{
"METABOLOMICS WORKBENCH":{"STUDY_ID":"ST000921","ANALYSIS_ID":"AN001510","VERSION":"1","CREATED_ON":"January 19, 2018, 1:56 pm"},

"PROJECT":{"PROJECT_TITLE":"Karenia brevis allelopathy compromises the lipidome, membrane integrity, and photosynthetic efficiency of competitors","PROJECT_TYPE":"Untargeted Lipidomics","PROJECT_SUMMARY":"Comparing effects on lipidome of phytoplankton competitors based on exposure to K. Brevis","INSTITUTE":"Georgia Institute of Technology","DEPARTMENT":"Chemistry","LABORATORY":"Fernández","LAST_NAME":"Hogan","FIRST_NAME":"Scott","ADDRESS":"901 Atlantic Drive, Atlanta, GA, 30332, USA","EMAIL":"srjhogan@gatech.edu","PHONE":"2156924657"},

"STUDY":{"STUDY_TITLE":"Karenia brevis allelopathy compromises the lipidome, membrane integrity, and photosynthetic efficiency of competitors","STUDY_TYPE":"Untargeted lipidomics","STUDY_SUMMARY":"Allelopathy, or the release of compounds that inhibit competitors, is a form of interference competition that is common among bloom-forming phytoplankton. Allelopathy is hypothesized to play a role in bloom propagation and maintenance and is well established in the red tide dinoflagellate Karenia brevis. K. brevis typically suppresses competitor growth through unknown mechanisms over the course of many days. When we investigated the effects of allelopathy on the lipidomes of two competing phytoplankton, Asterionellopsis glacialis and Thalassiosira pseudonana using nuclear magnetic resonance (NMR) spectroscopy and mass spectrometry (MS)- based metabolomics, we found that the lipidomes of both species were significantly altered, however A. glacialis maintained a more robust response whereas T. pseudonana saw significant alterations in fatty acid synthesis, cell membrane integrity, and a decrease in photosynthetic efficiency. Membrane- associated lipids were significantly suppressed for T. pseudonana exposed to allelopathy to the point of permeabilizing the cell membrane of living cells. The dominant mechanisms of K. brevis allelopathy appear to target lipid biosynthesis affecting multiple physiological pathways suggesting that exuded compounds have the ability to significantly alter competitor physiology and give K. brevis a competitive edge over sensitive species.","INSTITUTE":"Georgia Institute of Technology","DEPARTMENT":"Chemistry","LABORATORY":"Fernández","LAST_NAME":"Hogan","FIRST_NAME":"Scott","ADDRESS":"901 Atlantic Drive, Atlanta, GA, 30332, USA","EMAIL":"srjhogan@gatech.edu","PHONE":"2156924657","NUM_GROUPS":"4","TOTAL_SUBJECTS":"51"},

