Lok ma pat pat ma mako gwok pa yadi miyo pwony ma pire tek me tito kit ma jami ma kwo tiyo kwede, ento lok ma pire tek ma gi waco pud tye me temo. Kany, wa tiyo ki unbiased tandem mass spectrometry (MS/MS) analysis me kwedo metabolome pa kodi ma ki lwongo ni tobacco attenuated strains ki i kom yadi acel acel me oo i kom dul dano ki ler ma gi rwate kwede, ki wa tiyo ki wel mapol me mass spectrometric feature theories ma jenge i kom compound spectra ma tye i ngec. Yub me temo jami ma ki byeko ma mako gwok maber (OD) ki jami ma tye ka wot (MT). Lok ma mako jami ma kwo i kom yadi rwate ki lok pa OD, ento jemo i kom lok ma ki waco i kom MT i kom jami ma kwo ma gi camo lum. Cake i rwom ma piny nio wa i rwom ma lamal, lanyut me jasmonate onongo ki ngeyo ni obedo gin ma pire tek ma moko OD, kun lanyut me ethylene omiyo yub maber pa lagam pa lee ma camo lum ma ki coyo ki MS/MS molecular network.
Jami ma pat pat ma tye ki dul kom ma pat pat aye gubedo gin ma konyo i dongo pa yadi ki kabedo, tutwale i gwoko lukwor (1). Apokapoka ma mako jami ma kwo ma nonge i kom yadi okelo mwaka mapol me kwedo ngec matut i kom jami mapol ma kwako tic pa jami makwo, ki okelo lok mapol ma kwako gwok pa yadi, ma obedo dongo ki dongo pa jami ma kwo i kin jami ma kwo ki jami ma kwo. Kwedo ngec ma kitimo miyo pwony ma pire tek (2). Ento, lok ma mako gwok pa yadi ni pe olubo yoo ma ki ngeyo me tam ma ki tamo ni tye kakare, ma iye jami ma pire tek ma ki waco ni tye i rwom acel me nyamo lok (3) ki ki temo me temo me medo rwom me dongo lok ma lubo (4). Peko me tic ki diro gengo coko ngec i kom dul ma pat pat ki kwanyo kwan ma mako jami ma pat pat, ma gengo poro kin dul ma pire tek pi dongo rwom me kwan (5). Pek pa ngec ma mako metabolomics ki cente ma rwate me poro kit ma tic pa metabolic space tye ka wot kwede i kin dul yadi ma pat pat gengo dongo pa ngec me cayan i poto ni.
Dongo manyen i dul me tandem mass spectrometry (MS/MS) metabolomics twero nyutu alokaloka me metabolic i kin ki i kin dul kom ma kimiyo, ki twero ribbe ki yoo me kwan me kwano kit ma dul kom ma pat pat ni rwate kwede. Ngec macon ikom chemistry (5). Ribu pa diro ma malo i nyamo ki kwano jami miyo yoo ma miite me temo pi kare malac pa jami mapol ma ki waco ma lubu lok ma mako kwo pa jami ma kwo ki dongo pa jami makwo. Shannon (6) okelo lok ma mako ngec pi tyen mukwongo i pwonye ma pire tek i mwaka 1948, kun keto guti me kwano ngec ma mako cura, ma dong kitiyo kwede i dul mapol mapat ki tic kwede macon. I genomics, lok me ngec dong ki tiyo kwede maber me kwano ngec ma ki gwoko i rwom ma lamal (7). I kwan me transcriptomics, lok ma mako ngec nyamo alokaloka ma tye i transcriptome (8). I kwan mukato, wa tiyo ki yub me kwano lok ma mako ngec me metabolomics me tito ngec me metabolic pa rwom me kom i kom yadi (9). Kany, wa ribo tic ma jenge i kom MS/MS ki yub me kwano lok ma mako ngec, ma ki nyutu ki kit ma jami tiyo kwede i wel cente ma ki ngeyo, me poro gin ma pire tek ma ki waco i kom jami ma kwo ma gi camo lum.
Kit me gwoko yadi pol kare gi rwate ki twero poko ne i dul aryo: ma gi temo tito kit ma jami ma gi dongo i kom yadi gi poko kwede ma lubu tic me gwoko kom, ma calo gwoko maber (OD) (10), gin ma tye ka wot (MT) (11) ) ki kit ma nen kwede (12) lok, kun jo mukene gi yenyo kit ma jami ma tye i kom jami ma tye ka dongo kwede gudo kwede dongo pa yadi ki dongo pa jami ma pat pat, ma calo carbon: tam me rwom me cam (13), rwom me dongo (14), ki rwom me dongo ki apoka poka (15). Dul lok aryo ni tye i rwom me nyamo lok ma pat pat (4). Ento, lok aryo ma kwako tic me gwoke i rwom me tic aye loyo lok ma mako gwoke ma kelo ki gwoke: OD, ma tamo ni yat keto cente i kom gwoke ma wel ne tek ka mitte keken, me labolle, ka ki camo gi. tam pa MT waco ni pe tye gin mo ma twero loko kit ma metabolite ni loko kwede, ento metabolite ni loko labongo ngeyo, ma miyo twero me gengo "motion target" me lweny i kom ler ma gi camo lum. I lok mukene, lok aryo magi gi miyo tam ma pat pat i kom kit ma kom dano loko kwede ma time i nge lweny pa ler ma camo lum: wat ma tye i kin dongo pa kom ma tye ki tic me gwoke (OD) ki alokaloka ma pe ki doro (MT) (11 ).
Tam pa OD ki MT pe kwako alokaloka ma okelo ikom metabolome keken, ento bene adwogi me dongo pa jami ma kwo ki dongo pa jami magi, calo wel cente ma mite ki adwogi maber ma bino pi alokaloka magi i kabedo ma pat pat (16). Kadi bed ni lok aryo ni gi ngeyo tic me gwoko kom pa jami ma ki lwongo ni metabolites, ma twero bedo tek onyo pe, gin ma pire tek ma poko kin OD ki MT tye i kit ma alokaloka ma kelo alokaloka i kom dano. Lok ma ki waco i kom OD theory ni onongo jami mapol ma ki temo nio wa kombedi. Temo eni tye iye kwan ma mako tic pa kom ma pat pat ma tye i ot ma ki gwoko iye jami ma kwo ki kabedo ma tye iye, kacel ki alokaloka i rwom me dongo pa yadi (17-19). Ento, nio wa kombedi, pi peke pa yoo me tic ki yub me kwano wel ma mako jami makwo ma pat pat, apoka poka madit ma tye i kin jami aryo ni (ma en aye, kit ma jami makwo loko kwede) pud tye me temo. Kany, wamiyo lok ma kit meno.
Gin acel ma pire tek ma nyutu kit ma jami ma kwo gi dongo kwede i kom yat aye kit ma gi nen kwede i rwom weny ki i kom yat acel acel, wel dano ma pat pat nio wa i kom ler ma rom aroma (20). Aloka loka mapol i kom jami ma ki yubu ma pat pat ki twero neno ne i wel dano, kun apoka poka ma tek i kom jami ma pat pat pol kare ki gwoko i kom dul ler (20). Pi meno, kit ma jami dongo kwede i kom jami makwo obedo gin ma pire tek i kit ma gi tiyo kwede, ma nyutu kit ma gi twero loko kwede i kabedo ma pat pat, tutwale kabedo ma tye ki kero me donyo pa jami ma pat pat ki ler ma gi camo lum (21). Cake i kare ma Fraenkel's (22) ocoyo lok ma mako tyen lok me bedo tye pa jami ma kwo i kom yadi, rwate ki jami makwo ma pat pat ki ngeyo ni obedo gin ma pire tek me yero, ki rwate eni ki geno ni okonyo yadi i kare me dongo. Kit me tic pa kom (23). Apokapoka ma tye i kin dul ler ma pat pat i kom jami ma pat pat twero nyutu kit ma jami ma kwo gi gwoko kwede jami ma kwo i kom jami ma gi camo lum, ma calo dul ler aryo ni pol kare gi rwate marac i kin gii (24). Kadi bed ni twero bedo ber me gwoko gwok maber i kare weny, alokaloka ma bino i kare ma opore ma mako gwok miyo adwogi maber ma weko yadi poko jami ma pigi tego bot jami mukene (19, 24), ki gengo miti me bedo kacel. Bal ma ki keto (25). Medo ikom meno, yub pa jami ma ki lwongo ni metabolites ma pat pat ma aa ki ikom ler ma camo lum twero kelo poko ne i kin dano (26), ki twero nyutu lok ma mako alokaloka madit ikom jasmonic acid (JA) signal, ma twero bedo i kin dano. Lanyut me JA ma malo ki ma piny obedo lok ma ki loko i kin gwoke i kom ler ma camo lum ki pyem ki ler ma pat pat (27). Medo ikom meno, yoo me dongo pa jami makwo ma pat pat obi rweny oyotoyot ki loko i kare me dongo, ma obi kelo poko kin jami ma kwo ma rwate (28). Polymorphisms magi twero bedo ma ki keto oyoto me gamo alokaloka i kit ma ler ma camo lum gi loko kwede (29), ma nyutu ni alokaloka pa dul pa ler ma gi camo lum obedo gin ma pire tek ma weko kom dano bedo ma pat pat.
Kany, wa cobo peko ma lubo ni. (I) Kit ma ler ma camo lum ni loko kwede kit ma jami dongo kwede? (Ii) Ngo ma obedo ngec ma pire tek ma mako kit ma kom dano tiyo kwede ma ki twero kwano me temo gin ma ki byeko me gwoko kom pi kare malac? (Iii) Ka ki loko yub pa jami ma kwo i kom yat i yoo ma pat ki pa ngat ma tye ka jemo, ka tye kumeno, tic ango ma jami ma dongo i kom yat timo me yubu kit ma jami ma kwo tiyo kwede, ki jami mene ma konyo i kom kit ma yat ni twero gwoke kwede? (Iv) Kit ma lok ma ki waco ki lok mapol ma mako gwok twero bedo ma ki medo i kom rwom me kom weny, wapenye ni kit ma lagam pa jami makwo ni rwate kwede ki i poro ma tye i kin dul kom ki i kin dul kom ma pat pat? Me agiki, wa tye ka kwano kit ma pot yat ma ki lwongo ni tobacco nicotine nicotine ni tiyo kwede, ma obedo gin ma ki lwongo ni ecological metabolome ma tye ki metabolism ma rwom ne lamal, ki tiyo maber i kom ler ma gi camo lum, Lepidoptera Datura (Ms) (ma tek, ma pol kare ki camo) I kom Solanaceae ki Spodoptera (Storalist), obedo yat ma ki lwongo ni leaf worm "dul", ki yadi ma gi gwoko Solanaceae ki yadi mukene ma gi bedo i dul ki gang cam pa yadi. Wa poko MS/MS metabolomics spectrum ki wa kwanyo jami ma ki ngeyo me poro OD ki MT theories. Yub map me nyutu jami ma pire tek ma tye i kom dano. Kwedo ni ki medo i kom dul pa N. nasi ki kodi taba ma rwate kwede me medo kwedo apoka poka ma tye i kin jami ma miyo teko i kom yat ki kelo OD.
