High nutrient, low chlorophyll (HNLC) regions, where the availability of iron (Fe) limits primary production, comprise approximately 40% of the global ocean. Variability in Fe supply to these regions has the potential to impact Earth's climate by affecting the efficiency of the biological carbon pump, and thereby carbon dioxide uptake by the oceans. Characterizing Fe sources to HNLC regions is thus crucial for a better understanding of the connections and feedbacks between the ocean and climate change. This work addresses the question of Fe supply to two HNLC regions: the Southern Ocean and the subarctic northeast (NE) Pacific Ocean. In both regions, dissolved Fe (dFe) and the reduced form of iron, Fe(II), were measured in the water column. In the Southern Ocean, measurements were undertaken under the seasonal pack ice in the East Antarctic south of Australia. The results indicate that the sea ice represents a significant dFe source for the under-ice water column in spring, and that the Fe delivered from brine drainage and sea ice-melt likely contributes to the formation of the spring bloom at the ice edge. Shelf sediments were also found to supply dFe to the water column. Their effect was most pronounced near the shelf break and at depth, but offshore transport of Fe-enriched waters was also implicated. Fe(II) concentrations in spring were very low, most likely due to a lack of electron donors in the water column and limited solar radiation underneath the sea ice. Repeat measurements along a transect in the subarctic NE Pacific indicate that shelf sediments supply dFe and Fe(II) at depth, but their influence does not appear to extend offshore beyond several hundred kilometres. Episodic events such as the passage of sub-mesoscale eddies may transport subsurface waters a limited distance from the shelf break, supplying Fe(II) in a depth range where upwelling and deep mixing could bring it to the surface. Offshore, dFe shows little variability except in June 2012, where an aerosol deposition event is suspected to have increased dFe concentrations at depth. Fe(II) concentrations offshore are generally low, but show transient maxima at depth that likely result from remineralization processes in the oxygen deficient zone that stretches from ~600 to 1400 m depth in the subarctic NE Pacific. Elevated Fe(II) concentrations at depth were also observed in conjunction with the aerosol deposition event, which might indicate Fe(II) production associated with settling particles. However, the aerosol deposition event, which most likely stemmed from forest fires in Siberia, did not appear to trigger a phytoplankton bloom in surface waters, possibly due to a lack of Fe fertilization from the deposited material, or due to toxic effects on the resident phytoplankton community. Dust deposition from the atmosphere is considered a major Fe supply mechanism to remote HNLC regions, but the factors affecting Fe solubility of dust are poorly constrained. A laboratory experiment was conducted to test whether the presence of superoxide, a reactive oxygen species, enhances the dissolution of dust from different geographic source regions. The results indicate that superoxide may promote Fe solubilization from the dust sources tested, and that the effect of exposure to superoxide is on par with the Fe solubilizing effect of photochemical reactions. Given the possibility of widespread superoxide production by heterotrophic bacteria at all depths of the ocean, this finding suggests that significant Fe dissolution of dust particles could occur throughout the water column, not only in the well-lit surface layer. / Graduate / 0425 / 0996 / cschalle@uvic.ca

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.

