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Photosynthetic microbes provide food for nearly all other life in the ocean. Their metabolism produces organic molecules called metabolites that can leak out of cells, be intentionally excreted into seawater, or be released during cell death. Once outside the cell, these metabolites are the basis for specific interactions among microbes and determine community structure and activity. Yet, current understanding of metabolites in the ocean is limited by a historical lack of ability to measure them. The work proposed here will expand current knowledge of metabolite structures, concentrations, and production rates using recently developed analytical methods. These methods have already led to the discovery that homarine, a substituted pyridine first found in lobster in 1933, is the most abundant detectable metabolite in microbial communities of the North Pacific Ocean. While homarine is known as a predator deterrent, osmoprotectant, methyl donor, and antibiofouling agent, studies of its role in microbial community dynamics are lacking. The work proposed will clarify the role of homarine in the ocean’s microbial communities. This work will create an open-source metabolite database that will serve the broader field of metabolomics, a growing area in environmental, engineering, and medical sciences. This collaboration will also promote the careers of a graduate student and a postdoctoral researcher as well as an early career professor from an underrepresented group at a primarily undergraduate institution (PUI). Undergraduates from both institutions will contribute to project development and implementation, local cruises on the R.V. Carson, lab work, and dissemination of results. This research will be integrated into a curriculum-based research experience for undergraduates in a 200-level genetics course at the PUI, University of Puget Sound.&lt;/p&gt;
&lt;p&gt;The proposed work will carry out field studies and laboratory experiments to test the hypothesis that metabolites are quantitatively significant forms of carbon and nitrogen flowing through microbial communities. The identity, quantity, and production rates of metabolites will also be determined. For homarine, the enzymes and organisms responsible for its transformations will be determined. Specific proposed activities will 1) Quantify nitrogenous metabolite pools and their net production rates (particulate and dissolved) in phytoplankton cultures and in marine surface water communities; 2) Isolate homarine consuming heterotrophic bacteria and use mutagenesis techniques, transcriptomics, and stable isotope assisted metabolomics to annotate genes and characterize the biochemical reactions involved in the degradation of homarine; 3) Carry out incubations of stable isotope labeled homarine in phytoplankton cultures, heterotrophic bacterial cultures sensitive to homarine, and natural communities to quantitatively evaluate the effect of homarine on growth, track homarine through metabolic pathways, and determine the kinetics of homarine uptake; 4) Identify homarine consumers and biochemical pathways for homarine use in the environment by mining existing environmental metatranscriptomes for homarine catabolism genes. The combination of these approaches will provide better understanding of the flow of nitrogen containing metabolites through marine microbial ecosystems. Results from this work will be disseminated through peer reviewed open-source publications as well as presentations to the scientific community and the general public.&lt;/p&gt;
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	Name: Date
	Units: unitless
	Description: &lt;p&gt;Date of CTD casts  (UTC)&lt;/p&gt; 
http://lod.bco-dmo.org/id/dataset-parameter/997387.rdf
	Name: Cruise
	Units: unitless
	Description: &lt;p&gt;Cruise ID&lt;/p&gt; 
http://lod.bco-dmo.org/id/dataset-parameter/997388.rdf
