Plant defense theory hrang hrangte hian plant specialized metabolism pattern sawifiahna atan theoretical guidance pawimawh tak tak a pe a, mahse an key prediction te chu test a la ngai a ni. Hetah hian unbiased tandem mass spectrometry (MS/MS) analysis hmangin thlai chi hrang hrang atanga population leh a inzawm tlat chi hrang hranga zuk leh hmuam attenuated strain-te metabolome chu systematic takin kan chhui a, information-a compound spectra hmanga mass spectrometric feature theory tam tak kan process bawk. Optimal defense (OD) leh moving target (MT) theory-a prediction pawimawh tak takte test nan framework siam a ni. Plant metabolomics-a information component hi OD theory nen a inmil a, mahse herbivores-te avanga metabolomics dynamics chungchanga MT theory-in a sawi lawk ber chu a kalh a ni. Micro atanga macro evolutionary scale thlengin jasmonate signal chu OD tichiangtu ber nia hriat a ni a, ethylene signal chuan MS/MS molecular network-in herbivore-specific response annotated chu fine-tuning a pe thung.
Thilsiam hrang hrang nei special metabolites te hi thlai te chu boruak nena an insiamremnaah a tel ber a, a bik takin hmêlma venna kawngah an tel ber a ni (1). Thlai chi hrang hranga hmuh chhuah special metabolism chi hrang hrang mak tak hian kum sawm tam tak chhung chu a ecological functions hrang hrangah zirchianna thuk tak a siam a, plant defense theory list rei tak a siam bawk a, chu chu plant leh insect inzawmna evolutionary leh ecological development a ni. Empirical research hian kaihhruaina pawimawh tak a pe a (2). Mahse, heng plant defense theories te hian hypothetical deductive reasoning normative path an zawm lo va, chutah chuan key prediction te chu analysis level inangah an awm a (3) a, theoretical development cycle lo awm tur chu hmasawn zel turin experimental-in an test a ni (4). Technical limitations hian data lakkhawmna chu metabolic category bikteah a tihkhawtlai a, specialized metabolites te zirchianna kimchang tak a huam lo a, chu chuan theoretical development atana pawimawh tak inter-category comparisons a tikhawlo a ni (5). Metabolomics data kimchang tak leh plant group hrang hrangte inkara metabolic space-a processing workflow tehkhin theihna tur pawisa inang lo hian field scientific maturity a tikhawlo a ni.
Tandem mass spectrometry (MS/MS) metabolomics lama hmasawnna thar ber berte hian system clade pakhata species chhung leh species inkara metabolic inthlak danglamna chu kimchang takin a characterize thei a, computational methods nen pawh inzawmkhawmin heng complex mixtures te structural similarity hi chhut theih a ni bawk. Chemistry lama hriat hmasak (5). Analysis leh computing lama technology hmasawn tak takte inzawmkhawm hian metabolic diversity chungchanga ecological leh evolutionary theory-te’n an sawi lawk tam tak hun rei tak chhunga test theihna tur framework tul tak a pe a ni. Shannon (6) chuan kum 1948-a a seminal article-ah a vawi khat nan information theory a rawn hmang a, chu chuan information mathematical analysis atana lungphum a phum a, chu chu a hmanna hmasa tih loh chu kawng tam takah hman a ni tawh a ni. Genomics-ah chuan information theory chu hlawhtling takin hmangin sequence conservative information chu quantified a ni tawh (7). Transcriptomics research-ah chuan information theory hian transcriptome-a inthlak danglamna zawng zawng a zirchiang thin (8). Research hmasaah khan information theory statistical framework chu metabolomics-ah kan hmang a, chu chuan thlai tissue level-a metabolic expertise kan sawifiah a (9). Hetah hian MS/MS-based workflow leh information theory statistical framework, common currency-a metabolic diversity-a hriat lar tak nen kan inzawm khawm a, herbivores-te’n an tihchhuah metabolome-a plant defense theory-a key prediction-te chu kan khaikhin a ni.
Plant defense-a theoretical framework-te hi a tlangpuiin inzawmkhawm a ni a, chi hnih-ah then theih a ni: defense function-a innghat plant-specific metabolite-te insem darh dan sawifiah tumtute, chu chu optimal defense (OD) (10), moving target (MT) (11) ) Tin, a lan dan (12) theory te a ni a, mi dangte chuan resource availability inthlak danglamnain a nghawng dan mechanical explanation an zawng thung thanlenna leh specialized metabolites, carbon ang chi a khawlkhawm dan: nutrient balance hypothesis (13), growth rate hypothesis (14 ), leh growth and differentiation equilibrium hypothesis (15). Theory set pahnih hi analysis level hrang hrangah an awm a (4). Mahse, functional level-a defensive functions inrawlhna theory pahnih chuan plant constitutive leh inducible defense chungchanga inbiakna chu a thunun a: OD theory, chu chuan plant-te chuan an chemical defense man to tak takte chu a mamawh hunah chauh an invest tih a ngaihtuah a, entirnan, an ei hunah Grass animal-in a beih hunah, chuvangin, nakin lawka beih theih dan azirin, defensive function nei compound chu ruat a ni (10) MT hypothesis chuan directional metabolite inthlak danglamna axis a awm lo tih a rawt a, mahse metabolite chu random takin a inthlak a, chu chuan herbivores attacking metabolic “motion target” tihbuai theihna a siam a ni. Phên dangin sawi ila, heng theory pahnih hian herbivores-te beih hnua metabolic remodeling lo awm tur chungchangah hian sawi lawkna inthlau tak tak an siam a: defensive function (OD) nei metabolites unidirectional accumulation leh non-directed metabolic changes (MT) inzawmna (11 ).
OD leh MT hypothesis-ah hian metabolome-a induced changes chauh ni lovin, heng metabolites-te pungkhawm avanga ecological leh evolutionary consequences te pawh a tel a, chu chu ecological environment bik pakhata heng metabolic changes-te adaptive costs leh benefits te hi a ni (16). Hypothesis pahnih hian specialized metabolites-te defensive function chu hre chiang mah se, chu chu a man a to thei a, a man a to lo thei bawk a, mahse OD leh MT hypothesis thliar hrangtu key prediction chu induced metabolic changes directionality-ah a awm a ni. Tun thlenga OD theory prediction hian experimental attention a dawng nasa ber a ni. Heng test-ah hian greenhouse leh natural condition-a compound bik tissue hrang hrangte direct emaw indirect emaw defense function zir chianna te, thlai lo thanglian dan stage inthlak danglamna te zirchianna a tel a ni (17-19). Mahse, tun thleng hian eng organism pawh metabolic diversity global comprehensive analysis atana workflow leh statistical framework a awm loh avangin theory pahnih inkara danglamna prediction ber (chu chu metabolic changes kalna tur a ni) chu test a la ngai a ni. Hetah hian chutiang thlirletna chu kan pe a ni.
Plant-specific metabolites te ziarang langsar ber pakhat chu an structural diversity nasa tak chu level zawng zawngah plant pakhat, population atanga species inang thlenga an awm vang a ni (20). Specialized metabolites-a quantitative change tam tak chu population scale-ah hmuh theih a ni a, species level-ah erawh chuan qualitative difference chak tak chu a awm reng thung (20). Chuvangin, plant metabolic diversity hi functional diversity-a thil pawimawh ber a ni a, niche hrang hranga insiamrem theihna a lantir a, a bik takin special insect leh common herbivores-te invasion theihna hrang hrang nei niche-te chu a lantir a ni (21). Fraenkel-a (22)-a’n thlai chi hrang hrang metabolite awm chhan chungchanga groundbreaking article a ziah atang khan, insect hrang hrangte nena inzawmna chu selection pressure pawimawh takah ngaih a ni a, heng inzawmnate hian evolution lai khan thlai a siam niin an ngai. Metabolic kawng (23) a ni. Specialized metabolite hrang hranga interspecies danglamna hian herbivorous strategies laka constitutive leh inducible plant defense nena inzawm physiological balance a lantir thei bawk a, a chhan chu species pahnih hi negative takin an inzawm fo thin a ni (24). Engtik lai pawha invenna tha tak vawn reng chu a hlawkpui thei a, mahse, a hun taka invenna nena inzawm metabolic inthlak danglamna hian thlai te chu physiological investment dangte tana resource hlu tak tak an pek theih nan hlawkna chiang tak a pe a (19, 24), leh symbiosis mamawhna a pumpelh thei bawk. Collateral chhiatna (25) a ni. Chu bakah, heng insect herbivores-te avanga specialized metabolites reorganizations te hian population-ah destructive distribution a thlen thei bawk (26), chubakah jasmonic acid (JA) signal-a natural changes lian tham direct reading a lantir thei bawk a, chu chu population-ah chuan a awm reng thei bawk. JA signal sang leh hniam hi herbivores laka invenna leh chi hrang hrang nena inelna inkara trade-off a ni (27). Chu bakah, specialized metabolite biosynthetic pathways te hian evolution laiin hloh leh inthlak danglamna chak tak an tawk ang a, chu chuan chi hrang hrang inzawm tlat zingah patchy metabolic distribution a thlen ang (28). Heng polymorphisms te hi herbivore pattern danglam zel chhanna atan rang takin din theih a ni (29), chu chu herbivore community inthlak danglamna hi metabolic heterogeneity tichaktu pawimawh tak a ni tihna a ni.
