#METABOLOMICS WORKBENCH TruxalCarlson_20201022_124653 DATATRACK_ID:2221 STUDY_ID:ST001514 ANALYSIS_ID:AN002510 PROJECT_ID:PR001021 VERSION 1 CREATED_ON October 30, 2020, 11:03 am #PROJECT PR:PROJECT_TITLE Community metabolomes reflect taxon-specific fingerprints of phytoplankton in PR:PROJECT_TITLE the ocean PR:PROJECT_TYPE Marine Metabolomics PR:PROJECT_SUMMARY Phytoplankton transform inorganic carbon into thousands of biomolecules, PR:PROJECT_SUMMARY including polar metabolites that represent an important pool of labile fixed PR:PROJECT_SUMMARY carbon, nitrogen, and sulfur. Metabolite production is not identical among PR:PROJECT_SUMMARY phytoplankton, and the flux of these molecules through the microbial loop PR:PROJECT_SUMMARY depends on compound-specific bioavailability to a wider microbial community. Yet PR:PROJECT_SUMMARY relatively little is known about the diversity or concentration of polar PR:PROJECT_SUMMARY metabolites within marine plankton. Here we evaluate 313 metabolites in 21 PR:PROJECT_SUMMARY phytoplankton species and in natural marine particles across environmental PR:PROJECT_SUMMARY gradients to show that bulk community metabolomes reflect the phytoplankton PR:PROJECT_SUMMARY community on a chemical level. PR:INSTITUTE University of Washington PR:DEPARTMENT Oceanography PR:LABORATORY Ingalls Lab PR:LAST_NAME Heal PR:FIRST_NAME Katherine PR:ADDRESS 1501 NE Boat Street, Marine Science Building, Room G, Seattle, WA, 98195, USA PR:EMAIL kheal@uw.edu PR:PHONE 612-616-4840 #STUDY ST:STUDY_TITLE Community metabolomes reflect taxon-specific fingerprints of phytoplankton in ST:STUDY_TITLE the ocean ST:STUDY_TYPE Metabolomic survey of 21 phytoplankton species ST:STUDY_SUMMARY Phytoplankton transform inorganic carbon into thousands of biomolecules, ST:STUDY_SUMMARY including polar metabolites that represent an important pool of labile fixed ST:STUDY_SUMMARY carbon, nitrogen, and sulfur. Metabolite production is not identical among ST:STUDY_SUMMARY phytoplankton, and the flux of these molecules through the microbial loop ST:STUDY_SUMMARY depends on compound-specific bioavailability to a wider microbial community. Yet ST:STUDY_SUMMARY relatively little is known about the diversity or concentration of polar ST:STUDY_SUMMARY metabolites within marine plankton. Here we evaluate 313 metabolites in 21 ST:STUDY_SUMMARY phytoplankton species and in natural marine particles across environmental ST:STUDY_SUMMARY gradients to show that bulk community metabolomes reflect the phytoplankton ST:STUDY_SUMMARY community on a chemical level. ST:INSTITUTE University of Washington ST:DEPARTMENT Oceanography ST:LABORATORY Ingalls Lab ST:LAST_NAME Heal ST:FIRST_NAME Katherine ST:ADDRESS 1501 NE Boat Street, Marine Science Building, Room G, Seattle, WA, 98195, USA ST:EMAIL