#METABOLOMICS WORKBENCH TruxalCarlson_20200625_093715 DATATRACK_ID:2087 STUDY_ID:ST001410 ANALYSIS_ID:AN002360 PROJECT_ID:PR000966 VERSION 1 CREATED_ON June 26, 2020, 4:21 pm #PROJECT PR:PROJECT_TITLE Patterns in metabolite pools show that phytoplankton leave a taxon-specific PR:PROJECT_TITLE signature on particulate carbon: Surface samples from the North Pacific PR:PROJECT_TITLE Subtropical Gyre to North Pacific Transition Zone PR:PROJECT_TYPE Marine Metabolomics PR:PROJECT_SUMMARY In the surface ocean, carbon is fixed by phytoplankton and respired by the PR:PROJECT_SUMMARY entire marine community at an astonishingly high rate. At any point in time, the PR:PROJECT_SUMMARY difference between these two processes yields a carbon pool in surface particles PR:PROJECT_SUMMARY that is a combination of both freshly fixed and partially degraded material. On PR:PROJECT_SUMMARY a molecular level, we have a limited knowledge of the small molecules, or PR:PROJECT_SUMMARY metabolites, within this pool. Specific metabolites have been shown to be PR:PROJECT_SUMMARY responsible for fueling respiration, maintaining organismal interactions, and PR:PROJECT_SUMMARY transferring energy throughout the microbial community. Metabolomics, or the PR:PROJECT_SUMMARY direct observation and quantification of the small molecules that are the result PR:PROJECT_SUMMARY of cellular activity, provides an important lens through which we can begin to PR:PROJECT_SUMMARY assess the standing stocks of small compounds that likely fuel a great deal of PR:PROJECT_SUMMARY heterotrophic activity in the surface ocean. Here we describe community PR:PROJECT_SUMMARY metabolomes of particulate material into the North Pacific Ocean and compare the PR:PROJECT_SUMMARY metabolomes to a variety of phytoplankton grown in the lab. Using both targeted PR:PROJECT_SUMMARY and untargeted metabolomics, we identify metabolites in the particulate carbon PR:PROJECT_SUMMARY pool and explore their latitudinal and phylogenetic distributions. This analysis PR:PROJECT_SUMMARY reveals several compounds that have not been previously recognized as abundant PR:PROJECT_SUMMARY components of the marine organic carbon pool. We found that the community PR:PROJECT_SUMMARY metabolome showed distinct differences between the regimes that likely reflects PR:PROJECT_SUMMARY the phytoplankton community present. The community metabolome in surface waters PR:PROJECT_SUMMARY of the subtropical domain was remarkably consistent even when sampled weeks PR:PROJECT_SUMMARY apart, while the northern regions showed a patichier and less reproducible PR:PROJECT_SUMMARY community metabolome. Some individual compounds showed distinct patterns between PR:PROJECT_SUMMARY oceanographic regimes, including homarine, an