MetaboLights

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were identified by comparison to library entries of purified standards or recurrent unknown entities. Metabolon maintains a library based on authenticated standards that contains the retention time/index (RI), mass to charge ratio (m/z), and chromatographic data (including MS/MS spectral data) on all molecules present in the library. More than 4500 commercially available purified standard compounds have been acquired and registered into LIMS for analysis on all platforms for determination of their analytical characteristics. Additional mass spectral entries have been created for structurally unnamed biochemicals, which have been identified by virtue of their recurrent nature (both chromatographic and mass spectral). These compounds have the potential to be identified by future acquisition of a matching purified standard or by classical structural analysis.MetaboLightsPublicQ ExactiveAll methods utilize a Waters ACQUITY ultra-performance liquid chromatography (UPLC). Each reconstitution solvent contains a series of standards at fixed concentrations to ensure injection and chromatographic consistency. Method 1/Pos 1; One aliquot is analyzed using acidic positive ion conditions, chromatographically optimized for more hydrophilic compounds. In this method, the extract is gradient-eluted from a C18 column (Waters UPLC BEH C18-2.1x100 mm, 1.7 µm) using water and methanol, containing 0.05% perfluoropentanoic acid (PFPA) and 0.1% formic acid (FA).Method 2/Pos 2; A second aliquot is also analyzed using acidic positive ion conditions, but is chromatographically optimized for more hydrophobic compounds. In this method, the extract is gradient eluted from the aforementioned C18 column using methanol, acetonitrile, water, 0.05% PFPA and 0.01% FA, and is operated at an overall higher organic content.Method 3/Neg 1; A third aliquot is analyzed using basic negative ion optimized conditions using a separate dedicated C18 column. The basic extracts are gradient-eluted from the column using methanol and water, however with 6.5 mM Ammonium Bicarbonate at pH 8.Method 4/Neg 2; The fourth aliquot is analyzed via negative ionization following elution from a HILIC column (Waters UPLC BEH Amide 2.1x150 mm, 1.7 µm) using a gradient consisting of water and acetonitrile with 10mM Ammonium Formate, pH 10.8.How Does the Seed Pre-Germinative Metabolism Fight Against Imbibition Damage? Emerging Roles of Fatty Acid Cohort and Antioxidant Defence. 10.3389/fpls.2019.01505. PMID:31824536Alma BalestrazziUniversity of PaviaseedSeedGerminated or sprouted seedmass spectrometrySamples were prepared using the automated MicroLab STAR® system from Hamilton Company. Several recovery standards were added prior to the first step in the extraction process for QC purposes. Samples were extracted with methanol under vigorous shaking for 2 min (Glen Mills GenoGrinder 2000) to precipitate protein and dissociate small molecules bound to protein or trapped in the precipitated protein matrix, followed by centrifugation to recover chemically diverse metabolites. The resulting extract was divided into five fractions: two for analysis by two separate reverse phase (RP)/UPLC-MS/MS methods using positive ion mode electrospray ionization (ESI), one for analysis by RP/UPLC-MS/MS using negative ion mode ESI, one for analysis by HILIC/UPLC-MS/MS using negative ion mode ESI, and one reserved for backup. Samples were placed briefly on a TurboVap® (Zymark) to remove the organic solvent and stored overnight under nitrogen. In preparation for analysis, samples were then reconstituted in solvents compatible to each of the four analysis methods.Medicago truncatulahttps://www.ebi.ac.uk/metabolights/MTBLS181Anca Macovei. UNIPV. Department of Biology and Biotechnology 'L. Spallanzani', via Ferrata 9, 27100 Pavia, Italy. anca.macovei@unipv.it.Enrico Doria. UNIPV. Department of Biology and Biotechnology 'L. Spallanzani', via Ferrata 9, 27100 Pavia, Italy. enrico.doria@unipv.it.Carla Ferreri. CNR. Research Area of Bologna, Via P. Gobetti 101- 40129 Bologna (Italy). carla.ferreri@isof.cnr.it.Anna Larocca. Lipinutragen srl. Laboratorio di Lipidomica, Via P. Gobetti 101, 40129 Bologna. info@lipinutragen.it.Andrea Pagano. UNIPV. Department of Biology and Biotechnology 'L. Spallanzani', via Ferrata 9, 27100 Pavia, Italy. andrea.pagano01@universitadipavia.it.Susana Araújo. ITQB-NOVA. Avenida da Republica, EAN, 2780-157-Oeiras, Portugal. saraujo@itqb.unl.pt.Alma Balestrazzi. UNIPV. Department of Biology and Biotechnology 'L. Spallanzani', via Ferrata 9, 27100 Pavia, Italy. alma.balestrazzi@unipv.it.Raw data were extracted, peak-identified, and QC processed using Metabolon’s hardware and software. These systems are built on a web-service platform utilizing Microsoft’s .NET technologies, which run on high-performance application servers and fiber-channel storage arrays in clusters to provide active failover and load-balancing.