Biomineralization refers to the production of mineralized tissues by organisms. The fine control which organisms can exert over this process produces crystals with morphologies and properties contrasting to that of non-biogenic crystals and specifically altered to suit the required functional need. A key model system of biomineralization are a unicellular marine algae, coccolithophores, which produce calcium carbonate scales known as coccoliths. These coccoliths are comprised of arrangements of single crystals of calcite interlocked to form a plate-shaped structure. Coccoliths are developed intracellularly in a specialised compartment called the coccolith vesicle, before being extruded to the cell surface. In this work, two vital components of the coccolith biomineralization process are investigated - a soluble polysaccharide thought to act as a habit modifier and an insoluble organic scaffold known as a baseplate that provides the surface for nucleation and growth of the crystals. Whilst both these elements are thought to play a key part in the biomineralization process, the role of each is not fully understood. To investigate the effect of coccolith-associated polysaccharides (CAPs) on nucleation and polymorph selection, two systems that promote different polymorphs of calcium carbonate were utilised. In both systems, the intracrystalline polysaccharide fraction extracted from one species, Gephyrocapsa oceanica, was able to promote calcite nucleation in vitro, even under conditions favouring the kinetically-privileged polymorphs of calcium carbonate: vaterite and aragonite. As this property is not observed with CAPs extracted from its 'sister species', Emiliania huxleyi, the in vivo function of CAPs may differ between the two species. Both cryo-transmission electron microscopy (cryoTEM) and scanning electron microscopy (SEM) were used to determine the mechanism of calcite growth in the presence of G. oceanica CAPs, showing its impact on the forming amorphous calcium carbonate (ACC), decreasing the size of the particles and producing irregular, angular particles. Using cryo-electron tomography (cryoET), it was possible to create a 3D representation of the structure of the baseplate from the coccolithophore Pleurochrysis carterae, revealing its two-sided organisation. Examination of several stages of the coccolith growth process demonstrated the interlocking nature of the calcite crystals that make up the coccolith and the progression of the crystal morphologies over time, and the interaction of these crystals with the baseplate rim. Additionally, the effect of inhibiting carbonic anhydrase (CA), an enzyme involved in the regulation of carbonate species, revealed that inhibition of CA can affect coccolithogenesis as well as cell proliferation.

In this MSc thesis I determine if wind-generated bubbles elevated measurements of above-water normalized water-leaving radiance (nLw) and subsequent remote sensing estimates of particulate inorganic carbon (PIC) in a coccolithophore bloom on the Patagonian Shelf. Although no measurements were made of bubbles, shipboard wind speed was used as a proxy for bubble backscattering. An empirical orthogonal function (EOF) analysis was performed on nLw. The first EOF accounted for 95% of the variance, and was attributed to changes in spectral amplitude. Scores of the first EOF were positively correlated with flow-through PIC backscattering (bb′) > 5x10-4 m-1, indicating that above this threshold PIC was an optically active seawater constituent. There was only evidence for a bubble elevation of nLw at values of bb′ < 5x10-4 m-1 and wind speeds > 12.5 m s-1. There was no evidence for a bubble elevation of PIC estimated using the two-band PIC algorithm.

Phytoplankton dynamics and chemical characteristics of the euphotic zone were measured from 1998-2000 (an El Niño/La Niña cycle) at the 5 major stations along Line P. Near-shelf and offshore stations exhibited low seasonality in chlorophyll and moderate seasonality in particulate organic carbon (POC) production. During the 1998 El Niño, June was characterized by low chlorophyll and POC productivity due to nitrate depletion. In contrast, during the 1999 La Niña, and in 2000, higher POC productivity and nitrate occurred in June. During 1999, chlorophyll and POC productivity were similar to 1998 in late summer. Near-shelf biomass was highest in June and lowest in Feb. for the near-shelf stations. High nitrate, low chlorophyll (HNLC) stations had the highest chlorophyll in Feb. followed by June. The coccolithophore assemblage was usually numerically dominated by Emiliania huxleyi, particularly in June. Along the transect, coccolithophore abundance was much higher in June during the 1998 El Niño than in the 1999 La Niña, with Aug./Sept. abundance of both years being very low. Higher abundances were measured along the transect in June and the late summer of 2000 with sporadic ‘blooms’ of >1000 cells ml⁻¹ at some stations. Particulate inorganic carbon (PIC) production was high along the transect during June 1998, and low during both winters, June 1999 and during late summers of 1998 and 1999. There was an increase in diatom biomass and >20 µm POC production during the 1998 El Niño, specifically in the farthest offshore HNLC stations, yet diatoms were rarely found to dominate total phytoplankton biomass or production. However, there were some sporadic examples of anomalously high diatom biomass (carbon and abundance) as well as >20 µm POC production, specifically at P12 in Aug./Sept 2000. The same major diatom species were found throughout Line P (near-shelf, P16, and HNLC). Integrated silica production measured by ³²Si ranged from 0.2 to 4.7 mmol Si m⁻² d⁻¹ between 1999-2000. Silicic acid and nitrate were never limiting at all stations in Feb. and generally increased in concentration along Line P during all seasons.