abundant molecule that can PR:PROJECT_SUMMARY contribute up to 4% of the total particulate carbon pool in marine surface PR:PROJECT_SUMMARY waters. Glutamic acid and glutamine showed opposite patterns in the PR:PROJECT_SUMMARY oceanographic regimes, suggesting differences in community-level nitrogen PR:PROJECT_SUMMARY assimilation in these different regimes. Overall, this study offers a new PR:PROJECT_SUMMARY perspective into particulate carbon composition in oceanographic research, PR:PROJECT_SUMMARY reveals important carbon pools that may fuel the microbial loop, and suggests an PR:PROJECT_SUMMARY altered community-level nitrogen assimilation capacity over the North Pacific PR:PROJECT_SUMMARY transition zone. PR:INSTITUTE University of Washington PR:DEPARTMENT School of 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 206-616-4658 #STUDY ST:STUDY_TITLE Patterns in metabolite pools show that phytoplankton leave a taxon-specific ST:STUDY_TITLE signature on particulate carbon: Surface samples from the North Pacific ST:STUDY_TITLE Subtropical Gyre to North Pacific Transition Zone ST:STUDY_TYPE Marine metabolomics surface samples from the North Pacific Subtropical Gyre to ST:STUDY_TYPE North Pacific Transition Zone ST:STUDY_SUMMARY In the surface ocean, carbon is fixed by phytoplankton and respired by the ST:STUDY_SUMMARY entire marine community at an astonishingly high rate. At any point in time, the ST:STUDY_SUMMARY difference between these two processes yields a carbon pool in surface particles ST:STUDY_SUMMARY that is a combination of both freshly fixed and partially degraded material. On ST:STUDY_SUMMARY a molecular level, we have a limited knowledge of the small molecules, or ST:STUDY_SUMMARY metabolites, within this pool. Specific metabolites have been shown to be ST:STUDY_SUMMARY responsible for fueling respiration, maintaining organismal interactions, and ST:STUDY_SUMMARY transferring energy throughout the microbial community. Metabolomics, or the ST:STUDY_SUMMARY direct observation and quantification of the small molecules that are the result ST:STUDY_SUMMARY of cellular activity, provides an important lens through which we can begin to ST:STUDY_SUMMARY assess the standing stocks of small compounds that likely fuel a great deal of ST:STUDY_SUMMARY heterotrophic activity in the surface ocean. Here we describe community ST:STUDY_SUMMARY metabolomes of particulate material into the North Pacific Ocean and compare the ST:STUDY_SUMMARY metabolomes to a variety of phytoplankton grown in the lab. Using both targeted ST:STUDY_SUMMARY and untargeted metabolomics, we identify metabolites in the particulate carbon ST:STUDY_SUMMARY pool and explore their latitudinal and phylogenetic distributions. This analysis ST:STUDY_SUMMARY