The main goal of this study was to assess the impact that nitrate and light have on the relationship between the gene expression of the enzyme nitrate reductase and the incorporation of nitrate in the cosmopolitan diatom Thalassiosira pseudonana, both in laboratory experiments and in natural environments. Continuous cultures were grown at different nitrate (NO3-) concentrations (i.e., 60, 120, and 400 µM) to evaluate their effects on the expression levels of different genes of the nitrogen metabolic pathway (i.e., nitrate and ammonium transporters, nitrate and nitrite reductases, glutamine synthetases II and III). Semi-continuous cultures were grown under different irradiances (i.e., 50, 110, 200, and 320 µmol photon cm-2 s-1) to assess the influence of light intensity (irradiance) on the relationship between the expression of those genes, uptake, and assimilation of nitrate. The expression of all of the genes that were tested decreased significantly (p < 0.05) at the highest concentration of NO3- (i.e., 400 µM), with nitrate transporters showing the most pronounced change from 27.97 to 0.59 fold change cell-1 x 10-6, at 60 and 400 µM NO3- concentrations respectively. Ammonium transporters were detected at all concentrations of NO3-, suggesting that cells are always ready to metabolize ammonium. Growth was limited (µ = 0.99 d-1) by the low irradiance treatment, was maximum (µ = 2.04 d-1) at 200 µmol photon cm-2 s-1 and was inhibited (µ = 1.54 d-1) at the highest irradiance. These trends were reflected in gene expression and uptake rates, with minimum values at the lowest and highest irradiance levels. However, results from the enzymatic assay did not show any significant differences between treatments (p > 0.05). The trends observed in the enzymatic rates could be explained by the gene expression of NO3- reductase and the uptake and growth rates in a multiple regression analysis (R2 = 0.66, p < 0.05). The results of this study show that uptake is independent of gene expression, probably because of a decoupling between transcription and protein synthesis. Not all of the newly synthesized transcripts will inevitably be translated into proteins. And even if they were, there could be post-translational mechanisms preventing the enzymes to become active. This indicates that uptake can be independent of the expression. It was attempted to measure the expression of T. pseudonana genes involved in the metabolism of NO3- in natural diatom assemblages. The use of gene expression as a proxy for metabolic processes carried out by a phytoplankton assemblage in the field is limited and depends on environmental factors, since the current methods of assessing expression rely on genomic sequences that are particularly variable in phytoplankton. The assessment of gene expression provides a useful insight into physiological studies of phytoplankton, and it should be complemented with other measurements, such as the biomass and taxonomic composition of the assemblage for a more complete picture of marine ecosystem nutrient dynamics. / Graduate

Morphogenesis of the intricate patterns of diatom silica cell walls is a protein-guided process, yet to date only very few such silica biomineralization proteins have been identified. Therefore, it is currently unknown whether all diatoms share conserved proteins of a basal silica forming machinery, and whether unique proteins are responsible for the morphogenesis of species-specific silica patterns. To answer these questions, we extracted proteins from the silica of three diatom species (Thalassiosira pseudonana, Thalassiosira oceanica, and Cyclotella cryptica) by complete demineralization of the cell walls. Liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) analysis of the extracts identified 92 proteins that we name ‘soluble silicome proteins’ (SSPs). Surprisingly, no SSPs are common to all three species, and most SSPs showed very low similarity to one another in sequence alignments. In-depth bioinformatics analyses revealed that SSPs could be grouped into distinct classes based on short unconventional sequence motifs whose functions are yet unknown. The results from the in vivo localization of selected SSPs indicates that proteins, which lack sequence homology but share unconventional sequence motifs may exert similar functions in the morphogenesis of the diatom silica cell wall.