Me nongo map ma mako kit ma pot yat ma camo yat ma ki lwongo ni herbiourous tobacco ni twero loko kwede ki kit ma kome nen kwede, watiyo ki yoo ma ki yubu con me kwano ki kwano wel me coko ki poko ngec ma rwate ki MS/MS ma aa ki i kom yat ( 9). Yoo man ma pe ki poko ne (ma ki lwongo ni MS/MS) twero yubu jami ma pe rwate, ma ki twero tic kwede pi jami weny ma ki waco kany. Jami ma ki lwongo ni deconvoluted plant metabolites ni tye ma pat pat, ma tye ki jami ma ki lwongo ni metabolites miya mapol onyo alip mapol (ma romo 500-1000-s/MS/MS kany). Kany, wa tamo i kom kit ma kom dano tiyo kwede i yub me pwony me ngec, ki wa kwano kit ma kom dano pat pat kwede ki kit ma en tiyo kwede maber ma lubbe ki Shannon entropy me poko kin jami ma kwako kom dano. Nono yoo ma ki tiyo kwede con (8), wa kwano lanyut ma ki twero tic kwede me kwano kit ma metabolome pat pat kwede (Hj indicator), metabolic profilization (δj indicator) ki metabolic specificity pa metabolite acel (Si indicator). Medo ikom meno, wa tiyo ki Relative Distance Plasticity Index (RDPI) me kwano metabolome inducibility pa ler ma camo lum (lanyut 1A) (30). I kin yub me kwano wel man, wa tero MS/MS spectrum calo gin ma pire tek, ki wa tiyo ki wel MS/MS ma rwate i frequency distribution map, ki dong wa tiyo ki Shannon entropy me byeko apoka poka pa metabolome ki iye. Kit ma metabolome tye kwede ki pimo ki rwom ma rwate pa MS/MS spectrum acel. Pi meno, medo i wel pa dul mogo me MS/MS inge keto ler ma camo lum ki loko doko spectral inducibility, RDPI ki specialization, ma en aye, medo i δj index, pien jami mapol ma pat pat ki yubu ki Si index ma malo ki kelo. Dwoko piny pa Hj ma pat pat nyutu ni wel MS/MS ma ki yubu ni odok piny, onyo rwom me poko pa jami ni loko i yoo ma pe rom, kun dwoko piny peko ne. Ki ikom kwano Si index, twero bedo yot me nyutu MS/MS mene ma ki kelo ki ler mogo ma camo lum, ento, MS/MS mene ma pe gamo kelo ne, ma obedo lanyut ma pire tek me poko kin MT ki OD ma ki byeko.
(A) Jami ma ki kwano ma ki tiyo kwede pi ler ma camo lum (H1 onyo Hx) MS/MS data-inducibility (RDPI), apoka poka (Hj index), ngec (δj index) ki metabolite specificity (Si index). Medo i rwom me tic (δj) nyutu ni, i kin piny, jami mapol ma gi camo lum gubi kelo, kun dwoko piny i rwom ma pat pat (Hj) nyutu dwoko piny i yubu jami ma ki yubu onyo poko jami ma pe rom i map me poko ne. wel Si nyutu ka ce gin ma ki lwongo ni metabolite ni tye pi gin mo (kany, ma camo lum) onyo ki gwoko i rwom acel. (B) cal ma ki nyutu me lok ma mako gwoke kun kitiyo ki lok ma mako ngec. OD theory waco ni lweny pa ler ma camo lum obi medo jami ma gwoko kom, ki medo δj. I kare acel, Hj dwoko piny pien kit ma ki yubu kwede ni ki dwoko piny me bedo ki ngec ma mako kom. Lok me MT tito ni jemo pa ler ma camo lum obi kelo alokaloka ma pe rwate ki dul kom, ma obi medo Hj ma calo lanyut me medo ngec ma mako kom ki kelo poko pa Si ma pe ki ngeyo. Wa bene wa keto yub ma ki rubu, MT maber loyo, ma iye jami mogo ma tye ki rwom me gwoke ma malo obi mede (wel pa Si ma malo), kun mukene nyutu lagam ma pe ki ngeyo (wel pa Si ma lapiny).
Nono jami ma ki ngeyo, wa gonyo lok me OD me byeko ni alokaloka ma bino pi ler ma camo lum i kit ma pe ki kelo bi kelo (i) medo i kom kit ma kom dano dongo kwede (Si index) ma weko kit ma kom dano dongo kwede (δj index) Medo pa) dul kom mogo ma pat pat ma tye ki rwom me gwoke ma malo, ki (ii) dwoko piny rwom me kwo pa dano (Hj) loko rwom me poko jami ma mako kom dano me doko rwom me poko jami ma mako kom dano. I rwom pa gin acel ma ki lwongo ni metabolite, ki geno ni ki poko Si ma ki yubu, ma gin ma ki lwongo ni metabolite ni obi medo wel Si malube ki wel me gwoke (lanyut 1B). I kom lok man, wa tito lok me MT me waco ni jami ma kelo miti obi kelo (i) alokaloka ma pe rwate i kom jami ma ki yubu ma weko δj index dwoko piny, ki (ii) medo Hj index pi medo rwom me metabolic uncertainty. Onyo randomness, ma ki twero kwano ki Shannon entropy i kit me generalized diversity. Ma mako kit ma kom dano tiyo kwede, lok me MT obi tito kit ma Si poko kwede. Ki keto i tam ni jami mogo ma ki lwongo ni metabolites tye i te jami mogo ma pat pat, ki jami mukene pe tye i te jami mogo ma pat pat, ki rwom me gwoke gi cung i kom kabedo, wa bene wamoko tam me yubu kit me gwoke ma ki rubu, ma δj ki Hj ki poko gii i aryo i kom Si Mede i yoo weny, dul metabolite mogo keken, ma tye ki rwom me gwoke ma malo, ma bi medo rwom me gwoke ma malo, nongo Sire mukene bi bedo ki poko ma pe ki ngeyo (Figure a 1B).
Me temo lok ma ki loko i kom lok me gwoke i kom lok ma ki waco i kom lok me ngec, wa kelo ler ma camo lum ma ki lwongo ni expert (Ms) onyo generalist (Sl) i kom pot yat ma ki lwongo ni Nepenthes pallens (lanyut 2A). Nono MS/MS analysis, wa nongo 599 non-redundant MS/MS spectra (data file S1) ki i methanol ma ki kwanyo ki i kom pot yat ma ki kwanyo inge miyo cam bot caterpillar. Tic ki RDPI, Hj, ki δj me neno kit ma ki loko kwede ngec ma tye i MS/MS nyutu kit ma jami tye kwede (lanyut 2B). Gin ma tye ka time ni, kit ma ki waco kwede i lok ma ki coyo, ma calo ler ma ki lwongo ni caterpillars gi mede ki camo pot yat, rwom me tic pa jami ma gi tiyo kwede mede i kare: cawa 72 ingee ler ma camo lum ocamo, RDPI mede matek. Ka kiporo ki ma pe ki balo, Hj onongo odok piny tutwal, ma onongo tye pi medo rwom me tic pa jami ma kwako kom, ma onongo ki kwano ki δj index. Kit ma nen kwede ni rwate ki lok ma ki waco i OD theory, ento pe rwate ki lok ma pire tek ma ki waco i MT theory, ma ye ni alokaloka ma pe ki ngeyo (ma pe rwate) i rwom me metabolite kitiyo kwede me gwoko kom (lanyut 1B). Kadi bed ni jami ma ki lwongo ni oral secretion (OS) ki kit ma gi camo kwede cam pa ler aryo ni pat pat, camo gi okelo alokaloka marom i kit ma Hj ki δj gi tiyo kwede i kare me kac ma romo cawa 24 ki cawa 72. Apokapoka acel keken otimme i cawa 72 me RDPI. Ka kiporo ki ma oa ki i cam pa Ms, kit ma kom dano okelo kwede cam pa Sl onwongo tye malo.
(A) Yub me temo: ler ma camo lum ma ki lwongo ni common opego (S1) onyo ludiro (Ms) ki miyo cam ki pot yat ma ki kwanyo ki i pii, kun pi ler ma camo lum ma ki temo, OS pa Ms (W + OSMs) ki tiyo kwede me cobo peko ma ki keto i kom pot yat. S1 (W + OSSl) larvae onyo pii (W + W). Gin ma ki lwongo ni control (C) tye pot yat ma pe obale. (B) Inducibility (RDPI ka kiporo ki control chart), diversity (Hj index) ki specialization (δj index) index ma ki kwano pi special metabolite map (599 MS/MS; data file S1). Asterisks nyutu apoka poka madit i kin cam ma ki camo ki ler ma pe camo (Student's t-test ki paired t-test, *P<0.05 ki ***P<0.001). ns, pe pire tek. (C) Cawa me poko gin ma pire tek (bok ma bulu, amino acid, acid ma kwo ki cukal; karatac me ngec S2) ki jami ma ki lwongo ni special metabolite spectrum (bok makwar 443 MS/MS; karatac me ngec S1) ingee tic ki jami ma ki camo lum. Kala ma ki keto ni nyutu 95% me gen. Lakalatwe nyutu apokapoka ma pire tek i kin yat ki ma ki temo [kwadratic analysis of variance (ANOVA), ma lubu ki Tukey's honestly significant difference (HSD) pi apoka poka mapol, *P<0.05, **P<0.01 Ki *** P <0.001]. (D) Keto jami ma pat pat ma ki lwongo ni scatter plots ki jami ma pat pat ma ki lwongo ni metabolite (labol ma ki dwoko cen ki kit ma ki tiyo kwede ma pat pat).