reveals several compounds that have not been previously recognized as abundant ST:STUDY_SUMMARY components of the marine organic carbon pool. We found that the community ST:STUDY_SUMMARY metabolome showed distinct differences between the regimes that likely reflects ST:STUDY_SUMMARY the phytoplankton community present. The community metabolome in surface waters ST:STUDY_SUMMARY of the subtropical domain was remarkably consistent even when sampled weeks ST:STUDY_SUMMARY apart, while the northern regions showed a patchier and less reproducible ST:STUDY_SUMMARY community metabolome. Some individual compounds showed distinct patterns between ST:STUDY_SUMMARY oceanographic regimes, including homarine, an abundant molecule that can ST:STUDY_SUMMARY contribute up to 4% of the total particulate carbon pool in marine surface ST:STUDY_SUMMARY waters. Glutamic acid and glutamine showed opposite patterns in the ST:STUDY_SUMMARY oceanographic regimes, suggesting differences in community-level nitrogen ST:STUDY_SUMMARY assimilation in these different regimes. Overall, this study offers a new ST:STUDY_SUMMARY perspective into particulate carbon composition in oceanographic research, ST:STUDY_SUMMARY reveals important carbon pools that may fuel the microbial loop, and suggests an ST:STUDY_SUMMARY altered community-level nitrogen assimilation capacity over the North Pacific ST:STUDY_SUMMARY transition zone. ST:INSTITUTE University of Washington ST:DEPARTMENT School of 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 206-616-4658 #SUBJECT SU:SUBJECT_TYPE Other SU:SUBJECT_SPECIES Natural mixed marine microbial community #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 - FilterBlk_A Depth_m:NA | Vol_L:NA Latitude=NA; Longitude=NA; UTC=NA; Replicate=A; Type=Blk; RAW_FILE_NAME=170419_Blk_FilterBlk_A;170419_Blk_FilterBlk_A;170425_Blk_FilterBlk_A SUBJECT_SAMPLE_FACTORS - FilterBlk_B Depth_m:NA | Vol_L:NA Latitude=NA; Longitude=NA; UTC=NA; Replicate=B; Type=Blk; RAW_FILE_NAME=170419_Blk_FilterBlk_B;170419_Blk_FilterBlk_B;170425_Blk_FilterBlk_B SUBJECT_SAMPLE_FACTORS - S2C1_B Depth_m:15 | Vol_L:11 Latitude=23.496122; Longitude=-157.994013; UTC=2016-4-20T15;13;01; Replicate=B; Type=Smp; RAW_FILE_NAME=170419_Smp_S2C1_B;170419_Smp_S2C1_B;170425_Smp_S2C1_B SUBJECT_SAMPLE_FACTORS - S2C1_C Depth_m:15 | Vol_L:11 Latitude=23.496122; Longitude=-157.994013; UTC=2016-4-20T15;13;02; Replicate=C; Type=Smp; RAW_FILE_NAME=170419_Smp_S2C1_C;170419_Smp_S2C1_C;170425_Smp_S2C1_C SUBJECT_SAMPLE_FACTORS - S2C3_A Depth_m:15 | Vol_L:9.5 Latitude=23.596524; Longitude=-157.955307; UTC=2016-4-21T01;22;00; Replicate=A; Type=Smp; RAW_FILE_NAME=170419_Smp_S2C3_A;170419_Smp_S2C3_A;170425_Smp_S2C3_A SUBJECT_SAMPLE_FACTORS - S4C1_A Depth_m:15 | Vol_L:11.5 