Hetah hian a hnuaia harsatnate hi a bik takin kan chinfel sak a ni. (I) Engtin nge herbivorous insect hian thlai metabolome chu a siam thar leh? (Ii) Metabolic plasticity-a information component lian ber berte chu engte nge ni a, chu chu hun rei tak chhunga defense theory-in a sawi lawkte test nan quantified theih a ni? (Iii) Attacker-te tana danglam bik taka plant metabolome chu reprogram tur nge, chutiang a nih chuan, plant hormone hian metabolic response bik siamna kawngah eng chanvo nge a neih a, eng metabolite nge species specificity of defense-ah hian a thawhhlawk? (Iv) Defense theory tam takin an sawi lawk te hi biological tissue level zawng zawngah a zau thei a, chuvangin, internal comparison atanga interspecies comparison thlenga metabolic response lo awm chu engtiang chiahin nge a inmil tih kan zawt a. Chumi atan chuan tobacco nicotine hnah metabolome chu systematic takin kan zirchiang a, chu chu ecological model plant a ni a, specialized metabolism hausa tak a ni a, native herbivores pahnih, Lepidoptera Datura (Ms) ( Very aggressive, mainly eaten) On Solanaceae leh Spodoptera litoralis (Sl) te larvae lakah a tangkai hle “genus”, Solanaceae host plant leh genera leh chhungkaw dang host dangte nen Plant food. MS/MS metabolomics spectrum kan parse a, OD leh MT theory tehkhin nan information theory statistical descriptor kan la chhuak a. Key metabolites te identity hriat theihna turin specificity map siam rawh. He analysis hi N. nasi leh a kaihhnawih zuk leh hmuam chi hrang hrangte native population-ah pawh tihzauh a ni a, plant hormone signaling leh OD induction inkara covariance chu zirchian belh a ni.
Herbivorous tobacco leaf metabolome plasticity leh structure chungchanga map pum pui kan lak theih nan, a hmaa kan lo siam tawh analysis leh calculation workflow hmangin plant extract atanga high-resolution data independent MS/MS spectra kimchang takin kan khawlkhawm a, kan deconvolute a ( 9). He undifferentiated method (MS/MS tia koh) hian non-redundant compound spectra a siam thei a, chu chu hetia tarlan compound-level analysis zawng zawng atan hman theih a ni. Heng deconvoluted plant metabolites te hi chi hrang hrang an ni a, metabolite za tam tak atanga sang tam tak (hetah hian 500-1000-s/MS/MS vel) atanga siam an ni. Hetah hian information theory framework-ah metabolic plasticity kan ngaihtuah a, metabolic frequency distribution-a Shannon entropy hmangin metabolome-a diversity leh professionalism chu kan quantified a ni. A hmaa kan lo hman tawh formula (8) hmangin, metabolome diversity (Hj indicator), metabolic profile specialization (δj indicator) leh metabolite pakhat (Si indicator) metabolic specificity quantified nana hman theih tur indicator set kan chhut a. Chu bakah, Relative Distance Plasticity Index (RDPI) hmangin herbivores te metabolome inducibility kan teh a (Figure 1A) (30). He statistical framework chhungah hian MS/MS spectrum chu basic information unit angin kan en a, MS/MS relative abundance chu frequency distribution map-ah kan process a, chuta tang chuan Shannon entropy hmangin metabolome diversity kan estimate a ni. Metabolome specialization hi MS/MS spectrum pakhata specificity average atanga teh a ni. Chuvangin, herbivore induction hnua MS/MS class thenkhat tamna a punna chu spectral inducibility, RDPI leh specialization-ah a chang a, chu chu δj index a sanna a ni a, a chhan chu specialized metabolite tam zawk siam chhuah a nih vang leh Si index sang tak siam a nih vang a ni. Hj diversity index tihhniam hian MS/MS siam chhuah zat a tlahniam tih a tilang a, a nih loh leh profile frequency distribution chu kawng inang lo zawkah a inthlak a, chutih rualin a overall uncertainty a tihtlem bawk. Si index chhut dan hmang hian eng MS/MS nge herbivores thenkhatin an induce tih hriat theih a ni a, chu ai chuan eng MS/MS nge induction hi a chhang lo tih hriat theih a ni a, hei hi MT leh OD prediction thliar hrangna atana key indicator a ni.
(A) Herbivorous (H1 atanga Hx) MS/MS data-inducibility (RDPI), diversity (Hj index), specialization (δj index) leh metabolite specificity (Si index) atana hman statistical descriptor te. Degree of specialization (δj) tihpun chuan, a vaiin, herbivorous specific metabolites tam zawk siam chhuah a nih tur thu a tarlang a, diversity (Hj) tlahniam chuan metabolites siam chhuah tlahniam emaw, distribution map-a metabolites insem darh dan inang lo emaw a tilang thung. Si value hian metabolite chu condition pek tawh (hetah hian, herbivorous) atana specific a nih leh nih loh a teh a, a lehlamah chuan level inangah a awm reng em tih a teh a ni. (B) Information theory axis hmanga defense theory hrilhfiahna conceptual diagram. OD theory chuan herbivore attack hian defense metabolites a tipung dawn a, chu chuan δj a tipung dawn niin a sawi. Chutih rual chuan Hj chu a tlahniam a, a chhan chu profile chu metabolic information rinhlelhna tihtlem lam hawia insiam thar leh a nih vang a ni. MT theory chuan herbivores te beihna hian metabolome-ah non-directional change a thlen dawn niin a sawi a, chu chuan Hj chu metabolic information uncertainty tihpunna tilangtu atan a tipung ang a, Si chu random distribution a siam ang. Mixed model, MT tha ber kan rawt bawk a, chutah chuan defensive value sang zawk nei metabolite thenkhat chu a bik takin a pung ang (Si value sang), thenkhat erawh chuan random response (Si value hniam zawk) an lantir thung.
Information theory descriptor hmangin OD theory chu kan hrilhfiah a, uniduced constitutive state-a herbivore-induced special metabolite inthlak danglamna chuan (i) metabolic specificity (Si index) tihpunna a thlen ang a, chu chuan metabonomic specificity a thlen ang (δj index) The increase of) certain special metabolite groups with higher defense value, and (ii) the decrease of metabolome diversity (Hj) index metabolic frequency distribution chu leptin body distribution tam zawka a inthlak vang a ni. Metabolite pakhat level-ah chuan Si distribution ordered a awm beisei a ni a, chutah chuan metabolite chuan a defense value angin Si value a tisang ang (Figure 1B). Hemi kawngah hian MT theory chu excitation hian (i) metabolites-a non-directional changes a thlen ang a, chu chuan δj index a tihhniam ang a, (ii) metabolic uncertainty a san avangin Hj index a tisang ang tih sawi lawk turin MT theory kan sawifiah a ni. A nih loh leh randomness, chu chu Shannon entropy hmanga generalized diversity angin quantified theih a ni. Metabolic composition chungchangah chuan MT theory hian Si random distribution a sawi lawk ang. Metabolite thenkhat chu condition bik hnuaiah condition bik hnuaiah an awm a, condition dang chu condition bik hnuaiah an awm lo tih ngaihtuah chungin, an defense value chu environment-ah a innghat tih ngaihtuah chungin, mixed defense model kan rawt bawk a, chutah chuan δj leh Hj te chu Si Increase in all directions-ah pahnih-ah an insem a, metabolite group thenkhat chauh, defense value sang zawk nei chauhvin Si chu a bik takin an tipung ang a, thenkhat erawh chuan random an nei ang distribution (Figure 1B) a ni.
Information theory descriptor axis-a redefined defense theory prediction test theih nan Nepenthes pallens hnah-ah expert (Ms) emaw generalist (Sl) herbivore larvae kan enkawl a (Figure 2A). MS/MS analysis hmangin caterpillar chaw pek hnua leaf tissue lakkhawm methanol extract atanga non-redundant MS/MS spectra (data file S1) 599 kan la chhuak a. MS/MS configuration file-a information content reconfiguration dan hmuh theih nan RDPI, Hj, leh δj index hman hian pattern ngaihnawm tak tak a pholang a ni (Figure 2B). A tlangpuiin, information descriptor-in a sawi angin, caterpillar-te hian hnah an ei chhunzawm zel chuan, hun kal zelah metabolic reorganization zawng zawng degree a sang chho zel a ni: herbivore-in a ei hnu darkar 72-ah RDPI chu nasa takin a pung a ni. Undamaged control nena khaikhin chuan Hj chu nasa takin a tlahniam a, hei hi metabolic profile-a specialization degree a san vang a ni a, hei hi δj index hmanga quantified a ni. Hetianga thil thleng langsar tak hi OD theory-in a sawi lawk nen a inmil lo a, mahse MT theory-in a sawi lawk berte nen a inmil lo hle a, metabolite level-a random (non-directional) inthlak danglamna chu defensive camouflage atan hman a ni tih a ring (Figure 1B). Heng herbivore pahnih te hian oral secretion (OS) elicitor content leh feeding behavior hi inang lo mahse, an direct feeding hian darkar 24 leh darkar 72 chhunga an lakkhawm chhungin Hj leh δj lam hawia inthlak danglamna inang chiah a thlen a ni. A danglamna awm chhun chu RDPI darkar 72 chhung a ni. Ms feeding avanga lo awm nen khaikhin chuan Sl feeding avanga metabolism pum pui chu a sang zawk a ni.
(A) Experimental design: common pig (S1) emaw expert (Ms) herbivores te chu pitcher plant hnah desalinated hmanga chawm an ni a, simulated herbivory atan chuan Ms (W + OSMs) OS hmangin standardized leaf positions wound puncture an buaipui thung. S1 (W + OSSl) larva emaw tui (W + W) emaw a ni. Control (C) chu hnah chhia lo a ni. (B) Inducibility (RDPI control chart nena khaikhin chuan), diversity (Hj index) leh specialization (δj index) index te chu special metabolite map (599 MS/MS; data file S1) atana chhut a ni. Asterisk hian direct herbivore feeding leh control group te inkarah danglamna nasa tak a nei tih a tarlang (Student’s t-test with paired t-test, *P<0.05 leh ***P<0.001). ns, a pawimawh lo. (C) Simulated herbivory treatment hnua main (blue box, amino acid, organic acid leh sugar; data file S2) leh special metabolite spectrum (red box 443 MS/MS; data file S1) te time resolution index. Color band hian 95% confidence interval a kawk a ni. Asterisk hian treatment leh control inkara danglamna nasa tak a tarlang a [quadratic analysis of variance (ANOVA), chu chu post hoc multiple comparisons atan Tukey-a honestly significant difference (HSD) a ni a, *P<0.05, **P<0.01 And *** P <0.001]. (D) Scatter plot leh special metabolite profile (treatment hrang hrang hmanga sample repeated) te specialization.