Me temo ka ce alokaloka ma bino pi ler ma camo lum i rwom me metabolome ni nen i alokaloka i rwom pa metabolite acel acel, wa cako keto tamwa i kom metabolites ma ki kwano con i pot yat Nepenthes pallens ma tye ki teko me lweny i kom ler ma camo lum. Phenolic amides obedo hydroxycinnamamide-polyamine conjugates ma gi dongo i kare ma ler gi camo lum ki ki ngeyo ni gi dwoko piny tic pa ler (32). Wa yenyo jami ma okelo MS/MS ma rwate ki wa coyo rwom me tic gi (lanyut S1). Labongo ur, jami ma aa ki i phenol ma pe gi konyo i kom ler ma camo lum, calo chlorogenic acid (CGA) ki rutin, gi dwoko piny inge camo lum. Ka i poro, ler ma camo lum twero miyo phenol amides bedo matek. Camo cam pa ler aryo ni okelo rwom me tic ma rom ki pa phenolamides, ki kit man onongo nen atir pi de novo synthesis pa phenolamides. Gin acel-lu obi nen ka kitye ka temo 17-hydroxygeranyl nonanediol diterpene glycosides (17-HGL-DTGs) yoo, ma kelo wel madwong pa acyclic diterpenes ma tiyo maber i kom jami ma camo lum (33), ma Ms Feeding ki Sl okelo nyutu ma rom (Figure S1).
Peko ma twero bedo tye i temo cam pa ler ma camo lum aye apoka poka i wel camo pot yat ki kare me cam pa ler ma camo lum, ma miyo bedo tek me kwanyo adwogi marac ma bino pi ler ma camo lum. Me cobo peko pa ler ma camo lum ma mako kit ma pot yat gi loko kwede, wa temo miyo cam pa ler ma ki lwongo ni Ms ki Sl kun wa keto OS ma ki nongo nyen (OSM ki OSS1) i kom yat ma ki keto i pot yat ma rwate. Kit me timo man ki lwongo ni W + OS treatment, ki en dwoko rwom me tic ni piny kun ki keto kare ma opore me acaki me lagam ma ki nongo ki bot ler ma camo lum labongo kelo adwogi marac ma twero kelo apokapoka i rwom me rwenyo kom (lanyut 2A) (34). Nono MS/MS analysis ki calculation pipeline, wa nongo 443 MS/MS spectra (data file S1), ma orwate ki spectra ma ki nongo con ki i temo cam. Kwedo ngec ma ki nongo i kom MS/MS ni onyutu ni loko jami ma ki lwongo ni leaf-specialized metabolomes kun ki poro ler ma camo lum onyutu OS-specific inducements (lanyut 2C). I kom lok, ka kiporo ki yat OSS1, OSM okelo dongo pa metabolome i cawa 4. Ber me ngeyo ni ka kiporo ki jami ma ki nongo ki bot lee ma camo lum, kit ma kom dano tiyo kwede ma ki neno i kabedo ma tye ki dul aryo kun kitiyo ki Hj ki δj calo coordinates ki kit ma metabolome tye ka tic kwede me gamo kit ma ki miiyo kwede ler ma camo lum i kare ma okato. Mede ma rwate (lanyut 2D). I kare acel, wa kwano wel amino acids, organic acids ki cukal (data file S2) me kwedo ka ce medo ngec i kom metabolome ni tye pi loko kit ma carbon metabolism tye kwede i kom ler ma gi camo lum ( cal S2). Me gonyo lok man maber, wa medo ki neno kit ma kom dano dongo kwede i kom jami ma ki nyamo con ma ki lwongo ni phenolamide ki 17-HGL-DTG. OS-specific induction pa ler ma camo lum ki loko i kit ma ki loko kwede i kin phenolamide metabolism (lanyut S3). Phenolic amides ma tye ki coumarin ki caffeoyl moieties ki cwalo gi ki OSS1, kun OSMs gi kelo cwalo ferulyl conjugates. Pi 17-HGL-DTG yoo, apoka poka me OS ma ki kelo ki malonylation ki dimalonylation ma ki nongo (lanyut S3).
Ma lubo, wa kwano OS-induced transcriptome plasticity kun wa tiyo ki time-course microarray data set, ma nyutu tic ki OSMs me cango pot yat ma ki lwongo ni rosette i kom ler ma camo lum. Kit ma ki kwanyo kwede jami ni rwate ki kit ma ki tiyo kwede i kwan me metabolomics ni (35). Ka kiporo ki kit ma metabolome loko kwede ma metabolic plasticity tye ka mede i kare, waneno transient transcription bursts i pot yat ma ki kelo ki Ms, ma transcriptome inducibility (RDPI) ki specialization (δj) tye i 1. jami ma ki coyo ma mako jami ma ki coyo (lanyut S4). Dul pa gene ma mako kom (ma calo P450, glycosyltransferase, ki BAHD acyltransferase) gi nywako i yub me coko jami ma pat pat ma aa ki i dul kom ma aa ki i yub me kom, ma lubu kit ma kiloko kwede con. Ma calo kwan ma ki nyutu, yoo pa phenylalanine ni ki kwano. Kwedo ni moko ni gin ma pire tek i kom phenolamide metabolism tye ki OS-induced madwong i kom ler ma camo lum ka kiporo ki lum ma pe ki mako, ki gi rwate i kit ma gi nyutu kwede. Gin ma ki lwongo ni transcription factor MYB8 ki structural genes PAL1, PAL2, C4H ki 4CL ma tye malo i yoo man onyutu acaki me coc. Acyltransferases ma gi tiyo i yub me agiki pa phenolamide, ma calo AT1, DH29, ki CV86, nyutu rwom me dongo ma rii (lanyut S4). Gin ma ki neno malo ni nyutu ni acaki me tic pa transcriptome ki dong dongo pa metabolomics obedo gin ma ki keto kacel, ma twero bedo ni oa ki i yub me loc ma rwate ma cako tic me gwoko kom.
Lok ma ki loko i kom jami ma kwo i kom yadi tiyo calo gin ma doro jami ma gi camo yadi me loko kit ma yadi gi kwo kwede. Iyonge temo ler ma camo lum, wa pimo rwom me dongo pa dul kom ma pire tek i kom yadi ki waneno kit ma gi rwate kwede [Pearson correlation coefficient (PCC)> 0.4] (lanyut 3A). Kit ma onongo ki tamo kwede, jami ma miyo teko i kom yadi ma mako dongo pa jami ma kwo gikubbe i kin jami ma miyo teko i kom jami ma kwo. Medo ikom meno, kit ma kom dano tiyo kwede (Si index) ki keto i yub man me nyutu jami ma konyo kom ma ki nongo ki i yat ma pat pat. Kabedo aryo ma pire tek ma nyutu kit ma lee ma camo lum gi gamo kwede: acel tye i dul pa JA, ma JA (kit ma en tiyo kwede i kom JA-Ile) ki jami mukene ma aa ki i JA nyutu Si ma malo loyo; mukene ni ethylene (ET). Gibberellin onyutu rwom ma lapiny i kom ler ma camo lum, kun jami mukene ma konyo kom, calo cytokinin, auxin, ki abscisic acid, onongo tye ki rwom ma lapiny pi ler ma camo lum. Ka kiporo ki tic ki W + W keken, medo wel ma malo pa JA derivatives kun kitiyo ki OS application (W + OS) twero bedo ma ki loko me doko lanyut ma tek pa JAs. Labongo akalakala, OSM ki OSS1 ma tye ki jami ma pat pat ma gi kelo dongo pa JA ki JA-Ile. Ka i poro ki OSS1, OSM tye ma ki kelo ki OSMs, kun OSS1 pe medo rwom me gamo wang bur (lanyut 3B).
(A) Co-expression network analysis ma jenge i kom PCC ma ki kwano me herbivore-induced plant hormone accumulation kinetics simulation. Gin ma ki lwongo ni node ni nyutu gin ma ki lwongo ni hormone acel, ki dit pa node ni nyutu Si index ma rwate ki gin ma ki tiyo kwede i kin yat. (B) Medo pa JA, JA-Ile ki ET i pot yat ma oa ki i tic ma pat pat ma ki nyutu ki rangi ma pat pat: apricot, W + OSM; bulu, W + OSSl; macol, W + W; gray, C (control) . Asterisks nyutu apokapoka madit i kin yat ki yat ma ki miyo (two-way ANOVA ma lubo ki Tukey HSD post hoc multiple comparison, *** P <0.001). Kwedo lok ma mako (C)697 MS/MS (data file S1) i JA biosynthesis ki rwom me neno ma obale (irAOC ki irCOI1) ki (D)585 MS/MS (data file S1) i ETR1 ki ET signal ma obale Two simulated herbivore treatments triggered plant lines and empty vehicle controls (EV) plants. Asterisks nyutu apokapoka madit i kin W+OS ma ki miyo ki ma pe ki balo (two-way ANOVA ma lubo ki Tukey HSD post hoc multiple comparison, *P<0.05, **P<0.01 ki ***P<0.001). (E) cal ma ki poko ma nyutu apokapoka i kom tic ma pat pat. Kala ni nyutu jami ma pat pat ma ki loko gii; lanyut ni nyutu yoo me cang ma pat pat: triangle, W + OSS1; rectangle, W + OSM; guro C
Ma lubo, wa tiyo ki dul kom ma ki loko ma ki lwongo ni attenuated Nepenthes (irCOI1 ki sETR1) i rwom ma pire tek me JA ki ET biosynthesis (irAOC ki irACO) ki neno (irCOI1 ki sETR1) me nyamo kit ma jami aryo magi gi tiyo kwede i kom ler ma gi camo lum. Ma rwate ki temo macon, wamoko ni kelo herbivore-OS i kom yadi ma pe ki gin mo (EV) (lanyut 3, C onyo D) ki dwoko piny pa Hj index ma OSM okelo, kun δj index omede. Lagam ni nen maber loyo lagam ma OSS1 okelo. Graf ma tye ki lain aryo ma tiyo ki Hj ki δj calo coordinate nyutu deregulation ma ki ngeyo (lanyut 3E). Gin ma nen maber loyo tye ni i kom dul ma pe ki lanyut me JA, apoka poka me metabolome ki alokaloka ma aa ki i kom ler ma camo lum dong cok rweny woko (lanyut 3C). Ka i poro, neno pa ET ma pe loko i sETR1 yadi, kadi bed ni adwogi ne i kom alokaloka i kom jami ma camo jami ma kwo tye piny loyo pa JA, dwoko piny apokapoka i kin Hj ki δj i kin OSM ki OSS1 (lanyut 3D ki cal S5). . Man nyutu ni medo i kom tic ma pire tek me JA signal transduction, ET signal transduction bene tiyo calo yubu kit ma ler ma gi camo lum gamo kwede. Ma rwate ki tic me yubu ne maber ni, onongo pe tye alokaloka mo i kit ma metabolome inducibility ni tye kwede i sETR1 yat. Ki tung cing kany, ka kiporo ki yat sETR1, yat irACO okelo aloka loka ma rom ki pa ler ma camo lum, ento onyutu apokapoka madit i kin Hj ki δj i kin peko me OSM ki OSS1 (lanyut S5).