Latitude=28.143572; Longitude=-157.998665; UTC=2016-4-22T15;08;00; Replicate=A; Type=Smp; RAW_FILE_NAME=170419_Smp_S4C1_A;170419_Smp_S4C1_A;170425_Smp_S4C1_A SUBJECT_SAMPLE_FACTORS - S4C1_B Depth_m:15 | Vol_L:11.5 Latitude=28.143572; Longitude=-157.998665; UTC=2016-4-22T15;08;00; Replicate=B; Type=Smp; RAW_FILE_NAME=170419_Smp_S4C1_B;170419_Smp_S4C1_B;170425_Smp_S4C1_B SUBJECT_SAMPLE_FACTORS - S4C1_C Depth_m:15 | Vol_L:11.5 Latitude=28.143572; Longitude=-157.998665; UTC=2016-4-22T15;08;00; Replicate=C; Type=Smp; RAW_FILE_NAME=170419_Smp_S4C1_C;170419_Smp_S4C1_C;170425_Smp_S4C1_C SUBJECT_SAMPLE_FACTORS - S4C2_A Depth_m:15 | Vol_L:10 Latitude=28.140122; Longitude=-158.00368; UTC=2016-4-23T01;57;00; Replicate=A; Type=Smp; RAW_FILE_NAME=170419_Smp_S4C2_A;170419_Smp_S4C2_A;170425_Smp_S4C2_A SUBJECT_SAMPLE_FACTORS - S4C2_B Depth_m:15 | Vol_L:10 Latitude=28.140122; Longitude=-158.00368; UTC=2016-4-23T01;57;00; Replicate=B; Type=Smp; RAW_FILE_NAME=170419_Smp_S4C2_B;170419_Smp_S4C2_B;170425_Smp_S4C2_B SUBJECT_SAMPLE_FACTORS - S6C1_A Depth_m:15 | Vol_L:11 Latitude=32.583641; Longitude=-158.00027; UTC=2016-4-24T15;09;00; Replicate=A; Type=Smp; RAW_FILE_NAME=170419_Smp_S6C1_A;170419_Smp_S6C1_A;170425_Smp_S6C1_A SUBJECT_SAMPLE_FACTORS - S6C1_B Depth_m:15 | Vol_L:11 Latitude=32.583641; Longitude=-158.00027; UTC=2016-4-24T15;09;00; Replicate=B; Type=Smp; RAW_FILE_NAME=170419_Smp_S6C1_B;170419_Smp_S6C1_B;170425_Smp_S6C1_B SUBJECT_SAMPLE_FACTORS - S6C1_C Depth_m:15 | Vol_L:11 Latitude=32.583641; Longitude=-158.00027; UTC=2016-4-24T15;09;00; Replicate=C; Type=Smp; RAW_FILE_NAME=170419_Smp_S6C1_C;170419_Smp_S6C1_C;170425_Smp_S6C1_C SUBJECT_SAMPLE_FACTORS - S6C2_A Depth_m:15 | Vol_L:10.5 Latitude=32.699814; Longitude=-157.993171; UTC=2016-4-25T01;33;00; Replicate=A; Type=Smp; RAW_FILE_NAME=170419_Smp_S6C2_A;170419_Smp_S6C2_A;170425_Smp_S6C2_A SUBJECT_SAMPLE_FACTORS - S6C2_B Depth_m:15 | Vol_L:10.5 Latitude=32.699814; Longitude=-157.993171; UTC=2016-4-25T01;33;00; Replicate=B; Type=Smp; RAW_FILE_NAME=170419_Smp_S6C2_B;170419_Smp_S6C2_B;170425_Smp_S6C2_B SUBJECT_SAMPLE_FACTORS - U3_A Depth_m:15 | Vol_L:15.5 Latitude=34.533075; Longitude=-157.997757; UTC=2016-4-25T19;45;00; Replicate=A; Type=Smp; RAW_FILE_NAME=170419_Smp_U3_A;170419_Smp_U3_A;170425_Smp_U3_A SUBJECT_SAMPLE_FACTORS - U3_B Depth_m:15 | Vol_L:15 Latitude=34.533075; Longitude=-157.997757; UTC=2016-4-25T19;45;00; Replicate=B; Type=Smp; RAW_FILE_NAME=170419_Smp_U3_B;170419_Smp_U3_B;170425_Smp_U3_B SUBJECT_SAMPLE_FACTORS - U4_A Depth_m:15 | Vol_L:13.5 Latitude=35.493484; Longitude=-158.006387; UTC=2016-4-26T02;33;00; Replicate=A; Type=Smp; RAW_FILE_NAME=170419_Smp_U4_A;170419_Smp_U4_A;170425_Smp_U4_A SUBJECT_SAMPLE_FACTORS - U4_B Depth_m:15 | Vol_L:12 