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            <gco:CharacterString>Cite this dataset as: Duhamel, S., Diaz, J., Djaoudi, K., Waggoner, E. (2024) Hydrolysis rates from dissolved organic phosphorus (DOP) hydrolysis experiments with marine cyanobacterium Synechococcus laboratory cultures (WH8102 and WH5701) from 2018-2023. Biological and Chemical Oceanography Data Management Office (BCO-DMO). (Version 1) Version Date 2024-05-28 [if applicable, indicate subset used]. doi:10.26008/1912/bco-dmo.928984.1 [access date]</gco:CharacterString>
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        <gco:CharacterString>Dataset Description: &amp;lt;p&amp;gt;This dataset was utilized for&amp;amp;nbsp;Waggoner et al. (2024) Figure 2 and supplementary figure 2.&amp;amp;nbsp;See &amp;quot;Related Datasets&amp;quot; section on this page for other closely-related data from this study published in Waggoner et al. (2024).&amp;amp;nbsp; They are also listed under the&amp;amp;nbsp;BCO-DMO Project Page: https://www.bco-dmo.org/project/747715.&amp;lt;/p&amp;gt; Methods and Sampling: &amp;lt;p&amp;gt;&amp;lt;em&amp;gt;Synechococcus Growth&amp;lt;/em&amp;gt;–&amp;amp;nbsp;Axenic&amp;amp;nbsp;&amp;lt;em&amp;gt;Synechococcus&amp;lt;/em&amp;gt;&amp;amp;nbsp;WH8102 (open ocean strain) and WH5701 (coastal strain) were obtained from the National Center for Marine Algae and Microbiota (NCMA, Bigelow Laboratories, East Boothbay, Maine). Both strains were grown in batch culture using SN media (Waterbury&amp;amp;nbsp;&amp;lt;em&amp;gt;et al.&amp;lt;/em&amp;gt;&amp;amp;nbsp;1986) made with aged, filtered (0.2 µm), and autoclaved (120°C, 30 minutes) seawater from station ALOHA (A Long-term Oligotrophic Habitat Assessment). At the late-exponential phase, cultures were transferred in triplicate to one of two SN media: (1) +P&amp;lt;em&amp;gt;i&amp;lt;/em&amp;gt;&amp;amp;nbsp;(45 µmol L&amp;lt;sup&amp;gt;-1&amp;lt;/sup&amp;gt;&amp;amp;nbsp;KH&amp;lt;sub&amp;gt;2&amp;lt;/sub&amp;gt;PO&amp;lt;sub&amp;gt;4&amp;lt;/sub&amp;gt;, following Waterbury&amp;amp;nbsp;&amp;lt;em&amp;gt;et al.&amp;lt;/em&amp;gt;&amp;amp;nbsp;(1986)) and (2) -P&amp;lt;em&amp;gt;i&amp;lt;/em&amp;gt;&amp;amp;nbsp;(no KH&amp;lt;sub&amp;gt;2&amp;lt;/sub&amp;gt;PO&amp;lt;sub&amp;gt;4&amp;lt;/sub&amp;gt;&amp;amp;nbsp;added; P&amp;lt;em&amp;gt;i&amp;lt;/em&amp;gt;&amp;amp;nbsp;below detection limit). All cultures were incubated at 25°C on a 12h:12h light cycle at 130 µmol m&amp;lt;sup&amp;gt;-1&amp;lt;/sup&amp;gt;&amp;amp;nbsp;s&amp;lt;sup&amp;gt;-1&amp;lt;/sup&amp;gt;&amp;amp;nbsp;in sterile culture flasks with a vent cap (0.22 µm hydrophobic membrane).&amp;lt;/p&amp;gt;

&amp;lt;p&amp;gt;&amp;lt;em&amp;gt;DOP Hydrolysis&amp;lt;/em&amp;gt;– Representative phosphoester (P-ester) and polyphosphate (PolyP) hydrolysis rates were determined as in Diaz&amp;amp;nbsp;&amp;lt;em&amp;gt;et al.&amp;lt;/em&amp;gt;&amp;amp;nbsp;(2018). The nucleotide AMP was used as a model P-ester, 3-PolyP as a model PolyP, and ATP as a representative containing both P-ester and P-anhydride bonds. Two separate experiments were conducted with both&amp;amp;nbsp;&amp;lt;em&amp;gt;Synechococcus&amp;amp;nbsp;&amp;lt;/em&amp;gt;strains to determine DOP hydrolysis in the presence (whole cell) and absence (cell-free filtrate) of cells (Diaz&amp;amp;nbsp;&amp;lt;em&amp;gt;et al.