Metabolome level-a herbivore-induced remodeling hi metabolite pakhat zel level inthlak danglamnaah a lang em tih zirchian nan, a hmaa Nepenthes pallens hnah-a herbivore resistance proven nei metabolites zirchian tawhte kan ngaihtuah hmasa a ni. Phenolic amides hi hydroxycinnamamide-polyamine conjugates a ni a, insect-te herbivory process-ah a pung khawm a, insect performance a tihziaawm tih hriat a ni (32). MS/MS inmil precursor te kan zawng a, an cumulative kinetic curve te kan plot a (Figure S1). Mak lo tak maiin, phenol derivatives, herbivores laka invennaah direct-a inrawlh lo, chlorogenic acid (CGA) leh rutin te hi herbivory hnuah down-regulated an ni. Chumi danglamna chu herbivores te hian phenol amides te chu chak takin an siam thei a ni. Herbivore pahnih ei chhunzawm zel hian phenolamides excitation spectrum inang deuh vek a siam a, he pattern hi phenolamides de novo synthesis atan a langsar zual hle. Hetiang bawk hian 17-hydroxygeranyl nonanediol diterpene glycosides (17-HGL-DTGs) pathway kan chhui dawn a, hei hian anti-herbivore function tha tak nei acyclic diterpenes tam tak a siam chhuak a ni (33), chung zinga Ms Feeding with Sl chuan expression profile inang chiah a tichhuak a ni (Figure S1)).
Direct herbivore feeding experiment-a chhiatna awm thei chu herbivore-te hnah ei zat leh ei hun inang lo vang a ni a, hei hian hliam leh herbivore-te avanga herbivore-specific effect awm chu tihbo a harsa hle. Induced leaf metabolic response-a herbivore species specificity solve tha zawk nan, Ms leh Sl larvae te chaw pek dan chu kan simulate a, OS lakkhawm thar (OSM leh OSS1) chu standard puncture W of consistent leaf positions-ah hman nghal a ni. Hetiang tih dan hi W + OS treatment an ti a, tissue hloh rate emaw quantity emaw inthlauhna avanga confounding effects awm lovin herbivore-in a chhanna a siam chhuah tan hun dik tak siamin induction chu a standardise a ni (Figure 2A) (34). MS/MS analysis leh calculation pipeline hmangin MS/MS spectra 443 (data file S1) kan la chhuak a, chu chu direct feed experiment atanga a hmaa spectra kan lo dahkhawm tawhte nen a inzawm tlat a ni. He MS/MS data set-a information theory analysis-ah hian herbivores simulating hmanga leaf-specialized metabolomes reprogramming-ah hian OS-specific inducements a awm tih hmuhchhuah a ni (Figure 2C). A bik takin OSS1 enkawlna nena khaikhin chuan OSM hian darkar 4 chhungin metabolome specialization a tichak a ni. Hriat tur chu direct herbivore feeding experimental data set nena khaikhin chuan Hj leh δj te chu coordinate atan hmanga two-dimensional space-a hmuh theih metabolic kinetics leh hun kal zelah simulated herbivore treatment chhanna atana metabolome specialization directionality Increase consistent (Figure 2D). Chutih rual chuan, he metabolome expertise tihpunna tur targeted hi simulated herbivores te chhanna atana central carbon metabolism reconfiguration vang a ni em tih zirchian nan amino acids, organic acids leh sugars awm zat (data file S2) kan quantified a ( Figure S2). He pattern hi sawifiah tha zawk turin a hmaa kan sawi tawh phenolamide leh 17-HGL-DTG pathways te metabolic accumulation kinetics kan enfiah lehzual a. Herbivores te OS-specific induction chu phenolamide metabolism chhungah differential rearrangement pattern ah a inthlak a (Figure S3). Phenolic amides coumarin leh caffeoyl moieties awmna chu OSS1 hian a induce duh zawk a, OSMs erawh chuan ferulyl conjugates induction bik a tichhuak thung. 17-HGL-DTG pathway atan chuan downstream malonylation leh dimalonylation products hmanga differential OS induction hmuhchhuah a ni (Figure S3).
A dawtah chuan OS-induced transcriptome plasticity chu time-course microarray data set hmangin kan zirchiang a, hei hian herbivores-a rosette plant hnah enkawlna atana OSM hman dan a simulate a ni. Sampling kinetics hi a bul berah chuan he metabolomics study-a hman kinetics nen hian a inzawm tlat a ni (35). Metabolome reconfiguration nena khaikhin chuan metabolic plasticity chu hun kal zelah a bik takin a pung a, Ms-in a tihbuai hnah-ah transient transcription burst kan hmu a, chutah chuan transcriptome inducibility (RDPI) leh specialization (δj) chu 1-ah a awm a, darkar tam tak a pung a, he hun lai hian diversity (Hj) a sang hle a, BMP1 expression chu nasa takin a tlahniam a, chu chu relaxation a ni transcriptome specialization tih a ni (Figure S4). Metabolic gene chhungkaw (P450, glycosyltransferase, leh BAHD acyltransferase ang chi) te hian primary metabolism atanga lo chhuak structural unit atanga special metabolites an inzawmkhawmnaah an tel a, a hmaa kan sawi tawh angin high-specialization model hmasa ber chu an zawm a ni. Case study angin phenylalanine pathway chu an zirchiang a. Analysis hian phenolamide metabolism-a core genes te hi herbivores-ah hian unattracted plant nena khaikhin chuan OS-induced nasa tak an ni tih a nemnghet a, an expression pattern-ah pawh an inmil hle tih a nemnghet bawk. He pathway upstream-a transcription factor MYB8 leh structural genes PAL1, PAL2, C4H leh 4CL te hian transcription initiation hmasa tih an hmuchhuak a ni. Phenolamide final assembly-a chanvo nei acyltransferases, AT1, DH29, leh CV86 te hian upregulation pattern rei tak an nei a (Figure S4). A chunga kan hmuhte hian transcriptome specialization tan hma leh a hnu lama metabolomics specialization tihchakna hi coupled mode a ni tih a tilang a, hei hi defense response chak tak intan tirtu synchronous regulatory system vang pawh a ni thei.
Plant hormone signaling-a reconfiguration hian regulatory layer angin hna a thawk a, chu chuan herbivorous information chu a inzawm khawm a, thlai physiology chu a reprogram a ni. Herbivore simulation kan neih zawh hian key plant hormone category-te cumulative dynamics kan teh a, an inkara temporal co-expression kan hmuh theih nan [Pearson correlation coefficient (PCC)> 0.4] (Figure 3A). Beisei ang ngeiin biosynthesis nena inzawm plant hormone te hi plant hormone co-expression network chhungah an inzawm tlat a ni. Chu bakah, metabolic specificity (Si index) chu he network-ah hian map a ni a, hei hian enkawlna hrang hrang avanga plant hormone lo awmte a tarlang bawk. Herbivorous specific response area lian pahnih tarlan a ni a: pakhat chu JA cluster-ah a awm a, chutah chuan JA (a biologically active form JA-Ile) leh JA derivative dangte chuan Si score sang ber an nei a pakhat zawk chu ethylene (ET) a ni. Gibberellin hian herbivore specificity a pung tlem hle a, plant hormone dang, cytokinin, auxin, leh abscisic acid te chuan herbivore tan induction specificity a nei tlem hle thung. W + W chauh hman nen khaikhin chuan OS application (W + OS) hmanga JA derivatives peak value amplification hi a bul berah chuan JAs specific indicator chak takah a chantir thei a ni. Beisei loh takin elicitor content hrang hrang nei OSM leh OSS1 te hian JA leh JA-Ile te hi a accumulation inang vek tih hriat a ni. OSS1 nen a danglamna chu OSM hi OSMs hian a bik takin leh chak takin a tichhuak a, OSS1 erawh chuan basal wounds response a amplify lo thung (Figure 3B).
(A) Herbivore-induced plant hormone accumulation kinetics simulation hmanga PCC chhut dan hmanga co-expression network analysis. Node hian plant hormone pakhat a entir a, node lian tham tak hian treatment inkara plant hormone bik Si index a entir bawk. (B) JA, JA-Ile leh ET te chu hnah-ah a pungkhawm a, chu chu enkawlna hrang hrang avanga rawng hrang hranga tarlan a ni: apricot, W + OSM; blue, W + OSSl tih a ni a; dum, W + W tih a ni a; gray, C (thunun) . Asterisk hian enkawlna leh control inkarah danglamna nasa tak a nei tih a tarlang (two-way ANOVA hnuah Tukey HSD post hoc multiple comparison, *** P <0.001). JA biosynthesis leh impaired perception spectrum (irAOC leh irCOI1)-a (C)697 MS/MS (data file S1) leh ETR1-a (D)585 MS/MS (data file S1) te information theory analysis hmanga ETR1-a ET signal tihchhiat Simulated herbivore treatment pahnih hmangin plant line leh empty vehicle (EV) control plant te a tichhuak a ni. Asterisk hian W+OS treatment leh undamaged control inkarah danglamna nasa tak a awm tih a tarlang (two-way ANOVA hnuah Tukey HSD post hoc multiple comparison, *P<0.05, **P<0.01 leh ***P<0.001). (E) Specialization dodalna hmun hrang hranga graph hrang hrang. A rawng te hian genetically modified strain hrang hrang a entir a; chhinchhiahnate hian enkawlna kawng hrang hrang a entir a: triangle, W + OSS1; rectangle, W + OSM tih a ni a; circle C. a ni
A dawtah chuan JA leh ET biosynthesis (irAOC leh irACO) leh perception (irCOI1 leh sETR1) key step-a attenuated Nepenthes (irCOI1 leh sETR1) genetically modified strain hmangin heng plant hormone pahnih te hi herbivores-a metabolism kan zirchiang The relative contribution of reprogramming. Experiment hmasa nena inmil takin, empty carrier (EV) plant-a herbivore-OS induction (Figure 3, C to D) leh OSM vanga Hj index tlahniam zawng zawng chu kan nemnghet a, δj index erawh a sang thung. OSS1-in a trigger chhanna aiin a chhanna a langsar zawk. Hj leh δj te coordinate atana hman line hnih graph chuan specific deregulation chu a tarlang a (Figure 3E). Trend langsar ber chu JA signal nei lo strain-ah chuan herbivores-te avanga metabolome diversity leh specialization inthlak danglamna chu a bo vek tawh mai mai a ni (Figure 3C). Chumi danglamna chu sETR1 plant-a silent ET perception hian herbivorous metabolism inthlak danglamnaah nghawng pum pui chu JA signaling aiin a hniam zawk hle nachungin, OSM leh OSS1 excitations inkara Hj leh δj indices danglamna chu a tiziaawm a ni (Figure 3D leh Figure S5). . Hei hian JA signal transduction-a core function bakah hian ET signal transduction hian herbivores-te species-specific metabolic response fine-tuning hna a thawk bawk tih a tilang a ni. He fine-tuning function nena inmil hian sETR1 plant-a metabolome inducibility pumpuiah danglamna a awm lo. A lehlamah chuan sETR1 thlai nena khaikhin chuan irACO thlai hian herbivores-te avanga metabolic changes overall amplitude inang chiah chiah a thlen a, mahse OSM leh OSS1 challenge inkarah Hj leh δj score a inang lo hle thung (Figure S5).
