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The presented thesis includes three related projects, that are linked by a common interest in the evolution of eukaryotic organelles and machineries that import proteins into these compartments. The first project considers the possibility of peroxisomes (eukaryotic organelles known in aerobic organisms) being conserved in two related anaerobic protists: a free-living amoeba Mastigamoeba balamuthi and a parasite Entamoeba histolytica. The most important hint for the presence of peroxisomes was the discovery of proteins that are homologous to known components of the peroxisomal protein import machinery. The second project aims to characterize the unknown protein translocase of the inner membrane (TIM) in the mitosomes (extremely reduced mitochondria) of an anaerobic protozoan Giardia intestinalis. We have discovered an important subunit of the mitosomal translocase (Tim44), which usually tethers the Hsp70/PAM (presequence translocase-associated motor) complex to the TIM translocon. The last project shows that the protein translocase of the outer mitochondrial membrane in trypanosomatids is related to a typical eukaryotic channel Tom40. This finding is important because the absence of Tom40 was previously considered an ancestral feature of trypanosomatids.

Depuis les années 90, les études portant sur les différentes zones HNLC ont permis d'étudier les effets biologiques et biogéochimiques qu'entrainaient les enrichissements artificiels ou naturels en fer. Il est maintenant bien documenté que l'enrichissement en fer induit des blooms phytoplanctoniques et notamment des blooms de diatomées. En dehors des diatomées, très peu d'informations sont disponibles concernant les autres groupes de protistes et en particulier les protistes hétérotrophes consommateur du phytoplancton. Ce travail a été effectué dans un contexte de fertilisation naturelle en fer, dans la région des îles de Kerguélen (Océan Australe) pendant la campagne KEOPS 2 (Kerguelen Ocean and Plateau compared Study 2) lors de l'initiation du bloom phytoplanctonique et s'est focalisé en particulier sur les protistes hétérotrophes. Des approches moléculaires (tag-pyroséquençages 454) et morphologiques (microscopie) ont été utilisées afin de caractériser la structure des communautés de protistes dans la zone de référence HNLC et dans les différents blooms phytoplanctoniques. l'approche moléculaire a permis (i) de caractériser les communautés de protistes présentes (ii) de mettre en évidence des différences notables entre les structures de protistes dans la région HNLC et la région naturellement enrichie en fer, mais également entre les différents blooms. Les observations microscopiques ont révélé des tendances similaires entre les différentes régions mais aussi des liens significatifs entre les communautés microzooplanctoniques et leurs proies phytoplanctoniques. Les observations microscopiques ont également fournis des valeurs de biomasses des différents compatiments qui ont permis d'estimer le potentiel du microzooplancton en tant que consommateur du phytoplancton ou en tant que source nutritive pour le mésozooplancton. Ce travail représente la première étude caractérisant la communauté des protistes planctoniques dans son ensemble dans un contexte de fertilisation naturelle en fer. / Since the 90s, studies on different HNLC areas allowed to investigated the biological and biogeochemical effects due to artificial or natural iron-enrichment. It is now well documented that iron enrichment induced phytoplankton blooms and more specifically diatom blooms. With the exception of diatoms, very few information is available concerning other protists groups e. g. heterotrophic protists which are consumers of phytoplankton.This work was performed is a natural iron-fertilization context in the Kerguelen Island area (Southern Ocean) during the KEOPS 2 (Kerguelen Ocean and Plateau compared Study 2) cruise at the beginning of the phytoplankton bloom and focused specifically on heterotrophic protists. Molecular (tag-pyrosequencing 454) and morphological (microscopy) approaches were used to characterize the structure of protist communities in the HNLC reference area and in the phytoplankton blooms. The molecular approach allowed (i) to provide a complete picture of the protist communities (ii) to evidence significant differences in protists structures between HNLC and the naturally iron-fertilized area, but also between the different blooms. Microscopic observation revealed similar trends between regions but also significant links between microzooplanctonic communities and their phytoplankton preys. Microscopic observations also provided biomass values from different compartments allowing an estimation of the potential of microzooplankton as phytoplankton consumer or as a nutrient source for mesozooplankton. Above all, this work represents the first study characterizing the global planktonic protists community in the context of natural iron fertilization.

Tree canopies form the most important interface between the earth and the atmosphere, but their role as a potential habitat for eukaryotic microorganisms has been severely understudied. This thesis addresses the hitherto insufficient characterisation of protist diversity and community composition in forest soils and the canopy region, while in particular focusing on the prominent and mostly plant pathogenic Oomycota. Accordingly, several microhabitat compartments were sampled on the ground and in the canopy with the Leipzig Canopy Crane in two seasons over two years, and additionally in a palaeotropical rainforest in Papua New Guinea. Taxon-specific primers were applied to characterise the oomycete diversity in a high-throughput metabarcoding approach. In four chapters, I assess the initial description of oomycete communities in the canopy, their distribution in the forest ecosystem via air dispersal, their seasonality and functional diversity including the quantification of plant pathogens, and their diversity in tropical canopies. These results contribute to a more comprehensive understanding of oomycetes and their pathogenic lineages, their diversity, ecology, distribution, and ecosystem functioning.