&amp;lt;/em&amp;gt;&amp;amp;nbsp;2018, 2019). The +P&amp;lt;em&amp;gt;i&amp;lt;/em&amp;gt;&amp;amp;nbsp;and -P&amp;lt;em&amp;gt;i&amp;lt;/em&amp;gt;&amp;amp;nbsp;cultures were subsampled approximately every three days over ~20 days to obtain whole cell and cell-free DOP hydrolysis results along each phase of the cellular growth curve. To prepare cell-free filtrates,&amp;lt;em&amp;gt;&amp;amp;nbsp;&amp;lt;/em&amp;gt;the +P&amp;lt;em&amp;gt;i&amp;lt;/em&amp;gt;&amp;amp;nbsp;and -P&amp;lt;em&amp;gt;i&amp;lt;/em&amp;gt;&amp;amp;nbsp;culture aliquots were aseptically filtered (0.2 µm) at each subsampling day.&amp;amp;nbsp;To determine DOP hydrolysis rates, the production of P&amp;lt;em&amp;gt;i&amp;lt;/em&amp;gt;&amp;amp;nbsp;was measured over time. Aliquots (200 µL) of +P&amp;lt;em&amp;gt;i&amp;lt;/em&amp;gt;&amp;amp;nbsp;and -P&amp;lt;em&amp;gt;i&amp;lt;/em&amp;gt;&amp;amp;nbsp;treatments were amended with a single DOP substrate (3-PolyP, ATP or AMP; 20 µmol L&amp;lt;sup&amp;gt;-1&amp;lt;/sup&amp;gt;&amp;amp;nbsp;P, final concentration) in triplicate wells of a non-treated standard 96-well transparent microplate and incubated over 6-hours.&amp;amp;nbsp;An unamended control treatment to track P&amp;lt;em&amp;gt;i&amp;lt;/em&amp;gt;&amp;amp;nbsp;concentrations in the cultures over time were prepared in triplicate and monitored in parallel.&amp;amp;nbsp;Pi (or soluble reactive phosphorus) was measured using a standard colorimetric protocol (Hansen and Koroleff 1999) on a multimode plate reader (SpectraMax® M2, Molecular Devices). Absorbance read at 880 nm was calibrated using a standard curve of monopotassium phosphate (0, 0.5, 1, 2, 5, 10, 20, 40, 50, 75 µmol L&amp;lt;sup&amp;gt;-1&amp;lt;/sup&amp;gt;; KH&amp;lt;sub&amp;gt;2&amp;lt;/sub&amp;gt;PO&amp;lt;sub&amp;gt;4&amp;lt;/sub&amp;gt;). The average detection limit of P&amp;lt;em&amp;gt;i&amp;lt;/em&amp;gt;&amp;amp;nbsp;using this method, defined as three times the standard deviation of the triplicate blank measurements, was 0.125 ± 0.005 µmol L&amp;lt;sup&amp;gt;-1&amp;lt;/sup&amp;gt;. The calibration curve was prepared with 0.2 µm filtered ALOHA seawater with a P&amp;lt;em&amp;gt;i&amp;lt;/em&amp;gt;&amp;amp;nbsp;background concentration below the detection limit.&amp;lt;br /&amp;gt;
&amp;lt;br /&amp;gt;
&amp;lt;strong&amp;gt;Organism identifiers &amp;lt;/strong&amp;gt;(Life Science Identifier, LSID):&amp;lt;br /&amp;gt;
Synechococcus, urn:lsid:marinespecies.org:taxname:160572&amp;lt;/p&amp;gt;</gco:CharacterString>
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        <gmx:Anchor xlink:href="http://lod.bco-dmo.org/id/award/747714.rdf" xlink:title="OCE-1736967" xlink:actuate="onRequest">Funding provided by NSF Division of Ocean Sciences (NSF OCE) Award Number: OCE-1736967 Award URL: https://www.nsf.gov/awardsearch/show-award?AWD_ID=1736967</gmx:Anchor>
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        <gmx:Anchor xlink:href="http://lod.bco-dmo.org/id/award/747743.rdf" xlink:title="OCE-1737083" xlink:actuate="onRequest">Funding provided by NSF Division of Ocean Sciences (NSF OCE) Award Number: OCE-1737083 Award URL: https://www.nsf.gov/awardsearch/show-award?AWD_ID=1737083</gmx:Anchor>
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        <gmx:Anchor xlink:href="http://lod.bco-dmo.org/id/award/856540.rdf" xlink:title="OCE-2001212" xlink:actuate="onRequest">Funding provided by NSF Division of Ocean Sciences (NSF OCE) Award Number: OCE-2001212  Award URL: https://www.nsf.gov/awardsearch/show-award?AWD_ID=2001212</gmx:Anchor>
