Heterotrophic Prokaryotes Research Articles

Variability of ultraplankton composition and distribution in an oligotrophic coastal ecosystem of the NW Mediterranean Sea derived from a two-year survey at the single cell level.

Ultraplankton [heterotrophic prokaryotes and ultraphytoplankton (<10 μm)] were monitored weekly over two years (2009 & 2010) in a coastal area of the NW Mediterranean Sea. Six clusters were differentiated by flow cytometry on the basis of their optical properties, two heterotrophic prokaryote (HP) subgroups labelled LNA and HNA (low and high nucleic acid content respectively), Prochlorococcus, Synechococcus, autotrophic picoeukaryotes and nanoeukaryotes. HP represented an important component of the microbial assemblage over the survey with relatively small abundance variation through seasons. The carbon biomass ratio HP/ultraphytoplankton averaged 0.45, however this ratio exceeded 1 during spring. Ultraphytoplankton biomass made about 50% of the total autotrophic carbon estimates but this contribution increased up to 97% and 67% during the 2009 and 2010 spring periods respectively. Within ultraphytoplankton, nanoeukaryote represent the most important ultraphytoplankton group in terms of autotrophic carbon biomass (up to 70%). Picoeukaryote maximum abundance occurred in winter. Synechococcus was the most abundant population (maximum 1.2 x 10 5 cells cm-3) particularly in spring where it represented up to 54% of ultraphytoplankton carbon biomass. The warmer winter-spring temperatures and the lengthening of the stratification period created a favorable situation for the earlier appearance of Synechococcus and its persistence throughout summer, paralleling Prochlorococcus development. Prochlorococcus was dominant over summer and autumn with concentrations up to 1.0 × 10 5 cells cm-3. While the abundance of Synechococcus throughout survey was of the same order as that reported in western Mediterranean Sea, Prochlorococcus was more abundant and similar to the more typical oligotrophic and warm waters. The abundance variation of the ultraplankton components through the survey was relatable to variations in the hydrological and nutrient conditions.

Free-living chemoautotrophic and particle-attached heterotrophic prokaryotes dominate microbial assemblages along a pelagic redox gradient.

Using the anoxic Cariaco Basin as a natural laboratory, particle association of bacterial and archaeal taxa was assessed by iTag sequencing and qPCR gene assays of samples spanning an oxic-anoxic-euxinic gradient. A total of 10%-12% of all bacterial and archaeal cells were found in the particle-associated (PA) fraction, operationally defined as prokaryotes captured on 2.7µm membranes. Both redox condition and size fraction segregated bacterial taxa. Archaeal taxa varied according to redox conditions, but were similar between size fractions. Taxa putatively associated with chemoautotrophic sulfur oxidation and nitrification dominated the free-living (FL) fraction throughout the oxycline (<1-120µM O2 ) and upper anoxic layer. Bacteria in the oxycline's PA fraction included taxa known to be aerobic and anaerobic chemoorganotrophs. At shallow anoxic depths, PA taxa were primarily affiliated with anaerobic sulfate ( SO42-)-reducing lineages. PA fractions in the most sulfidic samples were dominated by taxa affiliated with CH4 oxidizing, fermenting and SO42- reducing lineages. Prevalence of particle-associated SO42--reducing taxa and abundant sulfur-oxidizing taxa in both size fractions across the oxic-anoxic interface is consistent with the cryptic sulfur cycling concept. Bacterial assemblage diversity in the PA fraction always exceeded the FL fraction except in the most oxic samples, whereas Archaeal diversity was not consistently different between size fractions. Our results suggest that these particle-associated and free-living bacterial assemblages are functionally different and that the interplay between particle microhabitats and surrounding geochemical regimes is a strong selective force shaping microbial communities throughout the water column.

Coastal Bacterioplankton Metabolism Is Stimulated Stronger by Anthropogenic Aerosols than Saharan Dust.

