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Phytoplankton growth is iron limited in at least 20% of the world’s oceans. Iron is an important nutrient required to synthesise enzymes necessary for photosynthesis, respiration, and nitrogen assimilation. Due to its low solubility in seawater, iron limitation of phytoplankton production has been the focus of much recent research. These organisms secrete ligands in order to solubilise the available iron, but not all of the iron dissolved in seawater is biologically available. In this study a molecular based approach was employed to investigate the acclimation of the marine haptophyte Coccolithus pelagicus to iron limitation. Using two dimensional electrophoresis, subtractive cDNA hybridisation, and RT real time PCR, changes in the proteome and in gene expression were examined. Iron limited cells were characterised by slower specific growth rates, lower chlorophyll a concentrations per unit biomass and less extensive calcification relative to iron replete cells. Addition of iron to iron limited cultures resulted in increased specific growth rates and increased chlorophyll a concentration per unit biomass. A subtracted cDNA library revealed seventeen identifiable sequences of which photosystem I protein E (PsaE), a fucoxanthin binding protein transcript, two chlorophyll binding proteins and a predicted membrane protein were shown to be up-regulated in iron-limited cells to varying extents. Two dimensional SDS PAGE revealed 11 differentially expressed proteins in iron limited cells and 1 highly expressed protein exclusive to iron replete cells. The potential utility of each of these as biomarkers of iron-limitation/iron sufficiency for natural populations of coccolithophorids like Coccolithus pelagicus is discussed.

Thesis (MSc)--Stellenbosch University, 2014. / ENGLISH ABSTRACT: Iron is an essential micronutrient for all phytoplankton and is found in seawater at picomolar-level concentrations. For the first time in South Africa, a technique that utilizes a flow injection analyser (FIA) coupled with a chemiluminescence reaction has been developed for the analysis of Fe in seawater samples. The developed method is an improvement on similar available methods and uses commercially available resin (IDA) as opposed to the one that requires synthesis in the laboratory. Furthermore, the method requires reduced reagent concentrations thereby providing better results in a cost-efficient and easy manner. The improvements resulted in better precision while eliminating the loss of resin through bleeding, a common problem when using 8-HQ resin as per prior methods. Method validation was performed using internationally calibrated reference material provided by GEOTRACES and the values obtained were within the error limits of certified range. An inter-laboratory calibration was also conducted as part of the verification of the system. Surface samples from the SANAE 51 cruise were analysed for dFe and TdFe. Subsequently, the method was implemented on the SANAE 53 voyage on board the SA Agulhas II, to assess trace metal sampling protocol for any contamination issues, as well as for the analyses of collected samples. Current results suggest some contamination during collection stages, but this is still to be verified by complementary data on macronutrients and chlorophyll. The method was successfully developed and implemented in a land based clean laboratory, as well as on board a vessel. / AFRIKAANSE OPSOMMING: Yster is „n noodsaaklike mikrovoedsel vir fitoplankton en word in seewater in pikomolêre konsentrasies gevind. Hierdie lae konsentrasies en potensiële besoedeling gedurende monsteropname vanaf „n skip se platform maak akkurate Fe-analise moeilik. Vir die eerste keer in Suid-Afrika is „n tegniek ontwikkel wat gebruik maak van „n vloei- inspuitinganaliseerder (VIA), met „n geassosieerde chemiluminessensiereaksie, om die analise van Fe in seewatermonsters uit te voer. In teenstelling met soortgelyke bestaande metodes wat labratorium-gesintetiseerde hars vereis, is die nuut-ontwikkelde metode „n verbetering wat gebruik maak van „n kommersieel-beskikbare hars (IDA). Verder vereis die metode verminderde reagenskonsentrasies wat sodoende beter resultate lewer op 'n koste-effektiewe en eenvoudiger wyse. Die verbeteringe het gelei tot verhoogde akkuraatheid en uitskakeling van die verlies van hars deur dreinering – „n algemene probleem wat ondervind is met die gebruik van 8-HQ hars in vorige metodes. Geldigheidsbepaling van die metode is met internasionaal-gekalibreerde verwysingsmateriaal, verskaf deur GEOTRACES, uitgevoer. Die waardes wat verkry is, was binne die foutgrense van die gesertifiseerde skaal. „n Interlaboratorium-kalibrasie is ook uitgevoer as deel van die verifikasie van die stelsel. Daarna is die metode geïmplementeer gedurende die SANAE 53 reis op die SA Agulhas II, om die spoormetaal-monsternemingprotokol vir enige besoedelingskwessies te evalueer, asook vir die ontleding van versamelde monsters. Huidige resultate dui op „n mate van besoedeling tydens die versamelingstadiums, maar dit moet nog geverifieer word deur aanvullende data van die totale oplosbare Fe, makrovoedingstowwe en chlorofil.