</td>Imbibition time pointPhenologyalma.balestrazzi@unipv.itFor imbibition experiments, Medicago truncatula seeds (Jemalong genotype ) were transferred to Petri dishes (diameter 90 mm) containing two filter papers moistened with 2.5 ml H2O and kept in a growth chamber at 22 °C under light conditions with photon flux density of 150 µmol m/s, photoperiod of 16/8 h and 70-80% relative humidity. M. truncatula seeds were collected at 0 h or dry seeds (DS), 2 h (W2) and 8 h (W8) after imbibition and at radicle protusion (WRD). After fresh weight (FW) measurement, samples were stored in liquid N2 for molecular and metabolomic analysis. Experiments were caried out twice. For each experiment, three replicates (20 seeds per replicate) were analysed. For each studied experimental conditions, the seed fresh weight (FW) was measured before sample were snap frozen into liquid N2 and stored at -80 ºC till further use. In paralel, similar seed samples with known fresh weight were used to estimate seed dry mass (after drying at 65 ºC, till reach a constant weight). For untargeted metabolomic analysis, DS, W2, W8 and WRD samples were grinded with mortar and pestle. Three biological replicates were made, in which each correspond to a pool of 20 seeds. Aproximately one hundred mg of frozen material was transferred to 1.5 ml tube and shipped (dry ice) to Metabolon.Metabolomicsultra-performance liquid chromatography-mass spectrometryImbibition damagelegume seedmetabolontandem mass spectrometryAntioxidant responseuntargeted metabolitesLipidomicsSeed germinationultra-performance liquid chromatography-mass spectrometryImbibition damagelegume seedmetabolontandem mass spectrometryAntioxidant responseuntargeted metabolitesLipidomicsSeed germinationA Thermo Scientific Q-Exactive high resolution/accurate mass spectrometer interfaced with a heated electrospray ionization (HESI-II) source and Orbitrap mass analyzer operated at 35,000 mass resolution was used. The MS analysis alternates between MS and data-dependent MSn scans using dynamic exclusion.pyridoxatesalicylate4-hydroxyphenylpyruvate1-palmitoyl-GPI (16:0)glycerophosphoethanolaminedeoxycarnitinecis-aconitatelinoleoyl-linoleoyl-glycerol (18:2|18:2) [1]thymidine1-palmitoyl-2-linoleoyl-digalactosylglycerol (16:0|18:2)2-hydroxyglutarate4-imidazoleacetatepyroglutaminetricarballylateglutamateshikimateorotate2'-deoxyadenosine1,2-dilinolenoyl-galactosylglycerol (18:3|18:3)cytidineglucuronatehomostachydrineN-acetyltyrosinebutyrylglutamine|isobutyrylglutaminemethionine sulfoxideadenosinechiro-inositollinoleoyl-linoleoyl-glycerol (18:2|18:2) [2]argininatecreatineN-formylphenylalaninexyloseglycylvaline2'-deoxycytidine1-palmitoyl-2-linoleoyl-GPI (16:0|18:2)benzoateoleoylcholinesedoheptuloseoleoyl-oleoyl-glycerol (18:1|18:1) [2]histaminealpha-ketoglutarategamma-glutamylglutamateoleoyl-linolenoyl-glycerol (18:1|18:3) [2]5-oxoprolineasparaginearabinoseN2,N2-dimethylguanosineglycerophosphoinositol1-palmitoyl-2-linoleoyl-GPC (16:0|18:2)1-palmitoyl-2-linoleoyl-GPE (16:0|18:2)cytosine1-palmitoyl-2-alpha-linolenoyl-GPC (16:0|18:3n3)2'-deoxyguanosinesalidrosidepantothenate (Vitamin B5)1-palmitoyl-2-linoleoyl-GPA (16:0|18:2)alaninethreonylphenylalanine2-hydroxypalmitatecys-gly, oxidizedoleoyl-oleoyl-glycerol (18:1|18:1) [1]UDP-glucose|UDP-galactoseglucosecitrateribose1-palmitoyl-GPC (16:0)3-hydroxy-3-methylglutaratenicotinamide ribosideprolylglycinedehydroascorbateN6-succinyladenosinearabonate|xylonateurateguanosinexanthineguanidinoacetatexanthosine1-linoleoyl-2-linolenoyl-digalactosylglycerol (18:2|18:3)lactatefumarate2-hydroxybehenatemethionine sulfoneN-acetylalaninetyrosylglycineergothioneinesulfatealpha-tocopherolcysteineapigeninpalmitoyl-oleoyl-glycerol (16:0|18:1) [1]succinimidepipecolate2-oxoadipateO-sulfo-L-tyrosinehomocitrullineargininepalmitoyl ethanolamideribulose|xylulosecystathioninetrans-aconitateribitolpalmitoyl-linolenoyl-glycerol (16:0|18:3) [2]methylphosphateN-acetyl-1-methylhistidinethreonateleucinepyridoxine (Vitamin B6)5'- GMPS-methylcysteineglycerophosphoglycerol3-deoxyoctulosonateagmatinegamma-glutamyltyrosinesoyasaponin IglycerolN-acetylvaline3-methylhistidine1,2-dilinoleoyl-GPC (18:2|18:2)lysinevalylleucinemethionineN-acetylmethionineallantoin1-methylguaninegulonateUMPN6-methyladenosinegamma-glutamyltryptophantyrosine1,2-dilinoleoyl-GPE (18:2|18:2)soyasaponin IIgamma-glutamylphenylalanine1-linoleoyl-GPC (18:2)1,2-dilinolenoyl-digalactosylglycerol (18:3|18:3)alpha-hydroxyisovalerateglucosaminatemaleatesuccinatemaltoseethanolamineN-delta-acetylornithine3-ureidopropionatesphinganineN-acetylglucosaminylasparaginephenylalaninepalmitoyl-oleoyl-glycerol (16:0|18:1) [2]phosphatefructose2-hydroxyoleateguanosine 3'-monophosphate (3'-GMP)1-palmitoyl-2-oleoyl-GPC (16:0|18:1)quinateS-carboxymethyl-L-cysteinepyruvate12,13-DiHOME5-methyluridine (ribothymidine)1-palmitoyl-GPE (16:0)3-hydroxyhexanoate2-hydroxymyristatenicotinateHEPES1-methyladeninecysteine-glutathione disulfideuracilmannitol|sorbitolbetainestigmasterolvalylglycinepyridoxalgluconateethylmalonateinosinedihomolinoleate (20:2n6)stachyosedihydrokaempferolisoleucinemyo-inositol2-methylcitrate|homocitratehomoserine (homoserine lactone)2-hydroxystearate1-palmitoyl-2-linolenoyl-GPA (16:0|18:3)4-hydroxybenzoatehypoxanthineglycinelinolenoyl-linolenoyl-glycerol (18:3|18:3) [2]isocitric lactone1-methylhistidinedigalactosylglycerolthiamin (Vitamin B1)glycerophosphoserinegamma-glutamylthreonine4-acetamidobutanoateglycerol 3-phosphateCMPcystinegamma-glutamylalaninemethylsuccinateN-acetylserinetyramineimidazole lactate5-methylcytidineallantoic acid2-hydroxyadipate2-oxoarginine3-methyl-2-oxobutyratehydroxyprolinekinetinguanosine 2'-monophosphate (2'-GMP)phosphocholine3-aminoisobutyrate13-HOTrE5-(2-Hydroxyethyl)-4-methylthiazolemalonate1,2-dilinoleoyl-GPA (18:2|18:2)glutaminearabitol|xylitololeoyl-linoleoyl-glycerol (18:1|18:2) [1]cholineferulateuridine 3'-monophosphate (3'-UMP)gamma-glutamylhistidinekynurenate4-hydroxy-2-nonenaltryptophan2,3-dihydroxyisovalerate1-methyladenosine2-isopropylmalate6-oxopiperidine-2-carboxylate3'-CMPgamma-tocopherol|beta-tocopherol1,2-dilinolenoyl-GPC (18:3|18:3)maltololeoyl-linoleoyl-glycerol (18:1|18:2) [2]orotidinebeta-alanine1-linoleoyl-2-linolenoyl-GPC (18:2|18:3)saccharopinelinolenoyl-linolenoyl-glycerol (18:3|18:3) [1]trizma acetateglycerategalactinollinoleoyl ethanolamideraffinosephytosphingosineNAD+histidine2-aminoheptanoategalactonateserinestearoyl-linoleoyl-glycerol (18:0|18:2) [1]N-acetylglutamineverbascosecysteinylglycinelinoleoyl-linolenoyl-glycerol (18:2|18:3) [2]nicotinamidetyrosolalanylleucinephenyllactate (PLA)S-adenosylhomocysteine (SAH)3-methyl-2-oxovaleratepalmitoyl-linoleoyl-glycerol (16:0|18:2) [2]beta-hydroxyisovalerateaspartatelinoleoyl-linolenoyl-glycerol (18:2|18:3) [1]spermidine2-hydroxybutyrate|2-hydroxyisobutyrateAMPbeta-sitosterolN-formylanthranilic acidpyridoxaminetaurineN-acetylphenylalaninesucroseerythronatexanthurenatenaringeninproline3-hydroxyoctanoatemalate4-methyl-2-oxopentanoateguaninehypotaurineoleate|vaccenate (18:1)cis-urocanate4-guanidinobutanoatecitrulline1-linoleoyl-GPA (18:2)1-linoleoyl-GPI (18:2)diacylglycerol (16:1|18:2 [2], 16:0|18:3 [1])stachydrinenicotianamine2'-deoxyuridineN-acetylasparagineuridineN-acetylglucosamine|N-acetylgalactosamine2-hydroxy-3-methylvalerateglycylisoleucinetrigonelline (N'-methylnicotinate)3-(4-hydroxyphenyl)lactate (HPLA)1-palmitoyl-2-linolenoyl-digalactosylglycerol (16:0|18:3)mannosestearoyl-linoleoyl-glycerol (18:0|18:2) [2]isoleucylglycine16-hydroxypalmitate3'-AMP1-palmitoyl-GPA (16:0)pseudouridineglycylleucineglutaminylleucineN-acetylhistidine3-phenylpropionate (hydrocinnamate)palmitoyl-linoleoyl-glycerol (16:0|18:2) [1]palmitoylcholineN-methylprolinethreonine9,10-DiHOMEadeninevalinethioprolineectoinegamma-glutamylvalineoleoyl ethanolamide1-oleoyl-2-linoleoyl-GPC (18:1|18:2)2-hydroxydecanoateerythritolgalactosylglycerolN-acetyl-beta-alanineDuring seed imbibition, lipids are engaged in membrane reorganization while facing free radical-mediated oxidative injury. In the present work, we explored changes in lipid components at different timepoints of imbibition (0.5, 2, 4, 6, and 8 h) in the legume Medicago truncatula, by combining biochemical approaches with targeted lipidomics and untargeted metabolomics. ROS and RNS (reactive oxygen and nitrogen species) accumulation was observed throughout the tested timepoints whereas lipid peroxidation increased at 4 h of imbibition. The seed response to oxidative damage was evidenced by a significant increase in tocopherols starting from 0.5 h of imbibition as well as by the reduction in total thiol content occurring at 2 h of imbibition. Since under physiological conditions, the proper functions of the cell membranes are strongly