kheal@uw.edu ST:PHONE 612-616-4840 #SUBJECT SU:SUBJECT_TYPE Other SU:SUBJECT_SPECIES Marine Plankton SU:GENDER Not applicable #SUBJECT_SAMPLE_FACTORS: SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Raw file names and additional sample data SUBJECT_SAMPLE_FACTORS - Nmar_1 Species:Nitrosopumilus maritimus | Strain:SCM1 | Broad taxon:Archaea Biovolume filtered_uL=0.11362; nmol C filtered=4116.666667; Short ID=Nmar; RAW_FILE_NAME=180130_Exp_1 180130_Smp_Exp_1 180131_Smp_Exp_1 SUBJECT_SAMPLE_FACTORS - Nmar_2 Species:Nitrosopumilus maritimus | Strain:SCM1 | Broad taxon:Archaea Biovolume filtered_uL=0.10396; nmol C filtered=3766.666667; Short ID=Nmar; RAW_FILE_NAME=180130_Exp_2 180130_Smp_Exp_2 180131_Smp_Exp_2 SUBJECT_SAMPLE_FACTORS - Nmar_3 Species:Nitrosopumilus maritimus | Strain:SCM1 | Broad taxon:Archaea Biovolume filtered_uL=0.09982; nmol C filtered=3616.666667; Short ID=Nmar; RAW_FILE_NAME=180130_Exp_3 180130_Smp_Exp_3 180131_Smp_Exp_3 SUBJECT_SAMPLE_FACTORS - 8501_B Species:Crocosphaera watsonii | Strain:8501 | Broad taxon:Cyanobacteria Biovolume filtered_uL=3.711258; nmol C filtered=15773; Short ID=8501; RAW_FILE_NAME=161114_Smp_8501_B 161114_Smp_8501_B 161118_Smp_8501_B SUBJECT_SAMPLE_FACTORS - 8501_C Species:Crocosphaera watsonii | Strain:8501 | Broad taxon:Cyanobacteria Biovolume filtered_uL=5.351031818; nmol C filtered=22742; Short ID=8501; RAW_FILE_NAME=161114_Smp_8501_C 161114_Smp_8501_C 161118_Smp_8501_C SUBJECT_SAMPLE_FACTORS - 8501_D Species:Crocosphaera watsonii | Strain:8501 | Broad taxon:Cyanobacteria Biovolume filtered_uL=3.515528571; nmol C filtered=14941; Short ID=8501; RAW_FILE_NAME=161114_Smp_8501.D 161114_Smp_8501.D 161118_Smp_8501_D SUBJECT_SAMPLE_FACTORS - 1314P_A Species:Prochlorococcus marinus | Strain:1314 | Broad taxon:Cyanobacteria Biovolume filtered_uL=0.3185; nmol C filtered=5898.2295; Short ID=1314P; RAW_FILE_NAME=170821_Smp_1314P_A 170821_Smp_1314P_A 170828_Smp_1314P_A SUBJECT_SAMPLE_FACTORS - 1314P_B Species:Prochlorococcus marinus | Strain:1314 | Broad taxon:Cyanobacteria Biovolume filtered_uL=0.34858; nmol C filtered=6455.239233; Short ID=1314P; RAW_FILE_NAME=170821_Smp_1314P_B 170821_Smp_1314P_B 170828_Smp_1314P_B SUBJECT_SAMPLE_FACTORS - 1314P_C Species:Prochlorococcus marinus | Strain:1314 | Broad taxon:Cyanobacteria Biovolume filtered_uL=0.31701; nmol C filtered=5870.54047; Short ID=1314P; RAW_FILE_NAME=170821_Smp_1314P_C 170821_Smp_1314P_C 170828_Smp_1314P_C SUBJECT_SAMPLE_FACTORS - As9601_A Species:Prochlorococcus marinus | Strain:AS9601 | Broad taxon:Cyanobacteria Biovolume filtered_uL=0.452556309; nmol C filtered=10475.84048; Short ID=As9601; RAW_FILE_NAME=170821_Smp_As9601_A 170821_Smp_As9601_A 170828_Smp_As9601_A SUBJECT_SAMPLE_FACTORS - As9601_B Species:Prochlorococcus marinus | Strain:AS9601 | Broad