Description: During two independent cruises in the north-eastern tropical Atlantic Ocean, we applied two different approaches to investigate the impact of diazotrophy on nitrogen stable isotope signatures in nitrate and particulate organic nitrogen (PON) of the food-web constituents. The first approach, used during the Poseidon cruise 348 in the Mauritanian upwelling, investigated the long-term influence of diazotrophy on the natural abundance of 15N-NO-3 and PON. The second approach, adopted during the Cape Verde field cruise, applied stable isotope tracer addition experiments. These served to determine the instantaneous transfer of diazotrophic N to the higher trophic level. Both approaches showed that N2 fixation was compatible with the pattern and the magnitude of the isotopic depletion of dissolved NO-3 during the Mauritanian upwelling cruise, as well as PON in zooplankton and phytoplankton during the Cape Verde cruises. An N-budget using 15N incorporation rates and diazotrophic N2 fixation rates showed that 6 % of the daily N2 fixation was potentially taken up by the mesozooplankton community. Direct grazing accounted for 56 % of gross mesozooplanktonic N incorporation, while 46 % occurred due to channelling through the microbial loop.

Description: During two simultaneous cruises in the Central Baltic Sea in July 2007 we applied a 15N tracer addition approach to assess the impact of cyanobacterial N2 fixation on mesozooplankton production in the Central Baltic Sea. We determined rates of diazotrophic 15N2 fixation, as well as uptake of diazotrophic derived 15N by mesozooplankton species. Diazotrophic 15N2 fixation rates were low representing pre-bloom situations. A first order estimate using a two source mixing model of natural δ15N-PON abundance revealed that diazotrophic fixed N contributed to 27 ± 8% to mesozooplankton biomass. Additionally, the application of stable isotope tracer showed that fixed 15N was detectable in the mesozooplankton fraction within 1 h after the onset of the incubation. On a daily basis, 5% up to 100% of newly fixed 15N and 14% of cyanobacteria standing stock were incorporated by mesozooplankton species in our experimental set-ups. By applying size fractionating experiments and the usage of different control treatments, we calculated that the majority of 15N transfer (67%) was mediated by the release of nitrogenous compounds and their channelling through the microbial loop towards the mesozooplankton community. Moreover, direct grazing on filamentous cyanobacteria accounted for 33% of gross 15N incorporation. Grazing in the experiments seemed to be largely influenced by cyanobacterial species dominating the community and by the abundance of Cladoceran species like Evadne. Overall, N2 fixing cyanobacteria are ecological more important as instantaneous sources of nitrogen for higher trophic levels of the Baltic Sea food web than previously assumed.

Description: Cyanobacteria blooms in the Baltic Sea appear after upwelling events, which transport phosphate-rich intermediate water to the surface. The growth potential of diazotrophic cyanobacteria in upwelled water was studied in a mesocosm (tank) experiment in summer 2007. An Anabaena bloom was only induced in the tanks filled with upwelled surface water but not in those filled with surface water from outside the upwelling cell and with intermediate water. The low initial cyanobacteria biomass in the intermediate water could not grow to bloom concentrations within three weeks. It is concluded that mixing of upwelled water with surrounding surface water forms a precondition for a cyanobacteria bloom. An additional mesocosm experiment conducted in 2009 revealed that mixing of intermediate water with surface water had the same stimulating effect on nitrogen fixation and cyanobacteria growth as artificial phosphate input. Phosphate input stimulates the growth of Nodularia and Anabaena more than that of Aphanizomenon. We suggest that the upwelled phosphate-rich intermediate water has to be mixed with the surface water containing physiologically “young” cyanobacteria biomass of at least 20 mg/m3 as an inoculum in order to initiate a cyanobacteria bloom.