Me nongo jami ma ki lwongo ni metabolites ma tye ki kony ma pire tek i kom kit ma ler ma camo lum gi gamo kwede ki me yubu kit ma gi yubu kwede kun wok ki i ET signals, watiyo ki yoo ma ki yubu con me MS/MS. Yoo man jenge i kom yoo me bi-clustering me dwoko rwom me metabolic family ki i MS/MS fragments [normalized dot product (NDP)] ki rwom me rwate ma jenge i kom neutral loss (NL). Gin ma ki lwongo ni MS/MS ma ki yubu ki ikom nyamo ET transgenic lines okelo 585 MS/MS (data file S1), ma ki cobo ne ki keto gi i MS/MS modules abiro (M) (Figure 4A) . Module mogo i kin jami eni tye ki jami ma ki ngeyo con: me labolle, M1, M2, M3, M4 ki M7 tye ki jami mapol ma ki lwongo ni phenol derivatives (M1), flavonoid glycosides (M2), acyl sugars (M3 ki M4), ki 17-HGL-DTG (M7). Medo ikom meno, ngec ma mako metabolic (Si index) pa metabolite acel acel i module acel acel ki kwano, ki poko pa Si ne ki twero neno ne maber. Ki lok macek, MS/MS spectra ma nyutu cam ma dwong ki genotype specificity ki nyutu ki Si ma malo, ki kurtosis statistics nyutu poko pa yer i tung lacuc. Gin acel ma ki poko ma ki lwongo ni lean colloid ni ki nongo i M1, ma i iye phenol amide onyutu dul Si ma malo loyo (lanyut 4B). Gin ma ki waco con ma ki lwongo ni herbivorous inducible 17-HGL-DTG i M7 onyutu Si score ma rwate, ma nyutu rwom ma rwate me apoka poka i kin dul OS aryo ni. Ka i poro, pol pa jami ma ki yubu ma pat pat, ma calo rutin, CGA, ki acyl sugars, tye i kin jami ma ki nongo ki Si ma piny loyo. Me neno maber kit ma jami tye kwede ki kit ma Si poko kwede i kin jami ma pat pat, ki yubu dul ma ki lwongo ni molecular network pi module acel acel (lanyut 4B). Gin ma pire tek ma ki waco i kom OD theory (ma ki coyo i cal 1B) tye ni dwoko rwom me metabolites ma pat pat inge camo lum omyera okel aloka loka acel i metabolites ma tye ki rwom me gwoke ma malo, tutwale ki medo rwom me ticgi (ma pe ki keto i rwom ma opore) Mode) Defensive metabolites ma ki waco ki MT theory. Pol pa jami ma ki lwongo ni phenol ma ki nongo i M1 gi tye ki wat ki dwoko piny tic pa ler (32). Ka ki poro wel Si ma tye i kom M1 i kin pot yat ma ki kelo ki pot yat ma tye i EV control plants i cawa 24, waneno ni metabolic specificity pa metabolites mapol inge winyo ma camo lum tye ki rwom me dongo ma pire tek (lanyut 4C). Medo pa wel Si ma ki nongo i defensive phenolamides keken, ento pe ki nongo medo i wel Si i phenols mukene ki jami ma pe ki ngeyo ma tye kacel i dul man. Man obedo labol ma ki ngeyo, ma kubbe ki lok me OD. Lok ma pire tek ma ki waco i kom alokaloka ma bino pi ler ma camo lum tye ma rwate. Me temo ka ce gin ma ki lwongo ni phenolamide spectrum ni oa ki i OS-specific ET, wa coyo metabolite Si index ki okelo apoka poka i kin OSM ki OSS1 i EV ki sETR1 genotypes (lanyut 4D). I sETR1, apoka poka ma ki kelo i kin OSM ki OSS1 odok piny tutwal. Yo me bi-clustering ni bene kitiyo kwede i kom ngec me MS/MS ma ki nongo ki ikom dul ma pe ki JA ma oromo me nongo MS/MS modules ma rwate ki JA-regulated metabolic specialization (lanyut S6).
(A) Adwogi me poko 585 MS/MS ma jenge i kom dul ma ki poko (NDP ma rom) ki rweny ma ki poko (NL ma rom) okelo module (M) ma rwate ki dul ma ki ngeyo, onyo ma pe ki ngeyo onyo ma pe ki loko maber. I nge dul acel acel, jami ma ki lwongo ni metabolite (MS/MS) ma pat pat (Si) ki nyutu. (B) Modular molecular network: Nodes nyutu MS/MS ki edges, NDP (red) ki NL (blue) MS/MS scores (cut-off,> 0.6). Gin ma ki lwongo ni graded metabolite specificity index (Si) ma ki keto kala ma lubu module (tung lacuc) ki ki keto i dul ma ki lwongo ni molecular network (tung lacuc). (C) Module M1 pa EV plant ma tye i constitutive (control) ki induced state (simulated herbivore) i cawa 24: cal ma ki lwongo ni molecular network diagram (Si value is the node size, defensive phenolamide is highlighted in blue). (D) M1 molecular network diagram me spectrum line sETR1 ma tye ki goro me EV ki ET: gin ma ki lwongo ni phenolic compound ma ki nyutu ki green circle node, ki apoka poka ma pire tek (P value) i kin W + OSM ki W + OSS1 ma ki tiyo kwede calo dit pa Node. CP, N-caffeoyl-tyrosine; CS, N-caffeoyl-spermidine; FP, N-ferulic acid ester-uric acid; FS, N-ferulyl-spermidine; CoP, N', N "-Coumarolyl-tyrosine; DCS, N', N"-dicaffeoyl-spermidine; CFS, N', N”-caffeoyl, feruloyl-spermidine; Lycium barbarum i wolfberry Son; Nick. O-AS, O-acyl sugar.
Wa medo rwom me nyamo lok ni ki i kom gin acel ma ki lwongo ni Nepenthes genotype me oo i kom dul dano ma gi kwo, ma alokaloka ma tek i kin dul ler ma gi camo lum ki rwom me metabolite ma pat pat ma ki waco con i kom dul dano ma gi kwo (26). Tii ki ngec man me mako 43 germplasms. Germplasms ni tye ki kodi 123 ma aa ki i N. pallens. Yat magi ki kwanyo ki i kom nyig ma ki nongo ki i kabedo ma pat pat i Utah, Nevada, Arizona, ki California (lanyut S7), wa kwano kit ma metabolome pat pat (kany ki lwongo ni wel dano) β kit ma pat pat) ki kit ma OSM okelo kwede. Ma rwate ki kwan mukato, waneno alokaloka mapol i kom Hj ki δj axes, ma nyutu ni germplasms tye ki apokapoka madit i kit ma gi gamo kwede jami ma gi camo lum (lanyut S7). Dul man poyo wiwa i kom jami ma ki neno con ma mako alokaloka ma bino pi ler ma camo lum, ki tye ka gwoko wel ma malo i kin dano acel (26, 36). Nono JA ki JA-Ile me temo rwom me rwate i kin Hj ki δj, wanongo ni tye rwate maber ma pire tek i kin JA ki metabolome β ma pat pat ki rwom me tic (lanyut S7). Man nyutu ni apokapoka ma bino pi ler ma camo lum ma ki nongo i kin dano twero bedo ni oa ki i kom ler ma camo lum.
Kwano mukato angec onyutu ni kodi taba pat pat i kit ki kit ma gi jenge kwede i kom jami ma miyo teko me kom. Ki geno ni alokaloka eni i lok ma mako ler ma camo lum ki kero me gwoke ki doro ki rwom pa kwidi, kwo pa yadi, ki wel cente ma mite me gwoke i kabedo ma ler moni dongo iye. Wa kwano kit ma pot yat loko kwede ma ki kelo ki ler ma camo lum ma ki lwongo ni Nicotiana ma aa ki i North America ki South America. Dul ler magi gi rwate ki Nepenthes North America, ma ki lwongo ni Nicolas Bociflo. La, N. nicotinis, Nicotiana n. lum ma ki lwongo ni attenuated grass, Nicotiana tabacum, linear tobacco, tobacco (Nicotiana spegazzinii) ki tobacco leaf tobacco (Nicotiana obtusifolia) (lanyut 5A) (37). Abic i kin dul magi, medo ki dul ma ki ngeyo maber N. please, obedo yadi ma dongo mwaka ki mwaka i dul pa petunia, ki obtusifolia N. obedo yadi ma dongo mwaka ki mwaka i dul pa Trigonophyllae (38). I ngeye, W + W, W + OSM ki W + OSS1 ma ki keto i kom ler abiro ni me pwonyo kit ma ler ni loko kwede i kom cam pa ler.
(A) Yat ma ki lwongo ni bootstrap phylogenetic tree ma jenge i kom gin ma twero time [pi nuclear glutamine synthesis (38)] ki poko pa dul ler abiro ma gi rwate (kala ma pat pat) (37). (B) cal ma ki nyutu ma pat pat pi kit ma kom ler abiro ma ki lwongo ni Nicotiana (939 MS/MS; data file S1). I rwom me dul ler, apoka poka pa jami makwo tye ki rwate marac ki rwom me tic ma pat pat. Kwedo lok ma mako dul ler ma pat pat ki rwom me tic ki jami ma pat pat ki dongo pa JA ki nyutu i cal 2. S9. kala, dul ma pat pat; triangle, W + OSS1; rectangle, W + OSM; (C) Nicotiana JA ki JA-Ile dynamics ki keto gi malube ki OS excitation amplitude (two-way ANOVA ki Tukey HSD post-multiple comparison, * P <0.05, ** P <0.01 ki * ** Pi poro W + OS ki W + W, P <0.001). Box plot me (D) apoka poka ki (E) apoka poka pa dul ler acel acel inge temo jami ma camo lum ki methyl JA (MeJA). Lakalatwe nyutu apokapoka ma pire tek i kin W + OS ki W + W onyo lanolin ki W (Lan + W) onyo Lan ki MeJA (Lan + MeJa) ki Lan control (kwano lok ma mako apoka poka, ma lubu ki Tukey's HSD post hoc multiple comparison , *P<0.05, **P<0.01 ki ***P<0.001).