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        <gmx:Anchor xlink:href="http://lod.bco-dmo.org/id/award/856541.rdf" xlink:title="OCE-1948042" xlink:actuate="onRequest">Funding provided by NSF Division of Ocean Sciences (NSF OCE) Award Number: OCE-1948042 Award URL: https://www.nsf.gov/awardsearch/show-award?AWD_ID=1948042</gmx:Anchor>
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Phosphorus (P) is an essential building block for life. Because P is in short supply over vast areas of the ocean, P availability may control biological productivity, such as photosynthesis and carbon fixation, which has implications for uptake of the greenhouse gas carbon dioxide and thus climate regulation. Marine microorganisms must satisfy their nutritional requirement for P by obtaining it from seawater, where P is present in a variety of chemical forms, from simple phosphate ions (Pi) to complex dissolved organic phosphorus (DOP) molecules. The concentration of DOP vastly exceeds Pi over most ocean areas, therefore DOP is a critically important source of P for marine microbial nutrition and productivity. However, much remains unknown about the contribution of specific DOP compounds to the P nutrition, productivity, and structure of marine microbial communities. In this project, the investigators will conduct field experiments in the Atlantic Ocean and perform a series of controlled laboratory studies with pure enzymes and microbial cultures to determine how and to what extent different DOP compounds are degraded to Pi in the marine environment. Furthermore, the contribution of these compound-specific DOP molecules to microbial P nutrition, carbon fixation, and community structure will be determined, thus advancing the current state of knowledge regarding the factors that control the activity and distribution of microbial species in the ocean, and the ocean?s role in the climate system. This project will support two female junior investigators, a postdoctoral researcher, and graduate and undergraduate students. The undergraduate students will be recruited from the Marine Sciences program at Savannah State University, an Historically Black Colleges and Universities. In addition, results will be incorporated into new hands-on K-12 educational tools to teach students about microbial P biogeochemistry, including a digital game and formal lesson plans with hands-on demos. These tools will be validated with K-12 educators and will be widely accessible to the public through various well-known online platforms. These activities will thus reach a broad audience including a significant fraction of underrepresented groups.&lt;/p&gt;
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http://lod.bco-dmo.org/id/dataset-parameter/929191.rdf
	Name: hydrolysis_rates_trip1
	Units: attomols per cel per hour (amol cell -1 hr -1)
	Description: &lt;p&gt;hydrolysis rate for triplicate culture flask #1. 0 indicates no hydrolysis measured.&lt;/p&gt; 
http://lod.bco-dmo.org/id/dataset-parameter/929192.rdf
	Name: hydrolysis_rates_trip2
	Units: attomols per cel per hour (amol cell -1 hr -1)
	Description: &lt;p&gt;hydrolysis rate for triplicate culture flask #2. 0 indicates no hydrolysis measured.&lt;/p&gt; 
http://lod.bco-dmo.org/id/dataset-parameter/929193.rdf