taxon:Cyanobacteria Biovolume filtered_uL=0.487486222; nmol C filtered=11284.40329; Short ID=As9601; RAW_FILE_NAME=170821_Smp_As9601_B 170821_Smp_As9601_B 170828_Smp_As9601_B SUBJECT_SAMPLE_FACTORS - As9601_C Species:Prochlorococcus marinus | Strain:AS9601 | Broad taxon:Cyanobacteria Biovolume filtered_uL=0.467084563; nmol C filtered=10812.14267; Short ID=As9601; RAW_FILE_NAME=170821_Smp_As9601_C 170821_Smp_As9601_C 170828_Smp_As9601_C SUBJECT_SAMPLE_FACTORS - MED4_D Species:Prochlorococcus marinus | Strain:MED4 | Broad taxon:Cyanobacteria Biovolume filtered_uL=0.390883343; nmol C filtered=5428.935323; Short ID=MED4; RAW_FILE_NAME=161114_Smp_MED4.D 161114_Smp_MED4.D 161118_Smp_MED4_D SUBJECT_SAMPLE_FACTORS - MED4_E Species:Prochlorococcus marinus | Strain:MED4 | Broad taxon:Cyanobacteria Biovolume filtered_uL=0.346499459; nmol C filtered=4812.492493; Short ID=MED4; RAW_FILE_NAME=161114_Smp_MED4_E 161114_Smp_MED4_E 161118_Smp_MED4_E SUBJECT_SAMPLE_FACTORS - MED4_F Species:Prochlorococcus marinus | Strain:MED4 | Broad taxon:Cyanobacteria Biovolume filtered_uL=0.395070736; nmol C filtered=5487.093558; Short ID=MED4; RAW_FILE_NAME=161114_Smp_MED4_F 161114_Smp_MED4_F 161118_Smp_MED4_F SUBJECT_SAMPLE_FACTORS - Nat_A Species:Prochlorococcus marinus | Strain:NATL2A | Broad taxon:Cyanobacteria Biovolume filtered_uL=0.062888292; nmol C filtered=1886.648745; Short ID=Nat; RAW_FILE_NAME=170821_Smp_Nat_A 170821_Smp_Nat_A 170828_Smp_Nat_A SUBJECT_SAMPLE_FACTORS - Nat_B Species:Prochlorococcus marinus | Strain:NATL2A | Broad taxon:Cyanobacteria Biovolume filtered_uL=0.054405; nmol C filtered=1632.15; Short ID=Nat; RAW_FILE_NAME=170821_Smp_Nat_B 170821_Smp_Nat_B 170828_Smp_Nat_B SUBJECT_SAMPLE_FACTORS - Nat_C Species:Prochlorococcus marinus | Strain:NATL2A | Broad taxon:Cyanobacteria Biovolume filtered_uL=0.054925488; nmol C filtered=1647.764655; Short ID=Nat; RAW_FILE_NAME=170821_Smp_Nat_C 170821_Smp_Nat_C 170828_Smp_Nat_C SUBJECT_SAMPLE_FACTORS - 7803_B Species:Synechococcus sp. | Strain:7803 | Broad taxon:Cyanobacteria Biovolume filtered_uL=2.20472; nmol C filtered=33039.88864; Short ID=7803; RAW_FILE_NAME=161114_Smp_7803_B 161114_Smp_7803_B 161118_Smp_7803_B SUBJECT_SAMPLE_FACTORS - 7803_F Species:Synechococcus sp. | Strain:7803 | Broad taxon:Cyanobacteria Biovolume filtered_uL=2.18928; nmol C filtered=35225.87859; Short ID=7803; RAW_FILE_NAME=161114_Smp_7803_F 161114_Smp_7803_F 161118_Smp_7803_F SUBJECT_SAMPLE_FACTORS - 8102_A Species:Synechococcus sp. | Strain:8102 | Broad taxon:Cyanobacteria Biovolume filtered_uL=0.708; nmol C filtered=47768.85007; Short ID=8102; RAW_FILE_NAME=161114_Smp_8102_A 161114_Smp_8102_A 161118_Smp_8102_A SUBJECT_SAMPLE_FACTORS - 8102_F Species:Synechococcus sp. | Strain:8102 | Broad taxon:Cyanobacteria Biovolume filtered_uL=0.75484; nmol C