Microbial nitrogen (N2) fixation supplies important nitrogen inputs to boreal peatlands, extremely oligotrophic ecosystems dominated by Sphagnum mosses. In this study, we coupled major and trace nutrient analyses and rate measurements to characterize interactions between N2 fixation and CH4 cycling at the S1 peat bog in Marcell Experimental Forest and the Zim bog (Minnesota, USA). Total dissolved inorganic nitrogen (NO3-+NO2-+NH4+) and phosphate were both consistently < 2 μM in the porewater of surface peat, indicating severe nutrient limitation. While dissolved Fe was fairly abundant (18-35 mM), Mo, V and Cu were scarce (2-40 nM), suggesting that alternative metalloenzymes containing Fe in place of other metals may be favored. Rates of diazotrophy measured by both 15N2 incorporation and the acetylene (C2H2) reduction assay (ARA) were 7-fold higher under anoxic vs. oxic incubations conducted at both 4°C and 25°C. No significant difference in N2 fixation rates measured by either method was observed with or without the amendment of 1% CH4 at 25 °C; however, a significant inhibitory effect by methane was seen at 4°C in material from the S1 bog hollows. Anoxic 15N2 incorporation was 3-4x higher in treatments lacking acetylene, suggesting that the ARA likely underestimates N2 fixation by inhibiting diazotrophs sensitive to C2H2. Aerobic methanotrophy was also inhibited by 1% C2H2 when incubated under oxic conditions. No observations for the production of ethane (C2H6) were detected during the ARA, a biomarker for alternative nitrogenase activity. Major differences in ARA rates were observed to vary locally within microhabitats and between two bogs. In June 2014, peat sampled from hollows incubated under anoxic conditions showed the highest ARA rates (94.9 ± 11.0 nmol C2H4 g-1 moss dry mass hr-1), while the lowest rates were observed in ix hummock samples incubated under oxic conditions (5.1 ± 0.8 nmol C2H4 g-1 moss dry mass hr-1) in the S1 bog (T3 site). Observed rates have the potential to be a function of oxygen concentrations and or water content. ARA rates in all microcosm treatments were significantly lower at Zim bog compared to the S1 bog. The developed conversion factor between the regression of 15N2 and ARA in this study was 3.9 and agrees with the theoretical conversion factor as well as previous studies of soils and forest mosses.

The overall aim of the study was to provide better insights to the ecological role and impact of cyanobacteria in Baltic Sea (BS) bay, coastal and open sea areas. Biomass and heterocyst development of diazotrophic, heterocystous cyanobacteria were monitored over several years simultaneously as physical parameters such as nutrients and temperature. Nitrogen fixation was estimated as well as its transfer in the BS food web. Even after decades of debate there is still controversy whether eutrophication of lakes and estuaries/coastal areas should be managed by reducing phosphorus only or also nitrogen. Central to this debate is whether nitrogen fixation by cyanobacteria can replace shortages of combined nitrogen quickly enough to make phosphorus the limiting nutrient and nitrogen removal pointless or even harmful. Also, it is not clear if available combined nitrogen inhibits heterocystous cyanobacterial nitrogen fixation and if it is used for their growth in situ. A large ecosystem-wide experiment started in Himmerfjärden bay in year 1997, where the N-loadings and release depth from a modern sewage treatment plant (STP), located in the inner part of the bay, were modulated. The STP creates a steep gradient of nutrients and stable nitrogen isotopes, which can be used to study uptake of combined nitrogen, as well as biomass development and primary productivity. A 35-year long data series was used to achieve good insights into phytoplankton development and primary productivity in the Baltic Sea over the last couple of decades. These in vivo long time series, based on monitoring data, in combination with shorter series (2-3 seasons, including measurements of colony stoichiometry and stable isotopes), have resulted in a unique meta-dataset, allowing for high-resolution observations into the role of the cyanobacteria in the Baltic Sea ecosystem. / <p>At the time of the doctoral defense, the following papers were unpublished and had a status as follows: Paper 3: Manuscript. Paper 4: Manuscript.</p>

