (2% and 10% of deep water) were tested at two incubation depths (deep chlorophyll-a maximum: DCM and bottom of the euphotic layer: BEL). The larger impact was obtained at DCM for the highest deep seawater addition (10%), with a development of diatoms and haptophytes, whereas 2% addition induced only a limited change of the phytoplankton community (relatively higher Prochlorococcus sp. abundance, but without significant shift of the assemblage). This study suggested that the OTEC plant would significantly modify the phytoplankton assemblage with a shift from pico-phytoplankton toward micro-phytoplankton only in the case of a discharge affecting the DCM and would be restricted to a local scale. Since the lower impact on the phytoplankton assemblage was obtained at BEL, this depth can be recommended for the discharge of the deep seawater to exploit the OTEC plant., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Nitrogen (N) is a limiting nutrient in vast regions of the world's oceans, yet the sources of N available to various phytoplankton groups remain poorly understood. In this study, we investigated inorganic carbon (C) fixation rates and nitrate (NO 3 - ), ammonium (NH 4 + ) and urea uptake rates at the single cell level in photosynthetic pico-eukaryotes (PPE) and the cyanobacteria Prochlorococcus and Synechococcus. To that end, we used dual 15 N and 13 C-labeled incubation assays coupled to flow cytometry cell sorting and nanoSIMS analysis on samples collected in the North Pacific Subtropical Gyre (NPSG) and in the California Current System (CCS). Based on these analyses, we found that photosynthetic growth rates (based on C fixation) of PPE were higher in the CCS than in the NSPG, while the opposite was observed for Prochlorococcus. Reduced forms of N (NH 4 + and urea) accounted for the majority of N acquisition for all the groups studied. NO 3 - represented a reduced fraction of total N uptake in all groups but was higher in PPE (17.4 ± 11.2% on average) than in Prochlorococcus and Synechococcus (4.5 ± 6.5 and 2.9 ± 2.1% on average, respectively). This may in part explain the contrasting biogeography of these picoplankton groups. Moreover, single cell analyses reveal that cell-to-cell heterogeneity within picoplankton groups was significantly greater for NO 3 - uptake than for C fixation and NH 4 + uptake. We hypothesize that cellular heterogeneity in NO 3 - uptake within groups facilitates adaptation to the fluctuating availability of NO 3 - in the environment.

Iron (Fe) is widely suspected as a key controlling factor of N2 fixation due to the high Fe content of nitrogenase and photosynthetic enzymes complex, and to its low concentrations in oceanic surface seawaters. The influence of Fe limitation on the recently discovered unicellular diazotrophic cyanobacteria (UCYN) is poorly understood despite their biogeochemical importance in the carbon and nitrogen cycles. To address this knowledge gap, we conducted culture experiments on Crocosphaera watsonii WH8501 growing under a range of dissolved Fe concentrations (from 3.3 to 403 nM). Overall, severe Fe limitation led to significant decreases in growth rate (2.6-fold), C, N and chlorophyll a contents per cell (up to 4.1-fold), N2 and CO2 fixation rates per cell (17- and 7-fold) as well as biovolume (2.2-fold). We highlighted a two phased response depending on the degree of limitation: (i) under a moderate Fe limitation, the biovolume of C. watsonii was strongly reduced, allowing the cells to keep sufficient energy to maintain an optimal growth, volume-normalized contents and N2 and CO2 fixation rates; (ii) with increasing Fe deprivation, biovolume remained unchanged but the entire cell metabolism was affected, as shown by a strong decrease in the growth rate, volume-normalized contents and N2 and CO2 fixation rates. The half-saturation constant for growth of C. watsonii with respect to Fe is twice as low as that of the filamentous Trichodesmium indicating a better adaptation of C. watsonii to poor Fe environments than filamentous diazotrophs. The physiological response of C. watsonii to Fe limitation was different from that previously shown on the UCYN Cyanothece sp, suggesting potential differences in Fe requirements and/or Fe acquisition within the UCYN community. These results contribute to a better understanding of how Fe bioavailability can control the activity of UCYN and explain the biogeography of diverse N2 fixers in ocean.