#METABOLOMICS WORKBENCH dagaul_20210715_105514 DATATRACK_ID:2747 STUDY_ID:ST001882 ANALYSIS_ID:AN003043 PROJECT_ID:000000 VERSION 1 CREATED_ON July 15, 2021, 4:41 pm #PROJECT PR:PROJECT_TITLE Predator Cues Target Signaling Pathways in Toxic Algal Metabolome Protocol PR:PROJECT_SUMMARY Early detection of predators is critical to the survival of all living PR:PROJECT_SUMMARY organisms. For phytoplankton, recognition and response to chemical cues from PR:PROJECT_SUMMARY predators, as evidence of predation risk, is particularly crucial. The PR:PROJECT_SUMMARY phytoplankton Alexandrium minutum upregulates its toxicity when exposed to PR:PROJECT_SUMMARY copepodamides, a suite of compounds released by copepod predators. However, how PR:PROJECT_SUMMARY A. minutum perceives these predatory cues and what metabolic pathways are PR:PROJECT_SUMMARY involved in initiating toxin induction remains unknown. In this study LC/MS and PR:PROJECT_SUMMARY NMR-based metabolomics uncovered subtle physiological responses of A. minutum to PR:PROJECT_SUMMARY copepodamides, including dysregulation of valine biosynthesis and enhancement of PR:PROJECT_SUMMARY butanoate metabolism and arginine biosynthesis. While we have yet to identify a PR:PROJECT_SUMMARY chemoreceptor directly activated by copepod cues, based on the results of PR:PROJECT_SUMMARY inhibition experiments detection of copepodamides appears to disrupt the PR:PROJECT_SUMMARY activity of serine/threonine phosphatases leading to increased jasmonic acid PR:PROJECT_SUMMARY biosynthesis and signaling, which leads to amplified gonyautoxin biosynthesis in PR:PROJECT_SUMMARY A. minutum. This study is an important step toward a better understanding of PR:PROJECT_SUMMARY chemosensory ecology of predator-prey interactions in phytoplankton. PR:INSTITUTE Georgia Institute of Technology PR:LAST_NAME Brown PR:FIRST_NAME Emily PR:ADDRESS 950 Atlantic Drive, Atlanta GA 30332 PR:EMAIL emily.brown@gatech.edu PR:PHONE 404-894-8424 #STUDY ST:STUDY_TITLE LC-MS for Predator Cues Target Signaling Pathways in Toxic Algal Metabolome ST:STUDY_TITLE Protocol ST:STUDY_SUMMARY Metabolomics investigation of the phytoplankton Alexandrium minutum with and ST:STUDY_SUMMARY without copepod cues in order to explore cell signaling involved in toxin ST:STUDY_SUMMARY induction. ST:INSTITUTE Georgia Institute of Technology ST:LAST_NAME Brown ST:FIRST_NAME Emily ST:ADDRESS 950 Atlantic Drive, Atlanta GA 30332 ST:EMAIL emily.brown@gatech.edu ST:PHONE 404-894-8424 #SUBJECT SU:SUBJECT_TYPE Other organism SU:SUBJECT_SPECIES Alexandrium minutum #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 - Aq_Control_01 Factor:no exposure RAW_FILE_NAME=Aq_Control_01 SUBJECT_SAMPLE_FACTORS - Aq_Control_02 Factor:no exposure RAW_FILE_NAME=Aq_Control_02 SUBJECT_SAMPLE_FACTORS - Aq_Control_03 Factor:no exposure RAW_FILE_NAME=Aq_Control_03 SUBJECT_SAMPLE_FACTORS - Aq_Control_04 Factor:no exposure RAW_FILE_NAME=Aq_Control_04 SUBJECT_SAMPLE_FACTORS - Aq_Control_05 Factor:no exposure RAW_FILE_NAME=Aq_Control_05 SUBJECT_SAMPLE_FACTORS - Aq_Control_06 Factor:no exposure RAW_FILE_NAME=Aq_Control_06 SUBJECT_SAMPLE_FACTORS - Aq_Control_07 Factor:no exposure RAW_FILE_NAME=Aq_Control_07 SUBJECT_SAMPLE_FACTORS - Aq_Control_08 Factor:no exposure RAW_FILE_NAME=Aq_Control_08 SUBJECT_SAMPLE_FACTORS - Aq_Control_09 Factor:no exposure RAW_FILE_NAME=Aq_Control_09 SUBJECT_SAMPLE_FACTORS - Aq_Control_10 Factor:no exposure RAW_FILE_NAME=Aq_Control_10 SUBJECT_SAMPLE_FACTORS - Aq_Control_11 Factor:no exposure RAW_FILE_NAME=Aq_Control_11 SUBJECT_SAMPLE_FACTORS - Aq_Control_12 Factor:no exposure RAW_FILE_NAME=Aq_Control_12 SUBJECT_SAMPLE_FACTORS - Aq_Control_13 Factor:no exposure RAW_FILE_NAME=Aq_Control_13 SUBJECT_SAMPLE_FACTORS - Aq_Control_14 Factor:no