filtered=47434.36605; Short ID=8102; RAW_FILE_NAME=161114_Smp_8102_F 161114_Smp_8102_F 161118_Smp_8102_F SUBJECT_SAMPLE_FACTORS - Cy_A Species:Cyclotella meneghiniana | Strain:338 | Broad taxon:Diatom Biovolume filtered_uL=7.0207956; nmol C filtered=66158.68808; Short ID=Cy; RAW_FILE_NAME=161107_Smp_Cy_A 161107_Smp_Cy_A 161110_Smp_Cy_A SUBJECT_SAMPLE_FACTORS - Cy_B Species:Cyclotella meneghiniana | Strain:338 | Broad taxon:Diatom Biovolume filtered_uL=2.285525455; nmol C filtered=21537.06991; Short ID=Cy; RAW_FILE_NAME=161107_Smp_Cy_B 161107_Smp_Cy_B 161110_Smp_Cy_B SUBJECT_SAMPLE_FACTORS - Cy_C Species:Cyclotella meneghiniana | Strain:338 | Broad taxon:Diatom Biovolume filtered_uL=4.9629762; nmol C filtered=46767.34847; Short ID=Cy; RAW_FILE_NAME=161107_Smp_Cy_C 161107_Smp_Cy_C 161110_Smp_Cy_C SUBJECT_SAMPLE_FACTORS - Np_A Species:Navicula pelliculosa | Strain:543 | Broad taxon:Diatom Biovolume filtered_uL=4.429566; nmol C filtered=50707.78032; Short ID=Np; RAW_FILE_NAME=170213_Smp_Np_A 170213_Smp_Np_A 170216_Smp_Np_A SUBJECT_SAMPLE_FACTORS - Np_C Species:Navicula pelliculosa | Strain:543 | Broad taxon:Diatom Biovolume filtered_uL=5.07693; nmol C filtered=58118.52699; Short ID=Np; RAW_FILE_NAME=170213_Smp_Np_C 170213_Smp_Np_C 170216_Smp_Np_C SUBJECT_SAMPLE_FACTORS - Np_F Species:Navicula pelliculosa | Strain:543 | Broad taxon:Diatom Biovolume filtered_uL=5.69769; nmol C filtered=65224.72243; Short ID=Np; RAW_FILE_NAME=170213_Smp_Np_F 170213_Smp_Np_F 170216_Smp_Np_F SUBJECT_SAMPLE_FACTORS - Pt_C Species:Phaeodactylum tricornutum | Strain:2561 | Broad taxon:Diatom Biovolume filtered_uL=52.49856; nmol C filtered=405673.6567; Short ID=Pt; RAW_FILE_NAME=170213_Smp_Pt_C 170213_Smp_Pt_C 170216_Smp_Pt_C SUBJECT_SAMPLE_FACTORS - Pt_F Species:Phaeodactylum tricornutum | Strain:2561 | Broad taxon:Diatom Biovolume filtered_uL=55.51368; nmol C filtered=428972.4816; Short ID=Pt; RAW_FILE_NAME=170213_Smp_Pt_F 170213_Smp_Pt_F 170216_Smp_Pt_F SUBJECT_SAMPLE_FACTORS - Pc55x_A Species:Pseudo-nitzschia pungens | Strain:Pc55x | Broad taxon:Diatom Biovolume filtered_uL=3.980689655; nmol C filtered=23670.0076; Short ID=Pc55x; RAW_FILE_NAME=170213_Smp_Pc55x_A 170213_Smp_Pc55x_A 170216_Smp_Pc55x_A SUBJECT_SAMPLE_FACTORS - Pc55x_B Species:Pseudo-nitzschia pungens | Strain:Pc55x | Broad taxon:Diatom Biovolume filtered_uL=4.154482759; nmol C filtered=24703.4175; Short ID=Pc55x; RAW_FILE_NAME=170213_Smp_Pc55x_B 170213_Smp_Pc55x_B 170216_Smp_Pc55x_B SUBJECT_SAMPLE_FACTORS - Pc55x_C Species:Pseudo-nitzschia pungens | Strain:Pc55x | Broad taxon:Diatom Biovolume filtered_uL=2.532631579; nmol C filtered=15059.55348; Short ID=Pc55x; RAW_FILE_NAME=170213_Smp_Pc55x_C 170213_Smp_Pc55x_C 170216_Smp_Pc55x_C SUBJECT_SAMPLE_FACTORS - To_A Species:Thalassiosira oceanica | Strain:1005 | Broad taxon:Diatom Biovolume