Les cycles biogéochimiques du carbone et des éléments biogènes (Si, N, P) ont été étudiés en lien avec la dynamique nutritionnelle du phytoplancton dans le système naturellement fertilisé des Kerguelen. Cette étude s'inscrit dans le programme KEOPS2, qui a ciblé le nord-est du plateau des Kerguelen au début de la saison productive. La comparaison avec une zone HNLC limitée en fer a confirmé certaines des précédentes observations réalisées au cours des expériences de fertilisations artificielles et naturelles : le fer stimule clairement la croissance du phytoplancton et plus particulièrement celle des diatomées. Les zones naturellement fertilisées se sont en effet caractérisées par des biomasses en chlorophylle a et en silice biogénique 3-10 fois supérieures à la zone non-fertilisée et, par de fortes vitesses de production de silice biogénique atteignant des valeurs rarement observées dans l'océan Austral. La zone HNLC s'est caractérisée quant à elle par une population nanoplanctonique principalement composée de nanoflagellés autotrophes non siliceux. Ces travaux soulignent l'importance d'étudier la composition spécifique des populations de diatomées pour comprendre leurs implications dans les cycles du C et du Si. Nos observations ont montré que les caractéristiques physiologiques des diatomées conditionnent directement l'export de matière en profondeur dans la région des Kerguelen. Cette idée a notamment été illustrée par l'étude de l'évolution saisonnière du bloom au sud-est du plateau. / Biogeochemical cycles of carbon, silicon, nitrogen and phosphorus were studied in relation to the nutritional dynamics of phytoplankton in the naturally iron-fertilized region of Kerguelen, in the Southern Ocean. This study was conducted in the framework of the KEOPS 2 program which took place in the northeastern part of the Kerguelen Plateau in early austral spring (October-November 2011). The comparison between this iron-fertilized region and an iron-limited HNLC (High Nutrient Low Chlorophyll) area confirmed some previous observations from artificial and natural fertilization experiments: iron availability clearly stimulates phytoplankton growth and especially diatom growth. Iron-fertilized regions were characterized by 3-10 fold higher chlorophyll a and biogenic silica biomasses than the iron-limited area, as well as higher biogenic silica production rates reaching values rarely observed in the Southern Ocean. The HNLC area was characterized by a nanoplanktonic assemblage mainly composed of non-siliceous autotrophic nanoflagellates. Our results highlight the importance of studying the specific composition of diatom assemblages to better understand their impact on the C and Si biogeochemical cycles. Our observations showed that physiological traits of diatoms directly drove matter export to depth in the Kerguelen region. This idea was illustrated through the seasonal evolution of the south-eastern bloom by combining our data with KEOPS 1 data. In this region, a shift in the diatom assemblage was observed in parallel to an evolution of the vertical flux of matter, and of uptake and particulate matter ratios Si:C:N.

Diatoms play a fundamental role in marine food webs, and significantly contribute to global primary production and carbon sequestration into the deep ocean. In many offshore areas of the open ocean, iron (Fe) input is low, and its availability often limits phytoplankton biomass. Recently, gene sequences encoding ferritin, a nearly ubiquitous iron storage and detoxifying protein, have been identified in pennate diatoms such as Pseudo-nitzschia, but not in other Stramenopiles (which include centric diatoms, brown algae and some protist plant parasites) or Cryptophyte relatives. Members of this genus readily bloom upon addition of iron to Fe-limited waters, and are known to produce the neurotoxin domoic acid. Until now, the reason for the success of pennate diatoms in the open ocean was uncertain; however, expressing ferritin would allow pennate species to store Fe after a transient input, using it to dominate Fe stimulated algal blooms. Here, the ferritin gene was cloned from the coastal pennate diatom Pseudonitzschia multiseries, overexpressed in Escherichia coli, and purified using liquid chromatography. The ferritin protein sequence appears to encode a non-heme, ferritinlike di-iron carboxylate protein, while gel filtration chromatography and SDS-PAGE indicate that this ferritin is part of the 24 subunit maxi-ferritins. Spectroscopically monitoring the addition of Fe(II) to a buffered ferritin solution shows that the P. multiseries protein demonstrates ferroxidase activity, binding iron and storing it as Fe(III) in excess of 600 equivalents per protein shell. In keeping with the typical stoichiometry of the ferroxidase reaction, oxygen (O₂) is consumed in a 2 Fe:O₂ratio while hydrogen peroxide is produced concurrently. iii Diatoms evolved from secondary endosymbiosis involving eukaryotic red algae; however, a broad phylogenetic comparison suggests that P. multiseries ferritin was likely acquired via lateral gene transfer from cyanobacteria – not from its ancestral endosymbionts. Until recently, no other ferritins have been identified in diatoms, and the protein characterized here is unique in that it seems to be derived from a prokaryotic organism yet it occurs in a marine eukaryote. These findings have direct implications for the success of pennate diatoms in both Fe rich coastal waters and upon Fe addition in the open ocean.

