Posted on Authorea 23 May 2021 | The copyright holder is the author/funder. All rights reserved. No reuse without permission. | https://doi.org/10.22541/au.162177776.64521316/v1 | This a preprint and has not been peer reviewed. Data may be preliminary. lcdtn h rnfro iatv opud ewe lnsi seta o nesadn plant-plant understanding compound. action. for this of essential of modes their is translocation understanding plants root-to-shoot and between the pesticides compounds indicates natural roots, bioactive which developing rye of interactions, by shoot, up transfer rye taken by rye the compounds the up the the Elucidating in of taken Among roots plants. detected were showed crops neighboring we rye was Furthermore, from several by wogonin spectrometry-based shoots. originating and synthesized their compounds mass into rye up (BXs) translocated untargeted took between benzoxinoids and plant compounds species that and plant bioactive demonstrated neighboring targeted of of results study, roots transfer Our present the species. the the weed elucidate understanding and In to in applied exudates. hints was important root metabolomics provides bioactive species plant of different fate between metabolites of Translocation Abstract [email protected] Email: +4522283382. Fors Tel: University, Aarhus Denmark. Agroecology, Slagelse, of Department Kudsk. Per Professor Correspondence: Denmark Lyngby, Kgs. 2800 221, b a Hazrati Hossein compounds plants bioactive neighboring of and transfer rye extensive the between unravel metabolomics spectrometry-based Mass natural developing interactions, plant-plant Elucidating understanding action. by compound. of for up this essential modes taken of is their compounds translocation plants understanding the root-to-shoot between and Among the Furthermore, pesticides compounds indicates plants. which shoots. bioactive neighboring shoot, their of from rye into originating transfer the translocated compounds the in benzoxinoids and up detected that species was took demonstrated plant wogonin plant results neighboring roots, rye rye Our of the the roots elucidate of species. by to roots weed up showed applied and taken we was crops were several rye metabolomics by and spectrometry-based synthesized rye mass (BXs) root between untargeted bioactive compounds of and bioactive fate targeted of the study, understanding transfer present in hints the important In provides species exudates. plant different between metabolites of Translocation Abstract 2021 23, May 3 2 1 Hazrati Hossein neighboring and rye plants between compounds bioactive of extensive transfer the unravel metabolomics spectrometry-based Mass ehia nvriyo Denmark of University Technical University Aarhus Universitet Aarhus eateto goclg,Aru nvriy Fors University, Aarhus Agroecology, of Department eateto itcnlg n imdcn,TcnclUiest fDnak S Denmark, of University Technical Biomedicine, and Biotechnology of Department a neSFomsgaard S Inge , 1 neFomsgaard Inge , a igDing Ling , 2 igDing Ling , b e Kudsk Per , 1 ø 3 se ,40 lgle Denmark Slagelse, 4200 1, gsvej n e Kudsk Per and , a 1 ø tfsPas Building Plads, ltofts ø se ,DK-4200 1, gsvej Posted on Authorea 23 May 2021 | The copyright holder is the author/funder. All rights reserved. No reuse without permission. | https://doi.org/10.22541/au.162177776.64521316/v1 | This a preprint and has not been peer reviewed. Data may be preliminary. aooiswsapidt lcdt h xesv rnfro iatv opud ewe y n several and me- rye spectrometry-based between Methods mass compounds and untargeted bioactive Material and of transfer have targeted species. extensive studies of weeds the no combination and metabolomics elucidate date, a crops untargeted to to an , Up applied using 2020). study was of targeted species al., present tabolomics class plant used et specific the between Lewerenz which a In compounds 2004, studies, or bioactive approach. al., metabolites several of et particular are transfer of Chiapusio two-way There shoot 2020, revealed into 2010). al., translocation