Nono yoo me dual cluster, wa nongo 9 modules me 939 MS/MS (data file S1). Kit ma ki yubu kwede MS/MS ma ki yubu ki yoo ma pat pat ni pat pat i kin dul ma pat pat i kin ler (lanyut S8). Neno Hj (ma ki lwongo ni species-level γ-diversity) ki δj nyutu ni species ma pat pat gi ribe i dul ma pat pat i kabedo me metabolic, ma apoka poka i kin species pol kare nen ma loyo excitation. Labongo kwanyo N. linear ki N. obliquus, gin nyutu jami mapol ma kelo alokaloka (lanyut 5B). Ka i poro, dul ler ma calo N. purpurea ki N. obtusifolia gi tye ki adwogi ma pe nen maber i kom yat, ento metabolome ni pat pat. Kit ma dul kom ma pat pat poko kwede i kom dul kom ma ki kelo ni okelo rwate marac i kin dul kom ma pat pat ki dul kom ma pat pat (PCC = -0.46, P = 4.9×10-8). Alokaloka ma OS okelo i rwom me JAs tye ki rwate maber ki rwom me metabolome, ki rwate marac ki metabolic gamma ma pat pat ma ki nyutu i kom ler acel acel (lanyut 5B ki cal S9). Ber me poyo ni dul ler ma ki lwongo ni "gin ma gamo lok" i cal 5C, calo Nepenthes nematodes, Nepenthes nepenthes, Nepenthes acute, ki Nepenthes attenuated, okelo lanyut ma pire tek i dakika 30. Two ma mako OS-specific JA ki JA-Ile ma cok cok ni, ma nongo kwidi mukene ma ki lwongo ni "signal unresponsive", calo Nepenthes mills, Nepenthes powdery ki N. obtusifolia nyutu JA-Ile Edge induction labongo OS specificity mo keken (lanyut 5C). I rwom me tic pa kom, ma calo ma kiloko iye malo ni, pi Nepenthes ma ki dwoko piny, jami ma gamo lanyut ni onyutu OS specificity ki omedo δj, kun dwoko piny Hj. Adwogi pa OS-specific priming ni pe ki nongo i kom ler ma ki poko calo ler ma pe tiyo ki lanyut (lanyut 5, D ki E). Jami ma ki lwongo ni OS-specific metabolites ki poko pol kare i kin ler ma gamo lanyut, ki dul lanyut eni gi rwate ki ler ma gamo lanyut ma goro, kun ler ma gamo lanyut goro nyutu ni pe gi jenge i kom gi (lanyut S8). Adwogi man nyutu ni OS-specific induction pa JAs ki OS-specific reconfiguration pa downstream metabolome gi rwate i rwom me ler.
Ma lubo, wa tiyo ki lanolin paste ma tye ki methyl JA (MeJA) me cango yadi me kwedo ka ce kit me ribbe ni ki gengo pi nongo pa JA ma ki keto ki JA ma aa ki woko, ma obi bedo i cytoplasm pa yadi. Rapid deesterification obedo JA. Wanongo ni kit ma rom ki alokaloka ma bino ki i kom ler ma gamo lanyut me doko ler ma pe gamo lanyut ma oa ki i miyo JA (lanyut 5, D ki E). Ki lok macek, tic ki MeJA odwoko rwom me tic pa ler ma ki lwongo ni linear nematodes, N. obliquus, N. aquaticus, N. pallens, ki N. mikimotoi, ma oweko δj omede ki Hj odok piny. N. purpurea onyutu medo pa δj, ento pe Hj. N. obtusifolia, ma onongo dong ki nyutu ni tye ka gwoko rwom ma piny pa JAs, bene gamo marac i kom yat me MeJA ma lubu kit ma metabolome loko kwede. Adwogi eni nyutu ni yubu JA onyo cwalo lanyut tye ma ki gengo i kom ler ma pe gamo lanyut. Me temo lok man, wa kwano dul ler angwen (N. pallens, N. mills, N. pink ki N. microphylla) ma ki kelo ki W + W, W + OSMs ki W + OSS1 Transcriptome (39). Ma rwate ki kit ma metabolome remodeling tye kwede, dul ler ni gipoke maber i kin transcriptome space, ma i kin gii N. attenuated onyutu OS-induced RDPI ma malo, kun N. gracilis onyutu ma piny loyo (lanyut 6A). Ento, kinongo ni rwom me coc ma ki kelo ki N. oblonga onwongo tye piny loyo i kin dul angwen ni, ma pat ki rwom ma malo pa N. oblonga ma ki nyutu i dul ler abiro. Kwano mukato angec onyutu ni dul pa gene ma kubbe ki lanyut me gwoke con, medo ki lanyut me JA, tito kit ma gwoke con ma ki kelo ki jami ma camo lum i dul ler ma ki lwongo ni Nicotiana (39). Ka kiporo yoo me cwalo ngec pa JA i kin dul ler angwen ni onyutu kit ma nen maber (lanyut 6B). Pol pa gene ma tye i yoo man, ma calo AOC, OPR3, ACX ki COI1, onyutu rwom ma malo me dongo i kom dul ler angwen eni. Ento, gene ma pire tek, JAR4, loko JA me doko gin ma tiyo i kom JA-Ile ma ki coko, ki rwom me coc ne tye piny tutwal, tutwale i N. mills, Nepenthes pieris ki N. microphylla. Medo ikom meno, coc pa gene mukene AOS keken aye pe kinongo i N. bifidum. Aloka loka eni i kit ma gene nyutu kwede twero bedo gin ma okelo kit ma jami nen kwede ma nok ma oa ki i JA ma nok i kom ler ma pe tiyo ki lanyut ki kelo N. gracilis.
(A) Ngec ma ki kwano i kom kit ma ki dwoko kwede rwom me tic pa ler ma ki lwongo ni tobacco ma ki kwanyo i nge dakika 30 ingee camo lum. RDPI ki kwano kun ki poro pot yat ma ki kelo ki OS ma camo lum ki yat ma ki gwoko. kala ni nyutu kodi ma pat pat, ki lanyut ni nyutu kit me cango ne ma pat pat. (B) Kwedo kit ma gene nyute kwede i yoo me JA i kin dul ler angwen. Yo ma ki yubu me JA ni ki nyutu i nget karatac ma ki nyutu ni. Kala ma pat pat nyutu yoo me tic ma pat pat. Lanyut nyutu ni tye apokapoka madit i kin W + OS ma ki miyo ki W + W ma ki temo (pi t-test pa latin kwan pi apokapoka ma tye i kin aryo, *P<0.05, **P<0.01 ki ***P<0.001). OPDA, 12-oxophytodienoic acid; OPC-8: 0,3-oxo-2(2′(Z)-pentenyl)-cyclopentane-1-octanoic acid.
I dul me agiki, wa kwano kit ma jami ma kwo ma pat pat ma gi loko kwo pa jami makwo ma pat pat twero bedo tek kwede i kom ler ma camo lum. Kwedo ngec mukato ni oketo cwinye i kom dul ler ma ki lwongo ni Nicotiana. Kit ma gi lweny kwede i kom Ms ki ler ma ki lwongo ni larvae pat pat tutwal (40). Kany, wa kwano rwate ma tye i kin labol man ki kit ma gi tiyo kwede. Nono kodi taba angwen ma malo ni, ki temo rwate ma tye i kin apoka poka ki rwom me tic pa jami ma gi camo lum ki teko pa yadi i kom Ms ki Sl, wanongo ni teko, apokapoka ki rwom me tic pa Sl weng tye maber, kun rwate i kin rwom me tic pa mon ma ryek ki rwom me tic tye magoro, ki rwate ki Sl pe pire tek (Figure 10). Ma lubu rwom me lweny pa S1, gin aryo ni N. chinensis ki N. gracilis, ma onongo ki nyutu ni gi nyutu rwom me JA signal transduction levels ki metabolome plasticity, onongo tye ki lagam ma pat pat i kom kelo ler ma camo lum, ki gin bene gunyutu teko ma malo marom. Buto.
I mwaka pyerabicel mukato angec, lok ma mako gwok pa yadi omiiyo yoo me kwan, ma jenge i kom ma lukwano gu tito wel ma dit me dongo ki tic pa jami ma gi dongo i kom yadi. Pol pa lok magi pe lubu yoo ma rwate me moko tam matek (41). Gi miiyo tam ma pire tek (3) i rwom acel me nyamo lok. Ka temo jami ma pire tek ma ki waco ni omi kare me nyamo jami mogo ma ki ngeyo, man obi miyo tic ni mede anyim. Bed ki cwak, ento kwer jo mukene (42). Ma ka meno, pwony manyen ni miyo lok ma ki waco i rwom ma pat pat me nyamo lok ki medo rwom manyen me tito lok (42). Ento, lok aryo ma ki waco i rwom me tic, MT ki OD, ki twero gonyo ne mayot calo gin ma pire tek ma ki waco i kom alokaloka ma bino pi ler ma camo lum: lok me OD ye ni alokaloka i "kabedo" me kom tye ki yoo ma malo. Lok me MT ni ye ni alokaloka eni obi bedo ma pe rwate ki yoo ma pat pat ki obi bedo i kabedo me tic pa kom, ki obi bedo ki jami ma gwoko kom ma malo. Kwedo macon me OD ki MT ma ki byeko ni ki temo kun kitiyo ki jami ma ki lwongo ni a priori "defense" compounds. Temo magi ma mako metabolite ni gengo kero me nyamo rwom ki kit ma metabolome loko kwede i kare me camo lum, ki pe ye ni ki temo i kin rwom me wel ma rwate me mito jami ma pire tek ma ki twero tero calo gin acel. Kany, watiyo ki diro manyen me metabolomics ma jenge ikom computational MS ki watimo deconvolution MS analysis i lok ma mako ngec me temo apoka poka ma tye i kin jami aryo ma ki waco i rwom me metabolomics me wilobo. Gin ma pire tek ma ki waco i lok man. Lok me ngec dong ki tiyo kwede i kabedo mapol, tutwale i lok ma mako jami makwo ki kit ma cam mol kwede (43). Ento, kit ma wangeyo kwede, man aye tic mukwongo ma ki tiyo kwede me tito lok ma mako jami ma kwo i kom yadi ki cobo peko ma mako kwo pa jami makwo ma mako alokaloka ma bino pi kare manok ma lubbe ki jami ma tye i kabedowa. I kom lok, kero pa yoo man tye i kero me poro kit ma jami tye kwede i kin ki i kin dul yadi me neno kit ma ler ma camo lum gu dongo kwede ki i dul ma pat pat me doko kit ma gi dongo kwede i kin dul ma pat pat i rwom me dongo ma pat pat. Kit ma kom tiyo kwede.