Herbivores te species-specific response-a thawhhlawk tak tak nei specialized metabolites te hriatchhuah nan leh ET signal hmanga an siam chhuah fine-tune theih nan a hmaa kan lo siam tawh structural MS/MS method kan hmang a. Hetiang method hian bi-clustering method hmangin MS/MS fragment [normalized dot product (NDP)] atanga metabolic family re-infer nan leh neutral loss (NL) atanga similarity score a innghat a ni. ET transgenic line-te thlirletna hmanga MS/MS data set siam chuan MS/MS 585 (data file S1) a siam a, chu chu MS/MS module lian pasarih (M)-a cluster-a dahin chinfel a ni (Figure 4A) Heng module thenkhat hi a hmaa characterized special metabolites hmanga densely packed an ni a: entirnan, M1, M2, M3, M4 leh M7 te hian phenol derivatives hrang hrang (M1), flavonoid glycosides (M2), acyl sugars (M3 And M4), leh 17-HGL-DTG (M7) te an pai tam hle. Chu bakah, module tina metabolite pakhat metabolic specific information (Si index) chu chhut a ni a, a Si distribution chu intuitive takin kan hmu thei bawk. A tawi zawngin, MS/MS spectra herbivory leh genotype specificity sang tak lantir te hi Si value sang tak nei an ni a, kurtosis statistics chuan tail corner dinglamah fur insem darh dan a tarlang bawk. Chutiang lean colloid distribution pakhat chu M1-ah hmuh a ni a, chutah chuan phenol amide hian Si fraction a nei sang ber a ni (Figure 4B). A hmaa kan sawi tawh ang khan M7-a herbivorous inducible 17-HGL-DTG hian Si score a nei tlem hle a, hei hian OS chi hnih inkarah hian differential regulation a nei tlem hle tih a tilang a ni. Chumi danglamna chu, constitutively produced specialized metabolites tam zawk, rutin, CGA, leh acyl sugars te chu Si score hniam ber zinga mi an ni. Special metabolites inkara structural complexity leh Si distribution te hi a tha zawka zirchian theih nan module tin tan molecular network siam a ni (Figure 4B). OD theory-a sawi lawk pawimawh tak (Figure 1B-a khaikhawm) chu herbivory hnua special metabolite-te reorganization hian defense value sang tak nei metabolite-te chu kawng khata inthlak danglamna a thlen tur a ni a, a bik takin an specificity tihpunna hmangin (random distribution nen a danglamna) Mode) MT theory-in a sawi lawk Defensive metabolite. M1-a phenol derivatives khawlkhawm tam zawk hi insect performance tlahniam nen functionally inzawm a ni (32). Darkar 24 chhunga EV control plant-a induced leaves leh constituent leaves inkara M1 metabolites-a Si value te kan khaikhin chuan herbivory insect hnua metabolite tam tak metabolic specificity chu nasa takin a pung tih kan hmu a (Figure 4C). Si value tihpunna bik hi defensive phenolamides-ah chauh hmuh a ni a, mahse he module-a awm dun phenol dang leh metabolite hriat lohte-ah chuan Si value tihpun erawh hmuhchhuah a ni lo. Hei hi specialized model a ni a, OD theory nen a inzawm tlat a ni. Herbivores-te’n metabolic change an neih dan tur hrilhlawkna ber chu a inmil vek a ni. Phenolamide spectrum-a he particularity hi OS-specific ET-in a thlen em tih test nan metabolite Si index kan plot a, EV leh sETR1 genotype-a OSM leh OSS1 inkara differential expression value kan siam a (Figure 4D ). sETR1-ah chuan OSM leh OSS1 inkara phenamide-induced difference chu nasa takin a tlahniam a ni. Bi-clustering method hi JA-regulated metabolic specialization nena inzawm MS/MS module lian ber berte infer turin JA tling lo strain-a MS/MS data lakkhawmah pawh hman a ni bawk (Figure S6).
(A) Shared fragment (NDP similarity) leh shared neutral loss (NL similarity) hmanga 585 MS/MS clustering result chuan module (M) chu compound chhungkaw hriat tawh nen a inmil a, a nih loh leh Metabolite composition hriat loh emaw, metabolized tha lo emaw a siam a ni. Module tin bulah hian metabolite (MS/MS) specific (Si) distribution tarlan a ni. (B) Modular molecular network: Node te hian MS/MS leh edge te, NDP (red) leh NL (blue) MS/MS score (cut-off,> 0.6) te a entir a ni. Graded metabolite specificity index (Si) chu module (khawi lamah nge) atanga rawng a siam a, molecular network (dinglamah)-ah map a ni. (C) EV plant chu darkar 24 chhunga constitutive (control) leh induced state (simulated herbivore)-a awm Module M1: molecular network diagram (Si value chu node size a ni a, defensive phenolamide chu blue-in a highlight a ni). (D) EV leh ET perception chhiatna nei spectrum line sETR1-a M1 molecular network diagram: green circle node-in a entîr phenolic compound, leh W + OSM leh W + OSS1 treatment-te Node size anga danglamna nasa tak (P value) te. CP, N-caffeoyl-tyrosine hmanga siam a ni a; CS, N-caffeoyl-spermidine hmanga siam a ni a; FP, N-ferulic acid ester-uric acid hmanga siam a ni a; FS, N-ferulyl-spermidine hmanga siam a ni a; A rilru a hah lutuk chuan a rilru a buai em em a, a rilru a hah lutuk chuan a rilru a buai em em bawk a. DCS, N’, N”-dicaffeoyl-spermidine; CFS, N’, N”-caffeoyl, feruloyl-spermidine, wolfberry Son-a Lycium barbarum, Nick.O-AS, O-acyl sugar te chu a awm a, a chhuahna tur hmun leh a hmanna tur hmun chu a inang lo hle.
Kan analysis chu attenuated Nepenthes genotype pakhat a\angin natural population-ah kan tizau lehzual a, chutah chuan natural population-ah herbivorous JA level leh specific metabolite level-a intraspecific inthlak danglamna nasa tak chu a hmain kan lo sawi tawh a ni (26). He data set hmang hian germplasm 43 cover rawh. Heng germplasm te hi N. pallens atanga lo chhuak thing chi hrang hrang 123 atanga siam a ni. Heng thlai te hi Utah, Nevada, Arizona, leh California-a native habitat hrang hranga thlai chi lakkhawm atanga lak a ni a (Figure S7), metabolome diversity (hetah hian population level tia koh) β diversity) leh OSM avanga specialization kan chhut a ni. Zirna hmasa nena inmil takin Hj leh δj axes-ah hian metabolic change hrang hrang kan hmu a, hei hian germplasm-te hian herbivores laka an metabolic response plasticity-ah danglamna nasa tak an nei tih a tilang (Figure S7). He pawl hian herbivores-te avanga JAs inthlak danglamna dynamic range chungchanga a hmaa thil hmuhchhuah tawhte a hriatchhuah tir a, population pakhatah pawh value sang tak a vawng reng a ni (26, 36). Hj leh δj inkara overall level correlation test nan JA leh JA-Ile hmangin JA leh metabolome β diversity leh specialization index inkarah hian positive correlation nasa tak a awm tih kan hmu chhuak a (Figure S7). Hei hian population level-a JAs induction-a herbivore-induced heterogeneity hmuhchhuah chu insect herbivores atanga thlan chhuah avanga key metabolic polymorphisms vang a ni thei tih a tilang.
Tun hmaa zirchianna hrang hrangah chuan zuk leh hmuam chi hrang hrangte hi chi leh induced leh constitutive metabolic defense-a innghahna (relative dependence)-ah a inang lo hle tih hmuhchhuah a ni. Heng anti-herbivore signal transduction leh defense capabilities inthlak danglamna hi insect population pressure, plant life cycle, leh species pakhat a lo thang lian na niche-a defense production costs te hian a thunun niin an ngai. North America leh South America-a piang Nicotiana chi paruk, herbivores-te’n leaf metabolome remodeling an tihchhuah dan inmil dan kan zirchiang a. Heng chi hrang hrangte hi Nepenthes North America nen hian an inzawm tlat a, chu chu Nicolas Bociflo a ni. La, N. nicotinis, Nicotiana n. attenuated grass, Nicotiana tabacum, linear zuk leh hmuam, zuk leh hmuam (Nicotiana spegazzinii) leh zuk leh hmuam hnah (Nicotiana obtusifolia) te (Figure 5A) (37). Heng zinga paruk, a chi chiang tak N. please te pawh hi petunia clade-a kum khata thlai chi an ni a, obtusifolia N. hi sister clade Trigonophyllae-a perennial an ni bawk (38). Chumi hnuah heng chi pasarih te hi W + W, W + OSM leh W + OSS1 induction te hmangin insect ei dan species-level metabolic rearrangement zirchian a ni.