al., et with and identify (Hazrati comparing et uptake to by metabilites root Krauss or aims the 2010, signi- 2016, show experiments profiling al., to 2019). most MS Hooft, metabolic approach et by al., the der spectrometry-based confirmed (Lee et mass van is be standards cases, (Pezzatti could and authentic features most which robustness (Misra generated in metabolites, its metabolomics putative Therefore, of as known/unknown 2013). spectrometry-based amount well al., mass vast and as et untargeted separation the Dunn polarities, in of from of versatility bottleneck its identification range ficant to wide and due global a annotation plants, for with from technique Metabolite dominant e.g. li- compounds the samples, Ultra-high-performance of currently biological 2012). detection is complex al., HRMS of sensitivity an et high profiling with their metabolite (Theodoridis hyphenated to coverage (UHPLC) due chromatography metabolic studies quid maximize metabolomics applied untargeted which often for mass selectivity, is High-resolution used metabolomics and 2010). routinely Untargeted Saghatelian, are and possible. metabo- (Vinayavekhin (HRMS) as the biomarkers spectrometers discover profile metabolites and globally on hypotheses to information detected generate aims much 2012). of to metabolomics coverage as al., Untargeted the gather et 2012). therefore, and Roberts al., and, lome et interest 2008, or (Vrhovsek of al., limited metabolites metabolites et is known of metabolites (Lu the groups and with metabolomics biologi- selected deals targeted non-targeted complex quantifying metabolomics approaches: using Targeted of and metabolomics identified understanding identifying biomarkers distinct two of to better and validation are refers a crop There metabolomics us several 2009). Targeted provides al., in untargeted. which et BXs research, (Cevallos-Cevallos of of systems cal field translocation selective a and of is represents development uptake species the Metabolomics root plant promote the other may in comparing it manner are shoots. species. because same their we weed demonstrated perspective the into Here in (2020) agronomic translocate herbicides. subsequently occur an al. been and BXs natural from et has plants of interest vetch Hazrati BXs translocation hairy particular recently, of and by of Just ability uptake up growth-inhibitory 2013). taken root The al., be al., whether can 2020). et Studying et BXs Frey al., (Schulz exuded 2019, et species root al., (Hazrati that plant et species (Wouters several (Hazrati herbivores plant 2019), on wheat other al., confirmed and et and (Cotton maize, 2016), microorganisms and rye, with al., pesticides interactions including et plant natural in roots, essential as Poaceae are act They the of 2009). that in transfer present metabolites the mainly heteroaromatic of are tryptophan-derived Elucidation are and shoots. (Davies (BXs) pesticides the Benzoxinoids natural into commercial inhibitors developing translocation scarce, growth for and are vital as plants 1999). is soil acting Caseley, between species the compounds plant before bioactive between from or 2008) of compounds metabolites uptake al., bioactive transfer root-exuded root the et absorb on other on Etzerodt must studies of However, plants specifically 2006, Moreover, growth 2004). Other al., 2019). al., or 2021). et al., et germination al., et (Chiapusio (Etzerodt et the Kudjordjie products Hazrati reduce 2021, 2004, breakdown or al., 2020, al., their inhibit et al., et secondary (Sikder directly et (Weir microbiome Root-exuded (Zhang can species beneficial herbivores plant allelochemicals 2006). are the and containing al., which shape pathogens exudates et or microbial metabolites, root 2018) (Bais against secondary al., compounds organisms including et defense other Hu compounds, as with act organic interaction can of plant metabolites mixture mediating complex in a essential exude roots Plant metabolites, Introduction Secondary