filtered_uL=10.10952; nmol C filtered=94030.34765; Short ID=To; RAW_FILE_NAME=161107_Smp_To_A 161107_Smp_To_A 161110_Smp_To_A SUBJECT_SAMPLE_FACTORS - To_B Species:Thalassiosira oceanica | Strain:1005 | Broad taxon:Diatom Biovolume filtered_uL=13.26875; nmol C filtered=123414.8313; Short ID=To; RAW_FILE_NAME=161107_Smp_To_B 161107_Smp_To_B 161110_Smp_To_B SUBJECT_SAMPLE_FACTORS - To_C Species:Thalassiosira oceanica | Strain:1005 | Broad taxon:Diatom Biovolume filtered_uL=12.20459; nmol C filtered=113516.9; Short ID=To; RAW_FILE_NAME=161107_Smp_To_C 161107_Smp_To_C 161110_Smp_To_C SUBJECT_SAMPLE_FACTORS - Tp_A Species:Thalassiosira pseudonana | Strain:1335 | Broad taxon:Diatom Biovolume filtered_uL=4.40222; nmol C filtered=133534.0977; Short ID=Tp; RAW_FILE_NAME=161107_Smp_Tp_A 161107_Smp_Tp_A 161110_Smp_Tp_A SUBJECT_SAMPLE_FACTORS - Tp_E Species:Thalassiosira pseudonana | Strain:1335 | Broad taxon:Diatom Biovolume filtered_uL=3.13927; nmol C filtered=95224.584; Short ID=Tp; RAW_FILE_NAME=161107_Smp_Tp_E 161107_Smp_Tp_E 161110_Smp_Tp_E SUBJECT_SAMPLE_FACTORS - Tp_F Species:Thalassiosira pseudonana | Strain:1335 | Broad taxon:Diatom Biovolume filtered_uL=6.36368; nmol C filtered=193031.5296; Short ID=Tp; RAW_FILE_NAME=161107_Smp_Tp_F 161107_Smp_Tp_F 161110_Smp_Tp_F SUBJECT_SAMPLE_FACTORS - 1771_A Species:Alexandrium tamarense | Strain:1771 | Broad taxon:Dinoflagellate Biovolume filtered_uL=0.45029728; nmol C filtered=4416.314582; Short ID=1771; RAW_FILE_NAME=170821_Smp_1771_A 170821_Smp_1771_A 170828_Smp_1771_A SUBJECT_SAMPLE_FACTORS - 1771_B Species:Alexandrium tamarense | Strain:1771 | Broad taxon:Dinoflagellate Biovolume filtered_uL=0.39780864; nmol C filtered=3901.529447; Short ID=1771; RAW_FILE_NAME=170821_Smp_1771_B 170821_Smp_1771_B 170828_Smp_1771_B SUBJECT_SAMPLE_FACTORS - 1771_C Species:Alexandrium tamarense | Strain:1771 | Broad taxon:Dinoflagellate Biovolume filtered_uL=0.23205504; nmol C filtered=2275.892177; Short ID=1771; RAW_FILE_NAME=170821_Smp_1771_C 170821_Smp_1771_C 170828_Smp_1771_C SUBJECT_SAMPLE_FACTORS - 1314_A Species:Amphidinium carterae | Strain:1314 | Broad taxon:Dinoflagellate Biovolume filtered_uL=0.56953488; nmol C filtered=8754.975631; Short ID=1314; RAW_FILE_NAME=170821_Smp_1314_A 170821_Smp_1314_A 170828_Smp_1314_A SUBJECT_SAMPLE_FACTORS - 1314_B Species:Amphidinium carterae | Strain:1314 | Broad taxon:Dinoflagellate Biovolume filtered_uL=0.5827728; nmol C filtered=8958.470924; Short ID=1314; RAW_FILE_NAME=170821_Smp_1314_B 170821_Smp_1314_B 170828_Smp_1314_B SUBJECT_SAMPLE_FACTORS - 1314_C Species:Amphidinium carterae | Strain:1314 | Broad taxon:Dinoflagellate Biovolume filtered_uL=0.66859104; nmol C filtered=10277.68179; Short ID=1314; RAW_FILE_NAME=170821_Smp_1314_C 170821_Smp_1314_C 170828_Smp_1314_C SUBJECT_SAMPLE_FACTORS - 