(A) Bootstrap phylogenetic tree chu maximum likelihood [nuclear glutamine synthesis atan (38)] leh Nicotiana chi hrang hrang inzawm tlat pasarih (a rawng hrang hrang) awmna hmuna an insem dan (37) atanga siam a ni. (B) Nicotiana chi pasarih (939 MS/MS; data file S1) metabolic profile atana specialized diversity scatter plot. Species level-ah chuan metabolome diversity hi specialization degree nen a inzawm tha lo hle. Metabolic diversity leh specialization leh JA accumulation inkara species-level correlation thlirletna chu Figure 2. S9-ah hian tarlan a ni. A rawng, chi hrang hrang; triangle, W + OSS1 tih a ni a; rectangle, W + OSM tih a ni a; (C) Nicotiana JA leh JA-Ile dynamics te hi OS excitation amplitude angin rank a ni (two-way ANOVA leh Tukey HSD post-multiple comparison, * P <0.05, ** P <0.01 leh * ** W + OS leh W + W tehkhin nan, P <0.001). Box plot-ah hian herbivorous leh methyl JA (MeJA) simulate hnua chi hrang hrang (D) diversity leh (E) specialization te tarlan a ni. Asterisk hian W + OS leh W + W emaw lanolin plus W (Lan + W) emaw Lan plus MeJA (Lan + MeJa) leh Lan control (two-way analysis of variance, chumi hnuah Tukey-a HSD post hoc multiple comparison , *P<0.05, **P<0.01 leh ***P<0.001) inkara danglamna nasa tak a tarlang a ni.
Dual cluster method hmangin MS/MS 939 (data file S1) awmna module 9 kan hmuchhuak a. Treatment hrang hrang hmanga MS/MS reconfigured composition hi species hrang hrang module hrang hrangah a inang lo hle a ni (Figure S8). Hj (hetah hian species-level γ-diversity tia sawi) leh δj te hmuh theihin metabolic space-ah hian species hrang hrangte chu group hrang hrangah an inzawm khawm tih a lang a, chutah chuan species-level division hi excitation aiin a langsar zawk tlangpui. N. linear leh N. obliquus tih lohah chuan induction effect dynamic range zau tak an nei a (Figure 5B). Chumi danglamna chu N. purpurea leh N. obtusifolia ang chi te hian enkawlnaah metabolic response a langsar lo hle a, mahse metabolome erawh a chi hrang hrang a ni. Induced metabolic response species-specific distribution hian specialization leh gamma diversity inkarah negative correlation nasa tak a siam a (PCC = -0.46, P = 4.9×10-8). OS-in a thlen JAs level inthlak danglamna hi metabolome specialization nen a inzawm tha hle a, species tinte’n metabolic gamma diversity an lantir nen pawh a inzawm tha lo hle bawk (Figure 5B leh Figure S9). Hriat tur chu Figure 5C-a tawngkam hmanga “signal response” tia sawi chi, Nepenthes nematodes, Nepenthes nepenthes, Nepenthes acute, leh Nepenthes attenuated te hian minute 30 chhungin chhinchhiahna langsar tak tak an nei a ni. Tun hnaia OS-specific JA leh JA-Ile hri leng te, bacteria dang “signal unresponsive” tia koh, Nepenthes mills, Nepenthes powdery leh N. obtusifolia te hian OS specificity awm lovin JA-Ile Edge induction chauh an lantir thung (Figure 5C). Metabolic level-ah chuan a chunga kan sawi tawh ang khan attenuated Nepenthes tan chuan signal-responsive substances te hian OS specificity an lantir a, δj chu nasa takin an tipung a, Hj erawh a ti tlem thung. He OS-specific priming effect hi signal non-reactive species anga classified species-ah hmuh a ni lo (Figure 5, D leh E). OS-specific metabolites te hi signal-responsive species te inkarah an insem tam zawk a, heng signal cluster te hi signal response chak lo zawk nei species te nen an cluster a, signal response chak lo species te erawh chuan interdependence an nei tlem zawk thung (Figure S8 ). He result hian OS-specific induction of JAs leh OS-specific reconfiguration of the downstream metabolome chu species level-ah a inzawm tih a tilang a ni.
A dawtah chuan methyl JA (MeJA) awmna lanolin paste hmangin thlai enkawl kan hmang a, heng coupling modes te hi exogenous JA hmanga JA hman a nih avangin a tihkhawtlai em tih kan zirchiang a, chu chu thlai cytoplasm-ah a awm dawn a ni. Rapid deesterification chu JA a ni. JA supply chhunzawm zel avanga signal-responsive species atanga signal-non-responsive species-a inthlak zauh zauh dan ang chiah kha kan hmu a (Figure 5, D leh E). A tawi zawngin, MeJA hmanga enkawlna hian linear nematode, N. obliquus, N. aquaticus, N. pallens, leh N. mikimotoi te metabolome chu nasa takin a reprogramme a, chu chuan δj nasa takin a tipung a, Hj pawh a tlahniam bawk. N. purpurea hian δj chauh a pung a, mahse Hj erawh a pung lo. Tun hmaa JAs level hniam tak tak a khawlkhawm thin N. obtusifolia pawhin MeJA enkawlnaah metabolome reconfiguration lamah a chhang tha lo hle bawk. Heng results te hian signal-unresponsive species ah hian JA production emaw signal transduction emaw chu physiologically restricted a nih thu a tarlang. He hypothesis test nan hian W + W, W + OSMs leh W + OSS1 Transcriptome (39) hmanga induced chi pali (N. pallens, N. mills, N. pink leh N. microphylla) te kan zirchiang a. Metabolome remodeling pattern nena inmil takin, transcriptome space-ah hian species te hi an inthen tha hle a, chung zingah chuan N. attenuated chuan OS-induced RDPI a nei sang ber a, N. gracilis erawh chuan a hniam ber thung (Figure 6A). Mahse, N. oblonga induced transcriptome diversity chu chi pali zingah a hniam ber tih hmuhchhuah a ni a, hei hi a hmaa N. oblonga metabolic diversity sang ber chi pasarih atanga hmuh tawh nen a danglam hle. Tun hmaa zirchianna hrang hrangah chuan JA signal telin defense signal hmasa nena inzawm gene set khat chuan Nicotiana chi hrang hranga herbivore nena inzawm elicitor-te’n defense response hmasa ber a thlen dan specificity a hrilhfiah tih hmuhchhuah a ni (39). Heng chi palite inkara JA signaling pathways te khaikhin chuan pattern ngaihnawm tak a awm tih hmuhchhuah a ni (Figure 6B). He pathway-a gene tam zawk, AOC, OPR3, ACX leh COI1 te hian heng chi paliah hian induction level sang tak an nei a ni. Mahse, key gene pakhat JAR4 chuan JA chu a biologically active form JA-Ile accumulated transcripts-ah a chantir a, a transcription level pawh a hniam hle a, a bik takin N. mills, Nepenthes pieris leh N. microphylla-ah te a ni. Tin, N. bifidum-ah hian gene dang AOS transcript chauh hmuhchhuah a ni lo. Heng gene expression inthlak danglamna te hi signal anergic species-a JA siam chhuah tlem leh N. gracilis induction avanga phenotypes nasa tak awm chhan a ni thei.
(A) Herbivory induction hnu minute 30-a sample lak, zuk leh hmuam chi hrang hrang, inzawm tlat pali te transcriptional response hmasa ber reprogramming chungchanga information theory analysis. RDPI hi herbivore OS induced hnah leh wound control nen khaikhin a ni. A rawng te hian chi hrang hrang a tarlang a, chhinchhiahna te hian enkawl dan hrang hrang a tarlang bawk. (B) Chi pali zinga JA signaling pathway-a gene expression thlirletna. Simplified JA path chu box plot bulah hian a lang. Color hrang hrang hian processing method hrang hrang a tarlang a. Asterisk hian W + OS treatment leh W + W control inkarah danglamna nasa tak a awm tih a tarlang (Student’s t-test for pairwise differences, *P<0.05, **P<0.01 leh ***P<0.001). OPDA, 12-oxophytodienoic acid hmanga siam a ni a; OPC-8: 0,3-oxo-2(2′(Z)-pentenyl)-cyclopentane-1-octanoic acid hmanga siam a ni.
A tawp berah chuan thlai chi hrang hrangte metabolome-a insect species-specific remodeling-in herbivores a do theih dan kan zirchiang a. Tun hmaa zirchiannaah khan Nicotiana chi hi an ngai pawimawh hle. Ms leh larvae laka an do theihna hi a inang lo hle (40). Hetah hian he model leh an metabolic plasticity inzawmna kan zirchiang a. A chunga zuk leh hmuam chi pali kan tarlan te hmang hian, leh herbivores-te avanga metabolome chi hrang hrang leh specialization leh thlai te Ms leh Sl laka invenna inzawmna te kan test a, generalist Sl All laka invenna, chi hrang hrang leh specialization te hi a inzawm tha hle tih kan hmu a, chutih laiin expert ladies laka invenna leh specialization inzawmna chu a chak lo hle a, diversity nena inzawmna chu a langsar lo hle bawk (Figure S10) a ni. S1 resistance chungchangah chuan attenuated N. chinensis leh N. gracilis te pahnih, a hmaa JA signal transduction level leh metabolome plasticity pahnih lantir tawhte hian herbivore induction-ah hian an chhanna a inang lo hle a, chutiang bawkin resistance sang tak an nei bawk. Sex hman dan.