interactions, Plant-plant spectrometry, compounds Mass Bioactive Metabolomics, Keywords: Keywords: 2 Posted on Authorea 23 May 2021 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.162177776.64521316/v1 — This a preprint and has not been peer reviewed. Data may be preliminary. h nltsit h ag ftesadr uv.Ngtv lcrsryinzto EI)wsue,adthe and used, (version Software was Analyst (ESI-) from (MRM). ionization signals mode electrospray monitoring the 3200 reaction Negative fit a multiple curve. the to with in standard diluted coupled operated the were was system of extracts spectrometry HPLC mass range The 1100 the CA). City, Agilent into Foster analytes an SCIEX, the using (AB LC-MS/MS spectrometer by mass analyzed QTRAP were LC-QqQ-MS extracts by plant 0.2 Material The Plant Then in hours. BXs analysis. six of qualitative solventQuantification for further the USA) for qualitative and Scientific, vials For to Thermo tubes, HPLC-QqQ-MS. transferred (SPD121P, Eppendorf by were SpeedVac MeOH/H into BXs a 80% transferred quantify of in was ml dryness to extraction until -20 vials of evaporated at glass 1ml Fi- was solvent stored into repeated. UHPLC-QTOF-MS, extraction and transferred were of by collected and collection ml analysis was supernatant One filter supernate and tube. syringe of centrifugation, 21 centrifuged Eppendorf 80% PTFE ml ultrasonication, and new and of 1.8 and a g min ml ˜ in pellet, 4500 45 nally, 1 collected the at for was to and Denmark) ultrasonicated ml) added Herlev, tube, 0.9 were was Holm, (˜ Eppendorf Samples and supernatant an Buch the added. into sequently, micro-centrifuge, was K transferred acid 1–14 was acetic (Sigma mg) twice). 1% (20 (repeated containing s material methanol 90 plant for ground mechanical rpm were a 1500 Homogenized using of Material Samples powder speed Plant @C. fine a from -80 a at Extraction was at to NJ) Metabolite material grounded (Metuchen, freezer was plant Spex a material from Harvested to shoot 2010 shoot. transferring and Genogrinder their root before disrupter lyophilized from nitrogen remove the to separated liquid and shaken and freeze-dried, the slightly were rye water, into harvest, roots distilled transferred At Plant sowing. with immediately other. after each washed weeks from five soil, separated harvested attached carefully were were Plants plants treatment. neighboring each and and for rye used were of replicates number Eight the study crop/weed the seeds, neighboring in ( the included a pea species of of plant germination seeds Neighboring ( ( 12 pot. After clover radish Alexandrian per and plants. rye, plants species, rye species: 8 crop target and the seven 4 as was around to pot thinned ring the were a h seedlings of neighboring 16/8 in center a sown the at in were greenhouse sown sand) a species were 57.2% in and conditions seeds controlled 22/18 silt, rye under of 28.4% Six clay, grown temperature were 11.5% a Plants matter, with organic medium. photoperiod (2.8% growth soil the field as loam used were sandy UHPLC- with filled in pots analysis One-liter for analysis Scientific the Fisher for experiment from MA) Pot purchased Billerica, was (Millipore, Ger- system water (Griesheim, Milli-Q Baker grade QTOF-MS. a from HPLC-MS from acid LC-MS/MS. obtained acetic acetonitrile, was and using methanol, water UK); MS-grade HPLC-grade (Loughborough, Scientific Scotland); many). methanol Fischer (Walkerburn, (HPLC)-grade from Rathburn chromatography isopropanol from liquid and purchased High-performance were 2017). acetonitrile al., 2- et and our elucidated). (Pedersen fully study in in fully not previous described a (structure not as process (HBOA-glc-hex) (structure obtained HBOA was (DIBOA-glc-hex) patenting of (DIMBOA-glc) derivative Glucopyranosyloxy-4-hydroxy-7-methoxy-1,4-benzoxazin-3-one DIBOA hexose on-going of double derivative and an hexose elucidated), of double (DIBOA-glc), part benzoxazin-3-one 2- (DIBOA), as 2,4-dihydroxy-1,4-benzoxazin-3-one (HMBOA-glc), obtained 3-one 2- were (HMBOA), 2- (HBOA-glc), 2-hydroxy-7-methoxy-1,4-benzoxazin-3-one standards d-glucopyranosyloxy-1,4-benzoxazin-3-one (HBOA), BXs 2-hydroxy-1,4-benzoxazin-3-one eight lab: Sigma-Aldrich. from following purchased were The (MBOA) 6-methoxy-benzoxazolin-2-one and (BOA) 2-Benzoxazolinone Chemicals iu sativum Pisum ahnssativus Raphanus 2 n w edseis oim( Lolium species: weed two and ) a de oteEpnoftb,adi a etiue o 2 eod.Extracts seconds. 