Oomycetes (Stramenopiles, protists) are among the most severe plant pathogens, comprising species with a high economic and ecologic impact on forest ecosystems. Their diversity and community structures are well studied in terrestrial habitats, but tree canopies as huge and diverse habitats have been widely neglected. A recent study highlighted distinct oomycete communities in the canopy stratum compared to the ground region of three temperate deciduous trees (Quercus robur, Tilia cordata, Fraxinus excelsior). While the communities from the two strata were distinct when taking oomycete abundances into account, they were rather similar when only OTU presence/absence was considered. It remains, however, unknown if this homogeneity in the OTU presence also leads to a functional homogenisation among microhabitats within the two strata ground and canopy. In this study, we supplemented functional traits to oomycete communities in the tree microhabitats, which were determined over a time period of 2 years with a metabarcoding approach. Our results showed that even though most oomycetes occurred in all microhabitats, a strong discrepancy between the strata and correspondingly the distribution of oomycete lifestyles could be observed. This pattern was constant over several seasons. Obligate biotrophic species, exclusively feeding on living host tissue, dominated the canopy region, implying tree canopies to be a hitherto neglected reservoir for parasitic protists. OTUs assigned to the genus Hyaloperonospora—parasites highly specialised on hosts that were not sampled—could be determined in high abundances in the canopy and the surrounding air, challenging the strict host dependencies ruled for some oomycetes. Our findings further contribute to the understanding of oomycete ecosystem functioning in forest ecosystems

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This thesis investigates the diversity of two silica-scaled protist groups, Paraphysomonadida and Thaumatomonadida by light and electron microscopical observations and sequencing (rDNA) on novel clonal cultures. Despite these groups of protist dominating pelagic, littoral as well as inland freshwater and soil habitats, they are taxonomically poorly understood to the extent that any progress in ecological theory is hampered. Now that environmental DNA sequencing is being carried out faster than we can characterise protists from culture it is important that we understand how molecular and physical diversity match up, especially because so many protists are morphospecies. Nearly one hundred isolates were cultured on which both morphological and molecular data was carried out in parallel to reveal around 50 new species of protist from eight different genera: two heterokont genera, Paraphysomonas and Incisomonas n. gen., and six cercozoan genera, Thaumatomonas, Allas, Reckertia, Thaumatospina n. gen., Cowlomonas n. gen., and Scutellomonas n. gen. These data make major contributions to taxonomy and understanding aspects of protist diversity where previously morphological diversity was heavily biased towards over- generalized morphotypes. This thesis quickly showed that gross lumping of morphospecies was true of Paraphysomonas, for which many of the isolates cultured herein might have been regarded as one species (not more than 20). The many cultured isolates exhibited varied cell and scale morphology, and by sequencing (rDNA), it was possible to see the evolution of scale morphology map on to trees. This marriage of molecular and morphological data made it possible to view distinct groups of species that shared scale detail that might have otherwise been overlooked had either method been used alone. This research has shed significant light on how scale morphology can be used as reliable taxonomic marker for protists, the insights of which can be applied to make taxonomic improvements to other silica-scaled protist groups.

Marine microbes are an essential component of global biogeochemical cycles. In oligotrophic marine surface waters, the phytoplankton, phototrophic, single-celled (on occasion, colonial) organisms, is often dominated by the picoplankton (cells <2 micrometers in size), which constitute the base of the marine food chain. The picophytoplankton is composed of three main groups of organisms: two genera of cyanobacteria, Prochlorococcus and Synechococcus, and a third group, the picoeukaryotes. Even though numerically less abundant than cyanobacteria, picoeukaryotes can contribute significantly to biomass and primary production in this size fraction. Furthermore, picoeukaryotes are a diverse group but this diversity is still underexplored and their ecological roles and physiology is poorly understood. Here uncultured protists are investigated using 18S rRNA gene clone libraries, phylogenetic analyses, specific fluorescence in situ hybridization (FISH) probes and other methods in tropical and subtropical waters. Gene sequences comprising a unique eukaryotic lineage, biliphytes, were identified in most samples, whether from high (30 degrees Celsius) or low (5 degrees Celsius) temperature waters. Sequences within this uncultured group have previously been retrieved from mid and high latitudes. Phycobilin-like fluorescence associated with biliphyte-specific FISH probed cells indicated they may be photosynthetic. Furthermore, the data indicated biliphytes are nanoplanktonic in size, averaging between 3.0 and 4.1 micrometers. Using the 18S rRNA gene, sequences belonging to a broadly distributed but uncultivated pico-prymnesiophytes were retrieved. We investigated the ecological importance of these natural pico-prymnesiophyte populations and field experiments showed that they could grow rapidly and contributed measurably to primary production. They also appear to form a large portion of global picophytoplankton biomass, with differing contributions in five biogeographical provinces, from tropical to high latitudes. Finally, the physiology of the picoeukaryote Micromonas was studied under a shift from medium to high light and UV radiation. Results showed that the growth of these photosynthetic cells was synchronized with the light: dark period. Forward angle side scatter and red autofluorescence from chlorophyll increased throughout the light period and decreased during the dark period. This is consistent with cell division occurring at the beginning of the dark period. Additionally, genes proposed to have roles in photoprotection were up-regulated under high light and UV, but not in controls.

The goal of this work was to examine the samples from archeological site in an attempt to identify human and/or animal intestinal parasite eggs. Another task was to detect parasitic protist antigens by ELISA test. Then the results were compared with other facts obtained from research of this locality.