DIMETHYLSULFONIOPROPIONATE ( DMSP ) AND DMSP - LYASE IN CNIDARIAN - ALGAL SYMBIOSES By Denise

Title of Dissertation: DIMETHYSULFONIOPROPIONATE (DMSP) AND DMSP-LYASE IN CNIDARIAN ALGAL SYMBIOSES. Denise Marie Yost, Doctor of Philosophy, 2010 Directed By: Dr. Carys L. Mitchelmore, University of Maryland Center for Environmental Science, Chesapeake Biological Laboratory Dimethylsulfoniopropionate (DMSP) is a multifaceted sulfur compound produced by several groups of marine phytoplankton, macroalgae and higher plants. Additionally, DMSP cleaving enzymes (most of which are thought to be DMSPlyases) are known to exist in many species of marine phytoplankton, macroalgae and bacteria. Endosymbiotic dinoflagellate microalgae (genus Symbiodinium) of reefbuilding corals produce high intracellular levels of DMSP. The existence of DMSPlyase(s) in Symbiodinium was, until recently, unknown. The function(s) of the DMSP/DMSP-lyase system in cnidarian-algal symbioses is poorly understood. Chapter one introduces coral symbioses, DMSP and the potential roles of the DMSP/DMSP-lyase system in cnidarian-algal symbioses. Chapter two describes the first evidence for in vivo DMSP-lyase activity in several isolated Symbiodinium strains, revealing varying levels of DMSP and DMSP-lyase activity. These results prompted further characterization of Symbiodinium DMSP-lyases. Enzyme assay optimization and substrate kinetics experiments found the measured activity of DMSP-lyase enzymes to be affected by permeabilization buffers, pH, temperature and potential oxidative stress effects (chapter three). Prior to investigations of fieldcollected intact corals, methods for DMSP analyses were optimized in the laboratory to address the inherent complexities of the coral holobiont. This work compared several preparation techniques for the analysis of particulate (algae only, DMSPp) and total (coral tissue and algae, DMSPt) DMSP in several species of stony corals (chapter four). Field-collected corals in chapter five showed DMSPp and DMSPt responses when exposed to the oxidative stressor, copper. The second field study (chapter six) describes how DMSPp and DMSPt concentrations within five prominent Bermudian corals changed with water depth. Finally, chapter seven presents a synthesis examination of the potential functional attributes and significance of the DMSP/DMSP-lyase system in cnidarian-algal symbioses. The factors influencing variable DMSP production and accumulation as well as differences in DMSP-lyase activity are discussed in light of methodological limitations, the biology and physiology of symbiont and coral, Symbiodinium phylotype and environmental variables. The results of this research highlight the existence of DMSP-lyases in Symbiodinium and provide insight into the partitioning of DMSP in cnidarian-algal symbioses, furthering our understanding of the production and potential turnover of DMSP while recognizing the limitations inherent in such investigations. DIMETHYLSULFONIOPROPIONATE (DMSP) AND DMSP-LYASE IN CNIDARIAN-ALGAL SYMBIOSES