Assessing the potential for zinc limitation of marine primary production: proteomic characterization of the low zinc stress response in marine diatoms

Abstract

Marine diatoms are abundant photoautotrophic algae that contribute significantly to photosynthetic carbon fixation and export throughout the oceans. Zinc is an important micronutrient in algal metabolism, with scarce dissolved concentrations in the upper euphotic zone reflecting high biological demand. In this thesis, I investigated the response of marine diatoms to Zn scarcity to characterize metabolic mechanisms used to combat Zn stress. I began by assaying the ability to metabolically substitute cobalt (Co) in place of Zn in four diatom species and found that enhanced abilities to use Co are likely an adaptation to high surface dCo:dZn ratios in the native environment. I next demonstrated that Zn/Co metabolic substitution in diatoms is not universal using culture studies of Chaetoceros neogracile RS19, which has an absolute Zn requirement. Using global proteomic analysis, I then identified and characterized diatom ZCRP-A and ZCRP-B, a putative Zn-chaperone and membrane-tethered Zn acquisition protein, respectively, as two proteins involved in the low-Zn response. I demonstrated that these proteins are widespread in marine phytoplankton and can be deployed as protein biomarkers of Zn stress in the field. I furthermore documented both the detection of ZCRPs in the Southern Ocean and the existence of Zn/Fe co-limitation within the natural phytoplankton population in Terra Nova Bay, demonstrating that Zn co-limitation can indeed occur in the field, even in high macronutrient waters. Lastly, I explored the relative demand of Zn and cadmium (Cd) within the Southern Ocean community using stable 67Zn and 110Cd tracers, documenting a high demand for both metals during the austral 2017-2018 summer season and investigating the cycling of these elements within this important region. Overall, this dissertation provides new information regarding Zn acquisition and homeostasis mechanisms within marine algae and demonstrates that Zn co-limitation in the field is not only possible, but detectable via protein biomarkers.