Phytoplankton dynamics and chemical characteristics of the euphotic zone were measured from 1998-2000 (an El Niño/La Niña cycle) at the 5 major stations along Line P. Near-shelf and offshore stations exhibited low seasonality in chlorophyll and moderate seasonality in particulate organic carbon (POC) production. During the 1998 El Niño, June was characterized by low chlorophyll and POC productivity due to nitrate depletion. In contrast, during the 1999 La Niña, and in 2000, higher POC productivity and nitrate occurred in June. During 1999, chlorophyll and POC productivity were similar to 1998 in late summer. Near-shelf biomass was highest in June and lowest in Feb. for the near-shelf stations. High nitrate, low chlorophyll (HNLC) stations had the highest chlorophyll in Feb. followed by June. The coccolithophore assemblage was usually numerically dominated by Emiliania huxleyi, particularly in June. Along the transect, coccolithophore abundance was much higher in June during the 1998 El Niño than in the 1999 La Niña, with Aug./Sept. abundance of both years being very low. Higher abundances were measured along the transect in June and the late summer of 2000 with sporadic ‘blooms’ of >1000 cells ml⁻¹ at some stations. Particulate inorganic carbon (PIC) production was high along the transect during June 1998, and low during both winters, June 1999 and during late summers of 1998 and 1999. There was an increase in diatom biomass and >20 µm POC production during the 1998 El Niño, specifically in the farthest offshore HNLC stations, yet diatoms were rarely found to dominate total phytoplankton biomass or production. However, there were some sporadic examples of anomalously high diatom biomass (carbon and abundance) as well as >20 µm POC production, specifically at P12 in Aug./Sept 2000. The same major diatom species were found throughout Line P (near-shelf, P16, and HNLC). Integrated silica production measured by ³²Si ranged from 0.2 to 4.7 mmol Si m⁻² d⁻¹ between 1999-2000. Silicic acid and nitrate were never limiting at all stations in Feb. and generally increased in concentration along Line P during all seasons.