Le modèle michaëlien d'acquisition d'azote, utilisé à l'origine pour représenter l'effet de la concentration d'un nutriment sur son taux d'absorption, s'avère insuffisant lorsque d'autres facteurs présentent des variations temporelles, à l'exemple du cycle diurne de lumière. Dans la littérature, les oscillations diurnes de l'acquisition d'azote sont généralement expliquées par la réponse du statut physiologique des microalgues au flux de photons et à la disponibilité en substrat. Cette thèse a pour but d'identifier dans quelle mesure le cycle cellulaire conditionne lui aussi la prise d'azote, et le cas échéant d'évaluer l'impact de cette propriété au niveau macroscopique. Pour cela, nous avons réalisé des expériences en cultures continues de la diatomée Thalassiosira weissflogii soumise à différentes conditions de lumière et d'azote. Notre étude met en évidence une diminution de l'acquisition d'azote lorsqu'une proportion significative de la population est en mitose. Il apparait également que les évènements mitotiques sont planifiés à la fois par les conditions de croissance et par une horloge interne. De plus, nos expériences montrent que les conséquences macroscopiques de ces propriétés individuelles sont fonction du degré de synchronisation de la population, et nous détaillons les facteurs agissant sur ce phénomène. Ensuite, nos hypothèses sont reprises dans une approche numérique afin de tester le gain de représentativité obtenu en prenant en compte le cycle cellulaire dans la modélisation de la croissance phytoplanctonique. Enfin, nous discutons de l'effet du cycle cellulaire sur l'acquisition d'azote en milieu marin et de ses possibles implications écologiques.

The sediment Core SWERUS-L2-2-PC1 (2PC) retrieved from the Chukchi Sea, Arctic Ocean sits in an oceanographically dynamic location at the Arctic-Pacific Ocean gateway. The 8.3 m-long core was retrieved in Herald Canyon at the marginal ice zone at 57 m depth. Core 2PC is well-positioned to record variability in inflow of Bering Sea Water (BSW) and Pacific Water (PW) in Herald Canyon. With the 2PC high sedimentation rate (200 cm/kyr), two independent age models (radiocarbon and palaeomagnetism) based on tephra age markers, and a richness in well-preserved siliceous sediment, validate 2PC as an outstanding sequence for applying diatom assemblage analysis as a proxy for ocean-climate change back to 4250 years BP, including the past few hundred years where global warming and sea ice decline is recorded by instrumental records. These characteristics make Core-2PC a useful record for investigating the role of PW on sea ice variability in the Chukchi Sea, both in the past and predicting the future. To investigate the impact of PW on ocean and sea ice conditions in the Chukchi Sea, diatom assemblage analysis was performed on 49 samples through the Late Holocene. The over-arching goal was to test the hypothesis, suggested by existing research on 2PC using benthic foraminifera Mg/Ca palaeothermometry, that the strength of PW inflow into the Chukchi Sea via Herald Canyon has varied on a time scales of ~500-1000 years in the past 4000 years. PW is slightly warmer than resident Arctic surface waters and is known to be an important control on Arctic sea-ice. The diatom assemblage approach assumes that there are recognizable differences between end-member diatom assemblages that are characteristic of PW versus Arctic Ocean type environments associated with extensive sea-ice conditions. The mapping of species in the Herald Canyon was used to test the idea of variability of sea-ice extent and the role of the Pacific Ocean forcings into the western Chukchi Sea. The results reveal diverse diatom assemblages throughout the past 4000 years in Herald Canyon, showing this core to be very useful for diatom palaeoclimate reconstructions. A total of 126 species with abundance &gt;1% are recognized. Several generalist species typically dominate assemblages especially Chaetoceros, ice-algae, marine-neritic and near ice or cold-water planktic centric diatoms. Distinct changes in stratigraphy are illustrated by changes in identified diatom assemblage zones. The 2PC diatom assemblages were contrasted with records from Chukchi-, Laptev-, East Siberian- and Bering Sea and North Pacific Ocean. At 2PC, sympagic (sea-ice related), planktic and neritic species abundance varies on time scales of ~500-1000 years. Importantly, there is a clear similarity between the timing of diatom assemblage changes and the 2PC benthic foraminifera Mg/Ca bottom water temperature (BWT) reconstruction. In particular, abundance changes in the warm water species Thalassionema nitzschioides, Shionodiscus oestrupii and Thalassionema simonsenii, tychoplanktic Paralia sulcata, Ice algae- and sympagic assemblages and cold-water indicators correspond best to BWT fluctuations shown by the Mg/Ca reconstruction. These oscillations are suggestive of changes in warmer PW inflow. Other aspects of the diatom data appear to correlate with colder and warmer climate events and suggest that changes in PW inflow amplified the effects of these events in the Chukchi Sea region through the Late Holocene in the Northern Hemisphere. It can thus, be concluded that diatoms from 2PC, support the palaeoceanographic reconstruction suggested by the benthic foraminifera Mg/Ca palaeothermometry and that variations in PW inflow through Herald Canyon is an important driver of sea ice variability on thousand-year time scales.