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Axenic cultures of the pico-prasinophyte Micromonas commoda<\/em> (CCMP 2709), the coccolithophore Emiliania huxleyi<\/em> (CCMP 2090), and the diatom Phaeodactylum tricornutum<\/em> (CCMP 2561) were obtained from the National Center for Marine Algae and Microbiota (Bigelow Laboratory for Ocean Sciences, East Boothbay, Maine). The cultures remained axenic throughout the experiments as determined by SYTO-staining and flow cytometric counting on a BD FACSJazz cell sorter; all cultures were free of bacteria during these experiments. Phytoplankton were grown in artificial sea water amended with L1 media with (P. tricornutum<\/em>) or without (M. commoda and E. huxleyi<\/em>) silica. The P source was added separately to achieve the desired growth conditions; Pi-replete media contained 36 \u00b5M PO\u2084\u00b3\u207b, the Pi-deficient condition received 0.1 \u00b5M PO\u2084\u00b3\u207b, and the phosphonate treatments received either 36 \u00b5M MPN or 2-AEPN. The Pi-deficient treatment (0.1 \u00b5M) represents a control for the low level of contaminating Pi measured in the phosphonate media; thus, an increase in growth in the MPN and 2-AEPN conditions above that measured in the Pi-deficient condition is due to phosphonate utilization. The potential for abiotic breakdown of phosphonate to Pi was investigated in media-only tubes exposed to the experimental temperature and light conditions for 10 days. Pi levels did not change throughout the experimental period (MPN average Pi = 0.11 \u00b5M \u00b1 0.02; 2-AEPN average Pi = 0.10 \u00b5M \u00b1 0.02), strongly supporting the notion of active enzymatic breakdown of phosphonates for growth. Natural P concentrations are much lower than those used in this study; the replete nutrient concentrations were used to support high cell yields necessary for analytical measurements. Cultures were acclimated to the four growth conditions described above as they had been maintained in each P treatment for a minimum of two transfers (20 days). Cultures were grown at 20\u00b0C in a 14h light\/10h dark cycle at ~100 \u00b5E m\u207b\u00b2 s\u207b\u00b9 with a starting concentration of ~1x10\u2074 cells mL\u207b\u00b9 in 25 mL culture volumes. Phytoplankton growth was monitored by fluorescence measurements using a Turner TD-700 fluorometer and cell counts analyzed by flow cytometry. Specific growth rates (\u03bc) were calculated from the linear regression of the natural log of cell counts during the exponential growth phase of cultures. Quadruplicate cultures were setup for each treatment; three replicates were harvested in the late exponential phase of growth for physiological measurements, while the fourth was used to monitor cell abundances later in the growth cycle.<\/p>\n Physiological measurements: Alkaline phosphatase content (APA) measurements were made by quantifying the hydrolysis of 6,8-difluoro-4-methylumbelliferyl phosphate using a Molecular Devices FilterMax F5 microplate reader. Abiotic substrate hydrolysis was accounted for in killed controls that were boiled and cooled prior to substrate addition, as well as in media-only controls. The fluorescent reference standard, 6,8-difluoro-4-methylcoumarin was used to calculate the rate of hydrolysis, which was then normalized to cell abundance to determine APA per cell.<\/p><\/div>","@type":"rdf:HTML"}],"http:\/\/purl.org\/dc\/terms\/description":[{"@value":" Three species of marine phytoplankton \u2013 Micromonas commoda<\/em>, Emiliania huxleyi<\/em>, and Phaeodactylum tricornutum<\/em> \u2013 were grown under four phosphorus (P) conditions. These include phosphate (Pi) replete and deplete conditions and the phosphonate conditions where cultures received either methylphosphonate (MPN) or 2-aminoethylphosphonate (2-AEPN) as the sole source of phosphorus at replete levels. Samples for cell abundance were collected throughout the experiment to monitor growth. In addition, particulate P, to calculate cellular P quota, as well as alkaline phosphatase activity was measured at a single time point when cells were in exponential growth.<\/p><\/div>","@type":"rdf:HTML"}],"http:\/\/www.w3.org\/2000\/01\/rdf-schema#label":[{"@value":"Phytoplankton phosphonate utilization","@type":"xsd:string"}],"http:\/\/ocean-data.org\/schema\/hasProcessingDescription":[{"@value":" Cell abundance data were processed from flow cytometry files by gating logic using Sortware software. Nutrient concentrations, both dissolved and particulate, were calculated based upon the instrument absorbance response to known concentration standards. Alkaline phosphatase activity was calculated based on fluorescence response of a known standard.<\/p>\n Any samples that were not collected are represented by 'nd' in the dataset.<\/p>\n BCO-DMO Processing:
\nCell samples (5 mL culture volume) for particulate P were collected onto precombusted 25 mm Whatman glass fiber filters, rinsed with 0.17 M Na\u2082SO\u2084, and stored frozen at -20\u00b0C until analysis. Determinations were made as previously described (Lomas et al. 2010). Briefly, filters were rinsed with 0.017 M MgSO\u2084, dried at 90\u00b0C, and combusted at 500\u00b0C for 2 h. Upon cooling, 0.2 M HCl was added and hydrolyzed at 80\u00b0C for 30 min. After cooling, mixed reagent was added, the samples were centrifuged, and absorbance was read at 885 nm using a Genesys 10 spectrophotometer.<\/p>\n
\n- renamed fields;
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Data reported are cell abundance, growth rate, cellular P quotas, and alkaline phosphatase activity.","@language":"en-US"}],"http:\/\/purl.org\/dc\/terms\/rights":[{"@id":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"http:\/\/ocean-data.org\/schema\/deprecated":[{"@value":"false","@type":"xsd:boolean"}],"http:\/\/purl.org\/dc\/terms\/bibliographicCitation":[{"@value":"Whitney, L., Lomas, M. (2020) Cell abundance, growth rate, cellular P quotas, and alkaline phosphatase activity from a laboratory experiment examining the response of three species of marine phytoplankton grown under different phosphorus (P) conditions. Biological and Chemical Oceanography Data Management Office (BCO-DMO). 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