#METABOLOMICS WORKBENCH omkar123_20220825_200544 DATATRACK_ID:3433 STUDY_ID:ST002282 ANALYSIS_ID:AN003727 VERSION 1 CREATED_ON 09-01-2023 #PROJECT PR:PROJECT_TITLE Arabdiopsis Root VOCs PR:PROJECT_SUMMARY The rhizosphere is a unique niche surrounding plant roots, where soluble and PR:PROJECT_SUMMARY volatile molecules mediate signaling between plants and the associated PR:PROJECT_SUMMARY microbiota. The preferred lifestyle of soil microbes is in the form of biofilms. PR:PROJECT_SUMMARY However, little is known about whether root VOCs (rVOCs) can influence soil PR:PROJECT_SUMMARY biofilms beyond the 2-10 mm rhizosphere zone influenced by soluble root PR:PROJECT_SUMMARY exudates. Here, we report that rVOCs shift the microbiome composition and growth PR:PROJECT_SUMMARY dynamics of complex soil biofilms. This signaling is evolutionarily conserved PR:PROJECT_SUMMARY from ferns to higher plants, which suggests its coevolution. The defense PR:PROJECT_SUMMARY phytohormone methyl jasmonate (MeJA) is present in rVOCs and drives this PR:PROJECT_SUMMARY bioactivity at nanomolar levels within a few hours. PR:INSTITUTE National University of Singapore PR:DEPARTMENT Biological Sciences PR:LABORATORY AESB Lab PR:LAST_NAME Kulkarni PR:FIRST_NAME Omkar PR:ADDRESS NA PR:EMAIL komkar21@gmail.com PR:PHONE 98668733 PR:DOI http://dx.doi.org/10.21228/M8R997 #STUDY ST:STUDY_TITLE Detection of Methyl jasmonate (MeJA) in Plant root VOCs ST:STUDY_SUMMARY Methyl jasmonate (MeJA) is a well-known plant hormone known for plant defense ST:STUDY_SUMMARY and plant-plant signaling. However, most of the studies are focussed on its ST:STUDY_SUMMARY aboveground presence and functions. Here we report that MeJA is also released by ST:STUDY_SUMMARY plant roots in a volatile form. More importantly, it is shown in Arabidopsis ST:STUDY_SUMMARY growing in natural conditions in soil. ST:INSTITUTE National University of Singapore ST:LAST_NAME Kulkarni ST:FIRST_NAME Omkar ST:ADDRESS Dept of Biological Sciences,Metabolites Biology Lab,, Science drive 4,Block S1A ST:ADDRESS #06-03 ST:EMAIL komkar21@gmail.com ST:PHONE 98668733 ST:SUBMIT_DATE 2022-08-25 #SUBJECT SU:SUBJECT_TYPE Plant SU:SUBJECT_SPECIES Arabidopsis thaliana SU:TAXONOMY_ID 3702 SU:AGE_OR_AGE_RANGE 2 weeks #SUBJECT_SAMPLE_FACTORS: SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Additional sample data SUBJECT_SAMPLE_FACTORS 4 tube10_jmt1B Genotype:jmt RAW_FILE_NAME=tube10_jmt1B.D SUBJECT_SAMPLE_FACTORS 5 tube11_jmt2A Genotype:jmt RAW_FILE_NAME=tube11_jmt2A.D SUBJECT_SAMPLE_FACTORS 5 tube12_jmt2B Genotype:jmt RAW_FILE_NAME=tube12_jmt2B.D SUBJECT_SAMPLE_FACTORS 6 tube13_jmt3A Genotype:jmt RAW_FILE_NAME=tube13_jmt3A.D SUBJECT_SAMPLE_FACTORS 6 tube14_jmt3B Genotype:jmt RAW_FILE_NAME=tube14_jmt3B.D SUBJECT_SAMPLE_FACTORS 4 tube9_jmt1A Genotype:jmt RAW_FILE_NAME=tube9_jmt1A.D SUBJECT_SAMPLE_FACTORS NA tube1 Genotype:NA RAW_FILE_NAME=tube1.D SUBJECT_SAMPLE_FACTORS NA tube2_emptypot Genotype:NA RAW_FILE_NAME=tube2_emptypot.D SUBJECT_SAMPLE_FACTORS 1 tube3_soil1A Genotype:NA RAW_FILE_NAME=tube3_soil1A.D SUBJECT_SAMPLE_FACTORS 1 tube4_soil1B Genotype:NA RAW_FILE_NAME=tube4_soil1B.D SUBJECT_SAMPLE_FACTORS 2 tube5_soil2A Genotype:NA RAW_FILE_NAME=tube5_soil2A.D SUBJECT_SAMPLE_FACTORS 2 tube6_soil2B Genotype:NA RAW_FILE_NAME=tube6_soil2B.D SUBJECT_SAMPLE_FACTORS 3 tube7_soil3A Genotype:NA RAW_FILE_NAME=tube7_soil3A.D SUBJECT_SAMPLE_FACTORS 3 tube8_soil3B Genotype:NA RAW_FILE_NAME=tube8_soil3B.D SUBJECT_SAMPLE_FACTORS 7 tube15_WT1A Genotype:WT RAW_FILE_NAME=tube15_WT1A.D SUBJECT_SAMPLE_FACTORS 7 tube16_WT1B Genotype:WT RAW_FILE_NAME=tube16_WT1B.D SUBJECT_SAMPLE_FACTORS 8 tube17_WT2A Genotype:WT RAW_FILE_NAME=tube17_WT2A.D SUBJECT_SAMPLE_FACTORS 8 tube18_WT2B Genotype:WT RAW_FILE_NAME=tube18_WT2B.D SUBJECT_SAMPLE_FACTORS 9 tube19_WT3A Genotype:WT RAW_FILE_NAME=tube19_WT3A.D SUBJECT_SAMPLE_FACTORS 9 tube20_WT3B Genotype:WT RAW_FILE_NAME=tube20_WT3B.D #COLLECTION CO:COLLECTION_SUMMARY Root VOCs and soil VOCs were trapped as described