Latitude=35.493484; Longitude=-158.006387; UTC=2016-4-26T02;33;00; Replicate=B; Type=Smp; RAW_FILE_NAME=170419_Smp_U4_B;170419_Smp_U4_B;170425_Smp_U4_B SUBJECT_SAMPLE_FACTORS - S8C1_A Depth_m:15 | Vol_L:10.5 Latitude=37.299396; Longitude=-158.000717; UTC=2016-4-26T15;10;00; Replicate=A; Type=Smp; RAW_FILE_NAME=170419_Smp_S8C1_A;170419_Smp_S8C1_A;170425_Smp_S8C1_A SUBJECT_SAMPLE_FACTORS - S9C1_A Depth_m:15 | Vol_L:11.5 Latitude=36.569491; Longitude=-158.003614; UTC=2016-4-27T15;12;00; Replicate=A; Type=Smp; RAW_FILE_NAME=170419_Smp_S9C1_A;170419_Smp_S9C1_A;170425_Smp_S9C1_A SUBJECT_SAMPLE_FACTORS - S9C1_B Depth_m:15 | Vol_L:11 Latitude=36.569491; Longitude=-158.003614; UTC=2016-4-27T15;12;00; Replicate=B; Type=Smp; RAW_FILE_NAME=170419_Smp_S9C1_B;170419_Smp_S9C1_B;170425_Smp_S9C1_B SUBJECT_SAMPLE_FACTORS - S9C2_A Depth_m:15 | Vol_L:10 Latitude=36.567248; Longitude=-157.999496; UTC=2016-4-28T03;43;00; Replicate=A; Type=Smp; RAW_FILE_NAME=170419_Smp_S9C2_A;170419_Smp_S9C2_A;170425_Smp_S9C2_A SUBJECT_SAMPLE_FACTORS - S9C2_B Depth_m:15 | Vol_L:10 Latitude=36.567248; Longitude=-157.999496; UTC=2016-4-28T03;43;00; Replicate=B; Type=Smp; RAW_FILE_NAME=170419_Smp_S9C2_B;170419_Smp_S9C2_B;170425_Smp_S9C2_B SUBJECT_SAMPLE_FACTORS - S11C1_A Depth_m:15 | Vol_L:10 Latitude=36.571243; Longitude=-157.995725; UTC=2016-4-29T06;40;00; Replicate=A; Type=Smp; RAW_FILE_NAME=170419_Smp_S11C1_A;170419_Smp_S11C1_A;170425_Smp_S11C1_A SUBJECT_SAMPLE_FACTORS - U6_A Depth_m:15 | Vol_L:10 Latitude=36.455193; Longitude=-157.957568; UTC=2016-4-29T10;00;00; Replicate=A; Type=Smp; RAW_FILE_NAME=170419_Smp_U6_A;170419_Smp_U6_A;170425_Smp_U6_A SUBJECT_SAMPLE_FACTORS - U6_B Depth_m:15 | Vol_L:10.5 Latitude=36.455193; Longitude=-157.957568; UTC=2016-4-29T10;00;00; Replicate=B; Type=Smp; RAW_FILE_NAME=170419_Smp_U6_B;170419_Smp_U6_B;170425_Smp_U6_B SUBJECT_SAMPLE_FACTORS - U7_A Depth_m:15 | Vol_L:10 Latitude=36.366833; Longitude=-157.972134; UTC=2016-4-29T11;14;00; Replicate=A; Type=Smp; RAW_FILE_NAME=170419_Smp_U7_A;170419_Smp_U7_A;170425_Smp_U7_A SUBJECT_SAMPLE_FACTORS - U7_B Depth_m:15 | Vol_L:10 Latitude=36.366833; Longitude=-157.972134; UTC=2016-4-29T11;14;00; Replicate=B; Type=Smp; RAW_FILE_NAME=170419_Smp_U7_B;170419_Smp_U7_B;170425_Smp_U7_B SUBJECT_SAMPLE_FACTORS - U8_A Depth_m:15 | Vol_L:10 Latitude=36.296547; Longitude=-157.991993; UTC=2016-4-29T12;15;00; Replicate=A; Type=Smp; RAW_FILE_NAME=170419_Smp_U8_A;170419_Smp_U8_A;170425_Smp_U8_A SUBJECT_SAMPLE_FACTORS - U8_B Depth_m:15 | Vol_L:10 Latitude=36.296547; Longitude=-157.991993; UTC=2016-4-29T12;15;00; Replicate=B; Type=Smp; RAW_FILE_NAME=170419_Smp_U8_B;170419_Smp_U8_B;170425_Smp_U8_B SUBJECT_SAMPLE_FACTORS - U9_A Depth_m:15 | Vol_L:10 Latitude=36.221701; Longitude=-157.998186; UTC=2016-4-29T13;19;00; Replicate=A; Type=Smp; RAW_FILE_NAME=170419_Smp_U9_A;170419_Smp_U9_A;170425_Smp_U9_A