Environ 30% du dioxyde de carbone produit par des activités humaines est absorbé par l’océan menant à une diminution de pH d’eau de mer et de l’état de saturation de carbonate de calcium (CaCO3). L’acidification des océans engendrera probablement de profonds changements dans les écosystèmes marins, en particulier chez les organismes marins calcifiants. Les coccolithophoridés produisent avec les foraminifères plus de 90% des carbonates pélagiques dans l’océan actuel. Les expériences de culture ont montré que la réponse des coccolithophoridés à l’acidification des océans varie au sein d’une même espèce ce qui complique l’estimation de l’impact global sur le cycle de carbone et des rétroactions sur le climat. En effet, la sensibilité des organismes et les réponses vis-à-vis de l’augmentation du CO2 dissous dans l’océan et donc de la diminution du pH de l’eau de mer sont différentes. Les conséquences de la calcification sur les coccolithophoridés sont encore très peu décrites et quantifiées. Les coccolithes sont formés à l’intérieur de la cellule dans une vésicule interne. Le pH à l’intérieur de cette vésicule est un paramètre central qui détermine la précipitation de la calcite et donc de la formation des coccolithes. Actuellement, le pH de la vésicule ne peut pas être précisément mesuré et c’est la mesure indirecte de paramètres géochimies qui nous permet d’estimer ces processus. La capacité de réguler le pH de la vésicule vis-à-vis des changements du pH d’eau de mer permet la précipitation de calcite et détermine l’adaptation potentielle de certainscoccolithophoridés à l’acidification des océans. Deux souches d’E. huxleyi ont été cultivées dans des cultures en batchs dilués dans trois conditions pH différentes afin d’évaluer les modalités de réponse aux variations du pH de l’eau de mer. Des paramètres physiologiques incluant le taux de croissance, le POC et le PIC et ont été examinés, en parallèle aux mesures de B/Ca et δ11B dans la calcite des coccolithes pour progresser sur la compréhension de ces mécanismes intracellulaire et sur l’existence d’une relation entre ces paramètres et le pH pour évaluer le potentiel de l’isotopie du bore comme proxy du paleo-pH. / About 30% of the carbon dioxide produced by human activities is absorbed by the ocean leading to a decrease of seawater pH and saturation state of calcium carbonate (CaCO3). The subsequent ocean acidification is likely to result in profound changes in marine ecosystems, in particular among the marine calcifiers. Coccolithophorides together with foraminifera produce more than 90% of the pelagic carbonate in the modern ocean. Culture experiments have shown that the response of coccolithophores to pH varies between and within species, thus complicating our understanding of the overall impact biological response on the carbon cycle and feedbacks on climate. Indeed, different sensitivities to increase dissolved CO2 and decrease seawater pH, and their consequences on calcification exist among coccolithophores, but they are still not fully described nor quantified. Calcareous coccoliths are formed inside the cell in an internal vesicle called the coccolith vesicle. The pH inside the coccolith vesicle would be a key parameter in determining calcite precipitation and therefore coccolith formation. Currently the coccolith vesicle pH cannot be accurately measured and thus estimates have to be based on indirect geochemical evidences. The capacity of the coccolith vesicle to regulate pH allowing for calcite precipitation could explain the resilience of somecoccolithophores to ocean acidification. To further explore this hypothesis, two strains of E. huxleyi were grown in batch cultures under three different pH conditions to assess their response to changing seawater pH. Physiological parameters including growth rate, POC and PIC were examine, in addition to assessing changes in the vesicle pH by measuring B/Ca and δ11B in coccolith calcite and evaluate the potential of boron for paleo-pH reconstruction.

Phytoplankton dynamics and chemical characteristics of the euphotic zone were measured from 1998-2000 (an El Niño/La Niña cycle) at the 5 major stations along Line P. Near-shelf and offshore stations exhibited low seasonality in chlorophyll and moderate seasonality in particulate organic carbon (POC) production. During the 1998 El Niño, June was characterized by low chlorophyll and POC productivity due to nitrate depletion. In contrast, during the 1999 La Niña, and in 2000, higher POC productivity and nitrate occurred in June. During 1999, chlorophyll and POC productivity were similar to 1998 in late summer. Near-shelf biomass was highest in June and lowest in Feb. for the near-shelf stations. High nitrate, low chlorophyll (HNLC) stations had the highest chlorophyll in Feb. followed by June. The coccolithophore assemblage was usually numerically dominated by Emiliania huxleyi, particularly in June. Along the transect, coccolithophore abundance was much higher in June during the 1998 El Niño than in the 1999 La Niña, with Aug./Sept. abundance of both years being very low. Higher abundances were measured along the transect in June and the late summer of 2000 with sporadic ‘blooms’ of >1000 cells ml⁻¹ at some stations. Particulate inorganic carbon (PIC) production was high along the transect during June 1998, and low during both winters, June 1999 and during late summers of 1998 and 1999. There was an increase in diatom biomass and >20 µm POC production during the 1998 El Niño, specifically in the farthest offshore HNLC stations, yet diatoms were rarely found to dominate total phytoplankton biomass or production. However, there were some sporadic examples of anomalously high diatom biomass (carbon and abundance) as well as >20 µm POC production, specifically at P12 in Aug./Sept 2000. The same major diatom species were found throughout Line P (near-shelf, P16, and HNLC). Integrated silica production measured by ³²Si ranged from 0.2 to 4.7 mmol Si m⁻² d⁻¹ between 1999-2000. Silicic acid and nitrate were never limiting at all stations in Feb. and generally increased in concentration along Line P during all seasons. / Science, Faculty of / Botany, Department of / Graduate