120 for centrifuged was it and tube, Eppendorf the to added was O ,ot( oat ), vn sativa Avena ° dyngt n eewtrdtruhasbirgto system. sub-irrigation a through watered were and (day/night) C rflu alexandrinum Trifolium ,sberna lvr( clover subterranean ), 3 oimmultiflorum Lolium β- d-glucopyranosyloxy-7-methoxy-1,4-benzoxazin- ° .Etat eefitrdtruha0.22 a through filtered were Extracts C. β- rflu subterraneum Trifolium ,hiyvth( vetch hairy ), d-glucopyranosyloxy-4-hydroxy-1,4- n Sinapis and ) ii villosa Vicia ° o 0mn Sub- min. 10 for C iai arvensis Sinapis ,Austrian ), ,fodder ), μ β- d- m β- . Posted on Authorea 23 May 2021 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.162177776.64521316/v1 — This a preprint and has not been peer reviewed. Data may be preliminary. l,DMO-l,HBAgc BAgc BAgchxadDBAgchx eeietfidadquan- and identified (DIBOA- BXs were glycoside DIBOA-glc-hex) six and and HBOA-glc-hex MBOA) HBOA-glc, and HMBOA HMBOA-glc, HBOA, plants DIMBOA-glc, BOA, neighboring glc, DIMBOA, of (DIBOA, BXs shoots aglycone and Six roots in BXs of on Quantification based compounds untargeted materials. from annotated supplementary data the raw the classify the Results in All to found 2016). used be al., was can et Feunang ClassyFire analysis (Djoumbou MetaboAn- metabolomics MS-DIAL by 2018). of from made al., InChIKey heatmaps were reported et clustering scaled) their the (Chong pareto hierarchical all 5.0 and The for version log-transformed reported Sweden). were to performed alyst intensities duplicate (Umea, was submitted (peak 17.0 removing (PCA) were metabolites version by analysis data normalized SIMCA component filtered metabolomics with Principle was untargeted compounds for file supplementary The method. annotated features Pareto data the the analysis. and The in curating statistical log-transformation by found to the 2017). quality be submitted list’s al., can feature before et the MS-DIAL data (DeFelice metabolite improve further of adducts to for utilized parameters and MS-DIAL was duplicates Detailed from MS-FLO MS exported S1). 2015). 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California, post-hoc using other Jolla with ANOVA files the (La one-way ABF with 8.4.2 a group plants, version each the Prism compare in Pad to BXs applied of quantification was 300 and test 325 biomass at at the flow compare temperature To and gas visualization temperature drying and gas eV, statistics, 30 sheath processing, at psi, Spectra Data B. energy 40 0% collision at capillary min, V, V, 21 pressure 120 65 B; gas at respectively. 0% at voltage nebulising 0% min, Fragmentor voltage follows: 18 min, L/min), ESI( skimmer as B; 1 in (8 V, set 100% B; collected were min, 4000 were 0% parameters 17.5 Data at min, QTOF-MS B; MS2. voltage Da. 0 100% data-dependent 75–1500 follows: min, and m/z as 15 MS1 of was B; scan range gradient 70% full The min, in acid. 10 acquired B; formic were 10% M min, 0.02 3 10 with 40 B; of acetonitrile at volume (B): set sample and was a Agilent formic oven and HSS an mL/min column An with 0.5 the software. of Masshunter of equipped rate by spectrometer flow controlled system a was mass UHPLC with system 1290 (QTOF) whole Infinity flight 1.8 The Agilent (C18) of source. an T3 time ion using electrospray performed quadrupole stream was 6545 dual-jet shoots Agilent and to roots coupled of UHPLC-QTOF-MS profiling using metabolite samples The standard plant according on of weighted based were profiling curves done standard metabolomic were the Untargeted Quantifications of to at points qualifier. 