dependent on the qualitative and quantitative balance of fatty acid residues in phospholipids, the investigation was expanded to the fatty acid cohort of M. truncatula seeds. Total saturated fatty acids (SFAs), monounsaturated fatty acids (MUFAs), polyunsaturated fatty acids (PUFAs), omega(ω)-3 and omega(ω)-6 fatty acids showed fluctuations during seed imbibition. The most remarkable finding was the profile of the ω-3 PUFA docosopentaenoic acid (DPA, 7 cis, 10 cis, 13 cis, 16 cis, and 19 cis-22:5) that showed a peak (up to 1.0% of the total fatty acid content) at 0.5 and 8 h of imbibition, concomitant with the peaks observed in tocopherol levels. It is possible that the observed changes in DPA alter the physical properties of membranes, as reported in animal cells, triggering signaling pathways relevant for the cell defense against oxidative injury. Furthermore, the content and balance between tocopherols and PUFAs is regarded as a determinant of storage stability. No enhancement in trans-fatty acids occurred throughout imbibition, suggesting for a proper antioxidant response carried by the seed. Fatty acids profiles were integrated with data from untargeted metabolomics showing changes in lipid sub-pathways, among which fatty acid amide, lyso-phospholipids, and phospholipid metabolism. The emerging lipid profiles and dynamics are discussed in view of the overall imbibition damage generated during M. truncatula seed imbibition.How Does the Seed Pre-Germinative Metabolism Fight Against Imbibition Damage? Emerging Roles of Fatty Acid Cohort and Antioxidant Defence.Doria Enrico E, Pagano Andrea A, Ferreri Carla C, Larocca Anna Vita AV, Macovei Anca A, Araújo Susana de Sousa SS, Balestrazzi Alma Aacido graso, E Vicotrat, acidos grasos, phospholipids, Compounds, DMP4, Physical, Vitamin-E Dragees, Metabonomics, Tocolion, nutcracker syndrome, Vita E, Snow-E Muscle, Peroxidation, MAGE-E1 antigen, trauma, Vita-Plus E, BB234005, Thiol, dmTAF[[II]]230, UnoVit, responsivity, oxigeno, Vitamin E-mp, Monounsaturated, docosapentaenoic acids, GEF-CK, Dal E, Esterified, Animalia, TFIID TAF250, Equivit E, cel, and osteosclerosis, N, O, Esterified Fatty, Oxygen-16, Tissue, Plant Embryo, composition, 4-heptanecarboxylic acid, E-Vitamin-Ratiopharm, c-ros-1, Oxygen, Aliphatic Acids, [OO], E Mulsin, Detulin, single organism signaling, Diagnostic Findings, close to, Vit E hydrosol, Thiols, dTAF[[II]]230, Mercaptans, Injury and Wounds, animalia, Di-n-propylessigsaeure, Mercapto Compounds, acid, Antioxidants, TAF200, integral to membrane, Endogenous, Physical Traumas, l(2)43Ca, Embial, Phosphatides, Fettsaeuren, Tocopharm, Expanded, Hydrovit E, DmeMCM4, Stickstoff, Vit. E Stada, Cis, CIS, 1270, DAMAGE, Hepatocellular carcinoma-associated protein 1, hypoplastic nose and midface, G18, Micorvit E, Acid, Vitamin Emp, CIS-1, dioxygen, Fatty Acid, VPA, Saturated Fatty Acid, whole organism, content, Uno-Vit, expanded, Ex, Plant Zygote, apparently low-set ears, 9030605P22Rik, Bio E, Lipid Peroxidations, fatty acids, nitrogen, Richtavit E, MCF3, Taf250, Koerper, Research-Related Injuries, Dermorelle, Acids, Clinical Finding, Auxina E, TAF230, Dioxygen, Vitamin E Natur, Saturated Fatty, big, Research Related Injuries, lipids, Esterified Fatty Acids, Endogenous Antioxidant, Sulfhydryl, number, E-948, antioxydant, Amide, Injuries and Wounds, large, DmelCG4114, Wounds, 2-propylpentanoic acid, Davitamon, Dpa, DPA, NCS, T22P11.60, Fatty Acids, Injury, dTAF[[II]]250, Eusovit, Tissues, cell, mcm4, Zygote, Aquasol E, AI847422, dTAF250, Mowivit Vitamin E, 22:5, combination of microcephaly, Antioxidant Activity, species, DPA TRANSCRIPTION FACTOR, n-DPA, Saeure, Vitamin E mp, Eplonat, Elaiosomes, RNA, Oxygen 16, body, E-ferol, Puncto E, E-Vicotrat, CG4114, Evion, whole body, RNS, BG:DS00004.13, compositionality, Unique E, oxygen, Cell, exophthalmos, 2-propylvaleric acid, dTAF230, near to, Vibolex, Fettsaeure, Mercaptan, Medicago, traumatic injury, Wounds and Injury, Medicago truncatulas, Trauma, Antioxidant Effect, TAF[[II]]250/230, ROS, oxygene, Mcm4, MCM4, OXYGEN MOLECULE, Vitamin E Sanum, Taf[[II]]250, Trans-Fatty Acids, O2, Togasan, di-n-propylacetic acid, T22P11_60, Sulfhydryl Compound, RGD1560259, storage, Saeuren, increased number, pyrene (narrow)., wound, azote, acide gras, Trans Fatty, Tocopa, l(2)01270, signalling process, Left renal vein entrapment syndrome, Compound, Mercapto, Togasan Vitamin E, vicinity of, structure, sequestering, E 948, 2-n-propyl-n-valeric acid, Vitazell, pyrene (narrow), Anti Oxidants, Activity, Wound, Lipidomic, Trans Fatty