Kwedo gin ma ki lwongo ni Principal component analysis (PCA) loko jami ma ki nongo ki i kom jami mapol me doko kabedo ma ki dwoko piny iye wek ki gony kit ma jami ni tye ka wot kwede, ki pol kare ki tiyo kwede calo yoo me yenyo jami ma ki nongo, ma calo deconvolution metabolome. Ento, dwoko piny rwom me rwom obi rwenyo dul ngec ma tye i kin ngec ma ki nongo, ki PCA pe twero miiyo ngec ma mako rwom ma mako jami ma rwate ki rwom me kwo, ma calo: kit ma ler ma camo lum gi loko kwede rwom me kwo i kabedo ma pat pat (me labolle, lonyo, poko ne) ki dwong pa jami ma kwo? Gin ango ma nyutu kit ma ler ma camo lum ni tye kwede? Me aa ki i neno pa jami ma pat pat, apoka poka ki kit ma twero kelo kwede, ngec ma tye i kom jami ma kwako pot yat ni ki poko, ki kinongo ni camo ler ma camo lum twero weko jami ma pat pat tiyo. Labongo akalakala, waneno ni, kit ma ki waco kwede i lanyut me ngec ma ki keto i tic, adwogi me tic pa kom tye ki rwate madit i nge lweny pa ler aryo ma gi camo lum (ma ki miyo cam i dyewor Sl) ki ladiro me Solanaceae Ms. Kadi bed ni kit me cam gi ki rwom me cam gi pat pat. Fatty acid-amino acid conjugate (FAC) initiator in OS (31). Nono tic ki jami ma camo lum me cano bur ma ki keto i kom dano, kit ma ki tiyo kwede ki jami ma camo lum bene onyutu kit ma rom. Kit me tic man ma ki ngeyo me nyutu kit ma yadi gi gamo kwede i kom ler ma camo lum kwanyo jami ma kelo apokapoka ma aa ki i alokaloka i kit ma ler ma camo lum gi camo kwede, ma kelo peko ma pat pat i kare ma pat pat (34). FAC, ma ki ngeyo ni en aye gin ma kelo OSM, dwoko piny JAS ki jami mukene ma miyo teko i OSS1, kun OSS1 dwoko piny tyen miya mapol (31). Ento, OSS1 okelo rwom me dongo pa JA ma rom ki OSM. Ki dong ki nyutu ni lagam pa JA i Nepenthes ma ki dwoko piny ni tye ma rwate ki OSM, ma FAC twero gwoko tic ne kede ka ki medo 1:1000 ki pii (44). Pi meno, ka kiporo ki OSM, kadi bed ni FAC i OSS1 tye piny tutwal, en romo kelo two JA ma oromo. Kwano mukato angec onyutu ni porin-like proteins (45) ki oligosaccharides (46) ki tiyo kwede calo lanyut me molecule me kelo lagam pa yadi i OSS1. Ento, pwod pe tye atir ka ce jami ma kelo two i OSS1 ni gi tye ki twero i kom dongo pa JA ma ki neno i kwan ma tye ka time ni.
Kadi bed ni tye kwan manok ma tito apoka poka ma bino pi tic ki ler ma camo lum ma pat pat onyo JA onyo SA (salicylic acid) (47), pe tye ngat mo ma obalo kit ler ma camo lum ma pat pat i kin lum ma ki pito ki adwogi ne i kom ler ma pat pat ma ki kwano. ngec ma kwako in. Kwedo man omedo ki moko ni dul kom ma tye i kin dul kom ma mako kom ki dul kom mukene ma pat ki JAs loko kit ma ler ma camo lum gi kelo kwede. I kom lok, wanongo ni ET ma OSM okelo onwongo dit kato ma OSS1 okelo. Kit man rwate ki jami mapol ma ki lwongo ni FAC i OSM, ma obedo gin ma mite ki ma oromo me kelo ET burst (48). I lok ma mako rwate i kin yadi ki ler ma camo lum, tic me nyutu pa ET i kom jami ma kwo i kom yadi pud tye ma rwate ki dul acel keken. Medo ikom meno, pol pa kwan ni otiiyo ki tic ki ET onyo jami ma okelo ne onyo jami ma gengo ne me kwan kit ma ET doro kwede, ma i kin gii tic ki jami ma ki kwanyo ki woko ni obi kelo adwogi marac mapol ma pe ki ngeyo. Ki ngeyo wa, kwan man nyutu rwom me acel ma dit me neno tic pa ET i tic ki ET me kelo ki neno jami ma ki loko i kom jami ma kwo ma obale me rwate ki kit ma jami tiyo kwede. ET ma ki lwongo ni Herbivore-specific ET induction twero loko kit ma metabolome gamo kwede. Gin ma pire tek loyo en aye loko jami ma ki lwongo ni ET biosynthesis (ACO) ki perception (ETR1) genes ma onyutu ni herbivore-specific de novo accumulation of phenolamides. Ki nyutu con ni ET twero yubu JA-ma okelo dongo pa nicotine kun tiyo ki putrescine N-methyltransferase (49). Ento, ki i neno me jami ma tiiyo, pe tye atir kit ma ET loko kwede rwom me tic pa phenamide. Medo i kom tic me cwalo ngec pa ET, kit ma kom dano loko kwede twero bedo S-adenosyl-1-methionine me doro keto cente i polyaminophenol amides. S-adenosyl-1-methionine tye ET ki gin ma rwate i kin polyamine biosynthetic pathway. Kit ma ET signal ni doro kwede rwom me phenolamide mito kwan mukene.
Pi kare malac, pi wel madwong pa jami ma pat pat ma pe ki ngeyo kit ma gi nen kwede, keto cwiny i kom dul jami ma pat pat ma pat pat pe twero moko alokaloka me kare me jami ma pat pat inge rwate pa jami makwo. I kare ni, malube ki ngec ma ki nongo, adwogi ma pire tek me nongo MS/MS spectrum ma jenge i kom jami ma pe ki poko kin dano tye ni ler ma camo lum ma camo onyo lubo ler ma camo lum gi mede ki dwoko piny kit ma kom gi nen kwede i pot yat kun gi medo rwom me tic gi. Medo pa jami ma ki lwongo ni metabolome ma ki kelo ki ler ma camo lum ni rwate ki medo pa jami ma ki coyo i kom jami ma ki coyo. Gin ma konyo tutwal i kom dul kom ma dit ni (ma tye ki wel Si ma malo) tye dul kom ma pat ma tye ki tic me camo lum ma ki ngeyo con. Labol man rwate ki lok ma ki waco i kom OD, ento lok ma ki waco i kom MT ma kubbe ki randomness me metabolome reprogramming pe rwate. Ento, ngec man bene rwate ki lok ma ki waco i kom dul ma ki rubu (MT maber loyo; cal 1B), pien jami mukene ma pe ki ngeyo ma tye ki tic me gwoke ma pe ki ngeyo twero lubo poko pa Si ma pe ki ngeyo.
Gin ma pire tek ma ki nongo ki i kwan man tye ni cake i rwom me dongo pa jami matino tino (yat acel ki yadi ma ki lwongo ni tabacco) me oo i rwom me dongo pa jami mapol (kwato taa ma rwate), rwom me dongo pa jami ma pat pat tye i "gwok maber loyo" Tye apokapoka madit i kero pa ler ma camo lum. Moore ki luwote (20) ki Kessler ki Kalske (1) gu moko tam gi me loko rwom adek me tic pa jami ma kwo ma Whittaker (50) opoko gii me doko alokaloka ma kelo ki kelo alokaloka i kare me jami makwo ma pat pat; lucoc magi pe gu coyo lok ma ki coyo ni kit me coko ngec ma mako kom dano mapol bene pe nyutu kit me kwano kit ma kom dano pat pat ki ikom ngec magi. I kwan man, alokaloka manok i kom poko tic pa Whittaker obi keto α-metabolic diversity calo apoka poka pa MS/MS spectra i kom gin ma ki pito, ki β-metabolic diversity calo basic intraspecific metabolism pa dul pa dano.
Lanyut me JA pire tek pi jami mapol ma mako ler ma camo lum. Ento, tye peko me temo ma rwate ki rwom me tic pa JA biosynthesis i kom dul kom ma pat pat, ki ka JA signal obedo kabedo ma rwate pi dul kom ma kelo peko i kom rwom ma malo me macroevolutionary pud pe ki ngeyo. Wa neno ni kit pa Nepenthes ma camo lum kelo rwom me metabolome ki apoka poka me metabolome i kin dul pa Nicotiana ki i kin dul pa Nicotiana ma gi rwate kwede tye ki rwate maber ki JA signaling. Medo ikom meno, ka JA signal ni obale, kit ma kom ler ma camo lum acel okelo kwede ki bi juku (lanyut 3, C ki E). Kit ma alokaloka i kit ma kom dano ma ki lwongo ni Nepenthes ni gi loko kwede pol ne tye ma lubu wel, alokaloka ma tye i kom kit ma kom dano tiyo kwede i kom jami ma ki lwongo ni metabolic β diversity ki specificity i kom jami ma ki kwano ni twero bedo ni oa ki i kom jami ma ki lwongo ni metabolite-rich compound categories. Dul jami ma ki rubu ni gi loyo dul kom ma ki lwongo ni metabolome profile ki gi kelo rwate maber ki lanyut pa JA.