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C, yeg oa RP-80 Polar Synergi m https://www.reifycs.com/AbfConverter/index.html n oA( MoNA and ) 4 μ a netdfrec u.Tetemperature The run. each for injected was L https://mona.fiehnlab.ucdavis.edu oun(hnmnx Macclesfield, (Phenomenex, column A ˚ μ .Tetmeaue ftecolumn the of temperatures The L. 1 stpcsetu,metabo- spectrum, isotopic ± oe ihamass a with modes ) ° n 2L/min, 12 and C eeused were ) .Spectra ). ° C Posted on Authorea 23 May 2021 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.162177776.64521316/v1 — This a preprint and has not been peer reviewed. Data may be preliminary. aidaogpatseis hssuyi h rtt opr h paeadtasoaino root-exuded of translocation and uptake the compare to BXs first root-exuded of the uptake is the if study unclear ( This was vetch it hairy species. 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All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.162177776.64521316/v1 — This a preprint and has not been peer reviewed. 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The plant reported all numerous plant. which rye absorb (2020), contain one could with al. species together et species plant Hazrati plant by other neighboring study species. previous plant that with neighboring showed growing by results spectrometry-based plants exuded Our mass rye been applied have of we likely present most Here roots compounds intended that have organic that approach. studies compounds exuded/decomposed metabolomics observed no BXs of However, untargeted shown and uptake 2020). an have the metabolomics al., studies using of et recent rhizosphere picture (Hazrati addition, the complete shoots their In a in the and provide 2019). to metabolite and al., translocated secondary unravel et be of to can Selmar uptake root 2007, root by al., uptake up et the taken show (Buer studies roots Several in transformation species. plant several in BXs P o l h etdBswsbten-. n ,adw ol o n infiatcorrelation significant a find not could we and 1, and -1.6 between was BXs tested the all for P ausfrteBstknu ynihoigpatrosadmn flavonoids, many and roots plant neighboring by up taken BXs the for values 6 P ausfrteBssuisi hseprmn are experiment this in studies BXs the for values vn sativa Avena ntepeetsuy aho the of each study, present the In . P au f- o3cudb taken be could 3 to -1 of value P 13.I h present the In =1-3). P au (Inoue value Posted on Authorea 23 May 2021 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.162177776.64521316/v1 — This a preprint and has not been peer reviewed. Data may be preliminary. ORC,G .20.Mtblmcaayi nfo cec:areview. a science: food in analysis Metabolomic D. M. community 2009. DANYLUK, microbial E., E. Technology, Soil ETXEBERRIA, G. I., RODRICK, 2015. J. & J. REYES-DE-CORCUERA, and D. M., location BURKE, J. spatial CEVALLOS-CEVALLOS, chemistry, & soil by and Y. followed identity, up S. species taken genus. STRAUSS, plant plant differentially with L., strongly are B. most Flavonoids correlates ANACKER, variation 2007. H., A. J. M. BURNS, DJORDJEVIC, Arabidopsis. & in root K. distances and long G. lipophilicity MUDAY, transported between Relationships S., barley. C. 1982. by BUER, A. chemicals A. exudates non-ionised EVANS, root of & of translocation role H. and The R. uptake BROMILOW, 2006. M. G., organisms. J. G. VIVANCO, other BRIGGS, & and S. plants GILROY, with G., L. interactions PERRY, rhizosphere L., in T. WEIR, P., H. BAIS, and Amfelt, Mette Andersen, for Denmark, Christian References of laboratory John University excellent Technical Aaron their Biomedicine, and support. for Dr. Biotechnology valuable Laursen thank of their Birgitte Department to the Bente wish from and Wang authors Heinrichson Xinhui The Kirsten thank assistance. to technical like that also relationships would We personal or interests financial paper. this competing Acknowledgments in known reported no work Aarhus have the Technology, they influence and may that Science declare of