In oligotrophic regions, such as the Mediterranean Sea, atmospheric deposition has the potential to stimulate heterotrophic prokaryote growth and production in surface waters, especially during the summer stratification period. Previous studies focused on the role of leaching nutrients from mineral particles of Saharan (S) origin, and were restricted to single locations at given times of the year. In this study, we evaluate the effect of atmospheric particles from diverse sources and with a markedly different chemical composition [S dust and anthropogenic (A) aerosols] on marine planktonic communities from three locations of the northwestern Mediterranean with contrasted anthropogenic footprint. Experiments were also carried out at different times of the year, considering diverse initial conditions. We followed the dynamics of the heterotrophic community and a range of biogeochemical and physiological parameters in six experiments. While the effect of aerosols on bacterial abundance was overall low, bacterial heterotrophic production was up to 3.3 and 2.1 times higher in the samples amended with A and S aerosols, respectively, than in the controls. Extracellular enzymatic activities [leu-aminopeptidase (AMA) and β-glucosidase (β-Gl)] were also enhanced with aerosols, especially from A origin. AMA and β-Gl increased up to 7.1 in the samples amended with A aerosols, and up to 1.7 and 2.1 times, respectively, with S dust. The larger stimulation observed with A aerosols might be attributed to their higher content in nitrate. However, the response was variable depending the initial status of the seawater. In addition, we found that both A and S aerosols stimulated bacterial abundance and metabolism significantly more in the absence of competitors and predators.

Flamingos and drought as drivers of nutrients and microbial dynamics in a saline lake

Waterbird aggregations and droughts affect nutrient and microbial dynamics in wetlands. We analysed the effects of high densities of flamingos on nutrients and microbial dynamics in a saline lake during a wet and a dry hydrological year, and explored the effects of guano on prokaryotic growth. Concentrations of dissolved organic carbon, total phosphorus and total nitrogen in the surface waters were 2–3 fold higher during the drought and were correlated with salinity. Flamingos stimulated prokaryotic heterotrophic production and triggered cascading effects on prokaryotic abundance, viruses and dissolved nitrogen. This stimulus of heterotrophic prokaryotes was associated with soluble phosphorus inputs from guano, and also from sediments. In the experiments, the specific growth rate and the carrying capacity were almost twice as high after guano addition than in the control treatments, and were coupled with soluble phosphorus assimilation. Flamingo guano was also rich in nitrogen. Dissolved N in lake water lagged behind the abundance of flamingos, but the causes of this lag are unclear. This study demonstrates that intense droughts could lead to increases in total nutrients in wetlands; however, microbial activity is likely constrained by the availability of soluble phosphorus, which appears to be more dependent on the abundance of waterbirds.

Spatio-temporal distribution pattern of the picocyanobacterium Synechococcus in lakes of different trophic states: a comparison of flow cytometry and sequencing approaches

In this study, we aimed to investigate the spatio-temporal distribution patterns of the picocyanobacterium Synechococcus, an important contributor to primary production in many freshwater lakes. Our study sites were four lakes with different trophic states within the Osterseen Lake District in Southern Germany. Flow cytometry counts and next-generation sequencing were used from April to October 2015 to analyse the occurrence of Synechococcus and heterotrophic prokaryotes in relation to physical and chemical habitat properties. Synechococcus was identified as the main representative of the autotrophic picoplankton, but cell counts varied widely. The Synechococcus taxa identified by flow cytometry were confirmed by sequencing data, but the comparison of the flow cytometry counts and sequence data revealed discrepancies for cells in the exponential phase. The eutrophic Lake Schiffhuettensee was dominated by algae and had the highest abundance of heterotrophic prokaryotes. The presence of distinct operational taxonomic units of Synechococcus varied seasonally and was lake-specific, indicating local niche adaptation. Our study sheds light on the ecology of these important primary producers in freshwater systems. Furthermore, the discrepancy observed with the direct comparison of the widely used methods of next-generation sequencing and flow cytometry should serve as a caveat for future data analysis.

The Response of Heterotrophic Prokaryote and Viral Communities to Labile Organic Carbon Inputs Is Controlled by the Predator Food Chain Structure.