<p>Discoveries in recent years have disclosed the importance of marine cyano-bacteria in the context of primary production and global nitrogen cycling. It is hypothesized here that microbial mats in tropical coastal habitats harbour a rich diversity of previously uncharacterized cyanobacteria and that benthic marine nitrogen fixation in coastal zones is substantial.</p><p>A polyphasic approach was used to investigate cyanobacterial diversity in three tropical benthic marine habitats of different characters; an intertidal sand flat and a mangrove forest floor in the Indian Ocean, and a beach rock in the Pacific Ocean. In addition, nitrogenase activity was measured over diel cycles at all sites. The results revealed high cyanobacterial diversity, both morphologically and genetically. Substantial nitrogenase activity was observed, with highest rates at daytime where heterocystous species were present. However, the three habitats were dominated by non-heterocystous and unicellular genera such as <i>Microcoleus</i>, <i>Lyngbya</i>, <i>Cyanothece</i> and a large group of thin filamentous species, identified as members of the Pseudanabaenaceae family. In these consortia nocturnal nitrogenase activities were highest and <i>nifH</i> sequencing also revealed presence of non-cyanobacterial potential diazotrophs. A conclusive phylogenetic analysis of partial nifH sequences from the three sites and sequences from geographi-cally distant microbial mats revealed new clusters of benthic potentially ni-trogen-fixing cyanobacteria. Further, the non-heterocystous cyanobacterium <i>Lyngbya majuscula</i> was subjected to a physiological characterization to gain insights into regulatory aspects of its nitrogen fixation. The data demon-strated that nitrogenase activity is restricted to darkness, which called upon a re-evaluation of its diazotrophic behaviour.</p>

Discoveries in recent years have disclosed the importance of marine cyano-bacteria in the context of primary production and global nitrogen cycling. It is hypothesized here that microbial mats in tropical coastal habitats harbour a rich diversity of previously uncharacterized cyanobacteria and that benthic marine nitrogen fixation in coastal zones is substantial. A polyphasic approach was used to investigate cyanobacterial diversity in three tropical benthic marine habitats of different characters; an intertidal sand flat and a mangrove forest floor in the Indian Ocean, and a beach rock in the Pacific Ocean. In addition, nitrogenase activity was measured over diel cycles at all sites. The results revealed high cyanobacterial diversity, both morphologically and genetically. Substantial nitrogenase activity was observed, with highest rates at daytime where heterocystous species were present. However, the three habitats were dominated by non-heterocystous and unicellular genera such as Microcoleus, Lyngbya, Cyanothece and a large group of thin filamentous species, identified as members of the Pseudanabaenaceae family. In these consortia nocturnal nitrogenase activities were highest and nifH sequencing also revealed presence of non-cyanobacterial potential diazotrophs. A conclusive phylogenetic analysis of partial nifH sequences from the three sites and sequences from geographi-cally distant microbial mats revealed new clusters of benthic potentially ni-trogen-fixing cyanobacteria. Further, the non-heterocystous cyanobacterium Lyngbya majuscula was subjected to a physiological characterization to gain insights into regulatory aspects of its nitrogen fixation. The data demon-strated that nitrogenase activity is restricted to darkness, which called upon a re-evaluation of its diazotrophic behaviour.