Acid, Metabonomic, GRO:0005339, 2610511A05Rik, injury, composed of, brl, gum hyperplasia, Tocovital, Membrane Tissues, Tier, Spondyvit, NH2(-), dipropylacetic acid, Saturated Fatty Acids, azanide, Esterified Fatty Acid, acide valproique, Valproinsaeure, Elaiosome, E948, Research-Related, increased, Anti Oxidant, seed, Tocopherol Bayer, EUNOVA Vitamin E, membrane region, Aliphatic, Malton E, free, Trans, retention, E Vitamin E, HCA1, Diaspores, Antioxidant, Anti-Oxidant Effect, Protein G18, Lasar, Vitamin E EVI MIRALE, truncatula, Membrane Tissue, SIGNS SYMPTOMS, amides, VitaminE EVIMIRALE, Vita-E, ribose nucleic acid, Monounsaturated Fatty Acids, Dal-E, DESC, Sanavitan S, ribonucleic acids, Ecoro, Bioweyxin, TAFII-250, TAF250/230, Anti-Oxidants, TAFII250, Abortosan, Renal Nutcracker Phenomenon, Vitamine E GNR, Sauerstoff, <new synonym>, Ribonukleinsaeure, Lipid, Depakene, pentosenucleic acids, Physical Trauma, Seed, Diaspore, Disauerstoff, antioxidants, MCM4_DROME, acide, acidum valproicum, Plant, acids, Aliphatic Acid, acido, CG17603, TAF[[II]], Vitamin E AL, Lipidome, Endogenous Antioxidants, Mowivit, enlarged, phospholipid metabolism, SR3-5, l(2)ey, Research-Related Injury, 7N, mMage-e1, Tocopherol, accessory, Vitagutt Vitamin E, d230, E-Mulsin, antoxidant, Vitamin E Dragees, Trans-Fatty, Vitamin E, dTAFII250, 8O, EfW1, supernumerary, presence, Symptoms and Signs, acido valproico, 43Ca, VitaE, dioxygene, acides gras, dmTAF1, Taf230, valproic acid, integral component of membrane, DMP-4, Anti-Oxidant, Animal, Finding, Anti-Oxidant Effects, Metabolomic, CG1616, Aggravating interaction, TAF250, study, reactivity, Vita Plus E, Taf200, Phospholipid, Taf1p, metazoa, Ephynal, Biosan, Traumas, great, region of membrane, Zygotes, VitaPlus E, amide, barrel medic, TAF, BACTS2, molecular oxygen, ahl4, membrane, Energy & Feritility, TAF[[II]]250, Saturated, DmelCG1616, dihydridonitrate(1-), Alpha-dystrobrevin-associated MAGE Protein, Lipidomes, l(3)84Ab, Monounsaturated Fatty Acid, Anti Oxidant Effect, Elex Verla, Monounsaturated Fatty, count in organism, Signs and Symptoms, Vitamin-E EVI-MIRALE, yeast nucleic acid, p230, Embryos, Metazoa, Peroxidations, whole membrane, TFIID, Biopto-E, Embryo, ribonucleic acid, a phospholipid derivative, Anti Oxidant Effects, transmembrane, TAF[[II]]230, 2-PROPYL-PENTANOIC ACID, Vitamin E Suspension, Antioxidans E-Hevert, TAF[II]250, Membrane, Plant Embryos, Injuries, nitrogeno, present in greater numbers in organism, Antioxidant Effects, Vitagutt, Uno Vit, DmelCG17603, DmMcm4, cleft palate, Plant Zygotes, SOCS, approaches, DmMCM4, response, Suppressor of cytokine signaling, C22:5, TAF1biochemical pathways, Saturated Fatty, acido graso, Anti Oxidants, Metabolic Process, Esterified Fatty Acids, Anti Oxidant Effect., acidos grasos, PhrB photolyase activity, Activity, Processes, P62, Endogenous Antioxidant, antoxidant, Metabolic Concepts, antioxydant, sci, GRO:0005339, photoreactivating enzyme activity, Metabolic Processes, pigmented epithelium, MAGE-E1 antigen, acides gras, Metabolism, Anti-Oxidant, HOW, How, Concepts, Saturated Fatty Acids, 3, stratum pigmentosa retinae, Esterified Fatty Acid, NUP96, Metabolism Concept, Anti-Oxidant Effects, Elaiosome, Phenomenon, epithelium, Metabolism Phenomena, l(3)j5D5, Esterified, 24B, Fatty Acids, Anti Oxidant, seed, MOS3, deoxyribocyclobutadipyrimidine pyrimidine-lyase activity, l(3)s2612, PRECOCIOUS, catabolism, Esterified Fatty, Aliphatic, stru, pigmented retina, F23A5.3, Plant Embryo, Metabolic Concept, Zygote, metabolic process resulting in cell growth, l(3)S053606, AI847422, CG10293, DNA cyclobutane dipyrimidine photolyase activity, SUPPRESSOR OF AUXIN RESISTANCE 3, l(3)j5B5, Aliphatic Acids, HCA1, Diaspores, Antioxidant, Antioxidant Activity, DmelCG10293, Anti-Oxidant Effect, Zygotes, biotransformation, Catabolism, F23A5_3, Elaiosomes, PRE, 0904/17, Saturated, degradation, Process, clone 2.39, deoxyribonucleic cyclobutane dipyrimidine photolyase activity, metabolism resulting in cell growth, Alpha-dystrobrevin-associated MAGE Protein, Antioxidants, Endogenous, Fettsaeuren, qkr, l(3)S090417, Anti-Oxidants, Concept, Metabolic Phenomena, Metabolism Concepts, Fettsaeure, MODIFIER OF SNC1, retinal pigment, SZ1, KH93F, Embryos, Antioxidant Effect, DAMAGE, Hepatocellular carcinoma-associated protein 1, deoxyribonucleic photolyase activity, Phenomena, dipyrimidine photolyase (photosensitive), secretion, Seed, metabolism, retinal pigment layer, Metabolic Phenomenon, who, Embryo, Acid, Anti Oxidant Effects, Diaspore, multicellular organism metabolic process, Fatty