exposure RAW_FILE_NAME=Aq_Control_14 SUBJECT_SAMPLE_FACTORS - Aq_Control_15 Factor:no exposure RAW_FILE_NAME=Aq_Control_15 SUBJECT_SAMPLE_FACTORS - Aq_Control_16 Factor:no exposure RAW_FILE_NAME=Aq_Control_16 SUBJECT_SAMPLE_FACTORS - Aq_Control_17 Factor:no exposure RAW_FILE_NAME=Aq_Control_17 SUBJECT_SAMPLE_FACTORS - Aq_Control_18 Factor:no exposure RAW_FILE_NAME=Aq_Control_18 SUBJECT_SAMPLE_FACTORS - Aq_Control_19 Factor:no exposure RAW_FILE_NAME=Aq_Control_19 SUBJECT_SAMPLE_FACTORS - Aq_Control_20 Factor:no exposure RAW_FILE_NAME=Aq_Control_20 SUBJECT_SAMPLE_FACTORS - Aq_COP_01 Factor:Copepodamide exposure RAW_FILE_NAME=Aq_COP_01 SUBJECT_SAMPLE_FACTORS - Aq_COP_02 Factor:Copepodamide exposure RAW_FILE_NAME=Aq_COP_02 SUBJECT_SAMPLE_FACTORS - Aq_COP_03 Factor:Copepodamide exposure RAW_FILE_NAME=Aq_COP_03 SUBJECT_SAMPLE_FACTORS - Aq_COP_04 Factor:Copepodamide exposure RAW_FILE_NAME=Aq_COP_04 SUBJECT_SAMPLE_FACTORS - Aq_COP_05 Factor:Copepodamide exposure RAW_FILE_NAME=Aq_COP_05 SUBJECT_SAMPLE_FACTORS - Aq_COP_06 Factor:Copepodamide exposure RAW_FILE_NAME=Aq_COP_06 SUBJECT_SAMPLE_FACTORS - Aq_COP_07 Factor:Copepodamide exposure RAW_FILE_NAME=Aq_COP_07 SUBJECT_SAMPLE_FACTORS - Aq_COP_08 Factor:Copepodamide exposure RAW_FILE_NAME=Aq_COP_08 SUBJECT_SAMPLE_FACTORS - Aq_COP_09 Factor:Copepodamide exposure RAW_FILE_NAME=Aq_COP_09 SUBJECT_SAMPLE_FACTORS - Aq_COP_10 Factor:Copepodamide exposure RAW_FILE_NAME=Aq_COP_10 SUBJECT_SAMPLE_FACTORS - Aq_COP_11 Factor:Copepodamide exposure RAW_FILE_NAME=Aq_COP_11 SUBJECT_SAMPLE_FACTORS - Aq_COP_12 Factor:Copepodamide exposure RAW_FILE_NAME=Aq_COP_12 SUBJECT_SAMPLE_FACTORS - Aq_COP_13 Factor:Copepodamide exposure RAW_FILE_NAME=Aq_COP_13 SUBJECT_SAMPLE_FACTORS - Aq_COP_14 Factor:Copepodamide exposure RAW_FILE_NAME=Aq_COP_14 SUBJECT_SAMPLE_FACTORS - Aq_COP_15 Factor:Copepodamide exposure RAW_FILE_NAME=Aq_COP_15 SUBJECT_SAMPLE_FACTORS - Aq_COP_16 Factor:Copepodamide exposure RAW_FILE_NAME=Aq_COP_16 SUBJECT_SAMPLE_FACTORS - Aq_COP_17 Factor:Copepodamide exposure RAW_FILE_NAME=Aq_COP_17 SUBJECT_SAMPLE_FACTORS - Aq_COP_18 Factor:Copepodamide exposure RAW_FILE_NAME=Aq_COP_18 SUBJECT_SAMPLE_FACTORS - Aq_COP_19 Factor:Copepodamide exposure RAW_FILE_NAME=Aq_COP_19 SUBJECT_SAMPLE_FACTORS - Aq_COP_20 Factor:Copepodamide exposure RAW_FILE_NAME=Aq_COP_20 SUBJECT_SAMPLE_FACTORS - Blank_SB_05 Factor:QC RAW_FILE_NAME=Blank_SB_05 SUBJECT_SAMPLE_FACTORS - Blank_SB_06 Factor:QC RAW_FILE_NAME=Blank_SB_06 SUBJECT_SAMPLE_FACTORS - Quality_Control_01 Factor:QC RAW_FILE_NAME=Quality_Control_01 SUBJECT_SAMPLE_FACTORS - Quality_Control_02 Factor:QC RAW_FILE_NAME=Quality_Control_02 SUBJECT_SAMPLE_FACTORS - Quality_Control_03 Factor:QC RAW_FILE_NAME=Quality_Control_03 SUBJECT_SAMPLE_FACTORS - Quality_Control_04 Factor:QC RAW_FILE_NAME=Quality_Control_04 SUBJECT_SAMPLE_FACTORS - Quality_Control_05 Factor:QC RAW_FILE_NAME=Quality_Control_05 SUBJECT_SAMPLE_FACTORS - Quality_Control_06 Factor:QC RAW_FILE_NAME=Quality_Control_06 SUBJECT_SAMPLE_FACTORS - Quality_Control_07 Factor:QC RAW_FILE_NAME=Quality_Control_07 #COLLECTION CO:COLLECTION_SUMMARY From culture (alexandrium minutum strain CCMP 113) CO:SAMPLE_TYPE Algae #TREATMENT TR:TREATMENT_SUMMARY Copepodamide que #SAMPLEPREP SP:SAMPLEPREP_SUMMARY Modified Bligh Dyer #CHROMATOGRAPHY CH:CHROMATOGRAPHY_SUMMARY Polar HILIC separation CH:CHROMATOGRAPHY_TYPE HILIC CH:INSTRUMENT_NAME Thermo Vanquish CH:COLUMN_NAME Waters Acquity BEH HILIC (150 x 2.1mm, 1.7um) #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME Thermo Orbitrap ID-X Tribrid MS:INSTRUMENT_TYPE Orbritrap/ion trap MS:MS_TYPE ESI MS:ION_MODE NEGATIVE MS:MS_COMMENTS Compound Discoverer MS:MS_RESULTS_FILE ST001882_AN003043_Results.txt UNITS:peak area Has m/z:Neutral masses Has RT:Yes RT units:Minutes #END