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THE DEVELOPMENT OF A SINGLE-CELL FIELD DIAGNOSTIC FOR NITROGEN LIMITATION IN HARMFUL ALGAE
Description:
This project proposes the development of an assay to identify cell-specific acetylglucosaminidase activity using the substrate ELF-NAG (ELF 97 N'-acetylglucosaminide). Preliminary data indicates this enzyme is nitrogen-regulated in Alexandrium and that this assay could be an important new tool for identifying and monitoring nitrogen nutrition in field populations of harmful algae. In short, this work addresses gaps in our knowledge of nutritional physiology in harmful species and in our ability to identify the nutritional factors that regulate bloom dynamics.
Objectives:
The research proposed here will test and further refine an assay for the N'-acetylglucosaminidase activity using the ELF-NAG substrate. Specific objectives are to: 1) Test for ELF-NAG labeling in a suite of HAB species. 2) Pursue a method for quantifying ELF-NAG labeling on a single-cell basis in a model Alexandrium species. 3) Relate the regulation of the ELF-NAG labeling to the physiological condition of the Alexandrium cells. 4) Further characterize the physiological function of the N'-acetylglucosaminidase enzyme. 5) Develop the assay for use on field populations of Alexandrium fundyesne. 6) Test the assay on field populations of the toxic dinoflagellate A. fundyense during nutrient addition experiments in limno-corrals