Factors controlling the community composition of marine heterotrophic prokaryotes include organic-C, mineral nutrients, predation, and viral lysis. Two mesocosm experiments, performed at an Arctic location and bottom-up manipulated with organic-C, had very different results in community composition for both prokaryotes and viruses. Previously, we showed how a simple mathematical model could reproduce food web level dynamics observed in these mesocosms, demonstrating strong top-down control through the predator chain from copepods via ciliates and heterotrophic nanoflagellates. Here, we use a steady-state analysis to connect ciliate biomass to bacterial carbon demand. This gives a coupling of top-down and bottom-up factors whereby low initial densities of ciliates are associated with mineral nutrient-limited heterotrophic prokaryotes that do not respond to external supply of labile organic-C. In contrast, high initial densities of ciliates give carbon-limited growth and high responsiveness to organic-C. The differences observed in ciliate abundance, and in prokaryote abundance and community composition in the two experiments were in accordance with these predictions. Responsiveness in the viral community followed a pattern similar to that of prokaryotes. Our study provides a unique link between the structure of the predator chain in the microbial food web and viral abundance and diversity.

Fronts at the Surface Ocean Can Shape Distinct Regions of Microbial Activity and Community Assemblages Down to the Bathypelagic Zone: The Azores Front as a Case Study

Oceanic fronts are widespread features which separate distinct water masses. They are well known to control the distribution of microbial communities in surface waters, although there is scarce information on their role in delimiting critical functions that microbes perform, and on whether their effects can be translated down into the dark ocean. Here we carried out the first study on the variability of hydrolysis of organic matter (extracellular enzymatic activities; EEA) across a permanent front (the Azores Front), coupled with changes in microbial assemblage composition, from the surface down to the bathypelagic zone. The front separated the study area (enclosed into the North Atlantic Subtropical Gyral Province) into two distinct latitudinal sub-regions with sharp differences in the abundance of autotrophic and heterotrophic microbial assemblages, as well as in the extracellular enzymes activities of glucosidases, alkaline phosphatase and leucine aminopeptidase. South of the front there was an abrupt decline in the abundance of picophytoplankton as well as in heterotrophic prokaryotes with high nucleic-acid content, but an increase in the abundance of prokaryotes with high side-scatter, an indication that cells were growing attached to particles. Concomitantly, there was also an increase in the aminopeptidase to glucosidase ratio, a proxy of higher degradation of proteinaceous material relative to carbohydrates. Interestingly, these sharp changes in microbial assemblages and enzymatic activities north and south of the front were translated down to the deep ocean. Our results suggest that permanent fronts, like the Azores Front, can act as ecological boundaries in the ocean (even within a biogeographical province), in terms of microbial community structure and biogeochemical cycling. Oriented studies on oceanic fronts down to the deep ocean will help to understand how the variability of these widely-extended hydrographic futures will impact microbial communities and carbon cycling in a future ocean affected by trends in global warming, de-oxygenation and acidification.

High dark carbon fixation in the tropical South China Sea

Dark carbon fixation (DCF) has been usually assumed to be insignificant in the study of microbial production and carbon cycling. In order to evaluate DCF distribution and its contribution to biogenic carbon flux, surface and vertical distribution of DCF, primary production (PP) and bacterial production (BP) were investigated in both offshore and coastal waters in the South China Sea (SCS). Surface DCF was ~ 0.058μgCL−1h−1 (~ 10% of PP), within the same order of magnitude as BP of ~ 0.047μgCL−1h−1 in the offshore waters. Integrated over the 1500m water column, DCF was ~ 196mgCm−2h−1, corresponding to ~ 384% of PP, and represented a newly produced source of organic matter. This suggested that DCF was an important microbial metabolic pathway in the SCS, which might support ~ 83% carbon demand of heterotrophic prokaryotes. Interestingly, the DCF was higher in the deep water (~ 0.140μgCL−1h−1) than the surface water (~ 0.089μgCL−1h−1). In addition to the different microbial community, this different vertical distribution of DCF was likely due to the nutrient status, as our nutrient enrichment experiment showed that the addition of glucose, ammonium and phosphate stimulated the DCF rates, especially the addition of glucose plus ammonium.