	Name: Station
	Units: unitless
	Description: &lt;p&gt;Cruise station number&lt;/p&gt; 
http://lod.bco-dmo.org/id/dataset-parameter/997389.rdf
	Name: LAT
	Units: decimal degrees
	Description: &lt;p&gt;Latitude, positive is North&lt;/p&gt; 
http://lod.bco-dmo.org/id/dataset-parameter/997391.rdf
	Name: LON
	Units: decimal degrees
	Description: &lt;p&gt;Longitude, negative is West&lt;/p&gt; 
http://lod.bco-dmo.org/id/dataset-parameter/997393.rdf
	Name: Cast
	Units: unitless
	Description: &lt;p&gt;Station cast number&lt;/p&gt; 
http://lod.bco-dmo.org/id/dataset-parameter/997394.rdf
	Name: PRS
	Units: db
	Description: &lt;p&gt;Pressure&lt;/p&gt; 
http://lod.bco-dmo.org/id/dataset-parameter/997395.rdf
	Name: Depth
	Units: meters
	Description: &lt;p&gt;Depth&lt;/p&gt; 
http://lod.bco-dmo.org/id/dataset-parameter/997397.rdf
	Name: TMP
	Units: degrees Celsius
	Description: &lt;p&gt;Temperature (ITS-90)&lt;/p&gt; 
http://lod.bco-dmo.org/id/dataset-parameter/997398.rdf
	Name: SAL
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	Description: &lt;p&gt;Salinity, Practical&lt;/p&gt; 
http://lod.bco-dmo.org/id/dataset-parameter/997399.rdf
	Name: OXY
	Units: milligrams per liter (mg/l)
	Description: &lt;p&gt;Oxygen, SBE 43&lt;/p&gt; 
http://lod.bco-dmo.org/id/dataset-parameter/997400.rdf
	Name: OXYsat
	Units: milligrams per liter (mg/l)
	Description: &lt;p&gt;Oxygen saturation, Garcia &amp;amp; Gordon&lt;/p&gt; 
http://lod.bco-dmo.org/id/dataset-parameter/997401.rdf
	Name: Beam_Attenuation
	Units: per meter
	Description: &lt;p&gt;Beam attenuation&lt;/p&gt; 
http://lod.bco-dmo.org/id/dataset-parameter/997402.rdf
	Name: Beam_Transmission
	Units: percent
	Description: &lt;p&gt;Beam transmission&lt;/p&gt; 
http://lod.bco-dmo.org/id/dataset-parameter/997403.rdf
	Name: Fluorescence
	Units: milligrams per cubic meter (mg/m^3)
	Description: &lt;p&gt;Fluorescence, WET Labs ECO-AFL/FL&lt;/p&gt; 
http://lod.bco-dmo.org/id/dataset-parameter/997404.rdf
	Name: PAR
	Units: micromoles photons per square meter per second (umol photons/m^2/s^1)
	Description: &lt;p&gt;PAR/ Irradiance, Biospherical/Licor&lt;/p&gt; 
http://lod.bco-dmo.org/id/dataset-parameter/997405.rdf
	Name: pH
	Units: unitless
	Description: &lt;p&gt;pH&lt;/p&gt; 
http://lod.bco-dmo.org/id/dataset-parameter/997406.rdf
	Name: filename_cnv
	Units: unitless
	Description: &lt;p&gt;Original SBE .cnv CTD profile file name with one meter bins&lt;/p&gt; 
http://lod.bco-dmo.org/id/dataset-parameter/997407.rdf
	Name: filename_csv
	Units: unitless
	Description: &lt;p&gt;Filename of .csv files converted from the Original SBE .cnv CTD profile files with one meter bins&lt;/p&gt; 
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                  <gco:CharacterString>- Loaded 65 CSV files from the one_meter_binned directory using filenames as resource names; missing values defined as empty strings and &amp;quot;nd&amp;quot;
- Concatenated all 65 resources into a single resource named ctd_all, mapping fields: year_month_day, Cruise, Station, Cast, LAT, LON, PRS, Depth, TMP, SAL, OXY, OXYsat, Beam.Attenuation, Beam.Transmission, Fluorescence, PAR, pH, and file_name; source file paths added as new column path_name
- Applied find/replace on path_name to strip directory path, retaining only the filename (e.g., RC0078_S3_C2.csv)
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- Reordered fields to move both filename fields to the end: Date, Cruise, Station, Cast, LAT, LON, PRS, Depth, TMP, SAL, OXY, OXYsat, Beam.Attenuation, Beam.Transmission, Fluorescence, PAR, pH, filename_cnv, filename_csv
- Renamed year_month_day to Date
- Renamed Beam.Attenuation to Beam_Attenuation and Beam.Transmission to Beam_Transmission by replacing dots with underscores
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- Corrected LAT values at rows 2013 and 2014 by negating them (0 - LAT)
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