Pien kit ma jami makwo pa kodi ma gi rwate kwede ni pat pat, jami ma gi loko i kom gi ki ngeyo gii maber, pi meno bedo me nyamo lok. Kit ma lok me ngec tiyo kwede i kom kit ma kom dano tiyo kwede nyutu ni cam ma ki camo i kom yadi medo apoka poka i kin kit ma kom dano loko kwede ki kit ma gi ngeyo kwede. Gin ma ki lwongo ni JA tye ka tic ma pire tek i lok man. Medo pa rwom me metabolome rwate ki OD ma ki byeko ki tye ki rwate maber ki JA signal, kun JA signal tye ki rwate marac ki metabolic gamma diversity. Labol magi nyutu ni kero me OD pa jami ma dongo dongo ki moko ki kit ma JA tye kwede, kadi bed ni i rwom me microevolutionary nyo i rwom me dongo ma dit. Temo ma ki lwongo ni Exogenous JA application ma gengo peko me JA biosynthesis nyutu ni kodi taba ma gi rwate ki gii ki twero poko gi i kin kodi ma gamo lanyut ki kodi ma pe gamo lanyut, kit ma calo kit gi me JA ki metabolome plasticity ma ki kelo ki ler ma camo lum. Jami ma pe gamo lok pe twero gamo lok pien pe gitwero kelo JA ma aa ki i kom gi ki dong gi tye ki peko me kom. Man twero bedo ni oa ki i alokaloka i gene mogo ma pire tek i JA signaling pathway (AOS ki JAR4 i N. crescens) me. Adwogi man nyutu ni kit ma ler ni gi dongo kwede twero bedo ni oa ki i alokaloka ma tye i kit ma kom gi winyo kwede ki kit ma gi gamo kwede.
Medo i kom rwate i kin yadi ki ler ma camo lum, yenyo kit ma jami makwo gi loko kwede rwate ki jami ma pire tek ma ki nongo i kwan ma mako kwo pa jami ma kwo i kabedowa ki dongo pa kit ma kom dano nen kwede. Ki medo wel ngec ma kinongo ki jami tic me MS me kare ni, temo tam i kom kit ma jami tiyo kwede i kom dano twero kato apoka poka pa dano acel acel/dul dano ki timo kwed i wilobo me nyutu jami ma pe ki tamo. I yub me kwedo jami mapol, gin ma pire tek ma ki poro kwede tye tam me yubu map ma twero tic me yenyo ngec. Pi meno, adwogi ma pire tek me rwate pa MS/MS metabolomics ma pe ki apokapoka ki lok me ngec tye ni en miyo yoo ma yot ma ki twero tic kwede me yubu map me neno kit ma metabolic tye kwede i rwom ma pat pat. En aye gin ma mite pi yoo man. Kwano lok ma mako dongo pa jami matino tino ki jami ma kwo i kin dano.
I rwom me macro-evolutionary, gin ma pire tek i kom lok ma mako dongo pa jami ma kwo ki ler ma ki lwongo ni Ehrlich ki Raven (51) tye me byeko ni apoka poka pa jami ma kwo i kin jami makwo ma pat pat aye okelo apoka poka i kin jami ma kwo. Ento, i mwaka pyerabic ni cake i kare ma ki coyo lok man, lok man pe ki temo (52). Man tye pi kit ma jami ma kwo gi nen kwede ma rom ki kit ma gi dongo kwede i kin yadi ma gibedo ka mabor. Gin ma pe nen ni ki tiyo kwede me keto yoo me nyamo gin ma ki miito. Kit me tic pa MS/MS ma tye i kare ni ma ki tiyo kwede ki pwony me ngec kwano kit ma MS/MS rwate kwede ki jami ma pe ki ngeyo (labongo yero jami ma ki yero con) ki loko MS/MS magi me doko MS/MS, pi meno i tic pa ludiro tic pa jami ma ki lwongo ni macro-evolutionary Models ni ki poro gi i rwom me poko kin jami. Jami ma nyutu wel ma yot. Kit me tiimo ne rom ki nyamo jami makwo, ma twero tic ki rwom me rwom me jami ma pat pat onyo kit ma gi loko kwede labongo byeko con.
I rwom me biochemical, yub me temo pa Firn ki Jones (53) nyutu ni kit ma kom dano tiyo kwede ki gwoko i rwom ma pat pat me miyo jami ma mitte me tic ki jami ma kwo ma onongo pe rwate nyo ma kiloko iye. Yoo me lok ma mako ngec miyo yoo ma weko alokaloka me dongo pa jami makwo ma time i kare me dongo pa jami makwo twero bedo ma ki kwano calo dul me yub me kwan ma ki yubu: loko kwo pa jami ma kwo ki i rwom ma lapiny me cito i rwom ma malo Gin ma ki gengo i kabedo ma kimiyo.
Me agiki ne, i acaki me kwan pa jami makwo, pwony ma pire tek ma mako gwok pa yadi ki oyubu, ki yoo ma ki moko ma ki ngeyo ni obedo yoo acel keken me dongo ngec me cayan. Man tye pi peko me diro me pimo metabolome weny. Kadi bed ni yoo ma ki tiyo kwede ki tam ma ki tamo ni ber tutwal i yero jami mukene ma kelo peko, kero gi me medo ngec wa i kom dul ma kwako jami makwo tye manok loyo yoo me kwano ma tye i kare ni i kare ni. Pi meno, lok ma pe kitwero byeko tye mabor ki rwom me ngec ma tye, pi meno yoo me byeko/temo me dongo i dul me kwan pe kitwero kwanyo woko (4). Wa neno ni tic me kwan me metabolomics ma ki kelo kany twero dwoko miti i kom jami manyen (kit ma) ki me agiiki (pingo) lok ma mako kit ma jami tiyo kwede, ki konyo i kare manyen me ngec ma ki ngeyo. Kare ni odwoko cen lok ma pire tek ma okelo miti i kom rwom me kwo pa dano macon.
Miiyo cam bot ler ma camo lum ki tiimo kun ki pito ler ma ki lwongo ni instar larva me aryo onyo Sl larva i kom pot yat acel ma rangi ne nen maleng ma tye ki yat acel ma tye ka dongo, ki yat 10 ma ki dwoko cen pi yat acel acel. Nyig ler ni ki mako ki jami ma ki mako, ki pot yat ma odong ni ki kwanyo cawa 24 ki 72 inge nongo two ki ki juku oyot, ki jami ma ki kwanyo ki i kom gi ni ki kwanyo.
Tem tic ki jami ma camo lum i yoo ma rwate. Yoo me tic kwede tye me tic ki jami ma ki yubu ki bongo me turo dul yat adek ma tye i dul kom yat adek ma dong odongo maber i kare ma yat ni tye ka dongo, ki cut cut i keto 1:5 Diluted Ms. Onyo tic ki cing ma ki yubu ki cing me keto S1 OS i bur ma ki turo. Kwany ki tic ki pot yat kit ma ki waco kwede malo ni. Tii ki yoo ma kiloko iye con me kwanyo jami ma konyo kom ki jami ma miyo teko i kom yat (54).
Pi tic ki JA ma woko, pot yat adek ma tye i kom yat abicel ma tye ka dongo i dul acel acel ki tiyo kwede ki 20μl me lanolin ma tye ki 150μg MeJA (Lan + MeJA), ki 20μl me lanolin ki yat ma ki lwongo ni wound treatment (Lan + W), onyo ki tiyo ki 20μl me lanolin ma calo gin ma ki tiyo kwede me gengo. Pot yat ni ki kwanyo i nge cawa 72 inge tic kwede, ki gwoko i pii ma ki lwongo ni liquid nitrogen, ki ki gwoko i -80°C nio ka ki tiyo kwede.
JA ki ET angwen ma ki loko, ma ki lwongo ni irAOC (36), irCOI1 (55), irACO ki sETR1 (48), ki nongo gi i dul kwan wa. irAOC onyutu ni rwom me JA ki JA-Ile odok piny, kun irCOI1 onongo pe winyo JAs. Ka kiporo ki EV, JA-Ile ma odongo omede. I yo acel-lu, irACO obi dwoko piny rwom me ET, ki ka kiporo ki EV, sETR1, ma pe winyo ET, obi medo rwom me ET.
Gin ma ki lwongo ni photoacoustic laser spectrometer (Sensor Sense ETD-300 real-time ET sensor) ki tiyo kwede me timo ET ma pe kelo peko. Cut cut ingee yat, nucu pa pot yat ni ki kwanyo ki keto i 4-ml ma ki umu ki gilaci, ki ki weko kabedo ma i wiye ni opong i kin cawa 5. I kare me pimo, ki lwoko kikopo acel acel ki yamo ma romo lita 2/cawa pi dakika 8, ma onongo okato ki i gin ma ki lwongo ni Sensor Sense me kwanyo CO2 ki pii.
Lok ma ki nongo ki i microarray ni ki coyo i (35) ki ki gwoko i National Center for Biotechnology Information (NCBI) Gene Expression Comprehensive Database (namba me donyo GSE30287). Ngec ma rwate ki pot yat ma oa ki i tic ki W + OSMs ki ma pe ki balo ki kwanyo pi kwan man. Gin ma ki lwongo ni raw intensity tye log2. Mapwod pi ki kwano wel, ki loko rwom me acaki ki ki dwoko piny me bedo 75th percentile kun kitiyo ki R software package.
Gin macon ma ki lwongo ni RNA sequencing (RNA-seq) pa ler ma ki lwongo ni Nicotiana ki nongo ki i NCBI Short Reading Archives (SRA), namba me yub ni tye PRJNA301787, ma ki coyo ki Zhou et al. (39) ki mede anyim kit ma ki waco kwede i (56). Lok ma ki nongo ma ki tiyo kwede ki W + W, W + OSM ki W + OSS1 ma rwate ki dul ler ma ki lwongo ni Nicotiana ki yero me nyamo i kwan man, ki ki tiyo kwede i yoo ma lubo ni: Mukwongo, kwan ma ki nongo ki i RNA-seq ki loko i kit me FASTQ. HISAT2 loko FASTQ me doko SAM, ki SAMtools loko SAM me doko BAM ma ki poko. StringTie ki tiyo kwede me kwano kit ma gene nyute kwede, ki kit me nyutu ne tye ni tye dul kom ma tye i kom dul kom alip acel acel i kom dul kom ma ki coyo ma ki lwongo ni sequenced transcription.
Acclaim chromatographic column (150 mm x 2.1 mm; particle size 2.2μm) ma ki tiyo kwede i kwedo ki 4 mm x 4 mm guard column tye ki jami acel. Gin ma ki lwongo ni binary gradient ni ki tiyo kwede i Dionex UltiMate 3000 Ultra High Performance Liquid Chromatography (UHPLC) system: 0 onyo 0.5 dakika, isocratic 90% A [deionized water, 0.1% (v/v) acetonitrile ki 0.05% formic acid], 10% Acetonitrile 0.05% formic acid); 0.5 onyo 23.5 dakika, rwom me rwom ma malo tye 10% A ki 90% B, kit meno; 23.5 onyo 25 dakika, isocratic 10% A ki 90% B. Kit ma mol kwede tye 400μl/min. Pi kwedo MS weny, ket gin ma ki lwongo ni column eluent i quadrupole ki time-of-flight (qTOF) analyzer ma tye ki electrospray source ma tiyo i positive ionization mode (capillary voltage, 4500 V; capillary outlet 130 V ; Drying temperature 200°C; drying airflow 10min/liter).
