Photosynthesis is responsible for fixing approximately 111 – 117 Pg of CO₂ into organic carbon each year, of which about half is performed by algae in the oceans. Over geological timescales, photosynthesis by algae was instrumental in transforming Earth’s atmosphere. Despite the integral role algae play in the carbon cycle, the interaction and feedbacks between CO₂ fixation by algae and atmospheric CO₂ is poorly understood. This thesis expands upon our current knowledge by tracing the evolution of the key enzyme of photosynthesis, Rubisco, in algae through geological history. It was found that Rubisco underwent adaptation during distinct periods corresponding with falling atmospheric CO₂. The pattern of adaptation hints at physiological adaptation to varying concentrations of atmospheric CO2 and possibly indicates the emergence of carbon concentrating mechanisms (CCMs). This adaptation was probed further within the red and chromist algae, identifying key residues within the Rubisco protein sequence that may influence its kinetic properties. This research also provided new measurements of Rubisco CO2 affinity within the haptophyte algae. Finally, the importance of HCO₃- use by phytoplankton in the modern ocean was explored. HCO₃- utilisation was modelled through signals retained within stable carbon isotopes of organic matter estimate the response to anthropogenic increases of CO₂. The results indicate that phytoplankton utilise a large proportion HCO₃- which shows little sensitivity to anthropogenic increases of CO₂, even when model predictions are extended to 2100. This thesis demonstrates how algae can respond to CO₂ levels over geological and anthropogenic time scales.

Coccolithophores are single-celled marine phytoplankton, which produce intricate calcium carbonate platelets or ‘coccoliths’. Emiliania huxleyi is the most abundant and widespread coccolithophore, and is one of the most productive calcifying species on earth, playing a key role in global carbon, carbonate and sulphur cycles. Despite much research into coccolithophore biology, the underlying function of their coccoliths is still unknown. The main aim of the research reported in this thesis was to examine the impact of calcification on metabolism in coccolithophores. Calcification is a significant global process, so it is important to discover what effect it has on the metabolism of cells. The major metabolites each have different costs and benefits to the cell, which will vary depending on the habitat and environmental conditions the cell is in. By comparing the metabolite profiles of different strains, including calcifying, non-calcifying, haploid and diploid cells, differences in metabolite composition and potential patterns related to cell type were investigated. Low molecular weight (LMW) metabolites were characterised using a combination of metabolomic techniques. In agreement with previous research, dimethylsulphoniopropionate (DMSP) was the most abundant compound, followed by mannitol and glycine betaine (GBT). Less abundant sugars, polyols and amino acids were also identified. Environmental factors were manipulated to investigate how the principal metabolites were affected by salinity, different light intensities and nutrient (phosphate and nitrate) limitation. The data revealed a striking difference between haploid and diploid cells of the same strain, with the haploid containing lower concentrations of most of the major metabolites. Thus it is proposed that haploid cells have a different osmoregulatory strategy from the diploid cells. A negative correlation was found between DMSP and mannitol, suggesting that mannitol has a dual function, not only as a major storage compound but also as a principal compatible solute. Untargeted metabolite profiling is becoming a popular tool to investigate phenotypes and varying environmental conditions. LC-ESI-QTOF-MS/MS analyses of a wide range of metabolites showed that it is an effective method to identify differences and similarities between E. huxleyi strains grown in different conditions. Strain and growth phase appear to be the more important factors in differentiating metabolite profiles. Surprisingly there were no obvious metabolite profiling differences between calcifying and non-calcifying cells. Untargeted analysis can, however, be used to identify the compounds that did display differences, and which may be important biomarkers, so warrant further investigation. A range of metabolite profiling techniques highlighted important differences between strains, which will hopefully lead onto further research into the metabolome of E. huxleyi, and the unravelling of important metabolic pathways. There has been little research into the LMW metabolites of E. huxleyi, and especially comparisons between strains. Thus the use of metabolomics is a novel way to investigate the difference between cell types and the possible functions of calcification.