Acid, antioxidants, RPE, Saturated Fatty Acid, biodegradation, Metabolic, photolyase activity, RGD1560259, Plant, Aliphatic Acid, Plant Zygote, Who/How, acide gras, Plant Embryos, p. pigmentosa retinae, Antioxidant Effects, phr A photolyase activity, fatty acids, Endogenous Antioxidants, DNA-photoreactivating enzyme, Plant Zygotes, qkr[93F], anon-EST:Liang-2.39, deoxyribonucleate pyrimidine dimer lyase (photosensitive), mMage-e1, pyrene (narrow), Anabolismacido graso, E Vicotrat, acidos grasos, phospholipids, Compounds, DMP4, Physical, Vitamin-E Dragees, Metabonomics, Tocolion, nutcracker syndrome, Vita E, Snow-E Muscle, Peroxidation, MAGE-E1 antigen, trauma, Vita-Plus E, BB234005, Thiol, dmTAF[[II]]230, UnoVit, DmelCG12298, responsivity, oxigeno, Vitamin E-mp, SCRAMBLED, docosapentaenoic acids, GEF-CK, Dal E, Esterified, Animalia, TFIID TAF250, Equivit E, cel, and osteosclerosis, N, O, Esterified Fatty, Oxygen-16, Tissue, Plant Embryo, composition, 4-heptanecarboxylic acid, E-Vitamin-Ratiopharm, c-ros-1, Oxygen, Aliphatic Acids, [OO], E Mulsin, Detulin, single organism signaling, Diagnostic Findings, close to, Vit E hydrosol, Thiols, dTAF[[II]]230, Mercaptans, Injury and Wounds, animalia, Di-n-propylessigsaeure, Mercapto Compounds, acid, Antioxidants, STRUBBELIG, TAF200, integral to membrane, Endogenous, Physical Traumas, l(2)43Ca, Embial, Phosphatides, Fettsaeuren, Tocopharm, Expanded, Hydrovit E, DmeMCM4, Stickstoff, Vit. E Stada, Cis, CIS, 1270, DAMAGE, Hepatocellular carcinoma-associated protein 1, hypoplastic nose and midface, G18, Micorvit E, Acid, Vitamin Emp, CIS-1, dioxygen, Fatty Acid, VPA, Saturated Fatty Acid, whole organism, content, Uno-Vit, expanded, Ex, Plant Zygote, apparently low-set ears, 9030605P22Rik, Bio E, Lipid Peroxidations, fatty acids, nitrogen, Richtavit E, MCF3, Taf250, Koerper, Research-Related Injuries, Dermorelle, Clinical Finding, Auxina E, TAF230, Dioxygen, Vitamin E Natur, Saturated Fatty, big, Research Related Injuries, lipids, Esterified Fatty Acids, Endogenous Antioxidant, Sulfhydryl, number, E-948, antioxydant, Amide, Injuries and Wounds, large, DmelCG4114, Wounds, 2-propylpentanoic acid, Davitamon, Dpa, DPA, NCS, T22P11.60, Fatty Acids, Injury, dTAF[[II]]250, Eusovit, Tissues, cell, mcm4, mei-1794, Zygote, Aquasol E, AI847422, dTAF250, Mowivit Vitamin E, 22:5, combination of microcephaly, Antioxidant Activity, species, DPA TRANSCRIPTION FACTOR, n-DPA, Saeure, Vitamin E mp, Eplonat, Elaiosomes, RNA, Oxygen 16, body, E-ferol, Puncto E, E-Vicotrat, CG4114, Evion, whole body, RNS, BG:DS00004.13, compositionality, Unique E, oxygen, Cell, exophthalmos, 2-propylvaleric acid, dTAF230, near to, Vibolex, Fettsaeure, Mercaptan, Medicago, traumatic injury, Wounds and Injury, Medicago truncatulas, Trauma, Antioxidant Effect, TAF[[II]]250/230, ROS, oxygene, Mcm4, MCM4, OXYGEN MOLECULE, Vitamin E Sanum, Taf[[II]]250, O2, Togasan, di-n-propylacetic acid, T22P11_60, Sulfhydryl Compound, RGD1560259, storage, Saeuren, increased number, Dub, pyrene (narrow)., wound, azote, acide gras, Tocopa, l(2)01270, signalling process, Left renal vein entrapment syndrome, Compound, Mercapto, Togasan Vitamin E, vicinity of, structure, sequestering, E 948, 2-n-propyl-n-valeric acid, Vitazell, pyrene (narrow), Anti Oxidants, Activity, Wound, Lipidomic, Metabonomic, GRO:0005339, 2610511A05Rik, injury, composed of, brl, gum hyperplasia, Tocovital, SUB, Membrane Tissues, Tier, Spondyvit, NH2(-), dipropylacetic acid, Saturated Fatty Acids, azanide, Esterified Fatty Acid, acide valproique, Valproinsaeure, Elaiosome, E948, KIF20A, Research-Related, increased, Anti Oxidant, seed, Tocopherol Bayer, EUNOVA Vitamin E, membrane region, Aliphatic, Malton E, retention, E Vitamin E, HCA1, Diaspores, Antioxidant, Anti-Oxidant Effect, Protein G18, Lasar, Vitamin E EVI MIRALE, truncatula, Membrane Tissue, SIGNS SYMPTOMS, amides, VitaminE EVIMIRALE, Vita-E, ribose nucleic acid, Dal-E, DESC, Sanavitan S, ribonucleic acids, Ecoro, Bioweyxin, TAFII-250, TAF250/230, Anti-Oxidants, TAFII250, Abortosan, Renal Nutcracker Phenomenon, Vitamine E GNR, Sauerstoff, <new synonym>, Ribonukleinsaeure, Lipid, Depakene, pentosenucleic acids, Physical Trauma, Seed, Diaspore, Disauerstoff, antioxidants, MCM4_DROME, acide, acidum valproicum, Plant, acids, Aliphatic Acid, acido, CG17603, TAF[[II]], Vitamin E AL, Lipidome, Endogenous Antioxidants, Mowivit, enlarged, phospholipid metabolism, SR3-5, l(2)ey, Research-Related Injury, 7N, mMage-e1, Tocopherol, accessory, Vitagutt Vitamin E, AT1G11140, d230, E-Mulsin, antoxidant, Vitamin E Dragees, Vitamin