	Name: hydrolysis_rates_trip3
	Units: attomols per cel per hour (amol cell -1 hr -1)
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&amp;lt;p&amp;gt;&amp;lt;em&amp;gt;DOP Hydrolysis&amp;lt;/em&amp;gt;– Representative phosphoester (P-ester) and polyphosphate (PolyP) hydrolysis rates were determined as in Diaz&amp;amp;nbsp;&amp;lt;em&amp;gt;et al.&amp;lt;/em&amp;gt;&amp;amp;nbsp;(2018). The nucleotide AMP was used as a model P-ester, 3-PolyP as a model PolyP, and ATP as a representative containing both P-ester and P-anhydride bonds. Two separate experiments were conducted with both&amp;amp;nbsp;&amp;lt;em&amp;gt;Synechococcus&amp;amp;nbsp;&amp;lt;/em&amp;gt;strains to determine DOP hydrolysis in the presence (whole cell) and absence (cell-free filtrate) of cells (Diaz&amp;amp;nbsp;&amp;lt;em&amp;gt;et al.&amp;lt;/em&amp;gt;&amp;amp;nbsp;2018, 2019). The +P&amp;lt;em&amp;gt;i&amp;lt;/em&amp;gt;&amp;amp;nbsp;and -P&amp;lt;em&amp;gt;i&amp;lt;/em&amp;gt;&amp;amp;nbsp;cultures were subsampled approximately every three days over ~20 days to obtain whole cell and cell-free DOP hydrolysis results along each phase of the cellular growth curve. To prepare cell-free filtrates,&amp;lt;em&amp;gt;&amp;amp;nbsp;&amp;lt;/em&amp;gt;the +P&amp;lt;em&amp;gt;i&amp;lt;/em&amp;gt;&amp;amp;nbsp;and -P&amp;lt;em&amp;gt;i&amp;lt;/em&amp;gt;&amp;amp;nbsp;culture aliquots were aseptically filtered (0.2 µm) at each subsampling day.&amp;amp;nbsp;To determine DOP hydrolysis rates, the production of P&amp;lt;em&amp;gt;i&amp;lt;/em&amp;gt;&amp;amp;nbsp;was measured over time. Aliquots (200 µL) of +P&amp;lt;em&amp;gt;i&amp;lt;/em&amp;gt;&amp;amp;nbsp;and -P&amp;lt;em&amp;gt;i&amp;lt;/em&amp;gt;&amp;amp;nbsp;treatments were amended with a single DOP substrate (3-PolyP, ATP or AMP; 20 µmol L&amp;lt;sup&amp;gt;-1&amp;lt;/sup&amp;gt;&amp;amp;nbsp;P, final concentration) in triplicate wells of a non-treated standard 96-well transparent microplate and incubated over 6-hours.&amp;amp;nbsp;An unamended control treatment to track P&amp;lt;em&amp;gt;i&amp;lt;/em&amp;gt;&amp;amp;nbsp;concentrations in the cultures over time were prepared in triplicate and monitored in parallel.&amp;amp;nbsp;Pi (or soluble reactive phosphorus) was measured using a standard colorimetric protocol (Hansen and Koroleff 1999) on a multimode plate reader (SpectraMax® M2, Molecular Devices). Absorbance read at 880 nm was calibrated using a standard curve of monopotassium phosphate (0, 0.5, 1, 2, 5, 10, 20, 40, 50, 75 µmol L&amp;lt;sup&amp;gt;-1&amp;lt;/sup&amp;gt;; KH&amp;lt;sub&amp;gt;2&amp;lt;/sub&amp;gt;PO&amp;lt;sub&amp;gt;4&amp;lt;/sub&amp;gt;). The average detection limit of P&amp;lt;em&amp;gt;i&amp;lt;/em&amp;gt;&amp;amp;nbsp;using this method, defined as three times the standard deviation of the triplicate blank measurements, was 0.125 ± 0.005 µmol L&amp;lt;sup&amp;gt;-1&amp;lt;/sup&amp;gt;. The calibration curve was prepared with 0.2 µm filtered ALOHA seawater with a P&amp;lt;em&amp;gt;i&amp;lt;/em&amp;gt;&amp;amp;nbsp;background concentration below the detection limit.&amp;lt;br /&amp;gt;
&amp;lt;br /&amp;gt;
&amp;lt;strong&amp;gt;Organism identifiers &amp;lt;/strong&amp;gt;(Life Science Identifier, LSID):&amp;lt;br /&amp;gt;
Synechococcus, urn:lsid:marinespecies.org:taxname:160572&amp;lt;/p&amp;gt;</gco:CharacterString>
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