Description: The filamentous and diazotrophic cyanobacterium Nodularia spumigena plays a major role in the productivity of the Baltic Sea as it forms extensive blooms regularly. Under phosphorus limiting conditions Nodularia spumigena has a high enzyme affinity for dissolved organic phosphorus (DOP) by production and release of alkaline phosphatase. Additionally, it is able to degrade proteinaceous compounds by expressing the extracellular enzyme leucine aminopeptidase. As atmospheric CO2 concentrations are increasing, we expect marine phytoplankton to experience changes in several environmental parameters including pH, temperature, and nutrient availability. The aim of this study was to investigate the combined effect of CO2-induced changes in seawater carbonate chemistry and of phosphate deficiency on the exudation of organic matter, and its subsequent recycling by extracellular enzymes in a Nodularia spumigena culture. Batch cultures of Nodularia spumigena were grown for 15 days aerated with three different pCO2 levels corresponding to values from glacial periods to future values projected for the year 2100. Extracellular enzyme activities as well as changes in organic and inorganic compound concentrations were monitored. CO2 treatment–related effects were identified for cyanobacterial growth, which in turn was influencing exudation and recycling of organic matter by extracellular enzymes. Biomass production was increased by 56.5% and 90.7% in the medium and high pCO2 treatment, respectively, compared to the low pCO2 treatment and simultaneously increasing exudation. During the growth phase significantly more mucinous substances accumulated in the high pCO2 treatment reaching 363 μg Gum Xanthan eq l−1 compared to 269 μg Gum Xanthan eq l−1 in the low pCO2 treatment. However, cell-specific rates did not change. After phosphate depletion, the acquisition of P from DOP by alkaline phosphatase was significantly enhanced. Alkaline phosphatase activities were increased by factor 1.64 and 2.25, respectively, in the medium and high compared to the low pCO2 treatment. In conclusion, our results suggest that Nodularia spumigena can grow faster under elevated pCO2 by enhancing the recycling of organic matter to acquire nutrients.

Description: Heterocystous cyanobacteria of the genus Nodularia form extensive blooms in the Baltic Sea and contribute substantially to the total annual primary production. Moreover,they dispense a large fraction of new nitrogen to the ecosystem when inorganic nitrogen concentration in summer is low. Thus, it is of ecological importance to know how Nodularia will react to future environmental changes, in particular to increasing carbon dioxide (CO2) concentrations and what consequences there might arise for cycling of organic matter in the Baltic Sea. Here, we determined carbon (C) and dinitrogen (N2) fixation rates, growth, elemental stoichiometry of particulate organic matter and nitrogen turnover in batch cultures of the heterocystous cyanobacterium Nodularia spumigena under low (median 315 μatm), mid (median 353 μatm), and high (median 548 μatm) CO2 concentrations. Our results demonstrate an overall stimulating effect of rising pCO2 on C and N2 fixation, as well as on cell growth. An increase in pCO2 during incubation days 0 to 9 resulted in an elevation in growth rate by 84±38% (low vs. high pCO2) and 40±25% (mid vs. high pCO2), as well as in N2 fixation by 93±35% and 38±1 %, respectively. C uptake rates showed high standard deviations within treatments and in between sampling days. Nevertheless, C fixation in the high pCO2 treatment was elevated compared to the other two treatments by 97% (high vs. low) and 44% (high vs. mid) at day 0 and day 3, but this effect diminished afterwards. Additionally, elevation in carbon to nitrogen and nitrogen to phosphorus ratios of the particulate biomass formed (POC : POP and PON: POP) was observed at high pCO2. Our findings suggest that rising pCO2 stimulates the growth of heterocystous diazotrophic cyanobacteria, in a similar way as reported for the non-heterocystous diazotroph Trichodesmium. Implications for biogeochemical cycling and food web dynamics, as well as ecological and socio-economical aspects in the Baltic Sea are discussed.

Description: Diazotrophic cyanobacteria often form extensive summer blooms in the Baltic Sea driving their environment into phosphate limitation. One of the main species is the heterocystous cyanobacterium Nodularia spumigena. N. spumigena exhibits accelerated uptake of phosphate through the release of the exoenzyme alkaline phosphatase that also serves as an indicator of the hydrolysis of dissolved organic phosphorus (DOP). The present study investigated the utilization of DOP and its compounds (e.g. ATP) by N. spumigena during growth under varying CO2 concentrations, in order to estimate potential consequences of ocean acidification on the cell's supply with phosphorus. Cell growth, phosphorus pool fractions, and four DOP-compounds (ATP, DNA, RNA, and phospholipids) were determined in three set-ups with different CO2 concentrations (341, 399, and 508 μatm) during a 15-day batch experiment. The results showed rapid depletion of dissolved inorganic phosphorus (DIP) in all pCO2 treatments while DOP utilization increased with elevated pCO2, in parallel with the growth stimulation of N. spumigena. During the growth phase, DOP uptake was enhanced by a factor of 1.32 at 399 μatm and of 2.25 at 508 μatm compared to the lowest pCO2 concentration. Among the measured DOP compounds, none was found to accumulate preferentially during the incubation or in response to a specific pCO2 treatment. However, at the beginning 61.9 ± 4.3% of the DOP were not characterized but comprised the most highly utilized fraction. This is demonstrated by the decrement of this fraction to 27.4 ± 9.9% of total DOP during the growth phase, especially in response to the medium and high pCO2 treatment. Our results indicate a stimulated growth of diazotrophic cyanobacteria at increasing CO2 concentrations that is accompanied by increasing utilization of DOP as an alternative P source.