River discharge leads to extensive phytoplankton blooms often observed in ocean color satellite images to extend far into the open ocean as high chlorophyll plumes. We investigated diversity, distribution and ecology of phytoplankton populations in the Mississippi River plume, both spatially and in the water column using molecular tools. A method was developed for the quantification of diatom/pelagophyte rbcL (large subunit of Ribulose-1,5-bisphosphate Carboxylase/Oxygenase) mRNA using quantitative PCR and applied to cultures and in the plume. The vertical structure of phytoplankton species in the Mississippi River plume was described by flow cytometry, pigments, rbcL mRNA and rbcL cDNA libraries. High productivity in the plume was associated with a large population of Synechococcus and elevated levels of cellular form IA rbcL mRNA. rbcL cDNA libraries indicated two vertically separated clades of Prochlorococcus (high-light and low-light adapted) in addition to a diverse group of prymnesiophytes and a microdiverse clade of prasinophytes, which may have dominated the SCM (Subsurface Chlorophyll Maximum). In situ sampling and satellite image analysis were used to estimate that the plume accounted for 41% and 13% of all surface water column ix productivity in the oligotrophic Gulf of Mexico, while covering less than 3% of its area. Coastally the plume is dominated by diatoms, which are replaced by a bloom of Synechococcus as the plume moves offshore. Diatoms as indicated by pigments and rbcL clone libraries again dominated the offshore, least productive plume. 15N uptake measurements indicated that rapid recycling of ammonium despite higher levels of nitrate primarily drives production in the offshore plume. rbcL mRNA levels and photosynthetic capacity displayed strong diel patters in three out of four time series sampled during the GRIST (Geochemical Rate/mRNA Integrated Study). In addition it was demonstrated that transcriptional regulation of the global nitrogen regulatory protein NtcA in Synechococcus WH7803 may involve a small cis-encoded anti-sense mRNA. Methods for the generation of large insert BAC (Bacterial Artificial Chromosome) from cultures and the environment were refined. Partial sequencing and genomic comparison of an ntcA containing BAC clone obtained from Synechococcus WH7803 indicated that ntcA is not part of a larger nitrogen assimilation operon in cyanobacteria.

La mer Méditerranée est une des zones océaniques les plus oligotrophes de la planète pour laquelle la fixation biologique d’azote a été proposée comme jouant un rôle important dans les flux de carbone et d’azote. Comme les organismes planctoniques potentiellement responsables de cette activité n’y ont jamais été étudiés, les objectifs de cette thèse étaient de caractériser la diversité (abondance et richesse spécifique), la distribution spatio-temporelle, et les facteurs de contrôle des diazotrophes, en s’axant particulièrement sur les cyanobactéries unicellulaires (UCYN). Pour cela, des hybridations in situ (TSAFISH)sur différentes fractions de taille en combinaison avec des comptages microscopiques et des analyses phylogénétiques sur les gènes 16S ADNr et nifH ont été réalisées, à la fois lors d’un cycle saisonnier à la station côtière SOMLIT de Marseille, et lors du transect Méditerranéen BOUM. Alors que les cyanobactéries filamenteuses Richelia intra cellularis et Trichodesmium sp. ont été détectées seulement ponctuellement dans le temps et dans l’espace, en très faible concentration (0.02 filament.ml-1), la communauté des cyanobactéries diazotrophes était dominée à 99,9% par du picoplancton hybridé avec la sonde Nitro821, spécifique aux UCYN. Ces cellules, de petite (0.7-1.5 μm) ou de grande (2.5-3.2 μm)taille, ont été retrouvées en faible concentration (1-6 cellule.ml-1) dans toute la Méditerranée et tout au long de l’année, à l’exception de l’été 2006 lorsque la concentration des petites cellules a atteint 5300 cellule.ml-1 au cours d’un évènement exceptionnel de pollution atmosphérique urbaine. Des efflorescences similaire sont été observées suite à des enrichissements en poussières sahariennes à une station côtière en Corse et au large au centre de la Méditerranée, simultanément avec des augmentations de l’activité de fixation d’azote.L’affiliation des petites cellules aux UCYN-A a été confirmée par phylogénie dans les eaux du large de la Méditerranée occidentale. Les librairies de clones nifH de la fraction de taille picoplanctonique étaient dominées par des séquences d’α-protéobactéries appartenant à un nouveau groupe marin de Bradyrhizobium. D’autres groupes de rhizobia et des γ-protéobactéries ont été détectés ponctuellement.Alors que l’absence de Trichodesmium sp. au large pourrait être liée aux faibles concentrations en phosphate et en fer, les facteurs inhibant le développement des UCYN-B et –C restent inconnus. Lesdiazotrophes picoplanctoniques (Bradyrhizobium, rhizobia, UCYN-A) pourraient avoir développé des stratégies spécifiques, telles que des associations avec des eucaryotes ou des particules inertes, et/ou la capacité de