Les coccolithophoridés sont des microalgues unicellulaires calcifiantes qui jouent un rôle important dans le cycle du carbone océanique via leurs processus cellulaires de photosynthèse (puits de CO2) et de calcification (source de CO2). L'espèce la plus abondante dans l'océan moderne est l'espèce cosmopolite Emiliania huxleyi, qui est caractérisée par une forte tolérance à de larges gammes de température, d'irradiance et de concentrations en nutriments. La distribution d'E. huxleyi qui s'étend des régions subarctiques aux régions subéquatoriales et des eaux eutrophes aux eaux oligotrophes en fait l'espèce de coccolithophoridés la plus étudiée jusqu'à présent. Cependant, sa réponse physiologique à des conditions environnementales clés comme la co-limitation en lumière et en nutriments reste peu étudiée, que ce soit in situ, où ces conditions sont rencontrées dans les niches profondes de coccolithophoridés des gyres oligotrophes, ou en laboratoire. J'ai ainsi réalisé des expériences de culture et une approche de modélisation numérique afin d'étudier la réponse physiologique d'E. huxleyi en conditions de limitation en nutriments et en lumière, avec pour objectif d'améliorer notre compréhension du contrôle environnemental des niches profondes de coccolithophoridés. / Coccolithophores are unicellular calcifying marine algae that play an important role in the oceanic carbon cycle via their cellular processes of photosynthesis (a CO2 sink) and calcification (a CO2 source). The most abundant coccolithophore species in the modern ocean is the cosmopolitan species Emiliania huxleyi that is characterized by a strong tolerance to a wide range of light, nutrient and temperature conditions. The distribution of Emiliania huxleyi from subarctic to subequatorial regions and from eutrophic to oligotrophic waters makes it the most widely studied coccolithophore species. However, its physiologic response under key environmental conditions such as the co-limitation of light and nutrients remains poorly investigated, both in the laboratory and in the field, such as in deep niches of coccolithophores in oceanic gyres. I conducted laboratory culture and numerical modeling experiments to understand the controls on the physiology of Emiliania huxleyi in low-nutrient and low-light conditions, with the aim of better understanding environmental controls on deep ecological niches of coccolithophores.

The work reported in this thesis applies a new isotope tracer, stable strontium isotopes (&delta;^88/86Sr), to address questions concerning changes in global climate that occur in response to continental weathering processes, and to constrain the modern marine geochemical Sr cycle. Stable Sr isotopes are a relatively new geochemical proxy, and as such their behavior needs to be understood in differing forms of marine calcium carbonate, the archives from which records of past stable Sr variability in the oceans can be constructed. Foraminifera, coccoliths and corals (both aragonite and high Mg calcite) acquire &delta;^88/86Sr values lighter than that of modern day seawater, (approximately 0.11, 0.05, 0.2 and 0.19 ‰ lighter than seawater at ~25°C respectively) providing a measureable offset which can be used to constrain the modern Sr outputs from the ocean and provide a better understanding of the modern Sr cycle. Using foraminifera as a sedimentary archive the first marine &delta;^88/86Sr record of seawater over the last two glacial cycles has been constructed, and used to investigate changing carbonate input and output over this 145 kyr period. Modelling of the large excursion of &delta;^88/86Sr to heavier values during Marine Isotope Stage (MIS) 3, reveals that this is more likely to be due to local changes in seawater or post-depositional alteration, rather then whole ocean changes. In the terrestrial environment &delta;&delta;88/86^Sr has been measured in the dissolved load of rivers from the Himalaya. It is found that, in general, rivers draining carbonate catchments possess lighter isotopic &delta;^88/86Sr values than those from rivers draining silicates. Covariations of either &delta;88/86Sr vs. &delta;30Si or &delta;^88/86Sr vs. 1/[Sr] can be used to distinguish between rivers draining different catchment areas.