Nitrogen fixation is no longer considered to be a minor factor of the nitrogen cycle in oceanic ecosystems. Recent geochemical and biological efforts have led to a significant increase in the estimated input of nitrogen to marine ecosystems by biological fixation, while molecular studies have increased our knowledge of the number and diversity of nitrogen fixers known to be active in the ocean. Although Trichodesmium spp. have long been viewed as the primary marine nitrogen fixers, recent efforts have shown that various members of the picoplankton community are also actively involved in nitrogen fixation. The relative abundance of different nitrogen fixers is an important ecosystem parameter since nitrogen fixers may differ significantly in their physiology, life history and ecology. Here we combine rate measurements and stable isotope natural abundance measurements to constrain the impact of N2 fixation in the waters north of Australia. Samples were collected in the Coral, Arafura, and East Timor Seas, thus spanning three distinct hydrographic regions. Our data show that Trichodesmium has a significant influence on the stable nitrogen isotope ratios of particulate and zooplankton biomass and suggest that Trichodesmium is a significant source of nitrogen for the pelagic ecosystem. Based on stable carbon isotope ratios, it is also likely that the pathways are indirect and nitrogen fixed by Trichodesmium enters the higher trophic levels via decomposition as dissolved organic and inorganic nitrogen. Picocyanobacteria showed high diazotrophic activity at some stations, but unlike Trichodesmium, their N2 fixation rate was not reflected in the stable N isotope ratios of particulate and zooplankton biomass. Our results suggest an important N contribution to biomass by diazotrophs in the Coral Sea, Arafura Sea and East Timor Sea.

Ce travail s'inscrit dans le cadre du projet OUTPACE visant à caractériser les eaux du Pacifique tropical Sud-Ouest (WTSP) en termes de stocks et flux biogéochimiques, et de diversité biologique des diazotrophes le long d'un transect longitudinal Ouest-Est. Il est le fruit de la combinaison étroite de deux approches, l'une expérimentale et l'autre basée sur la modélisation, dans le but d'étudier le rôle de la diazotrophie dans la dynamique planctonique et les cycles biogéochimiques des eaux de surface du WTSP. Les analyses de plusieurs grandeurs mesurées lors de la campagne, et notamment la production primaire (PP) et du temps de turnover du phosphate inorganique dissous (DIP), ont permis d'observer un gradient Ouest-Est de productivité et de disponibilité nutritive, étroitement lié à la variabilité spatiale des taux de fixation de N$_2$. L'utilisation d'un modèle biogéochimique mécaniste (implémenté dans la plateforme Eco3M) incluant explicitement deux compartiments de diazotrophes, couplé à un modèle physique 1D vertical, a permis de mettre en évidence le fait que l'absence/présence de diazotrophie permettait d'expliquer le contraste observé entre les régions de l'Ouest de l'archipel Mélanésien (WMA) et de l'Ouest de la gyre du Pacifique Sud (WGY). Les résultats du modèle ont montré que les organismes non diazotrophes bénéficiaient de l'apport d'azote nouveau apporté par la fixation de N$_2$, et que la production planctonique de surface dépendait significativement de l'activité des diazotrophes, cette dernière contrôlée à l'Ouest par la disponibilité en phosphate et à l'Est par la disponibilité en fer. / This work is part of the OUTPACE project which aimed to characterize the western tropical south Pacific (WTSP) in terms of biogeochemical stocks and fluxes and biological diversity of diazotrophs along a West-East longitudinal transect. This work combines an experimental with a modeling approach in order to study the role of diazotrophy in the planktonic dynamics and biogeochemical cycles of the WTSP surface waters. The values measured during the campaign, iespecially those of primary production (PP) and dissolved inorganic phosphate (DIP) turnover time, revealed a West-East gradient of productivity and nutrient availability, closely related to the spatial variability of N$_2$ fixation rates. The use of a mechanistic biogeochemical model (implemented in the Eco3M platform) explicitly including two compartments of diazotrophs and coupled with a vertical 1D physical model, allowed to highlight the fact that the absence / presence of diazotrophy could explain the contrast between the western regions of the Melanesian Archipelago (WMA) and the west of the south Pacific gyre (WGY). ). The model results showed that non-diazotrophic organisms benefited from the new nitrogen supply provided by nitrogen fixers, and that the surface planktonic production depended significantly on diazotroph activity, which is controlled by the phosphate availability in the west and by the iron availability in the east of the WTSP.