Kum sawmruk kalta chhung khan plant defense theory hian theoretical framework a pe a, chuta tang chuan zirchiangtute chuan plant specialized metabolites te evolution leh function tam tak an lo sawi lawk tawh a ni. Heng theory tam zawk hian strong inferences kalphung pangngai an zawm lo (41). Analysis level inangah key prediction (3) an rawt a ni. Key predictions testing hian theory bikte chu zirchian theih a nih hunah chuan hei hian The field is moving forward a siam ang. thlawp ni mah la, midangte chu hnawl rawh (42). Chu ai chuan theory thar hian analysis level hrang hrangah prediction a siam a, descriptive considerations layer thar a belh bawk (42). Mahse, functional level-a theory pahnih rawt, MT leh OD te hi awlsam takin herbivores-te’n specialized metabolic changes an neihte hrilhlawkna pawimawh tak angin sawifiah theih a ni: OD theory chuan specialized metabolic “space”-a inthlak danglamna hi directional sang tak a ni tih a ring MT theory chuan heng inthlak danglamna te hi non-directional leh randomly located in metabolic space ah a awm dawn niin a ngai a, defense value sang tak metabolites a nei duh hle bawk. Tun hmaa OD leh MT prediction enfiahna te chu a priori “defense” compound set te tak te hmangin an lo test tawh a ni. Heng metabolite-centric test te hian herbivory chhunga metabolome reconfiguration zau zawng leh trajectory thlirlet theihna a tikhawlo a, statistical framework mumal tak chhunga test neihna tur hian a pum puia ngaihtuah theih tur key prediction mamawh a phal lo Quantify changes in plant metabolome. Hetah hian computational MS hmanga metabolomics lama technology thar kan hmang a, global metabolomics level-a rawtna pahnih inthliarna test nan information theory descriptor-te general currency-a deconvolution MS analysis kan ti a ni. He theory-a prediction pawimawh ber chu. Information theory hi kawng tam takah hman a ni tawh a, a bik takin biodiversity leh nutrient flow research-ah te hman a ni tawh (43). Mahse, kan hriat dan chuan hei hi thlai chi hrang hrangte metabolic information space sawifiahna atana hman hmasak ber a ni a, environmental cues chhanna atana hun eng emaw chen chhunga metabolic inthlak danglamna nena inzawm ecological problems chinfelna atana hman hmasak ber a ni. A bik takin, he method hian a theihna chu thlai chi hrang hrang chhung leh chi hrang hrang inkara pattern tehkhin theihnaah a awm a, chu chuan herbivores te chu chi hrang hrang atanga inter-species macroevolutionary pattern ah evolution level hrang hranga an lo inthlak danglam dan a zirchiang thei a ni. Thilsiam (metabolism) a awm.
Principal component analysis (PCA) hian multivariate data set chu dimensionality reduction space-ah a chantir a, chutiang chuan data-a main trend chu sawifiah theih a ni a, chuvangin data set parse nan exploratory technique atan hman a ni tlangpui a, chu chu deconvolution metabolome ang chi hi a ni. Mahse, dimensionality tihtlem hian data set-a information content thenkhat a hloh dawn a, PCA hian ecological theory nena inzawm tlat characteristics chungchangah quantitative information a pe thei lo va, chu chu: how herbivores reconfigure diversity in specialized fields (for example, richness, distribution) And abundance) metabolites? Eng metabolite nge herbivore pakhat induced state hriltu? Specificity, diversity leh inducibility thlirna atang chuan leaf-specific metabolite profile-a information awm chu a chhe vek a, herbivores ei hian specific metabolism a tichak thei tih hmuhchhuah a ni. Beisei loh takin, information theory indicators implemented-a tarlan angin, a chhuak metabolic situation chu herbivore pahnih (zan lama chaw ei generalist Sl) leh Solanaceae expert Ms. te beihna hnuah overlap lian tak a nei tih kan hmu a. Although their feeding behavior and concentration are significantly different. OS-ah hian fatty acid-amino acid conjugate (FAC) hmanga siam a ni (31). Standardized puncture hliam enkawlna atana herbivore OS hman a nih avangin simulated herbivore treatment pawhin hetiang bawk hian a lantir bawk. He standardized procedure, herbivore attack-a thlai-te chhanna simulating hian herbivore-te ei leh in lama inthlak danglamna avanga thil buaithlak tak takte chu a ti bo a, chu chuan hun hrang hrangah chhiatna degree hrang hrang a thlen thin (34). OSM thlentu ber nia hriat FAC hian OSS1-a JAS leh plant hormone dangte chhanna a tihtlem a, OSS1 erawh chuan a let za tam takin a tihtlem thung (31). Mahse, OSS1 hian OSM nena khaikhin chuan JA accumulation level inang chiah a thlen a ni. Tun hmain attenuated Nepenthes-a JA response hi OSM-ah a sensitive hle tih hmuhchhuah a ni tawh a, chutah chuan FAC hian tui nen 1:1000-a diluted pawh nise a activity chu a vawng reng thei a ni (44). Chuvangin OSM nena khaikhin chuan OSS1-a FAC hi a hniam hle nachungin JA hri leng tling tur induce turin a tawk a ni. Tun hmaa zirchianna hrang hrangah chuan porin ang chi protein (45) leh oligosaccharides (46) te hi OSS1-a plant defense response tichhuak turin molecular clues atan hman theih a ni tih hmuhchhuah a ni. Mahse, OSS1-a heng elicitor-te hi tuna zirchiannaa JA pungkhawmnaah hian mawhphurtu an ni em tih erawh hriat chian a la ni lo.
Herbivore hrang hrang emaw exogenous JA emaw SA (salicylic acid) hman avanga differential metabolic fingerprints awm dan sawifiahna zirchianna tlemte chauh awm mahse (47), plant grass network-a herbivore species-specific perturbation leh a specific-a nghawng a neih dan chu tumahin an tibuai lo The overall impact of metabolism is systematically studied. mimal chanchin. He analysis hian JAs tih loh plant hormone dang nena internal hormone network inzawmna hian herbivores-te avanga metabolic reorganization specificity a siam tih a nemnghet lehzual a ni. A bik takin OSM avanga ET lo awm chu OSS1 avanga ET lo awm aiin a tam zawk tih kan hmuchhuak a ni. He mode hi OSM-a FAC content tam zawk nen a inmil a, hei hi ET burst trigger na tur condition tul leh tling tak a ni (48). Thlai leh herbivores inkara inzawmna chungchangah chuan ET hian plant-specific metabolite dynamics-a signaling function hi a la awm fo a, compound group pakhat chauh a target a ni. Chu bakah, zirchianna tam zawkah chuan ET emaw a hmahruaitu emaw, inhibitor hrang hrang emaw exogenous application hmangin ET regulation an zirchiang a, chung zingah chuan heng exogenous chemical application hian non-specific side effect tam tak a siam dawn a ni. Kan hriat dan chuan he zirchianna hian plant metabolome dynamics coordinate turin impired transgenic plant siam leh hriatchhuahna atana ET hmanna kawnga ET chanvo chu large-scale systematic examination hmasa ber a ni. Herbivore-specific ET induction hian a tawpah chuan metabolome response chu a modulate thei a ni. A pawimawh ber chu ET biosynthesis (ACO) leh perception (ETR1) genes transgenic manipulation hmanga herbivore-specific de novo accumulation of phenolamides hmuhchhuah a ni. Tun hmain ET hian putrescine N-methyltransferase a regulate-in JA-induced nicotine accumulation chu a fine-tune thei tih hmuhchhuah a ni tawh bawk (49). Mahse, mechanical lam atanga thlir chuan ET hian phenamide induction hi engtin nge a fine-tune tih erawh a chiang lo. ET signal transduction function bakah hian metabolic flux chu S-adenosyl-1-methionine-ah shunt theih a ni bawk a, hei hian polyaminophenol amides-a investment a tidanglam thei a ni. S-adenosyl-1-methionine hi ET a ni a, polyamine biosynthetic pathway-a Common intermediate a ni. ET signal hian phenolamide level a tihregulate dan hi zirchian belh a ngai a ni.
Hun rei tak chhung chu, structure hriat loh metabolite bik tam tak a awm avangin, metabolic category bik ngaihvenna nasa tak chuan biological interaction hnua metabolic diversity-a temporal changes chu khauh takin a zirchiang thei lo. Tunah hian information theory analysis atanga chhut chuan unbiased metabolites hmanga MS/MS spectrum acquisition atanga rah chhuah ber chu herbivores ei emaw, herbivores simulating emaw hian leaf metabolome metabolic diversity pum pui a tihtlem chhunzawm zel a, chutih rualin a degree of specialization a tisang zel a ni. Hetianga herbivores-te avanga metabolome specificity rei lo te chhunga tihpun hi transcriptome specificity synergistic increase nen a inzawm a ni. He metabolome specialization lian zawk (Si value sang zawk nei)-a thawhhlawk ber feature chu a hmaa herbivorous function hriat lar tak nei special metabolite hi a ni. He model hi OD theory prediction nen a inmil a, mahse metabolome reprogramming randomness nena inzawm MT prediction hi a inmil lo. Mahse, he data hi mixed model (best MT; Figure 1B) prediction nen pawh a inmil hle a, a chhan chu uncharacterized metabolite dang, defense function hriat loh nei te chuan random Si distribution an la zawm thei tho a ni.
He zirchianna hian a rawn man lehzual dan ngaihnawm tak chu micro-evolution level (single plant leh tobacco population) atanga evolutionary scale lian zawk (closely related tobacco species) thlengin, evolutionary organization level hrang hrangte chu “best defense”-ah an awm a, herbivores te theihnaah hian danglamna nasa tak a awm a ni. Moore leh a thawhpuiten an sawi. (20) leh Kessler leh Kalske (1) te chuan Whittaker (50)-a’n a tir lama a thliar hrang biodiversity functional level pathum chu chemical diversity constitutive leh induced temporal changes-ah chantir turin independent takin an rawt a these authors neither summarized Metabolome data lian tham tak tak lakkhawmna atana kalphung pawh hian heng data atanga metabolic diversity chhut dan tur a tarlang lo. He zirchiannaah hian Whittaker-a functional classification-a siamthatna tenau te chuan α-metabolic diversity chu plant pakhata MS/MS spectra chi hrang hrang anga ngaih a ni ang a, β-metabolic diversity chu population group pakhat Space-a intraspecific metabolism bulpui anga ngaih a ni ang a, γ-metabolic diversity chu species inang chiah chiah thlirletna tihzauh a ni ang.
JA signal hi herbivore metabolic response hrang hrang atan a pawimawh hle. Mahse, metabolome diversity-a JA biosynthesis intraspecific regulation-in a thawhhlawkzia quantitative testing khauh tak a awm lo a, JA signal hi macroevolutionary scale sang zawka stress-induced metabolic diversification atana general site a nih leh nih loh chu hriat chian a la ni lo. Nepenthes herbivorous-a herbivorous nature hian metabolome specialization a thlen tih kan hmu a, Nicotiana chi population chhungah leh Nicotiana chi hrang hrang inzawm tlat zingah metabolome specialization danglamna chu JA signaling nen systematic takin a inzawm tha hle tih kan hmu bawk. Tin, JA signal a chhiat chuan genotype herbivore pakhatin a tihbuai metabolic specificity chu a cancel bawk ang (Figure 3, C leh E). Naturally attenuated Nepenthes population-te metabolic spectrum inthlak danglamna hi a tam zawk chu quantitative a nih avangin, he analysis-a metabolic β diversity leh specificity inthlak danglamna hi a tam zawk chu metabolite-rich compound category-te excitation chak tak vang a ni thei. Heng compound class te hian metabolome profile thenkhat an thunun a, JA signals te nen positive correlation an siam thin.
