Extracellular phosphatase activity of natural plankton studied with ELF97 phosphate: fluorescence quantification and labelling kinetics

Environmental Microbiology
Jirí NedomaK Simek

Abstract

ELF(R)97 phosphate (ELFP) is a phosphatase substrate which produces ELF(R)97 alcohol (ELFA), a fluorescent water-insoluble product, upon hydrolysis. We studied the kinetics of ELFA precipitation in freshwater samples at levels of total plankton and single phytoplankton cells, and tested the suitability of ELFP for measurement of surface-bound algal extracellular phosphatases. Samples from acidic Plesné Lake (pH approximately 5; high phosphatase activity) and eutrophic Rímov reservoir (pH approximately 7-10; moderate phosphatase activity) were incubated with ELFP for 5-300 min, fixed with HgCl2 and filtered through polycarbonate filters. Relative fluorescence of filter-retained ELFA precipitates was quantified with image analysis. Time-courses of ELFA formation exhibited lag periods followed by finite periods of linear increase. In Plesné Lake, lag-times were shorter (1-18 min) and rates of increase in ELFA fluorescence higher (by approximately 2 orders of magnitude) than in Rímov reservoir (lag-times 30-200 min). Similar patterns of ELFA formation kinetics were also observed in Plesné Lake samples in cuvette spectrofluorometer measurements (which failed in Rímov reservoir). Linear regression of seasonal data on rates of increas...Continue Reading

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Citations

Mar 28, 2012·Journal of Microbiological Methods·Melissa Blakely Peacock, Raphael M Kudela
Jan 11, 2012·Proceedings of the National Academy of Sciences of the United States of America·Caio G PereiraRafael S Oliveira
Sep 9, 2005·Microbiology and Molecular Biology Reviews : MMBR·Jakob Pernthaler, Rudolf Amann
Feb 19, 2011·Annual Review of Marine Science·Michael W LomasGer van den Engh
Jul 22, 2008·Journal of Microbiological Methods·Solange DuhamelJirí Nedoma
Dec 4, 2008·Cytometry. Part a : the Journal of the International Society for Analytical Cytology·Solange DuhamelJirí Nedoma
May 23, 2014·Cytometry. Part a : the Journal of the International Society for Analytical Cytology·Daniel Diaz-de-QuijanoMarisol Felip
Apr 20, 2016·Journal of Environmental Sciences (China)·Shuangshuang LiDunhai Li
Sep 6, 2014·Microbial Ecology·Pavel RychteckýJaroslav Vrba
Oct 6, 2007·American Journal of Physiology. Gastrointestinal and Liver Physiology·Yasutada AkibaJonathan D Kaunitz
Sep 1, 2003·The New Phytologist·Dagmara SirováJaroslav Vrba
Feb 22, 2018·Analytical Chemistry·Mathias GiraultJean-Christophe Baret

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