Esta tese apresenta o registro de variações paleoceanográficas na superfície e em oceano profundo na porção oeste do Atlântico Sul, através de evidências de nanoplâncton calcário, durante os últimos 200.000 anos. Os principais parâmetros avaliados foram relativos à paleoprodutividade primária, através de estimativas de abundância e taxa de acumulação do nanoplâncton calcário, índices paleoceanográficos de produtividade, variação de carbonato de cálcio, contribuição relativa das espécies de nanoplâncton para o carbonato de cálcio total e análises de dados geoquímicos complementares. Os dados derivam de três testemunhos coletados na Bacia de Santos entre 1900 e 2200 metros de profundidade. Os resultados demonstram que as mudanças paleoceanográficas observadas no período analisado devem estar relacionadas, concomitantemente, a variações na dinâmica do oceano na camada superficial e em oceano profundo. As evidências de paleoprodutividade primária e a subsequente deposição do nanoplâncton calcário apontam que ambas são dependentes de condições favoráveis em superfície e no fundo, respectivamente. Estas condições ocorreram durante o período de aquecimento das águas superficiais no oeste do Atlântico Sul e a retração de águas profundas de origem sul devido à intensificação da célula de revolvimento meridional do Atlântico. Os últimos 200.000 anos foram divididos em sete intervalos definidos pela variação de paleoprodutividade primária com base no registro de nanoplâncton calcário. / This thesis presents a view of the paleoceanographic changes that occurred in the surface and deep ocean in the western South Atlantic by means of calcareous nannoplankton during the last 200,000 years. Data was obtained from three sediment cores collected in the Santos Basin between 1900 and 2200 meters water depth. Estimates of primary paleoproductivity from calcareous nannoplankton abundance and accumulation rates, paleoproductivity proxies calcium carbonate content, the relative contribution of nannoplankton species to total calcium carbonate and complementary geochemical data analyzes were performed. The results demonstrate that paleoceanographic changes observed in the analyzed period may be related to changes in ocean dynamics at surface and deep ocean. Primary paleoproductivity record and the subsequent calcareous nannoplankton deposition indicate that these are dependent on surface and bottom conditions, respectively. These conditions were recorded during sea surface temperature warming in the western South Atlantic and retraction of southern-sourced deep waters due to strengthening of the Atlantic meridional overturning circulation. The last 200,000 years were divided into seven intervals defined by the changes in primary paleoproductivity record of calcareous nannoplankton.

Esta tese apresenta o registro de variações paleoceanográficas na superfície e em oceano profundo na porção oeste do Atlântico Sul, através de evidências de nanoplâncton calcário, durante os últimos 200.000 anos. Os principais parâmetros avaliados foram relativos à paleoprodutividade primária, através de estimativas de abundância e taxa de acumulação do nanoplâncton calcário, índices paleoceanográficos de produtividade, variação de carbonato de cálcio, contribuição relativa das espécies de nanoplâncton para o carbonato de cálcio total e análises de dados geoquímicos complementares. Os dados derivam de três testemunhos coletados na Bacia de Santos entre 1900 e 2200 metros de profundidade. Os resultados demonstram que as mudanças paleoceanográficas observadas no período analisado devem estar relacionadas, concomitantemente, a variações na dinâmica do oceano na camada superficial e em oceano profundo. As evidências de paleoprodutividade primária e a subsequente deposição do nanoplâncton calcário apontam que ambas são dependentes de condições favoráveis em superfície e no fundo, respectivamente. Estas condições ocorreram durante o período de aquecimento das águas superficiais no oeste do Atlântico Sul e a retração de águas profundas de origem sul devido à intensificação da célula de revolvimento meridional do Atlântico. Os últimos 200.000 anos foram divididos em sete intervalos definidos pela variação de paleoprodutividade primária com base no registro de nanoplâncton calcário. / This thesis presents a view of the paleoceanographic changes that occurred in the surface and deep ocean in the western South Atlantic by means of calcareous nannoplankton during the last 200,000 years. Data was obtained from three sediment cores collected in the Santos Basin between 1900 and 2200 meters water depth. Estimates of primary paleoproductivity from calcareous nannoplankton abundance and accumulation rates, paleoproductivity proxies calcium carbonate content, the relative contribution of nannoplankton species to total calcium carbonate and complementary geochemical data analyzes were performed. The results demonstrate that paleoceanographic changes observed in the analyzed period may be related to changes in ocean dynamics at surface and deep ocean. Primary paleoproductivity record and the subsequent calcareous nannoplankton deposition indicate that these are dependent on surface and bottom conditions, respectively. These conditions were recorded during sea surface temperature warming in the western South Atlantic and retraction of southern-sourced deep waters due to strengthening of the Atlantic meridional overturning circulation. The last 200,000 years were divided into seven intervals defined by the changes in primary paleoproductivity record of calcareous nannoplankton.