authors School The Graduate interest the competing at of (28180) Declaration project a Denmark. AU), by (GSST, funded University was study This utilization shoot. agrochemicals. future and Funding and natural for root essential as rye composition is Furthermore, compounds both mechanisms the originating bioactive in translocation compounds detected the species. that root-to-shoot was up of and plant flavonoid, showed took absorption phytotoxic different roots we root a plant the is among Meanwhile, rye Elucidating which varied Wogonin, the that were plants. indicated shoots. neighboring plants metabolomics from to tested untargeted from them in The results weed BXs translocated the and metabolomic. absorbed absorbed and experiment crops spectrometry-based of this root several in mass concentration plants and their applying neighboring the rye by by all between revealed that BXs demonstrated compounds been metabolomics targeted bioactive has from plants of results neighboring transfer as extensive study, species increase present may the shoot In plant in wogonin has to wogonoside It of Conclusion Conversion cancer shoot. of 2013). rye growth effects. al., the the inhibitory et inhibiting Yu in growth by 2018, their bioactivity wogonin deglycosylation its al., into undergoes enhances et covert wogonoside (Wang likely in However, and Wogonoside of cells found deglycosylation wogonin. metabolism wogonin root. that that than microbiome the reported assume concentration in or been We higher present reactions detected. much stresses wogonoside not enzymatic abiotic a the was and through wogonoside in from biotic and originates found against shoot, was shoot compound rye defense wogonoside rye the a in root, as detected rye act was et the they wogonin Lin that In 2018, likely anti- al., very plants. as et therefore, such in Kim is, effects 2018, It biological al., various et 2012). have (Wang al., humans wogonoside, in glucuronide, anti-angiogenesis its and and anti-inflammatory, Wogonin cancerogenic, shoots. Lolium in BXs 20 o PLANTS, AoB , 557-566. 7. ln physiology, Plant 7 145 nuRvPatBiol, Plant Rev Annu etcd Science, Pesticide , 478-490. 13 rnsi odSine& Science Food in Trends , 57 495-504. , 233-66. Posted on Authorea 23 May 2021 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.162177776.64521316/v1 — This a preprint and has not been peer reviewed. Data may be preliminary. RUS . IGR .&HLEDR .21.L–ihrslto Si niomna analysis: environmental in MS resolution LC–high unknowns. 2010. of identification J. the to HOLLENDER, screening & target wogonin from H. of effect SINGER, Anti-inflammatory M., mouse. 2018. KRAUSS, the T. S. in KIM, rhinitis & allergic G. ovalbumin-induced KANG, R.I.), in S., Y. responses CHOI, barley. allergic H., in on J. bases JUNG, exudate uptake amine A., the Root K. of affecting KIM, factors shoots Physicochemical to N., 2018. 1998. translocation CHERVET, H. and M. R. D., roots BROMILOW, ERB, by & MANZO, K. & B., CHAMBERLAIN, microbiota. K. J., rhizosphere LI, INOUE, the SCHLAEPPI, M., shaping by A., YE, defense G. and X., growth M. ZHANG, Communications, on in feedbacks S., HEIJDEN, plant-soil products drive CADOT, DER natural metabolites VAN M., of A. T., Role C. STEINGER, 2019. ROBERT, P. L., KUDSK, HU, & species. B. plant MELANDER, between S., competition. interactions ac- belowground I. interspecific in FOMSGAARD, to alteration H., response unveil HAZRATI, a metabolomics as Targeted flavonoids 2021. of P. exudation Interactions, KUDSK, root & and S. cumulation I. FOMSGAARD, H., Translo- and HAZRATI, Uptake Benzoxazinoids: Plants. Root-Exuded benzoxa- Neighboring 2020. allelochemial P. in KUDSK, the cation & to S. sativa I. FOMSGAARD, Avena bacteria. H., of HAZRATI, root-colonizing Tolerance by 1996. 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Microbiome, 6 , 2226-2247. , aua rdc Com- Product Natural 7-16. 