Tim MS / MS fragment analysis (ma ki lwongo ni MS / MS) ma pe rwate ki gin ma ki nongo me nongo ngec ma mako kit ma kom dano tiyo kwede. Tam me yoo me MS/MS ma pe ki poko jenge i kom lok ada ni quadrupole tye ki wang dirica ma dit [pi meno, nen ni wel ma ki keto i rwom me pek (m/z) ducu calo dul kom]. Pi tyen lok eni, pien gin ma ki lwongo ni Impact II onongo pe twero yubu CE tilt, kitimo kwedo mogo ma pat pat kun ki tiyo ki wel ma ki medo i kom collision-induced dissociation collision energy (CE). Ki lok macek, mukwongo nyam gin ma ki nyutu ni ki UHPLC-electrospray ionization/qTOF-MS kun kitiyo ki kit me mass spectrometry acel (kit ma ki poko kwede ma nok ma ki kelo ki in-source fragmentation), ma ki kwanyo ki i m/z 50 nio wa i 1500 i rwom me 5 Hz. Tii ki nitrogen calo yamo ma rwate pi MS/MS analysis, ki tim pimo ma pat pat i kom teko ma pat pat ma bino pi rwate: 20, 30, 40, ki 50 eV. I kare me pimo ne, quadrupole ni tye ki wang ma dit loyo me poko pek pa jami, cake ki m/z 50 nio wa i 1500. Ka m/z ki lac pa kom ma i anyim ki keto i 200, rwom me pek pa jami ni ki cako tic kene ki jami ma tiyo kwede ki 0 Da. Yeny pi dul ma pek ma calo i kit me pek acel. Tii ki sodium formate (50 ml isopropanol, 200 μl formic acid ki 1 ml 1M NaOH aqueous solution) me pimo pek pa jami. Nono yoo pa Bruker ma ki lwongo ni high-precision calibration algorithm, gin ma ki coyo ni ki keto i nge tic ki rwom me rwom ma malo i kare ma kimiyo. Tii ki tic me cwalo jami ma ki lwongo ni Data Analysis v4.0 software (Brook Dalton, Bremen, Germany) me loko karatac ma ki coyo ni i NetCDF. Gin ma ki lwongo ni MS/MS ni ki gwoko i database ma ki lwongo ni MetaboLights (www.ebi.ac.uk) ki namba me donyo. MTBLS1471.
MS/MS assembly ki twero tiimo ne ki i kin MS1 ​​ki MS/MS ma nyutu rwom me teko ma lapiny ki ma malo ki cik manyen ma ki keto. R script ni ki tiyo kwede me nongo rwom me poko kin jami ma ki yubu, ki C# script (https://github.com/MPI-DL/indiscriminant-MS-MS-assembly-pipeline) ki tiyo kwede me keto cik ni i tic.
Me dwoko piny bal ma pi adaa ma oa ki i koko ma tye i ngeye ki rwate ma pi adaa ma oa ki i nongo jami mogo ma ki lwongo ni m/z i kom labol manok keken, wa tiyo ki "filled peak" ma tye i R package XCMS (pi yubu background noise correction) Myero ki ti kwede me loko "NA" (undetected peak) intensity. Ka kitiyo ki tic me pongo rwom ma malo, tye wel mapol ma ki lwongo ni "0′′ i kin jami ma ki nongo ma obi gudo rwom me rwate. Ki dong, wa poro adwogi me tic ki ngec ma kinongo ka kitiyo ki tic ma opong ki rwom ma malo ki ka pe kitiyo ki rwom ma malo ma opong, ki kwano rwom me koko ma tye i ngeye malube ki rwom ma ki loko, ki dong ki loko rwom magi ki 0" wel ma ki kwano me angec. Wan bene wa tamo i kom jami ma tek ne kato wel me angec tyen adek ki waneno gi calo "rwom ma malo kikome." Pi kwano PCC, ki kwano m/z keken pa gin ma ki nyutu (MS1) ki dul ma tye ki rwom ma romo aboro ma tye kakare.
Ka rwom me rwom me rwom ma lamal i kom gin ma ki nyutu ni tye ki rwate ki rwom ma piny pa rwom acel-lu ma tye ka rwate ki teko ma lapiny onyo ma malo, ki rwom ni pe ki keto calo isotope peak ki CAMERA, ki twero medo gonyo ne. I ngeye, ki kwano jami weny ma twero bedo tye i kin 3 s (kare me gwoko ma ki byeko pi gwoko ma malo), kitimo kwedo i kom rwate. Ka wel m/z tye piny loyo wel ma okwongo ki poko MS/MS time i kabedo acel ma ki nyutu iye i kin jami ma ki nongo ki iye, en aye ki ngeyo ni obedo poko.
Malube ki cik aryo magi ma yot ni, wa kwanyo dul ma ki keto ma tye ki wel m/z ma dit loyo m/z pa gin ma ki ngeyo ni, ki malube ki kama gin ma ki nyutu ni nen iye ki dul ma ki waco ni. Ki bene twere me yero jami maber ma ki yubu ki jami mapol ma ki yubu i MS1 calo jami ma ki yero, ma weko ki kelo jami ma pe rwate ki MS/MS. Me dwoko piny rwom me ngec man, ka NDP ma rom pa jami ma ki neno ni okato 0.6, ki gi tye i chromatogram "pcgroup" ma ki coyo ki CAMERA, wa bi keto gi kacel. Me agiki ne, wa ribo adwogi me CE angwen ma mako gin ma okwongo ki dul ne i dul ma ki poko gii me agiki kun wa yero gin ma ki lwongo ni intensity peak ma malo loyo i kin peaks weny ma tye ki m/z acel i kom collision energy ma pat pat. Tyelo tyelo me tic ma lubo ni jenge i kom tam me composite spectrum ki kwako kit ma CE ma pat pat ma mite me medo rwom me poko poko, pien poko mogo ki twero nongo gi i te teko me rwate.
RDPI (30) onwongo ki tiyo kwede me kwano kit ma kom dano twero tic kwede. Kit ma metabolic spectrum diversity (Hj index) ki nongo ki i dwong pa MS/MS precursors kun kitiyo ki Shannon entropy me MS/MS frequency distribution kun kitiyo ki equation ma ki waco ki Martínez et al. (8). Hj = −∑i = 1mPijlog2(Pij) kan ma Pij rwate ki rwom me i-th MS/MS i j-th sample (j = 1, 2,..., m) (i = 1, 2, …, m) t).
Metabolic specificity (Si index) ki gonyo calo nyutu pa MS/MS ma kimiyo ma lubu kit ma pol pa gin ma ki nyutu ni tye kwede. MS/MS specificity ki kwano ni Si = 1t (∑j = 1tPijPilog2PijPi)
Tii ki yoo ma lubo ni me pimo metabolome-specific δj index pa j acel acel, ki rwom me MS/MS specificity δj = ∑i = 1mPijSi
MS/MS spectra ki keto i dul aryo, ki kit ma rom kwede ki kwano malube ki wel aryo ni. Mukwongo, kun itiyo ki NDP ma ki ngeyo (ma ki ngeyo ni cosine correlation method), tii ki equation ma lubo ni me nongo rwom me rwate i kin spectra NDP = (∑iS1 & S2WS1, iWS2, i) 2∑iWS1, i2∑iWS2, i2 kan ma S1 ki S2 Ma rwate, pi spectrum 1 ki 2, ki WS2, ki i2 nyutu pek ma jenge i kom rwom ma malo ma apoka poka pa i-th common peak i kin spectra aryo ni tye ma nok loyo 0.01 Da. Pek ne ki kwano kit man: W = [peak intensity] m [quality] n, m = 0.5, n = 2, kit ma MassBank owaco kwede.
Yo me aryo me keto wel ma ki keto i tic, ma kwako nyamo NL ma ki poko i kin MS/MS. Me agiki, watiyo ki 52 NL ma ki nongo kare ki kare i kare me MS fragmentation process kacel, ki NL ma ki ngeyo maber (data file S1) ma onongo dong ki coyo con pi MS/MS spectrum pa secondary metabolites pa dul Nepenthes ma goro ( 9, 26). Yub gin ma ki lwongo ni binary vector me 1 ki 0 pi MS/MS acel acel, ma rwate ki ma tye ki ma pe tye pa NL mogo. Malube ki rwom me rwate pa Euclidean, rwom me rwate pa NL ki kwano pi dul acel acel me binary NL vectors.
Me timo dual clustering, watiyo ki R package DiffCoEx, ma jenge i kom medo pa Weighted Gene Co-expression Analysis (WGCNA). Nono NDP ki NL scoring matrices me MS/MS spectra, watiyo ki DiffCoEx me kwano comparative correlation matrix. Binary clustering ki tiimo kun ki keto "cutreeDynamic" parameter me method = "hybrid", cutHeight = 0.9999, deepSplit = T, ki minClusterSize = 10. R source code pa DiffCoEx ki kwanyo ki i file mukene 1 pa Tesson et al. (57); Gin ma mite me R WGCNA software package ni ki twero nongo ne i https://horvath.genetics.ucla.edu/html/CoexpressionNetwork/Rpackages/WGCNA.
Me timo MS/MS molecular network analysis, wa kwano paired spectral connectivity ma lubu NDP ki NL similarity types, ki wa tiyo ki Cytoscape software me neno network topology kun wa tiyo ki organic layout i CyFilescape yFiles layout algorithm extension application.
Tii ki R version 3.0.1 me timo kwan ma mako wel ma ki nongo. Kit ma ki kwano kwede ki kwano kun kitiyo ki two-way analysis of variance (ANOVA), ki lubu ki Tukey's honestly significant difference (HSD) post-hoc test. Me temo apoka poka ma tye i kin yat ma ki camo ki yat ma pe ki tiyo kwede, poko kin dul aryo ma ki kwanyo ma tye ki apoka poka acel ki kwano kun kitiyo ki t test pa latin kwan.
Pi jami ma ki medo i kom pwony man, nen http://advances.sciencemag.org/cgi/content/full/6/24/eaaz0381/DC1
Man obedo gin ma dano twero donyo iye ma ki poko i te cik me Creative Commons Attribution-Non-Commercial License, ma ye ni ki ti kwede, poko ne ki dwoko ne i yoo mo keken, ka ce tic kwede me agiki pe pi nongo cente ki lok ni tic ma ki coyo ni tye kakare. Ka neno.
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