SUBJECT_SAMPLE_FACTORS - U9_B Depth_m:15 | Vol_L:10 Latitude=36.221701; Longitude=-157.998186; UTC=2016-4-29T13;19;00; Replicate=B; Type=Smp; RAW_FILE_NAME=170419_Smp_U9_B;170419_Smp_U9_B;170425_Smp_U9_B SUBJECT_SAMPLE_FACTORS - S12C1_A Depth_m:15 | Vol_L:10 Latitude=33.090833; Longitude=-158.002214; UTC=2016-4-30T15;05;00; Replicate=A; Type=Smp; RAW_FILE_NAME=170419_Smp_S12C1_A;170419_Smp_S12C1_A;170425_Smp_S12C1_A SUBJECT_SAMPLE_FACTORS - S12C1_B Depth_m:15 | Vol_L:10 Latitude=33.090833; Longitude=-158.002214; UTC=2016-4-30T15;05;00; Replicate=B; Type=Smp; RAW_FILE_NAME=170419_Smp_S12C1_B;170419_Smp_S12C1_B;170425_Smp_S12C1_B SUBJECT_SAMPLE_FACTORS - S13C1_A Depth_m:15 | Vol_L:10 Latitude=29.700608; Longitude=-157.999422; UTC=2016-5-01T15;02;00; Replicate=A; Type=Smp; RAW_FILE_NAME=170419_Smp_S13C1_A;170419_Smp_S13C1_A;170425_Smp_S13C1_A SUBJECT_SAMPLE_FACTORS - S13C1_B Depth_m:15 | Vol_L:10 Latitude=29.700608; Longitude=-157.999422; UTC=2016-5-01T15;02;00; Replicate=B; Type=Smp; RAW_FILE_NAME=170419_Smp_S13C1_B;170419_Smp_S13C1_B;170425_Smp_S13C1_B SUBJECT_SAMPLE_FACTORS - S14C1_A Depth_m:15 | Vol_L:10 Latitude=26.283176; Longitude=-157.998902; UTC=2016-5-01T15;02;00; Replicate=A; Type=Smp; RAW_FILE_NAME=170419_Smp_S14C1_A;170419_Smp_S14C1_A;170425_Smp_S14C1_A SUBJECT_SAMPLE_FACTORS - S14C1_B Depth_m:15 | Vol_L:10 Latitude=26.283176; Longitude=-157.998902; UTC=2016-5-01T15;02;00; Replicate=B; Type=Smp; RAW_FILE_NAME=170419_Smp_S14C1_B;170419_Smp_S14C1_B;170425_Smp_S14C1_B SUBJECT_SAMPLE_FACTORS - S14C1_C Depth_m:15 | Vol_L:10 Latitude=26.283176; Longitude=-157.998902; UTC=2016-5-01T15;02;00; Replicate=C; Type=Smp; RAW_FILE_NAME=170419_Smp_S14C1_C;170419_Smp_S14C1_C;170425_Smp_S14C1_C SUBJECT_SAMPLE_FACTORS - April1718AqExtracts_1 Depth_m:NA | Vol_L:NA Latitude=NA; Longitude=NA; UTC=NA; Replicate=1; Type=Pool; RAW_FILE_NAME=170419_Poo_April1718AqExtracts_1;170419_Poo_April1718AqExtracts_1 SUBJECT_SAMPLE_FACTORS - April1718AqExtracts_2 Depth_m:NA | Vol_L:NA Latitude=NA; Longitude=NA; UTC=NA; Replicate=2; Type=Pool; RAW_FILE_NAME=170419_Poo_April1718AqExtracts_2;170419_Poo_April1718AqExtracts_2 SUBJECT_SAMPLE_FACTORS - April1718AqExtracts_3 Depth_m:NA | Vol_L:NA Latitude=NA; Longitude=NA; UTC=NA; Replicate=3; Type=Pool; RAW_FILE_NAME=170419_Poo_April1718AqExtracts_3;170419_Poo_April1718AqExtracts_3 SUBJECT_SAMPLE_FACTORS - April21AqExtracts_1 Depth_m:NA | Vol_L:NA Latitude=NA; Longitude=NA; UTC=NA; Replicate=1; Type=Pool; RAW_FILE_NAME=170419_Poo_April21AqExtracts_1;170419_Poo_April21AqExtracts_1 SUBJECT_SAMPLE_FACTORS - April21AqExtracts_2 Depth_m:NA | Vol_L:NA Latitude=NA; Longitude=NA; UTC=NA; Replicate=2; Type=Pool; RAW_FILE_NAME=170419_Poo_April21AqExtracts_2;170419_Poo_April21AqExtracts_2 