Temperature regulation of marine heterotrophic prokaryotes increases latitudinally as a breach between bottom-up and top-down controls.

Planktonic heterotrophic prokaryotes make up the largest living biomass and process most organic matter in the ocean. Determining when and where the biomass and activity of heterotrophic prokaryotes are controlled by resource availability (bottom-up), predation and viral lysis (top-down) or temperature will help in future carbon cycling predictions. We conducted an extensive survey across subtropical and tropical waters of the Atlantic, Indian and Pacific Oceans during the Malaspina 2010 Global Circumnavigation Expedition and assessed indices for these three types of controls at 109 stations (mostly from the surface to 4,000m depth). Temperature control was approached by the apparent activation energy in eV (ranging from 0.46 to 3.41), bottom-up control by the slope of the log-log relationship between biomass and production rate (ranging from -0.12 to 1.09) and top-down control by an index that considers the relative abundances of heterotrophic nanoflagellates and viruses (ranging from 0.82 to 4.83). We conclude that temperature becomes dominant (i.e. activation energy >1.5eV) within a narrow window of intermediate values of bottom-up (0.3-0.6) and top-down 0.8-1.2) controls. A pervasive latitudinal pattern of decreasing temperature regulation towards the Equator, regardless of the oceanic basin, suggests that the impact of global warming on marine microbes and their biogeochemical function will be more intense at higher latitudes. Our analysis predicts that 1°C ocean warming will result in increased biomass of heterotrophic prokaryoplankton only in waters with <26°C of mean annual surface temperature.

Assessing ultraphytoplankton and heterotrophic prokaryote composition by flow cytometry in a Mediterranean lagoon

In the eutrophic Ghar El Melh Lagoon (GML, Tunisia), the distribution of heterotrophic prokaryotes, pico- and nanophytoplankton was studied at five stations in November 2012 at the single cell level, along with environmental factors. Flow cytometry analysis of ultraplankton (<10μm) resolved (i) two heterotrophic prokaryote groups, low and high nucleic acid contents (LNA and HNA, respectively), and (ii) eight to nine ultraphytoplankton groups (cryptophyte-like cells, two nanoeukaryote subgroups, two picoeukaryote subgroups and three Synechococcus-like cells subgroups). Prochlorococcus was not detected. According to redundancy analysis (RDA), a significant difference was found in the distribution of the ultraplankton between stations (F=2.61, p<0.05); maximum proliferations of heterotrophic prokaryotes were observed in the inner parts of the lagoon at stations 3, 4 and 5 affected by urban, agricultural and industrial discharges. Ultraphytoplankton concentrations were the highest near the outlet of the lagoon at stations 1 and 2 influenced by freshwater outflow and oligotrophic Mediterranean water inflow, respectively. At station 1, the large ultraphytoplankton concentration derives from the high abundance of cryptophyte-like cells favoured by the freshwater outflow whereas at station 2, the input of oligotrophic Mediterranean water enhanced the abundance of Synechococcus and picoeukaryotes at the expense of nanoeukaryotes. Two trophic regimes were thus differentiated in GML.

Corrigendum: Depth Dependent Relationships between Temperature and Ocean Heterotrophic Prokaryotic Production

XA, XM, and JG were funded by the Malaspina expedition 2010 (grant no. CSD2008-00077) and HOTMIX (grant no. CTM2011-30010-C02-02) projects, the latter being co-financed with FEDER funds.