"SUBJECT":{"SUBJECT_TYPE":"Plankton","SUBJECT_SPECIES":"Thalassiosira pseudonana;Karenia brevis"},
"SUBJECT_SAMPLE_FACTORS":[
{
"Subject ID":"-",
"Sample ID":"AgC2",
"Factors":{"Class":"Control"}
},
{
"Subject ID":"-",
"Sample ID":"AgC10",
"Factors":{"Class":"Control"}
},
{
"Subject ID":"-",
"Sample ID":"AgC11",
"Factors":{"Class":"Control"}
},
{
"Subject ID":"-",
"Sample ID":"AgC6",
"Factors":{"Class":"Control"}
},
{
"Subject ID":"-",
"Sample ID":"AgC1",
"Factors":{"Class":"Control"}
},
{
"Subject ID":"-",
"Sample ID":"AgC3",
"Factors":{"Class":"Control"}
},
{
"Subject ID":"-",
"Sample ID":"AgC5",
"Factors":{"Class":"Control"}
},
{
"Subject ID":"-",
"Sample ID":"AgC4",
"Factors":{"Class":"Control"}
},
{
"Subject ID":"-",
"Sample ID":"AgC8",
"Factors":{"Class":"Control"}
},
{
"Subject ID":"-",
"Sample ID":"AgC15",
"Factors":{"Class":"Control"}
},
{
"Subject ID":"-",
"Sample ID":"AgC9",
"Factors":{"Class":"Control"}
},
{
"Subject ID":"-",
"Sample ID":"AgC7",
"Factors":{"Class":"Control"}
},
{
"Subject ID":"-",
"Sample ID":"AgC12",
"Factors":{"Class":"Control"}
},
{
"Subject ID":"-",
"Sample ID":"AgT2",
"Factors":{"Class":"treatment"}
},
{
"Subject ID":"-",
"Sample ID":"AgT15",
"Factors":{"Class":"treatment"}
},
{
"Subject ID":"-",
"Sample ID":"AgT8",
"Factors":{"Class":"treatment"}
},
{
"Subject ID":"-",
"Sample ID":"AgT7",
"Factors":{"Class":"treatment"}
},
{
"Subject ID":"-",
"Sample ID":"AgT10",
"Factors":{"Class":"treatment"}
},
{
"Subject ID":"-",
"Sample ID":"AgT9",
"Factors":{"Class":"treatment"}
},
{
"Subject ID":"-",
"Sample ID":"AgT5",
"Factors":{"Class":"treatment"}
},
{
"Subject ID":"-",
"Sample ID":"AgT12",
"Factors":{"Class":"treatment"}
},
{
"Subject ID":"-",
"Sample ID":"AgT6",
"Factors":{"Class":"treatment"}
},
{
"Subject ID":"-",
"Sample ID":"AgT4",
"Factors":{"Class":"treatment"}
},
{
"Subject ID":"-",
"Sample ID":"AgT3",
"Factors":{"Class":"treatment"}
},
{
"Subject ID":"-",
"Sample ID":"AgT11",
"Factors":{"Class":"treatment"}
},
{
"Subject ID":"-",
"Sample ID":"AgT1",
"Factors":{"Class":"treatment"}
},
{
"Subject ID":"-",
"Sample ID":"TpC2",
"Factors":{"Class":"Control"}
},
{
"Subject ID":"-",
"Sample ID":"TpC7",
"Factors":{"Class":"Control"}
},
{
"Subject ID":"-",
"Sample ID":"TpC5",
"Factors":{"Class":"Control"}
},
{
"Subject ID":"-",
"Sample ID":"TpC10",
"Factors":{"Class":"Control"}
},
{
"Subject ID":"-",
"Sample ID":"TpC8",
"Factors":{"Class":"Control"}
},
{
"Subject ID":"-",
"Sample ID":"TpC3",
"Factors":{"Class":"Control"}
},
{
"Subject ID":"-",
"Sample ID":"TpC15",
"Factors":{"Class":"Control"}
},
{
"Subject ID":"-",
"Sample ID":"TpC11",
"Factors":{"Class":"Control"}
},
{
"Subject ID":"-",
"Sample ID":"TpC6",
"Factors":{"Class":"Control"}
},
{
"Subject ID":"-",
"Sample ID":"TpC4",
"Factors":{"Class":"Control"}
},
{
"Subject ID":"-",
"Sample ID":"TpC12",
"Factors":{"Class":"Control"}
},
{
"Subject ID":"-",
"Sample ID":"TpC9",
"Factors":{"Class":"Control"}
},
{
"Subject ID":"-",
"Sample ID":"TpC13",
"Factors":{"Class":"Control"}
},
{
"Subject ID":"-",
"Sample ID":"TpT8",
"Factors":{"Class":"treatment"}
},
{
"Subject ID":"-",
"Sample ID":"TpT5",
"Factors":{"Class":"treatment"}
},
{
"Subject ID":"-",
"Sample ID":"TpT13",
"Factors":{"Class":"treatment"}
},
{
"Subject ID":"-",
"Sample ID":"TpT10",
"Factors":{"Class":"treatment"}
},
{
"Subject ID":"-",
"Sample ID":"TpT2",
"Factors":{"Class":"treatment"}
},
{
"Subject ID":"-",
"Sample ID":"TpT9",
"Factors":{"Class":"treatment"}
},
{
"Subject ID":"-",
"Sample ID":"TpT4",
"Factors":{"Class":"treatment"}
},
{
"Subject ID":"-",
"Sample ID":"TpT7",
"Factors":{"Class":"treatment"}
},
{
"Subject ID":"-",
"Sample ID":"TpT3",
"Factors":{"Class":"treatment"}
},
{
"Subject ID":"-",
"Sample ID":"TpT6",
"Factors":{"Class":"treatment"}
},
{
"Subject ID":"-",
"Sample ID":"TpT15",
"Factors":{"Class":"treatment"}
},
{
"Subject ID":"-",
"Sample ID":"TpT11",
"Factors":{"Class":"treatment"}
}
],
"COLLECTION":{"COLLECTION_SUMMARY":"Briefly, diatoms Thalassiosira pseudonana strain CCMP 1335 and Asterionellopsis glacialis strain CCMP 137 were grown in silicate-amended L1 media in artificial seawater (Instant Ocean, 35 ppt). Karenia brevis strain CCMP 2228 was cultured in similar conditions above with L1 media-amended artificial seawater. All cultures were maintained at 21 ˚C with a 12:12 light/dark cycle and an irradiance of 100-145 µmol/m2s in a Percival incubator (Biospherical Instrument QSL2100).","SAMPLE_TYPE":"Cell extract"},