Amah nena inzawm tlat zuk leh hmuam chi hrang hrangte biochemical mechanism a danglam em avangin, metabolites te hi qualitative aspect-ah chiang taka hriatchhuah a nih avangin analytical zawk a ni. Information theory-in captured metabolic profile a process dan chuan herbivorous induction hian metabolic gamma diversity leh specialization inkara trade-off a tizual tih a tarlang. He trade-off-ah hian JA signal hian hmun pawimawh tak a chang a ni. Metabolome specialization tihpun hi main OD prediction nen a inmil a, JA signal nen a inzawm tha hle a, JA signal erawh chu metabolic gamma diversity nen a inzawm tha lo thung. Heng model te hian thlai OD capacity hi JA plasticity atanga teh a ni ber tih a tarlang a, microevolutionary scale emaw evolutionary scale lian zawk emaw pawh nise. JA biosynthesis defects circumvent tu exogenous JA application experiment-ah chuan zuk leh hmuam chi hrang hrang inzawm tlat te chu signal-responsive leh signal-non-responsive species-ah an thliar hrang thei tih a tarlang belh a, chu chu an mode of JA leh metabolome plasticity herbivores induced ang bawkin. Signal non-responsive species te hian endogenous JA an siam theih loh avangin an chhang thei lo va, chuvangin physiological limitation an nei a ni. Hei hi JA signaling pathway-a key gene thenkhat (N. crescens-ah AOS leh JAR4) of. He result hian heng interspecies macroevolutionary patterns te hi internal hormone perception leh responsiveness inthlak danglamna avanga lo awm ber a ni thei tih a tilang chiang hle.
Thlai leh herbivores inkara inzawmna bakah hian metabolic diversity exploration hi biological adaptation to the environment leh complex phenotypic traits evolution zirchiannaa theoretical advancement pawimawh zawng zawng nen a inzawm vek a ni. Tunlai MS instrument hmanga data an hmuh tam chhoh zel avangin metabolic diversity chungchanga hypothesis testing hian mimal/category metabolite danglamna a paltlang thei tawh a, global analysis hmangin beisei loh pattern a pholang thei ta a ni. Large-scale analysis kalpui danah hian metaphor pawimawh tak chu data chhui nan hman theih tur map awmze nei tak tak ngaihtuah chhuah hi a ni. Chuvangin, tuna unbiased MS/MS metabolomics leh information theory inzawmkhawmna result pawimawh tak chu taxonomic scale hrang hranga metabolic diversity browse theihna tur map siamna atana hman theih tur metric awlsam tak a pe a ni. Hetiang hmanrua atana thil tul bulpui ber a ni. Micro/macro evolution leh khawtlang nunphung zir chianna.
Macro-evolutionary level-ah chuan Ehrlich leh Raven (51) te plant-insect co-evolution theory bulpui ber chu interspecies metabolic diversity danglamna hi plant lineage hrang hrang awm chhan a nih thu sawi lawk hi a ni. Mahse, he seminal work hi tihchhuah a nih atanga kum sawmnga chhung hian he hypothesis hi test a ni tlem hle (52). Hei hi a chhan lian ber chu hla tak atanga thlai chi hrang hranga metabolic characteristic tehkhin theih phylogenetic characteristics vang a ni. Rarity hi target analysis methods anchor nan hman theih a ni. Tuna information theory hmanga processed MS/MS workflow hian metabolite hriat loh (metabolite thlan hmasak loh) te MS/MS structural similarity chu a quantified a, heng MS/MS te hi MS/MS set ah a chantir a, chutiang chuan professional metabolism ah Heng macro-evolutionary model te hi classification scale ah tehkhin a ni. Statistical indicator awlsam tak tak te. Hetiang kalphung hi phylogenetic analysis nen a inang a, sequence alignment hmangin diversification emaw character evolution rate emaw chu a hmaa sawi lawk ngai lovin quantified theih a ni.
Biochemical level-ah chuan Firn leh Jones (53) te screening hypothesis chuan metabolic diversity chu level hrang hrangah vawng rengin, a hmaa inzawm lo emaw, substituted metabolites te biological activity exert turin raw materials pe turin a awm tih a tarlang. Information theory methods hian framework a pe a, chutah chuan heng metabolite-specific evolutionary transition te hi metabolite specialization chhunga thlengte hi evolutionary screening process ruahman zingah hian quantified theih a ni: biologically active adaptation from low specificity to high specificity Inhibited metabolites of a given environment.
A vaiin, molecular biology hun hmasa lamah khan plant defense theory pawimawh tak tak siam a ni a, deductive hypothesis-driven methods hi science lama hmasawnna awmchhun anga ngaih a ni. Hei hi a chhan lian ber chu metabolome pumpui tehna technical limitation vang a ni. Hypothesis-driven methods hi causal mechanism dang thlan nan a tangkai hle nachungin, biochemical network chungchanga kan hriatthiamna tihhmasawn theihna chu tunlai data-intensive science-a computational method hman mek aiin a tlem zawk a ni. Chuvangin, theory sawi lawk theih lohte chu data awmsa huam chhungah a hla hle a, chuvangin research field-a hmasawnna hypothetical formula/test cycle chu tihtawp theih a ni lo (4). Heta metabolomics-a computational workflow rawn luh hian tun hnaia (engtin nge) leh a tawp ber (engvangin nge) metabolic diversity chungchanga ngaihvenna a tiharh leh thei tih kan hmu lawk a, theoretically guided data science hun thar a siam thei bawk. Chu hun lai chuan thlah hmasate rilru hnehtu theory pawimawh tak takte chu a zirchiang leh a.
Direct herbivore feeding hi rose par chi khat Pale-colored pitcher plant hnah pakhatah second instar larva emaw Sl larva emaw enkawl a ni a, thlai khatah plant replicate 10 a awm a ni. Insect larvae te chu clamp hmangin an clamp a, a hnah tissue la awm chu an kai atanga darkar 24 leh 72 hnuah an la khawm a, quick-frozen an ni a, metabolites te chu an la chhuak a ni.
Herbivorous treatment chu synchronized sang tak angin simulate rawh. A tih dan chu fabric pattern wheel hmangin fabric garland thanna stage-ah thlai hnah pathum fully expanded midrib sir lehlamah hling row thum chu puncture a, 1:5 Diluted Ms. Or use gloved fingers hmangin puncture wound-ah S1 OS dah nghal a ni. A chunga kan sawi tawh ang khan hnah pakhat chu lakkhawm leh process rawh. A hmaa kan sawi tawh ang khan primary metabolites leh plant hormones te chu lakchhuah nan hmang ang che (54).
Exogenous JA application atan chuan chi hrang hranga rose blooming plant paruk zinga pakhat theuh petiole hnah pathum chu lanolin paste 20μl, 150μg MeJA (Lan + MeJA) awmna, leh 20μl lanolin plus wound treatment (Lan + W) hmanga enkawl a ni a, a nih loh leh control atan 20μl pure lanolin hman a ni bawk. A hnah chu enkawl a nih hnu darkar 72-ah lakkhawm a ni a, liquid nitrogen-ah snap-frozen a ni a, hman thlengin -80°C-ah dah a ni.
Kan research group-ah hian JA leh ET transgenic line pali, irAOC (36), irCOI1 (55), irACO leh sETR1 (48) te chu hmuhchhuah a ni tawh a. irAOC hian JA leh JA-Ile level a tlahniam nasa hle a, irCOI1 erawh chuan JAs a sensitive lo thung. EV nen khaikhin chuan JA-Ile accumulation a pung a ni. Chutiang bawkin irACO hian ET siamchhuahna a ti tlem dawn a, EV nena khaikhin chuan ET laka insensitive lo sETR1 hian ET siamchhuahna a tipung dawn a ni.
Photoacoustic laser spectrometer (Sensor Sense ETD-300 real-time ET sensor) hmangin ET tehna chu non-invasive-in an ti thei a ni. Treatment an neih zawh veleh a hnah chanve chu an tan a, 4-ml sealed glass vial-ah an dah a, darkar 5 chhungin headspace chu a khawlkhawm theih nan an dah a ni. Measurement neih chhung hian vial tin chu boruak thianghlim litre 2/hour stream hmangin minute 8 chhung an flush a, chu chu a hmain Sensor Sense-in CO2 leh tui paih chhuahna atana catalyst a pek hmangin a lo kal tawh a ni.
Microarray data hi a tir chuan (35)-ah tihchhuah a ni a, National Center for Biotechnology Information (NCBI) Gene Expression Comprehensive Database (accession number GSE30287)-ah dah a ni. He zirchianna atan hian W + OSMs treatment avanga hnah lo awm leh undamaged control nena inmil data lakchhuah a ni. Raw intensity chu log2 a ni. Statistical analysis hmain baseline chu R software package hmangin a 75th percentile-ah convert leh normalized a ni.
Nicotiana chi hrang hrangte RNA sequencing (RNA-seq) data original chu NCBI Short Reading Archives (SRA) atanga lak chhuah a ni a, project number chu PRJNA301787 a ni a, hei hi Zhou et al. (39) leh (56)-a tarlan angin kalpui rawh. He zirchiannaah hian Nicotiana chi hrang hrang nena inmil W + W, W + OSM leh W + OSS1 hmanga raw data process te chu analysis atan thlan a ni a, hetiang hian process a ni: A hmasain raw RNA-seq reading te chu FASTQ format-ah an chantir a ni. HISAT2 hian FASTQ chu SAM ah a convert a, SAMtools hian SAM file te chu sorted BAM file ah a convert bawk. StringTie hi gene expression chhut nan hman a ni a, a expression method chu sequenced transcription fragment maktaduai khatah base fragment sang khatah fragment a awm tih hi a ni.