449_A Species:Heterocapsa triquetra | Strain:449 | Broad taxon:Dinoflagellate Biovolume filtered_uL=0.33903; nmol C filtered=4462.579749; Short ID=449; RAW_FILE_NAME=170821_Smp_449_A 170821_Smp_449_A 170828_Smp_449_A SUBJECT_SAMPLE_FACTORS - 449_B Species:Heterocapsa triquetra | Strain:449 | Broad taxon:Dinoflagellate Biovolume filtered_uL=0.48124; nmol C filtered=6334.523314; Short ID=449; RAW_FILE_NAME=170821_Smp_449_B 170821_Smp_449_B 170828_Smp_449_B SUBJECT_SAMPLE_FACTORS - 449_C Species:Heterocapsa triquetra | Strain:449 | Broad taxon:Dinoflagellate Biovolume filtered_uL=0.55139; nmol C filtered=7257.95976; Short ID=449; RAW_FILE_NAME=170821_Smp_449_C 170821_Smp_449_C 170828_Smp_449_C SUBJECT_SAMPLE_FACTORS - 2021_A Species:Lingulodinium polyedra | Strain:2021 | Broad taxon:Dinoflagellate Biovolume filtered_uL=2.75917726; nmol C filtered=26427.31696; Short ID=2021; RAW_FILE_NAME=170821_Smp_2021_A 170821_Smp_2021_A 170828_Smp_2021_A SUBJECT_SAMPLE_FACTORS - 2021_B Species:Lingulodinium polyedra | Strain:2021 | Broad taxon:Dinoflagellate Biovolume filtered_uL=5.11555492; nmol C filtered=48996.63145; Short ID=2021; RAW_FILE_NAME=170821_Smp_2021_B 170821_Smp_2021_B 170828_Smp_2021_B SUBJECT_SAMPLE_FACTORS - 2021_C Species:Lingulodinium polyedra | Strain:2021 | Broad taxon:Dinoflagellate Biovolume filtered_uL=1.53228256; nmol C filtered=14676.15636; Short ID=2021; RAW_FILE_NAME=170821_Smp_2021_C 170821_Smp_2021_C 170828_Smp_2021_C SUBJECT_SAMPLE_FACTORS - 2090_A Species:Emiliania huxleyi | Strain:2090 | Broad taxon:Haptophyte Biovolume filtered_uL=0.29131; nmol C filtered=3813.921528; Short ID=2090; RAW_FILE_NAME=161107_Smp_2090_A 161107_Smp_2090_A 161110_Smp_2090_A SUBJECT_SAMPLE_FACTORS - 2090_B Species:Emiliania huxleyi | Strain:2090 | Broad taxon:Haptophyte Biovolume filtered_uL=0.33255; nmol C filtered=4353.791699; Short ID=2090; RAW_FILE_NAME=161107_Smp_2090_B 161107_Smp_2090_B 161110_Smp_2090_B SUBJECT_SAMPLE_FACTORS - 2090_C Species:Emiliania huxleyi | Strain:2090 | Broad taxon:Haptophyte Biovolume filtered_uL=0.381324; nmol C filtered=4992.347815; Short ID=2090; RAW_FILE_NAME=161107_Smp_2090_C 161107_Smp_2090_C 161110_Smp_2090_C SUBJECT_SAMPLE_FACTORS - 371_A Species:Emiliania huxleyi | Strain:371 | Broad taxon:Haptophyte Biovolume filtered_uL=0.365805; nmol C filtered=4789.170869; Short ID=371; RAW_FILE_NAME=161107_Smp_371_A 161107_Smp_371_A 161110_Smp_371_A SUBJECT_SAMPLE_FACTORS - 371_B Species:Emiliania huxleyi | Strain:371 | Broad taxon:Haptophyte Biovolume filtered_uL=0.380920909; nmol C filtered=4987.070491; Short ID=371; RAW_FILE_NAME=161107_Smp_371_B 161107_Smp_371_B 161110_Smp_371_B SUBJECT_SAMPLE_FACTORS - 371_C Species:Emiliania huxleyi | Strain:371 | Broad taxon:Haptophyte Biovolume filtered_uL=0.294256364; nmol C filtered=3852.445988; Short