SUBJECT_SAMPLE_FACTORS - April21AqExtracts_3 Depth_m:NA | Vol_L:NA Latitude=NA; Longitude=NA; UTC=NA; Replicate=3; Type=Pool; RAW_FILE_NAME=170419_Poo_April21AqExtracts_3;170419_Poo_April21AqExtracts_3 SUBJECT_SAMPLE_FACTORS - QC_Half1 Depth_m:NA | Vol_L:NA Latitude=NA; Longitude=NA; UTC=NA; Replicate=1; Type=Pool; RAW_FILE_NAME=170419_Poo_QC_Half1;170419_Poo_QC_Half1 SUBJECT_SAMPLE_FACTORS - QC_Half2 Depth_m:NA | Vol_L:NA Latitude=NA; Longitude=NA; UTC=NA; Replicate=2; Type=Pool; RAW_FILE_NAME=170419_Poo_QC_Half2;170419_Poo_QC_Half2 SUBJECT_SAMPLE_FACTORS - QC_Half3 Depth_m:NA | Vol_L:NA Latitude=NA; Longitude=NA; UTC=NA; Replicate=3; Type=Pool; RAW_FILE_NAME=170419_Poo_QC_Half3;170419_Poo_QC_Half3 SUBJECT_SAMPLE_FACTORS - QC_Half4 Depth_m:NA | Vol_L:NA Latitude=NA; Longitude=NA; UTC=NA; Replicate=4; Type=Pool; RAW_FILE_NAME=170419_Poo_QC_Half4;170419_Poo_QC_Half4 SUBJECT_SAMPLE_FACTORS - QC_Full1 Depth_m:NA | Vol_L:NA Latitude=NA; Longitude=NA; UTC=NA; Replicate=1; Type=Pool; RAW_FILE_NAME=170419_Poo_QC_Full1;170419_Poo_QC_Full1 SUBJECT_SAMPLE_FACTORS - QC_Full2 Depth_m:NA | Vol_L:NA Latitude=NA; Longitude=NA; UTC=NA; Replicate=2; Type=Pool; RAW_FILE_NAME=170419_Poo_QC_Full2;170419_Poo_QC_Full2 SUBJECT_SAMPLE_FACTORS - QC_Full3 Depth_m:NA | Vol_L:NA Latitude=NA; Longitude=NA; UTC=NA; Replicate=3; Type=Pool; RAW_FILE_NAME=170419_Poo_QC_Full3;170419_Poo_QC_Full3 SUBJECT_SAMPLE_FACTORS - QC_Full4 Depth_m:NA | Vol_L:NA Latitude=NA; Longitude=NA; UTC=NA; Replicate=4; Type=Pool; RAW_FILE_NAME=170419_Poo_QC_Full4;170419_Poo_QC_Full4 SUBJECT_SAMPLE_FACTORS - TruePoo_Half1 Depth_m:NA | Vol_L:NA Latitude=NA; Longitude=NA; UTC=NA; Replicate=1; Type=Pool; RAW_FILE_NAME=170425_Poo_TruePoo_Half1 SUBJECT_SAMPLE_FACTORS - TruePoo_Half2 Depth_m:NA | Vol_L:NA Latitude=NA; Longitude=NA; UTC=NA; Replicate=2; Type=Pool; RAW_FILE_NAME=170425_Poo_TruePoo_Half2 SUBJECT_SAMPLE_FACTORS - TruePoo_Half3 Depth_m:NA | Vol_L:NA Latitude=NA; Longitude=NA; UTC=NA; Replicate=3; Type=Pool; RAW_FILE_NAME=170425_Poo_TruePoo_Half3 SUBJECT_SAMPLE_FACTORS - TruePoo_Half4 Depth_m:NA | Vol_L:NA Latitude=NA; Longitude=NA; UTC=NA; Replicate=4; Type=Pool; RAW_FILE_NAME=170425_Poo_TruePoo_Half4 SUBJECT_SAMPLE_FACTORS - TruePoo_Full1 Depth_m:NA | Vol_L:NA Latitude=NA; Longitude=NA; UTC=NA; Replicate=1; Type=Pool; RAW_FILE_NAME=170425_Poo_TruePoo_Full1 SUBJECT_SAMPLE_FACTORS - TruePoo_Full2 Depth_m:NA | Vol_L:NA Latitude=NA; Longitude=NA; UTC=NA; Replicate=2; Type=Pool; RAW_FILE_NAME=170425_Poo_TruePoo_Full2 SUBJECT_SAMPLE_FACTORS - TruePoo_Full3 Depth_m:NA | Vol_L:NA Latitude=NA; Longitude=NA; UTC=NA; Replicate=3; Type=Pool; RAW_FILE_NAME=170425_Poo_TruePoo_Full3 SUBJECT_SAMPLE_FACTORS - TruePoo_Full4 Depth_m:NA | Vol_L:NA