Fungi associated with chimney and sulfide samples from a South Mid-Atlantic Ridge hydrothermal site: Distribution, diversity and abundance

Deep-sea hydrothermal vent ecosystems are known to support remarkably diverse microbial communities, ranging from chemoautotrophic prokaryotes to heterotrophic prokaryotes and microeukaryotes. While fungi have generally been identified as an important component of various microbial communities in the environment, little is known about the species richness and abundance of such microorganisms in deep-sea hydrothermal vent ecosystems. In this study, a combined culture-dependent and culture-independent sequence-based approach was used to investigate fungal distribution and diversity at a deep-sea hydrothermal vent site located at the Mid-Atlantic Ridge of the South Atlantic Ocean. Sequence analyses showed that the fungal community was dominated by members of the Ascomycota and the Basidiomycota. Several new phylotypes (28 of 65 total fungal OTUs and 2 of 19 culturable fungal phylotypes) were identified, contributing to the literally unknown diversity of fungi in this understudied habitat. The fungal community structures in the chimney samples were distinct from those in three sulfide samples. The qPCR results revealed that fungal LSU rRNA gene copy numbers ranged from 5.88×105 to 6.77×106 copies/gram rock (wet weight), and the Ascomycota was significantly more abundant 2–3 orders) than the Basidiomycota. Our findings provide new insights into the diversity and abundance of fungi in deep-sea hydrothermal vent ecosystems, which increases our knowledge and understanding of the fungal diversity in deep-sea environments.

Comparison of community structures between particle-associated and free-living prokaryotes in tropical and subtropical Pacific Ocean surface waters

The subtropical and tropical regions of the Pacific Ocean are less productive than other oceanic regions. Although particle association should be an important strategy for heterotrophic prokaryotes to survive in such environments, we have little information on particle-associated (PA) prokaryotes in these regions. The specific aim of this study was to determine bacterial and archaeal community structures in the PA assemblage in comparison to the free-living (FL) assemblage in the North Pacific Subtropical Gyre, the South Pacific Subtropical Gyre, and an eastern equatorial region of the Pacific Ocean. Community profiles and phylogenetic identities were obtained by denaturing gradient gel electrophoresis, 454-pyrosequencing, and cloning followed by Sanger sequencing of 16Sr RNA gene amplicons. The distribution patterns of some abundant groups in three regions and two lifestyles (PA and FL) are shown in this study. Also, the PA community structures of bacteria differed from the FL ones and exhibited higher diversity than the FL ones, while the archaeal community structures did not show significant differences between PA and FL assemblages. We found that specific phylotypes of Gammaproteobacteria and Flavobacteria were abundant in PA bacterial assemblages, suggesting that they prefer to attach and consume particulate organic matter. In summary, the surface seawater PA assemblages represent very different bacterial and archaeal community structures between three different oceanic regions, each of which had distinct PA and FL community structures. These results imply that environmental factors determine microbial community structures.

Ecoenzyme activity ratios reveal interactive effects of nutrient inputs and UVR in a Mediterranean high-mountain lake

To understand how atmospheric dust deposition and ultraviolet radiation (UVR) can affect remote, freshwater ecosystems through changes in their microbial metabolism, it is important to have tools that allow us detecting alterations and anticipating potential shifts in the functioning of microbial communities. Ecoenzyme activities (EA) are easy to measure and their ratios can be used to assess system microbial metabolism of freshwater bodies, thus evaluating the effects of global change stressors. We carried out an in situ full factorial experiment to determine how the interaction between the addition of C and P, and UVR affect the microbial metabolism of a Mediterranean high-mountain lake. Overall, activities of five ecoenzymes involved in the degradation of C-compounds and in the acquisition of N and P revealed that, under natural conditions, the growth of heterotrophic prokaryotes was dependent on organic compounds released by algae, which is consistent with a higher constraint of bacterial carbon production by C than by P or N, as suggested by EA ratios. Accordingly, the addition of a labile C source did not lead to any significant response of microbial communities, but the addition of P provoked a clear change in the microbial metabolism of the lake, promoting the growth of phytoplankton and leading heterotrophic prokaryotes to be more constrained by P, and to a lesser extent by N, in relation to C. UVR played a secondary role, probably because microbial communities inhabiting high-mountain lakes possess several evolutionary adaptations to high UVR levels. Changes in the microbial metabolism of our model lake under different scenarios of nutrient inputs and UVR can therefore be evaluated by EA ratios.