ID=371; RAW_FILE_NAME=161107_Smp_371_C 161107_Smp_371_C 161110_Smp_371_C SUBJECT_SAMPLE_FACTORS - 1545_A Species:Micromonas pusilla | Strain:1545 | Broad taxon:Prasinophyte Biovolume filtered_uL=0.557926221; nmol C filtered=65905.03488; Short ID=1545; RAW_FILE_NAME=170821_Smp_1545_A 170821_Smp_1545_A 170828_Smp_1545_A SUBJECT_SAMPLE_FACTORS - 1545_B Species:Micromonas pusilla | Strain:1545 | Broad taxon:Prasinophyte Biovolume filtered_uL=0.571037924; nmol C filtered=67453.85477; Short ID=1545; RAW_FILE_NAME=170821_Smp_1545_B 170821_Smp_1545_B 170828_Smp_1545_B SUBJECT_SAMPLE_FACTORS - 1545_C Species:Micromonas pusilla | Strain:1545 | Broad taxon:Prasinophyte Biovolume filtered_uL=0.536432713; nmol C filtered=63366.11429; Short ID=1545; RAW_FILE_NAME=170821_Smp_1545_C 170821_Smp_1545_C 170828_Smp_1545_C SUBJECT_SAMPLE_FACTORS - 3430_A Species:Ostreococcus lucimarinus | Strain:3430 | Broad taxon:Prasinophyte Biovolume filtered_uL=0.169351485; nmol C filtered=33870; Short ID=3430; RAW_FILE_NAME=180227_Smp_3430_A 180227_Smp_3430_A 180302_Smp_3430_A SUBJECT_SAMPLE_FACTORS - 3430_B Species:Ostreococcus lucimarinus | Strain:3430 | Broad taxon:Prasinophyte Biovolume filtered_uL=0.172402985; nmol C filtered=34481; Short ID=3430; RAW_FILE_NAME=180227_Smp_3430_B 180227_Smp_3430_B 180302_Smp_3430_B SUBJECT_SAMPLE_FACTORS - 3430_C Species:Ostreococcus lucimarinus | Strain:3430 | Broad taxon:Prasinophyte Biovolume filtered_uL=0.175519802; nmol C filtered=35104; Short ID=3430; RAW_FILE_NAME=180227_Smp_3430_C 180227_Smp_3430_C 180302_Smp_3430_C #COLLECTION CO:COLLECTION_SUMMARY Axenic phytoplankton were cultured in controlled laboratory settings and CO:COLLECTION_SUMMARY harvested under exponential growth using a gentle vacuum filtration onto 47 mm CO:COLLECTION_SUMMARY Durapore filters (pore size 0.2 µm). Samples were flash frozen in liquid N2 and CO:COLLECTION_SUMMARY stored at -80°C until extraction for metabolites. CO:SAMPLE_TYPE Cultured plankton cells CO:STORAGE_CONDITIONS Described in summary #TREATMENT TR:TREATMENT_SUMMARY Media, light, and temperature were chosen for optimal growth of each TR:TREATMENT_SUMMARY phytoplankton species. In short, axenic phytoplankton were cultured in TR:TREATMENT_SUMMARY controlled laboratory settings and harvested under exponential growth using a TR:TREATMENT_SUMMARY gentle vacuum filtration onto 47 mm Durapore filters (pore size 0.2 µm). TR:TREATMENT_SUMMARY Samples were flash frozen in liquid N2 and stored at -80°C until extraction. In TR:TREATMENT_SUMMARY addition to samples, media blanks corresponding to each media type were TR:TREATMENT_SUMMARY harvested and served as a matrix blank to each corresponding phytoplankton TR:TREATMENT_SUMMARY sample. #SAMPLEPREP SP:SAMPLEPREP_SUMMARY Each sample was extracted using a modified