Experimental Approaches and Protocols for Functional Genomics of Coccolithophorid Phytoplankton

• Main Author: Hockin, Nicola
• Other Authors: Faculty of Science
• Format: Thesis
• Language: unknown
• Published: University of Plymouth 2007
• Subjects: Ocean acidification
• Online Access: http://hdl.handle.net/10026.2/334

Coccoiitliophores play a crucial role in the marine carbon cycle. The formation of Calcite (CaC03) by coccolithophores in surface waters and its subsequent removal by sinking has important implications for the flux of inorganic carbon into the oceans. To understand the impact of global change, such as ocean acidification, on calcification it is important to understand the underlying biological mechanisms involved. Despite extensive research into coccolithophore calcification, the cellular mechanisms of this process remain poorly understood. Molecular techniques now hold great potential to further our understanding in this field. The application of such techniques is, however, severely limited by the lack of a transformation system for these organisms. The aim of this study was to develop such a system. The parameters for the microparticle bombardment of the coccolithophores Emiliania huxleyi and Pleurochrysis carterae were optimised, providing a mechanism for the introduction of DNA and dyes to these cells. The sensitivity of both species to the herbicide, Basta®, was also demonstrated by decreasing growth in media and on agarose plates, with increasing Basta® concentration. E. huxleyi and P. carterae were bombarded with the bar gene, which confers resistance to Basta® and grown in selective media containing the herbicide to select for transformed cells. Although the results of the transformation were inconclusive, this study provides extensive ground work for the transformation of coccolithophores and has the potential to open up completely new routes of research in this field. Faculty of Science, Marine Biological Association, Plymouth