E, dTAFII250, 8O, EfW1, supernumerary, presence, Symptoms and Signs, CG12298, acido valproico, 43Ca, Free Radical, VitaE, dioxygene, acides gras, dmTAF1, Taf230, valproic acid, integral component of membrane, DMP-4, Anti-Oxidant, Animal, Finding, Anti-Oxidant Effects, Metabolomic, CG1616, Aggravating interaction, TAF250, study, reactivity, Vita Plus E, Taf200, Phospholipid, Taf1p, metazoa, Ephynal, Biosan, Traumas, great, region of membrane, Zygotes, VitaPlus E, SRF9, amide, barrel medic, TAF, BACTS2, molecular oxygen, ahl4, membrane, Energy & Feritility, TAF[[II]]250, Saturated, DmelCG1616, dihydridonitrate(1-), Alpha-dystrobrevin-associated MAGE Protein, Lipidomes, l(3)84Ab, Anti Oxidant Effect, Elex Verla, STRUBBELIG-RECEPTOR FAMILY 9, count in organism, Signs and Symptoms, Vitamin-E EVI-MIRALE, yeast nucleic acid, p230, Embryos, Metazoa, Peroxidations, whole membrane, TFIID, Biopto-E, Embryo, ribonucleic acid, a phospholipid derivative, Anti Oxidant Effects, transmembrane, TAF[[II]]230, 2-PROPYL-PENTANOIC ACID, Vitamin E Suspension, Antioxidans E-Hevert, TAF[II]250, Membrane, Plant Embryos, Injuries, nitrogeno, present in greater numbers in organism, Antioxidant Effects, Vitagutt, Uno Vit, DmelCG17603, DmMcm4, cleft palate, Plant Zygotes, SOCS, approaches, DmMCM4, response, Suppressor of cytokine signaling, C22:5, T19D16.8, SCM, TAF1biochemical pathways, Saturated Fatty, acido graso, Anti Oxidants, Metabolic Process, Esterified Fatty Acids, Anti Oxidant Effect., acidos grasos, PhrB photolyase activity, Activity, Processes, P62, Endogenous Antioxidant, antoxidant, Metabolic Concepts, antioxydant, sci, GRO:0005339, photoreactivating enzyme activity, Metabolic Processes, pigmented epithelium, MAGE-E1 antigen, acides gras, Metabolism, Anti-Oxidant, HOW, How, Concepts, Saturated Fatty Acids, 3, stratum pigmentosa retinae, Esterified Fatty Acid, NUP96, Metabolism Concept, Anti-Oxidant Effects, Elaiosome, Phenomenon, epithelium, Metabolism Phenomena, l(3)j5D5, Esterified, 24B, Fatty Acids, Anti Oxidant, seed, MOS3, deoxyribocyclobutadipyrimidine pyrimidine-lyase activity, l(3)s2612, PRECOCIOUS, catabolism, Esterified Fatty, Aliphatic, stru, pigmented retina, F23A5.3, Plant Embryo, Metabolic Concept, Zygote, metabolic process resulting in cell growth, l(3)S053606, AI847422, CG10293, DNA cyclobutane dipyrimidine photolyase activity, SUPPRESSOR OF AUXIN RESISTANCE 3, l(3)j5B5, Aliphatic Acids, HCA1, Diaspores, Antioxidant, Antioxidant Activity, DmelCG10293, Anti-Oxidant Effect, Zygotes, biotransformation, Catabolism, F23A5_3, Elaiosomes, PRE, 0904/17, Saturated, degradation, Process, clone 2.39, deoxyribonucleic cyclobutane dipyrimidine photolyase activity, metabolism resulting in cell growth, Alpha-dystrobrevin-associated MAGE Protein, Antioxidants, Endogenous, Fettsaeuren, qkr, l(3)S090417, Anti-Oxidants, Concept, Metabolic Phenomena, Metabolism Concepts, Fettsaeure, MODIFIER OF SNC1, retinal pigment, SZ1, KH93F, Embryos, Antioxidant Effect, DAMAGE, Hepatocellular carcinoma-associated protein 1, deoxyribonucleic photolyase activity, Phenomena, dipyrimidine photolyase (photosensitive), secretion, Seed, metabolism, retinal pigment layer, Metabolic Phenomenon, who, Embryo, Acid, Anti Oxidant Effects, Diaspore, multicellular organism metabolic process, Fatty Acid, antioxidants, RPE, Saturated Fatty Acid, biodegradation, Metabolic, photolyase activity, RGD1560259, Plant, Aliphatic Acid, Plant Zygote, Who/How, acide gras, Plant Embryos, p. pigmentosa retinae, Antioxidant Effects, phr A photolyase activity, fatty acids, Endogenous Antioxidants, DNA-photoreactivating enzyme, Plant Zygotes, qkr[93F], anon-EST:Liang-2.39, deoxyribonucleate pyrimidine dimer lyase (photosensitive), mMage-e1, pyrene (narrow), AnabolismfalseHow Does the Seed Pre-germinative Metabolism Fight against Imbibition Damage? Emerging Roles of Fatty Acid Cohort and Antioxidant DefenceDuring seed imbibition, lipids are engaged in membrane reorganization while facing free radical-mediated oxidative injury. In the present work, we explored changes in lipid components at different timepoints of imbibition (0.5, 2, 4, 6, and 8 h) in the legume Medicago truncatula, by combining biochemical approaches with targeted lipidomics and untargeted metabolomics. ROS and RNS (reactive oxygen and nitrogen species) accumulation was observed throughout the tested timepoints whereas lipid peroxidation increased at 4 h of imbibition. The seed response to oxidative damage was evidenced by a significant increase in tocopherols starting from 0.5 h of imbibition as well as by the