Analysis-a hman Acclaim chromatographic column (150 mm x 2.1 mm; particle size 2.2μm) leh 4 mm x 4 mm guard column te hi material inang atanga siam a ni. Dionex UltiMate 3000 Ultra High Performance Liquid Chromatography (UHPLC) system-ah hian a hnuaia binary gradient hi hman a ni: minute 0 atanga 0.5 chhung, isocratic 90% A [deionized water, 0.1% (v/v) acetonitrile leh 0.05% formic acid], 10% B (Acetonitrile leh 0.05% formic acid) Minute 0.5 atanga 23.5 chhungin gradient phase chu 10% A leh 90% B a ni a; 23.5 atanga minute 25 chhung, isocratic 10% A leh 90% B. Flow rate chu 400μl/min a ni. MS analysis zawng zawng atan column eluent chu quadrupole leh time-of-flight (qTOF) analyzer-ah inject tur a ni a, electrospray source hmanga thuam, positive ionization mode-a thawk (capillary voltage, 4500 V; capillary outlet 130 V ; Drying temperature 200°C; drying airflow 10 liters/min).
Data atanga thil pawimawh lo emaw, thliar hran theih loh emaw MS / MS fragment analysis (hetah hian MS / MS tia sawi tur) ti la, overall detectable metabolic profile chungchanga structural information hmuh theih a ni. Indiscriminate MS/MS method concept hi quadrupole hian mass isolation window lian tak a neih vang a ni [chuvangin, mass-to-charge ratio (m/z) signal zawng zawng chu fragment angin ngai rawh]. Hemi avang hian Impact II instrument hian CE tilt a siam theih loh avangin independent analysis engemawzat chu collision-induced dissociation collision energy (CE) value tihpun hmangin an ti a ni. A tawi zawngin, sample chu UHPLC-electrospray ionization/qTOF-MS hmangin single mass spectrometry mode (in-source fragmentation atanga lo chhuak low fragmentation conditions) hmangin enfiah hmasa la, m/z 50 atanga 1500 thleng repetition frequency 5 Hz-ah scan rawh. MS/MS analysis atan nitrogen chu collision gas atan hmang la, a hnuaia collision-induced dissociation voltage hrang hrang pali: 20, 30, 40, leh 50 eV-ah independent measurement ti rawh. Measurement process chhung zawng hian quadrupole hian mass isolation window lian ber a nei a, m/z 50 atanga 1500 thleng a nei a, front body m/z leh isolation width experiment chu 200-a dah a nih chuan instrument operating software leh 0 Da hmangin mass range chu automatic-in a activate a ni. Single mass mode ang bawkin mass fragment te chu scan rawh. Mass calibration atan sodium formate (50 ml isopropanol, 200 μl formic acid leh 1 ml 1M NaOH aqueous solution) hmang ang che. Bruker-a high-precision calibration algorithm hmangin data file chu hun bituk chhunga spectrum average run hnuah calibrate a ni. Data Analysis v4.0 software (Brook Dalton, Bremen, Germany) export function hmangin raw data file te chu NetCDF format ah convert rawh. MS/MS data set chu open metabolomics database MetaboLights (www.ebi.ac.uk) ah accession number nen save a ni tawh bawk. MTBLS1471 a ni.
MS/MS assembly hi MS1 leh MS/MS quality signal te inkara correlation analysis hmanga collision energy hniam leh sang leh rules hman thar hmanga tihhlawhtlin theih a ni. R script hi product-a precursor distribution-a correlation analysis tihhlawhtlin nan hman a ni a, C# script (https://github.com/MPI-DL/indiscriminant-MS-MS-assembly-pipeline) hmangin rules kalpui a ni bawk.
Sample tlemte chauh a m/z feature thenkhat hmuhchhuah avanga background noise avanga false positive error leh false correlation awmte tihtlem nan R package XCMS (background noise correction atan) Should be used to replace the “NA” (undetected peak) intensity a “filled peak” function kan hmang a ni. Fill peak function hman a nih chuan data set-ah hian correlation calculation nghawng thei tur “0′′ intensity value tam tak a la awm a, chutah chuan, filled peak function hman a nih lai leh filled peak function hman loh huna data processing result kan hmuhte chu kan khaikhin a, average corrected estimated value atanga background noise value kan chhut a, chutah chuan heng 0 intensity value te hi calculated background value hmangin kan thlak leh bawk a chakna chu background value aiin a let thum aia tam feature-te chauh a ngaihtuah a, “true peaks” anga ngaih a ni. PCC chhut dan atan chuan sample precursor (MS1) leh fragment data set-a peak dik tak pariat tal nei m/z signal chauh ngaihtuah a ni.
Sample pumpuiah precursor quality feature intensity chu collision energy hniam emaw sang emaw hnuaia awm quality feature khata intensity tihtlem nen a inzawm tlat a, chu feature chu CAMERA-in isotope peak anga a label loh chuan sawifiah belh theih a ni. Tichuan, s 3 chhunga precursor-product pair awm thei zawng zawng (peak retention atana estimated retention time window) chhut chhuakin, correlation analysis chu tih a ni. m/z value chu precursor value aiin a hniam a, MS/MS fragmentation chu a chhuahna precursor nen data set-a sample awmna hmun khatah a awm chauhvin fragment anga ngaih a ni.
Heng dan awlsam pahnih hmang hian, precursor hriat chhuah m/z aia m/z value tam zawk nei fragment tarlan te chu kan paih chhuak a, precursor lo langna sample position leh fragment tarlan te a zirin kan paih chhuak bawk. MS1 mode-a in-source fragment tam tak siam chhuah quality features te chu candidate precursor atan thlan theih a ni bawk a, chu chuan redundant MS/MS compound a siam chhuak thei bawk. He data redundancy tihtlem nan hian spectra-te NDP similarity 0.6 aia tam a nih a, CAMERA-in a annotated chromatogram “pcgroup”-a tel an nih chuan kan merge ang. A tawp berah chuan precursor leh fragment nena inzawm CE result pali zawng zawng chu final deconvoluted composite spectrum-ah kan dah khawm vek a, collision energie hrang hranga m/z value inang nei candidate peak zawng zawng zingah intensity peak sang ber kan thlang a ni. A hnu lama processing step te hi composite spectrum concept atanga siam niin, fragmentation probability tihpun nan CE condition hrang hrang mamawh te ngaihtuah a ni a, a chhan chu fragment thenkhat chu collision energy bik hnuaiah chauh hmuhchhuah theih a nih vang a ni.
RDPI (30) hmangin metabolic profile inducibility an chhut a. Metabolic spectrum diversity (Hj index) hi Martínez et al. (8) tih a ni. Hj = −∑i = 1mPijlog2(Pij) chutah chuan Pij chu j-th sample-a i-th MS/MS relative frequency (j = 1, 2,..., m) (i = 1, 2, ..., m) t) nen a inmil a ni.
Metabolic specificity (Si index) tih hi MS/MS pek tawh pakhata sample ngaihtuah mek inkara frequency nena inzawm expression identity tihna a ni. MS/MS specificity chu Si = 1t (∑j = 1tPijPilog2PijPi) anga chhut a ni.
A hnuaia formula hmang hian j sample tin metabolome-specific δj index tehna hmang la, MS/MS specificity average δj = ∑i = 1mPijSi teh rawh
MS/MS spectra hi pahnih pahnih in align a ni a, score pahnih atanga inmilna chu chhut a ni. Pakhatnaah chuan standard NDP (cosine correlation method tia hriat bawk) hmangin a hnuaia equation hmang hian spectra NDP = (∑iS1 & S2WS1, iWS2, i) 2∑iWS1, i2∑iWS2, i2 where S1 leh S2 Correspondingly, spectrum 1 leh spectrum 2, bakah WS1, i leh WS2, i spectra pahnih inkara i-th common peak danglamna chu 0.01 Da aia tlem a nih thu peak intensity atanga chhut rit a entir a ni. A rit zawng chu hetiang hian chhut a ni: W = [peak intensity] m [quality] n, m = 0.5, n = 2, MassBank-in a rawt angin.
Scoring method pahnihna hman a ni a, chu chu MS/MS inkara NL insem dan thlirletna a ni. Chumi atan chuan MS fragmentation process-a kan tawn fo thin NL list 52 te chu tandem-in kan hmang a, a hmaa Nepenthes chi chak lo zawkte secondary metabolite-te MS/MS spectrum atana annotate tawh NL chiang zawk (data file S1) te chu kan hmang bawk ( 9, 26 ). MS/MS tin tan binary vector 1 leh 0 siam la, NL thenkhat current leh non-existent nen a inmil tur a ni. Euclidean distance similarity atanga chhut chuan binary NL vector pair tin tan NL similarity score chu chhut a ni.
Dual clustering tih nan R package DiffCoEx kan hmang a, hei hi Weighted Gene Co-expression Analysis (WGCNA) extension atanga siam a ni. MS/MS spectra-a NDP leh NL scoring matrices hmangin DiffCoEx hmangin comparative correlation matrix kan chhut a. Binary clustering hi “cutreeDynamic” parameter chu method = “hybrid”, cutHeight = 0.9999, deepSplit = T, leh minClusterSize = 10-a dah a ni a, DiffCoEx-a R source code hi file 1 dang atanga Tesson et al. (57) leh; R WGCNA software package mamawh chu https://horvath.genetics.ucla.edu/html/CoexpressionNetwork/Rpackages/WGCNA ah hian hmuh theih a ni.
MS/MS molecular network analysis tih theih nan NDP leh NL similarity type hmangin paired spectral connectivity kan chhut a, Cytoscape software hmangin CyFilescape yFiles layout algorithm extension application-ah organic layout hmangin network topology kan hmu thei a ni.
Data chungchangah statistical analysis tih nan R version 3.0.1 hmang ang che. Statistical significance chu two-way analysis of variance (ANOVA) hmangin an teh a, chu chu Tukey-a honestly significant difference (HSD) post-hoc test hmangin an zirchiang leh a ni. Herbivorous treatment leh control inthlauhna zirchian nan hian variance inang sample group pahnih te two-tailed distribution chu Student’s t test hmangin an zirchiang a.
He thuziak atana supplementary materials hriat duh chuan http://advances.sciencemag.org/cgi/content/full/6/24/eaaz0381/DC1 ah hian en theih a ni
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