photosynthèse, pour acquérir le carbone nécessaire au soutien du processus de la diazotrophie / The Mediterranean Sea is one of the most oligotrophic marine areas on earth where nitrogenfixation has been formally believed to play an important role in carbon and nitrogen fluxes. Although thisview is under debate, the diazotrophs responsible for this activity have still not been investigated. The aimsof this PhD were to characterise the diversity (abundance and species richness) and the spatio-temporaldistribution of diazotrophs, as well as factors controlling their development, with a particular focus onunicellular cyanobacteria. A combination of microscopic counts with size fractionated in situ hybridization(TSA-FISH), and 16S rDNA and nifH phylogenies were done, either over a year and a half seasonal cycle atthe coastal SOMLIT station off Marseilles, and across the entire Mediterranean Sea during the BOUMtransect. Low concentrations of diazotrophic cyanobacteria were detected and this community wasdominated at 99.9% by picoplankton hybridized with Nitro821 probe, specific for unicellular diazotrophiccyanobacteria (UCYN). Among filamentous cyanobacteria, only 0.02 filament ml-1 of Richeliaintracellularis and Trichodesmium sp. were detected sporadically in time and space. Small (0.7-1.5 μm) andlarge (2.5-3.2 μm) Nitro821-targeted cells were recovered in low concentrations (1-6 cell ml-1) across theentire Mediterranean Sea and all the year long, except over a month period in summer 2006 whenconcentrations of small cells reached 5300 cell ml-1, during an exceptionally high urban pollution event.Similar blooms of small and large cells were reported after Saharan dust inputs off Corsica and at open Sea,simultaneously with increases in N2 fixation rates. The affiliation of the small Nitro821-targeted cells toUCYN-A was confirmed by 16S and nifH phylogenies offshore in the western Mediterranean Sea. Rhizobiasequences, including the ones of a new marine group of Bradyrhizobium, were dominating nifH clonelibraries from picoplanktonic size fractions. A few sequences of γ-proteobacteria were also detected incentral Mediterranean Sea. While low phosphate and iron concentrations could explain the absence ofTrichodesmium sp. offshore, the factors that prevent the development of UCYN-B and C remain unknown.It is proposed that the dominating Mediterranean picoplankters (Bradyrhizobium, rhizobia, UCYN-A)probably developed specific strategies, such as associations with protists or particles, and/or photosyntheticactivity, to acquire carbon for sustaining diazotrophy.

With a volcanic origin, Lake Kivu is deep and meromictic, and shows a very particular limnology and some astonishing features. The data available on its limnology and phytoplanktic communities are limited, dispersed or outdated. This is the first deep, long term study (2002-2004) on limnology and phytoplankton ecology of Lake Kivu, combining different techniques: HPLC analysis of marker pigments, flow cytometry, light, epifluorescence and electron microscopy. Lake Kivu combines a relatively shallow euphotic layer (~18m) usually smaller than its mixed layer (20 – 60 m), and with a weak thermal gradient in the mixolimnion. With an annual average chlorophyll a in the mixed layer of 2.2 mg m-3 and low nutrient levels in the euphotic zone, the lake is clearly oligotrophic. Concerning its phytoplanktonic composition, the most common species were the pennate diatoms Nitzschia bacata Hust. and Fragilaria danica (Kütz.) Lange Bert., and the cyanobacteria Planktolyngbya limnetica (Lemm.) Komárková-Legnerová and Cronberg and Synechococcus sp. Diatoms were the dominant group in the lake, particularly during the dry season episodes of deep mixing. During the rainy season, the stratified water column, with high light and lower nutrient availability, favoured dominance of cyanobacteria. Phycoerythrin-rich phototrophic picoplankton and heterotrophic bacteria cell numbers were constantly high, with relatively subtle spatial, seasonal and vertical variations. In open lake waters, where allochthonous carbon inputs are most probably inconsequential, HNA heterotrophic bacteria abundance is strongly correlated with chlorophyll a. Recent investigations revealed an increasing methane production in the deep waters during the past three decades, leading to an accumulation of gas and the subsequent lowering of the energy required to trigger a devastating release of gasses. The role of primary producers and the probable changes on the export ratio of the organic matter into deep waters after the introduction of the planktivorous sardine from Lake Tanganyika Limnothrissa miodon, is discussed. The actual primary production, 0.71 g C m-2 d-1 (~ 260 g C m-2 y-1), doesn’t seem substantially different from some punctual measurements made in the past, which discards the hypothesis of anthropogenic eutrophication. We believe that the export ratio of organic matter into the deep waters was largely affected by biological changes that could explain part of the methane increase in the past 30 years in Lake Kivu.