"TREATMENT":{"TREATMENT_SUMMARY":"To expose diatoms to competition with allelopathic K. brevis, K. brevis was co-cultured with each of the two diatom species (n=14 per species). K. brevis was grown inside a permeable dialysis membrane to allow for exchange of exuded allelopathic compounds without direct interaction of K. brevis and diatom cells, which were grown in flasks in which the dialysis tubes were placed. Control cultures consisted of dialysis membranes (molecular weight cutoff, 50 kDa) filled with L1 media diluted to conditions similar to that of exponential growth phase K. brevis (n = 15 per diatom species) in place of diatom species. This co-culture experiment was halted once competitor cultures reached exponential growth stage, which was 6 d for T. pseudonana and 8 d for A. glacialis, after which diatom cells were filtered onto GF/C filters (Whatman #1922-110, muffled at 450 ˚C for 3 h) and dipped into liquid nitrogen to quench intracellular metabolism."},

"SAMPLEPREP":{"SAMPLEPREP_SUMMARY":"To separate polar and lipid intracellular metabolites, dried extracts were dissolved in a biphasic mixture of 9:10:15 water/methanol/chloroform. The more lipophilic layer was removed and washed twice with 9:10 water/methanol. Lipid extracts were reconstituted in 200 μL 2-propanol. Quantitative metabolomics data were acquired using a Waters Xevo G2 QTOF mass spectrometer."},

"CHROMATOGRAPHY":{"CHROMATOGRAPHY_TYPE":"Reversed phase","INSTRUMENT_NAME":"Waters Synapt G2 QTOF","COLUMN_NAME":"Waters Acquity BEH C18 (50 x 2.1mm, 1.7um)","FLOW_GRADIENT":"0-1 min, 70% B; 1-3 min, 75% B; 3-6 min, 80% B; 6-10 min, 90% B; 10-14 min, 100% B.","FLOW_RATE":".3 mL/min","COLUMN_TEMPERATURE":"60","SOLVENT_A":"water: acetonitrile (40:60) +10 mM ammonium formate + 0.1% formic acid","SOLVENT_B":"10% acetonitrile in 2-propanol +10 mM ammonium formate + 0.1% formic acid"},

"ANALYSIS":{"ANALYSIS_TYPE":"MS"},

"MS":{"MS_COMMENTS":"-","INSTRUMENT_NAME":"Waters Synapt G2 QTOF","INSTRUMENT_TYPE":"QTOF","MS_TYPE":"ESI","ION_MODE":"NEGATIVE","CAPILLARY_VOLTAGE":"2.0","SOURCE_TEMPERATURE":"90°C","DESOLVATION_GAS_FLOW":"600 L/h","DESOLVATION_TEMPERATURE":"250 °C","MS_RESULTS_FILE":"ST000921_AN001510_Results.txt UNITS:Normalized Abundance"}

}