Distributions and relationships of virio- and picoplankton in the epi-, meso- and bathypelagic zones of the Western Pacific Ocean.

Virio- and picoplankton mediate important biogeochemical processes and the environmental factors that regulate their dynamics, and the virus-host interactions are incompletely known, especially in the deep sea. Here we report on their distributions and relationships with environmental factors at 21 stations covering a latitudinal range (2-23° N) in the Western Pacific Ocean. This region is characterized by a complex western boundary current system. Synechococcus, autotrophic picoeukaryotes, heterotrophic prokaryotes and virus-like particles (VLPs) were high (<2.4 × 102-6.3 × 104, <34-2.8 × 103, 3.9 × 104-1.3 × 106 cells mL-1 and 5.1 × 105-2.7 × 107 mL-1, respectively), and Prochlorococcus were low (<2.3 × 102-1.0 × 105 cells mL-1) in the Luzon Strait and the four most southerly stations, where upwelling occurs. Covariations in the abundances of VLPs with heterotrophic and autotrophic picoplankton, and their correlation (i.e. r2 = 0.63 and 0.52, respectively) suggested a strong host dependence in the epi- and mesopelagic zones. In the bathypelagic zone, only abiotic factors significantly influenced VLPs abundance variation (r2 = 0.12). This study shows that the dynamics of virio- and picoplankton in this Western Pacific are controlled by suite of complex and depth-dependent relationship among physical and biological factors that in turn link the physical hydrography of the western boundary current system with microbial-mediated biogeochemical processes.

Glycerol metabolism in hypersaline environments.

Glycerol is a key compound for the understanding of the microbiology of hypersaline environments. At the highest salt concentrations the main or even sole primary producer is the green unicellular alga Dunaliella, which uses photosynthetically produced glycerol as osmotic stabilizer and compatible solute. Glycerol can be expected to be a major carbon source available to the heterotrophic communities of Archaea and Bacteria in hypersaline ecosystems. Use of Dunaliella has even been explored for the commercial production of glycerol. This article reviews our current understanding of glycerol metabolism in Dunaliella and of the ways glycerol can be degraded by heterotrophic prokaryote communities under aerobic and under anaerobic conditions. Dunaliella-derived glycerol may also be the key toward long-term survival of heterotrophic prokaryotes in fluid inclusions within salt crystals.

Increasing CO2 changes community composition of pico- and nano-sized protists and prokaryotes at a coastal Antarctic site

Ocean acidification is a globally recognised phenomenon, but little is known of its impacts on Antarctic marine microbes. Here we report on the community response of pico- and nanophytoplankton, heterotrophic nanoflagellates (HNF) and prokaryotes ( Archaea and Bacteria ) to elevated CO 2 during 3 minicosm experiments over the 2008/2009 summer at Davis Station, Antarctica. Coastal seawater was incubated in 650 l minicosms ( n = 6) for ≤ 12 d at CO 2 concentrations ranging from preindustrial levels to those predicted for 2100 and beyond. The abundance of pico- and nano-sized protists and prokaryotes were determined by flow cytometry, using chlorophyll autofluorescence to discriminate the phytoplankton, SYBR-Green to stain the prokaryotes and LysoTracker Green stain to discriminate the HNF. While the effects on nanophytoplankton abundance were inconclusive, our results show that increasing CO 2 can alter the composition of the microbial community in Antarctic coastal waters. Our 3 experiments consistently showed lower concentrations of HNF and higher abundances of picophytoplankton and prokaryotes in treatments exposed to elevated CO 2 . While the mechanism remains to be confirmed, our study suggests that CO 2 may reduce the mortality of picoplankton by HNF grazing. Our results indicate that changes in the composition of Antarctic microbial communities may occur within the concentration range of 750 to 1118 ppm CO 2 , potentially impacting the Antarctic food web through reduced food availability.