reduction in total thiol content occurring at 2 h of imbibition. Since under physiological conditions, the proper functions of the cell membranes are strongly dependent on the qualitative and quantitative balance of fatty acid residues in phospholipids, the investigation was expanded to the fatty acid cohort of Medicago truncatula seeds. Total saturated fatty acids (SFAs), monounsaturated fatty acids (MUFAs), polyunsaturated fatty acids (PUFAs), omega-3 and omega-6 fatty acids showed fluctuations during seed imbibition. The most remarkable finding was the profile of the omega-3 PUFA docosopentaenoic acid (DPA, 7 cis, 10 cis, 13 cis, 16 cis, 19 cis-22:5) that showed a peak (up to 1.0% of the total fatty acid content) at 0.5 and 8 h of imbibition, respectively, concomitant with the peaks observed in tocopherol levels. It is possible that the observed changes in DPA alter the physical properties of membranes, as reported in animal cells, triggering signaling pathways relevant for the cell defence against oxidative injury. Furthermore, the content and balance between tocopherols and PUFAs is regarded as a determinant of storage stability. No enhancement in trans-fatty acids occurred throughout imbibition, suggesting for a proper antioxidant response carried by the seed. Fatty acids profiles were integrated with data from untargeted metabolomics showing changes in lipid sub-pathways, among which fatty acid amide, lyso-phospholipids and phospholipid metabolism. The emerging lipid profiles and dynamics are discussed in view of the overall imbibition damage generated during M. truncatula seed imbibition.2019-10-142018-07-13MTBLS181MTBLC15428MTBLC17822MTBLC45441MTBLC45696MTBLC15356MTBLC17755MTBLC17376MTBLC16163MTBLC16668MTBLC15891MTBLC16919MTBLC17521MTBLC16119MTBLC36575MTBLC27897MTBLC18344MTBLC10072MTBLC28044MTBLC25998MTBLC18186MTBLC15760MTBLC15999MTBLC17385MTBLC46215MTBLC21626MTBLC68561MTBLC133536MTBLC35391MTBLC30653MTBLC26986MTBLC22653MTBLC25094MTBLC16927MTBLC17964MTBLC15753MTBLC17023MTBLC87998MTBLC16811MTBLC16680MTBLC132188MTBLC49033MTBLC45658MTBLC16449MTBLC40992MTBLC138368MTBLC16682MTBLC21557MTBLC30835|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|MTBLC67119MTBLC18222MTBLC22605|MTBLC17151MTBLC17261MTBLC59640|MTBLC28800MTBLC28721|MTBLC27353MTBLC23098MTBLC17268MTBLC32817MTBLC28757MTBLC17505MTBLC24149MTBLC29864|MTBLC30911MTBLC17306MTBLC37684MTBLC16634MTBLC17164MTBLC17992MTBLC28586MTBLC24266MTBLC16196|MTBLC88670MTBLC1148|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|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:15428CHEBI:17822CHEBI:45441CHEBI:45696CHEBI:15356CHEBI:17755CHEBI:17376CHEBI:16163CHEBI:16668CHEBI:15891CHEBI:16919CHEBI:17521CHEBI:16119CHEBI:36575CHEBI:27897CHEBI:18344CHEBI:10072CHEBI:28044CHEBI:25998CHEBI:18186CHEBI:15760CHEBI:15999CHEBI:17385CHEBI:46215CHEBI:21626CHEBI:68561CHEBI:133536CHEBI:35391CHEBI:30653CHEBI:26986CHEBI:22653CHEBI:25094CHEBI:16927CHEBI:17964CHEBI:15753CHEBI:17023CHEBI:87998CHEBI:16811CHEBI:16680CHEBI:132188CHEBI:49033CHEBI:45658CHEBI:16449CHEBI:40992CHEBI:138368CHEBI:16682CHEBI:21557CHEBI:30835|CHEBI:36458CHEBI:27487CHEBI:70959CHEBI:17645CHEBI:29016CHEBI:18211CHEBI:4828CHEBI:14321CHEBI:28300CHEBI:18295CHEBI:27570CHEBI:58148CHEBI:73685CHEBI:16437CHEBI:26271CHEBI:133469CHEBI:35280CHEBI:24741CHEBI:90344CHEBI:30818CHEBI:70958CHEBI:5757CHEBI:44673CHEBI:133181CHEBI:15728CHEBI:57969CHEBI:16344CHEBI:28116CHEBI:24898CHEBI:15689CHEBI:89228CHEBI:16530CHEBI:35932CHEBI:48430CHEBI:60645CHEBI:37084CHEBI:25017CHEBI:91315CHEBI:21565CHEBI:27266CHEBI:28635CHEBI:17431CHEBI:16610CHEBI:25520CHEBI:143243CHEBI:4047CHEBI:50621CHEBI:133622CHEBI:133032CHEBI:82966CHEBI:82968CHEBI:133028CHEBI:82969CHEBI:68848CHEBI:16010CHEBI:17234CHEBI:24298CHEBI:78320CHEBI:32816CHEBI:30769CHEBI:45969CHEBI:32805CHEBI:32806CHEBI:30916CHEBI:26806CHEBI:37154CHEBI:18300CHEBI:6650CHEBI:139374CHEBI:16802CHEBI:33508CHEBI:22599CHEBI:17113CHEBI:37654CHEBI:69438CHEBI:33805CHEBI:15963CHEBI:33942CHEBI:18066|CHEBI:67119CHEBI:18222CHEBI:22605|CHEBI:17151CHEBI:17261CHEBI:59640|CHEBI:28800CHEBI:28721|CHEBI:27353CHEBI:23098CHEBI:17268CHEBI:32817CHEBI:28757CHEBI:17505CHEBI:24149CHEBI:29864|CHEBI:30911CHEBI:17306CHEBI:37684CHEBI:16634CHEBI:17164CHEBI:17992CHEBI:28586CHEBI:24266CHEBI:16196|CHEBI:88670CHEBI:1148|CHEBI:50129CHEBI:17084CHEBI:37035CHEBI:37098CHEBI:32585CHEBI:133172CHEBI:59270CHEBI:65101CHEBI:55328CHEBI:19660CHEBI:143096CHEBI:76980CHEBI:72663CHEBI:72665CHEBI:72641CHEBI:741548CHEBI:133677CHEBI:133693CHEBI:71464CHEBI:64032CHEBI:71466CHEBI:133223CHEBI:38083CHEBI:57518CHEBI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