7 , 59. ora of Journal Posted on Authorea 23 May 2021 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.162177776.64521316/v1 — This a preprint and has not been peer reviewed. Data may be preliminary. al .OtmzdCmon eedn Sprmtr o BXs for parameters Profiling MS Metabolic dependent Exudates Compound Optimized Root Black 1. 2020. Table Against Cultivars F. Tobacco WANG, Susceptible & and J. Disease. Resistant Z. WANG, Between Shank C. Mechanisms Y., WANG, Defense ZHENG, enhance & Distinct to C., Suggests S. approach FENG, C. promising C., YUAN, a ZHANG, Y., hydrolase: WANG, glycoside T., with CALWAY, wogonoside Z., potential potential. and ZHEN, anticancer A baicalin H., of transfer: ZHANG, Pretreatment product Z., 2013. natural ZHANG, Horizontal C., YU, 2017. phytopharmaceuticals. H. in contaminants KIMA, alkaloidal & of source M. NOWAK, M., YAHYAZADEH, elei oeta.Teprmtr o h eodtasto r o eotdhr eas hywr only were they because here reported not Log are 2. transition Table second CXP: identification. energy, the compound collision for confirm CE: parameters to potential, The entrance used cell potential. CEP: exit potential, entrance cell EP: potential, declustering DP: physiological and Biochemical 2004. VASS B. & herbivores. P. J. insect I. and GERSHENZON, O. benzoxazinoids B., counter-defense: C. BLANCHETTE, herbivore J. C., M. F. of J. WOUTERS, VIVANCO, Deglycosylation 7 2018. & allelochemicals. by C.-S. S.-W. mediated YUAN, PARK, mechanisms & L., L. T. multiple CHEN, WEIR, of F., potential. quantification LU, anticancer rapid C.-F., its enhances ZHANG, the wogonoside & J.-Y., for WAN, R. method C.-Z., VIOLA, beverages. WANG, metabolomics and L., fruits targeted CAPUTI, in P., versatile phenolics of FRANCESCHI, A classes M., 2012. GASPEROTTI, F. D., MATTIVI, MASUERO, U., VRHOVSEK, VAN- M., deconvolution KANAZAWA, metabolomics. 30 Untargeted MS/MS K., 2010. data-independent A. SAGHATELIAN, IKEDA, & MS-DIAL: VINAYAVEKHIN, N. B., 2015. HIGGINS, M. analysis. ARITA, Y., metabolome & comprehensive MA, O. for T., FIEHN, KIND, J., T., DERGHEYNST, CAJKA, H., TSUGAWA, BA1419-0- 1 2 -2 -1.2 -2 -22 -2 -16 -2 -20 -0 -16.52 -14 -2 -8 -2 -22 -24.42 0 -2 -42 -2 -28 -7 -5 -10.5 -14 -23.28 -40 -2 -29.43 -40 -16 -20 -12 -30 -8.5 -23.81 -35 -16 -11 -36 -9.5 -16 -12 -22.67 -10.5 -9.5 -15 -28.83 -30 149 149 164 -45 -9.5 -4.5 -10 -35 -10.5 -45 134 134 164 164 372 134 -50 -30 -40 -80 123 DIMBOA-glc 194 DIMBOA 180 342 MBOA 504 (V) 108 164 CXP 42 164 DIBOA-glc-hex (V) 194 CE 356 DIBOA-glc (V) DIBOA CEP 164 HMBOA-glc (V) 326 134 EP 488 HMBOA (V) DP HBOA-glc-hex (m/z) HBOA-glc Q3 HBOA (m/z) Q1 BOA Compound , nt30.1.1-24. Unit O . 03DBAgc-. 58114415 -1.6 DIBOA-glc 6043 1.2 BOA Log Compound P rnir nPatScience, Plant in Frontiers au fsuisBsfo uCe database PubChem from BXs studies of value no Rep, Oncol P 30 , uCe I opudLog Compound CID PubChem 2411-8. auemethods, Nature 11 , gi odChem, Food Agric J 472-479. , ora fcne eerhadtherapeutics, and research cancer of Journal 1352. 10 12 Phytomedicine, , 523-526. 60 htceityReviews, Phytochemistry , 8831-40. O .G 06 ln ees and defense Plant 2016. G. D. AO, ˜ p 34 urPoo o Biol, Mol Protoc Curr , 21. uCe CID PubChem 14 15 , , S594-S599. 1127-1151. Chapter Posted on Authorea 23 May 2021 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.162177776.64521316/v1 — This a preprint and has not been peer reviewed. Data may be preliminary. Log *Log P P au hudb mle hntersnl lcsd om(IO-l n HBOA-glc). and (DIBOA-glc form glycoside single their than smaller be should value au sntrpre nPbhmdtbs.Det h eoepeec ntersrcue their structure, their in presence hexose the to Due database. PubChem in reported not is value MO . 523HO-l-e * * 77195052 14605136 * -1.3 4480305 -1.3 * HBOA-glc-hex DIBOA-glc-hex -1.6 HMBOA-glc HBOA-glc DIMBOA-glc 152213 322636 10772 0.3 0.3 2358 28495 1.1 HMBOA HBOA 0 0 MBOA DIMBOA DIBOA Log Compound P uCe I opudLog Compound CID PubChem 11 p uCe CID PubChem Posted on Authorea 23 May 2021 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.162177776.64521316/v1 — This a preprint and has not been peer reviewed. Data may be preliminary. 12