Diatoms play a fundamental role in marine food webs, and significantly contribute to global primary production and carbon sequestration into the deep ocean. In many offshore areas of the open ocean, iron (Fe) input is low, and its availability often limits phytoplankton biomass. Recently, gene sequences encoding ferritin, a nearly ubiquitous iron storage and detoxifying protein, have been identified in pennate diatoms such as Pseudo-nitzschia, but not in other Stramenopiles (which include centric diatoms, brown algae and some protist plant parasites) or Cryptophyte relatives. Members of this genus readily bloom upon addition of iron to Fe-limited waters, and are known to produce the neurotoxin domoic acid. Until now, the reason for the success of pennate diatoms in the open ocean was uncertain; however, expressing ferritin would allow pennate species to store Fe after a transient input, using it to dominate Fe stimulated algal blooms. Here, the ferritin gene was cloned from the coastal pennate diatom Pseudonitzschia multiseries, overexpressed in Escherichia coli, and purified using liquid chromatography. The ferritin protein sequence appears to encode a non-heme, ferritinlike di-iron carboxylate protein, while gel filtration chromatography and SDS-PAGE indicate that this ferritin is part of the 24 subunit maxi-ferritins. Spectroscopically monitoring the addition of Fe(II) to a buffered ferritin solution shows that the P. multiseries protein demonstrates ferroxidase activity, binding iron and storing it as Fe(III) in excess of 600 equivalents per protein shell. In keeping with the typical stoichiometry of the ferroxidase reaction, oxygen (O₂) is consumed in a 2 Fe:O₂ratio while hydrogen peroxide is produced concurrently. iii Diatoms evolved from secondary endosymbiosis involving eukaryotic red algae; however, a broad phylogenetic comparison suggests that P. multiseries ferritin was likely acquired via lateral gene transfer from cyanobacteria – not from its ancestral endosymbionts. Until recently, no other ferritins have been identified in diatoms, and the protein characterized here is unique in that it seems to be derived from a prokaryotic organism yet it occurs in a marine eukaryote. These findings have direct implications for the success of pennate diatoms in both Fe rich coastal waters and upon Fe addition in the open ocean.

Diatoms play a fundamental role in marine food webs, and significantly contribute to global primary production and carbon sequestration into the deep ocean. In many offshore areas of the open ocean, iron (Fe) input is low, and its availability often limits phytoplankton biomass. Recently, gene sequences encoding ferritin, a nearly ubiquitous iron storage and detoxifying protein, have been identified in pennate diatoms such as Pseudo-nitzschia, but not in other Stramenopiles (which include centric diatoms, brown algae and some protist plant parasites) or Cryptophyte relatives. Members of this genus readily bloom upon addition of iron to Fe-limited waters, and are known to produce the neurotoxin domoic acid. Until now, the reason for the success of pennate diatoms in the open ocean was uncertain; however, expressing ferritin would allow pennate species to store Fe after a transient input, using it to dominate Fe stimulated algal blooms. Here, the ferritin gene was cloned from the coastal pennate diatom Pseudonitzschia multiseries, overexpressed in Escherichia coli, and purified using liquid chromatography. The ferritin protein sequence appears to encode a non-heme, ferritinlike di-iron carboxylate protein, while gel filtration chromatography and SDS-PAGE indicate that this ferritin is part of the 24 subunit maxi-ferritins. Spectroscopically monitoring the addition of Fe(II) to a buffered ferritin solution shows that the P. multiseries protein demonstrates ferroxidase activity, binding iron and storing it as Fe(III) in excess of 600 equivalents per protein shell. In keeping with the typical stoichiometry of the ferroxidase reaction, oxygen (O₂) is consumed in a 2 Fe:O₂ratio while hydrogen peroxide is produced concurrently. iii Diatoms evolved from secondary endosymbiosis involving eukaryotic red algae; however, a broad phylogenetic comparison suggests that P. multiseries ferritin was likely acquired via lateral gene transfer from cyanobacteria – not from its ancestral endosymbionts. Until recently, no other ferritins have been identified in diatoms, and the protein characterized here is unique in that it seems to be derived from a prokaryotic organism yet it occurs in a marine eukaryote. These findings have direct implications for the success of pennate diatoms in both Fe rich coastal waters and upon Fe addition in the open ocean. / Science, Faculty of / Earth, Ocean and Atmospheric Sciences, Department of / Graduate