Depth Dependent Relationships between Temperature and Ocean Heterotrophic Prokaryotic Production

Marine prokaryotes play a key role in cycling of organic matter and nutrients in the ocean. Using a unique dataset (>14,500 samples), we applied a space-for-time substitution analysis to assess the temperature dependence of prokaryotic heterotrophic production (PHP) in epi- (0–200 m), meso- (201–1,000m) and bathypelagic waters (1,001–4,000 m) of the global ocean. Here, we show that the temperature dependence of PHP is fundamentally different between these major oceanic depth layers, with an estimated ecosystem-level activation energy (Ea) of 36 ± 7 kJ mol–1 for the epipelagic, 72 ± 15 kJ mol–1 for the mesopelagic and 274 ± 65 kJ mol–1 for the bathypelagic realm. We suggest that the increasing temperature dependence with depth is related to the parallel vertical gradient in the proportion of recalcitrant organic compounds. These Ea predict an increased PHP of about 5, 12 and 55% in the epi-, meso- and bathypelagic ocean, respectively, in response to a water temperature increase by 1oC. Hence, there is indication that a major thus far underestimated feedback mechanism exists between future bathypelagic ocean warming and heterotrophic prokaryotic activity.

Heterotrophic bacterial production and metabolic balance during the VAHINE mesocosm experiment in the New Caledonia lagoon

Abstract. Studies investigating the fate of diazotrophs through the microbial food web are lacking, although N2 fixation can fuel up to 50 % of new production in some oligotrophic oceans. In particular, the role played by heterotrophic prokaryotes in this transfer is largely unknown. In the frame of the VAHINE (VAriability of vertical and tropHIc transfer of diazotroph derived N in the south wEst Pacific) experiment, three replicate large-volume (∼ 50 m3) mesocosms were deployed for 23 days in the new Caledonia lagoon and were intentionally fertilized on day 4 with dissolved inorganic phosphorus (DIP) to stimulate N2 fixation. We specifically examined relationships between heterotrophic bacterial production (BP) and N2 fixation or primary production, determined bacterial growth efficiency and established carbon budgets. BP was statistically higher during the second phase of the experiment (P2: days 15–23), when chlorophyll biomass started to increase compared to the first phase (P1: days 5–14). Phosphatase alkaline activity increased drastically during the second phase of the experiment, showing adaptations of microbial populations after utilization of the added DIP. Notably, among autotrophs, Synechococcus abundances increased during P2, possibly related to its capacity to assimilate leucine and to produce alkaline phosphatase. Bacterial growth efficiency based on the carbon budget (27–43 %), was notably higher than generally cited for oligotrophic environments and discussed in links with the presence of abundant species of bacteria expressing proteorhodopsin. The main fates of gross primary production (particulate + dissolved) were respiration (67 %) and export through sedimentation (17 %). BP was highly correlated with particulate primary production and chlorophyll biomass during both phases of the experiment but was slightly correlated, and only during P2 phase, with N2 fixation rates. Heterotrophic bacterial production was strongly stimulated after mineral N enrichment experiments, suggesting N-limitation of heterotrophic bacteria across the experiment. N2 fixation rates corresponded to 17–37 % of the nitrogen demand of heterotrophic bacteria. Our results suggest that most of the diazotroph-derived nitrogen fuelled the heterotrophic bacterial community through indirect processes generating dissolved organic matter and detritus, like mortality, lysis and grazing of both diazotrophs and non-diazotrophs.

Draft Genome Sequence of Limnobacter sp. Strain CACIAM 66H1, a Heterotrophic Bacterium Associated with Cyanobacteria.

Ecological interactions between cyanobacteria and heterotrophic prokaryotes are poorly known. To improve the genomic studies of heterotrophic bacterium-cyanobacterium associations, the draft genome sequence (3.2 Mbp) of Limnobacter sp. strain CACIAM 66H1, found in a nonaxenic culture of Synechococcus sp. (cyanobacteria), is presented here.