Nitrogen (N) fixation is fueling planktonic production in a multitude of aquatic environments. In meso-and poly-haline estuaries, however, the contribution of N by pelagic N 2 fixation is believed to be insignificant due to the high input of N from land and the presumed absence of active N 2 -fixing organisms. Here we report N 2 fixation rates, nifH gene composition and nifH gene transcript abundance for key diazotrophic groups over 1 year in two contrasting, temperate, estuarine systems: Roskilde Fjord (RF) and the Great Belt (GB) strait. Annual pelagic N 2 fixation rates averaged 17 and 61 mmol N m À 2 per year at the two sites, respectively. In RF, N 2 fixation was mainly accompanied by transcripts related to heterotrophic (for example, Pseudomonas sp.) and photoheterotrophic bacteria (for example, unicellular diazotrophic cyanobacteria group A). In the GB, the first of two N 2 fixation peaks coincided with a similar nifH-expressing community as in RF, whereas the second peak was synchronous with increased nifH expression by an array of diazotrophs, including heterotrophic organisms as well as the heterocystous cyanobacterium Anabaena. Thus, we show for the first time that significant planktonic N 2 fixation takes place in mesohaline, temperate estuaries and that the importance of heterotrophic, photoheterotrophic and photosynthetic diazotrophs is clearly variable in space and time.

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Bacterioplankton play a key role in marine waters facilitating processes important for carbon cycling. However, the influence of specific bacterial populations and environmental conditions on bacterioplankton community performance remains unclear. The aim of the present study was to identify drivers of bacterioplankton community functions, taking into account the variability in community composition and environmental conditions over seasons, in two contrasting coastal systems. A Least Absolute Shrinkage and Selection Operator (LASSO) analysis of the biological and chemical data obtained from surface waters over a full year indicated that specific bacterial populations were linked to measured functions. Namely, Synechococcus (Cyanobacteria) was strongly correlated with protease activity. Both function and community composition showed seasonal variation. However, the pattern of substrate utilization capacity could not be directly linked to the community dynamics. The overall importance of dissolved organic matter (DOM) parameters in the LASSO models indicate that bacterioplankton respond to the present substrate landscape, with a particular importance of nitrogenous DOM. The identification of common drivers of bacterioplankton community functions in two different systems indicates that the drivers may be of broader relevance in coastal temperate waters.

We used continuous measurements of dissolved oxygen (DO) in dark bottles to characterise patterns of the dark respiration rate (Rdark) for three marine phytoplankton monocultures and in natural-water samples from two marine coastal systems. Furthermore, patterns of ecosystem community respiration rate were determined from open-water changes in DO in a fjord and in a lake. We considered two models of Rdark to describe temporal changes in DO: constant Rdark and decreasing Rdark; increasing Rdark. In addition, the effect of incubation time on Rdark was investigated in bottle incubations. Constant Rdark was observed in short-term (12-h) bottle incubations in natural-water samples from two marine coastal systems. Declining Rdark was observed in marine phytoplankton cultures and open-water measurements in a lake. Increasing Rdark was observed in open-water measurements in a fjord, particularly during summer. Long-term (120-h) bottle incubations in natural-water samples showed an increase in Rdark after 48 and 72h. We show that the conventional expectation of constant rates of respiration in darkness is far from typical, because non-linear changes are common under both controlled experimental conditions, as well as for open-water measurements of ecosystem respiration.

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