Bligh-Dyer extraction. Briefly, SP:SAMPLEPREP_SUMMARY filters were cut up and put into 15 mL teflon centrifuge tubes containing a SP:SAMPLEPREP_SUMMARY mixture of 100 µm and 400 µm silica beads. Heavy isotope-labeled internal SP:SAMPLEPREP_SUMMARY standards were added along with ~2 mL of cold aqueous solvent (50:50 SP:SAMPLEPREP_SUMMARY methanol:water) and ~3 mL of cold organic solvent (dichloromethane). The samples SP:SAMPLEPREP_SUMMARY were shaken on a FastPrep-24 Homogenizer for 30 seconds and chilled in a -20°C SP:SAMPLEPREP_SUMMARY freezer repeatedly for three cycles of bead-beating and a total of 30 minutes of SP:SAMPLEPREP_SUMMARY chilling. The organic and aqueous layers were separated by spinning samples in a SP:SAMPLEPREP_SUMMARY centrifuge at 4,300 rpm for 2 minutes at 4°C. The aqueous layer was removed to SP:SAMPLEPREP_SUMMARY a new glass centrifuge tube. The remaining organic fraction was rinsed three SP:SAMPLEPREP_SUMMARY more times with additions of 1 to 2 mL of cold 50:50 methanol:water. All aqueous SP:SAMPLEPREP_SUMMARY rinses were combined for each sample and ~2mL of cold dichloromethane was added SP:SAMPLEPREP_SUMMARY to the combined aqueous layer. Tubes were shaken and centrifuged at 4,300 rpm SP:SAMPLEPREP_SUMMARY for 2 minutes at 4°C. The aqueous layer was removed to a new glass vial and SP:SAMPLEPREP_SUMMARY dried under N2 gas. The remaining organic layer in the bead beating tubes was SP:SAMPLEPREP_SUMMARY transferred into the glass centrifuge tube and the bead beating tube was rinsed SP:SAMPLEPREP_SUMMARY two more times with cold organic solvent. The combined organic rinses were SP:SAMPLEPREP_SUMMARY centrifuged, transferred to a new glass vial, and dried under N2 gas. Dried SP:SAMPLEPREP_SUMMARY aqueous fractions were re-dissolved in 380 µL of water. Dried organic fractions SP:SAMPLEPREP_SUMMARY were re-dissolved in 380 µL of 1:1 water:acetonitrile. 20 µL of SP:SAMPLEPREP_SUMMARY isotope-labeled injection standards in water were added to both fractions. Media SP:SAMPLEPREP_SUMMARY blanks were extracted alongside samples as methodological blanks. SP:PROCESSING_STORAGE_CONDITIONS On ice SP:EXTRACTION_METHOD Bligh-Dyer SP:EXTRACT_STORAGE -80℃ #CHROMATOGRAPHY CH:CHROMATOGRAPHY_SUMMARY See attached summary CH:CHROMATOGRAPHY_TYPE HILIC CH:INSTRUMENT_NAME Waters Acquity I-Class CH:COLUMN_NAME SeQuant ZIC- pHILIC (150 x 2.1mm, 5um) CH:METHODS_FILENAME Ingalls_Lab_LC_Methods.txt #ANALYSIS AN:ANALYSIS_TYPE MS AN:ANALYSIS_PROTOCOL_FILE Ingalls_Lab_MS_Methods.txt #MS MS:INSTRUMENT_NAME Thermo Q Exactive HF hybrid Orbitrap MS:INSTRUMENT_TYPE Orbitrap MS:MS_TYPE ESI MS:ION_MODE NEGATIVE MS:MS_COMMENTS See attached protocol. MS:MS_RESULTS_FILE ST001514_AN002510_Results.txt UNITS:Adjusted and normalized peak areas Has m/z:Yes Has RT:Yes RT units:Seconds #END