here (Schulz-Bohm et al., CO:COLLECTION_SUMMARY 2018). Briefly, 2 Tenax cartridges were fitted into the side arms of the glass CO:COLLECTION_SUMMARY pots in such a way that their opening was exposed toward the plant roots/soil. CO:COLLECTION_SUMMARY VOCs were sampled for 40 hours and immediately analyzed by thermal CO:COLLECTION_SUMMARY desorption-gas chromatography‒mass spectrometry. CO:SAMPLE_TYPE Plant #TREATMENT TR:TREATMENT_SUMMARY No specific treatment. #SAMPLEPREP SP:SAMPLEPREP_SUMMARY After 40 hours of VOC trapping, Tenax cartridges were immediately subjected to SP:SAMPLEPREP_SUMMARY thermal desorption. #CHROMATOGRAPHY CH:CHROMATOGRAPHY_SUMMARY Sample preparation and injection were performed using the fully automated CH:CHROMATOGRAPHY_SUMMARY Gerstel MPS-2 autosampler and Gerstel MAESTRO software. Volatile compounds were CH:CHROMATOGRAPHY_SUMMARY adsorbed on a Tenax TA tube. A thermal desorption unit (TDU) was used to CH:CHROMATOGRAPHY_SUMMARY thermally desorb the volatiles in splitless mode at 230°C for 10 min. To ensure CH:CHROMATOGRAPHY_SUMMARY that the volatiles released from the TDU were quantitatively trapped, a cooled CH:CHROMATOGRAPHY_SUMMARY injection system-programmed temperature vaporizer (CIS-PTV) was used. The CIS CH:CHROMATOGRAPHY_SUMMARY was heated from 80°C to 230°C at a rate of 12°C/s with the split valve closed CH:CHROMATOGRAPHY_SUMMARY during sample injection into the GC inlet. Analyses of volatile compounds were CH:CHROMATOGRAPHY_SUMMARY performed on an Agilent 7890B GC coupled to a 5977B quadruple mass spectrometer. CH:CHROMATOGRAPHY_SUMMARY Separation of compounds was performed on a DB-FFAP column (60 m x 250 µm x 0.25 CH:CHROMATOGRAPHY_SUMMARY µm, Agilent Technologies, Middleburg, OI, USA). Helium was used as the carrier CH:CHROMATOGRAPHY_SUMMARY gas at a flow rate of 1.9 ml/min, and solvent vent mode was used. The inlet CH:CHROMATOGRAPHY_SUMMARY temperature was 250°C. The oven program was as follows: initial temperature of CH:CHROMATOGRAPHY_SUMMARY 50°C held for 1 min, increased to 230°C at the rate of 10°C/min and held for CH:CHROMATOGRAPHY_SUMMARY 20 min. The temperature of the ion source and transfer line was 250°C. CH:INSTRUMENT_NAME Agilent 7890B CH:COLUMN_NAME Agilent DB-FFAP CH:CHROMATOGRAPHY_TYPE GC #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME Agilent 5977B MS:INSTRUMENT_TYPE Single quadrupole MS:MS_TYPE EI MS:MS_COMMENTS The mass spectrometer was in electron ionization mode with an ionization energy MS:MS_COMMENTS of 70 eV, scan range of 40-300 m/z and solvent delay of 3.75 minutes. Analysis MS:MS_COMMENTS was performed in single ion monitoring (SIM) mode by monitoring the ions at 83, MS:MS_COMMENTS 151.1, and 224.1 with a dwell time of 150 ms. Mass Hunter Qualitative Analysis MS:MS_COMMENTS was used to extract and integrate peak spectra. The peak area of these ions was MS:MS_COMMENTS considered for the relative quantification of MeJA among different samples. MS:ION_MODE UNSPECIFIED #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS ion intensity peak area MS_METABOLITE_DATA_START Samples tube10_jmt1B tube11_jmt2A tube12_jmt2B tube13_jmt3A tube14_jmt3B tube9_jmt1A tube1 tube2_emptypot tube3_soil1A tube4_soil1B tube5_soil2A tube6_soil2B tube7_soil3A tube8_soil3B tube15_WT1A tube16_WT1B tube17_WT2A tube18_WT2B tube19_WT3A tube20_WT3B Factors Genotype:jmt Genotype:jmt Genotype:jmt Genotype:jmt Genotype:jmt Genotype:jmt Genotype:NA Genotype:NA Genotype:NA Genotype:NA Genotype:NA Genotype:NA Genotype:NA Genotype:NA Genotype:WT Genotype:WT Genotype:WT Genotype:WT Genotype:WT Genotype:WT Methyl Jasmonate 51380.3500 12718.9000 15363.3500 33962.7200 581408.6500 2347.0800 11247.8800 14200.1900 42016.5100 20850.5350 8040.0100 603018.8700 117534.9900 13982.3400 168327.7200 6365.8400 271639.5400 6804.8800 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name pubchem_id inchi_key kegg_id other_id other_id_type ri ri_type moverz_quant Methyl Jasmonate 5281929 C11512 METABOLITES_END #END