Latitude=NA; Longitude=NA; UTC=NA; Replicate=4; Type=Pool; RAW_FILE_NAME=170425_Poo_TruePoo_Full4 #COLLECTION CO:COLLECTION_SUMMARY Samples for particulate metabolites from cruise KOK1606 were collected aboard CO:COLLECTION_SUMMARY the Ka’imikai-O- Kanoloa research vessel from April 20 to May 2, 2016, all at CO:COLLECTION_SUMMARY 15 m. At each sampling location, single, duplicate, or triplicate filters were CO:COLLECTION_SUMMARY collected using either niskins attached to a conductivity, temperature, depth CO:COLLECTION_SUMMARY array (CTD) or the underway intake. Samples (10-15 L) were collected into CO:COLLECTION_SUMMARY polycarbonate carboys, filtered onto 147 mm 0.2 μm PTFE filters using CO:COLLECTION_SUMMARY peristaltic pumps, polycarbonate filter holders, and Masterflex PharMed BPT CO:COLLECTION_SUMMARY tubing (Cole-Parmer). Filters were flash frozen in liquid nitrogen and stored at CO:COLLECTION_SUMMARY -80°C until extraction. Blank PTFE filters were extracted alongside samples as CO:COLLECTION_SUMMARY methodological blanks. CO:SAMPLE_TYPE Suspended Marine Particulate Matter CO:STORAGE_CONDITIONS Described in summary #TREATMENT TR:TREATMENT_SUMMARY No treatment - this was a study of the of natural marine microbial population in TR:TREATMENT_SUMMARY the surface ocean from the North Pacific Subtropical Gyre to North Pacific TR:TREATMENT_SUMMARY Transition Zone. #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 50:50 methanol:water. All aqueous SP:SAMPLEPREP_SUMMARY rinses were combined for each sample and dried down under N2 gas. The remaining SP:SAMPLEPREP_SUMMARY organic layer was transferred into a clean glass centrifuge tube and the SP:SAMPLEPREP_SUMMARY remaining bead beating tube was rinsed two more times with cold organic solvent. SP:SAMPLEPREP_SUMMARY The combined organic rinses were centrifuged, transferred to a new tube, and SP:SAMPLEPREP_SUMMARY dried under N2 gas. Dried aqueous fractions were re-dissolved in 380 µL of SP:SAMPLEPREP_SUMMARY water. Dried organic fractions were re-dissolved in 380 µL of 1:1 SP:SAMPLEPREP_SUMMARY water:acetonitrile. 20 µL of isotope-labeled injection standards in water were SP:SAMPLEPREP_SUMMARY added to both fractions. Blank filters were extracted alongside samples as SP:SAMPLEPREP_SUMMARY methodological blanks. SP:PROCESSING_STORAGE_CONDITIONS On ice SP:EXTRACTION_METHOD Bligh-Dyer SP:EXTRACT_STORAGE -80℃ #CHROMATOGRAPHY CH:CHROMATOGRAPHY_SUMMARY See attached protocol 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 CH_Ingalls_Lab_LC_Methods.txt;MS_Ingalls_Lab_MS_Methods.txt #ANALYSIS AN:ANALYSIS_TYPE MS #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 ST001410_AN002360_Results.txt UNITS:Adjusted and normalized peak areas Has m/z:Yes Has RT:Yes RT units:Seconds #END