Thlaici kilvennak theory phunphun nih thingram a hleiin a ummi thilri thlennak phunglam fianternak caah a biapi mi theory lamhruainak an pek, asinain an chimchungnak biapi cu hneksak a hau rih. Hika ah, kannih nih thleidannak a ngei lomi tandem mass spectrometry (MS/MS) hlathlainak kha kan hman i, thingram pakhat cio in milu le aa pehtlaimi phun hna tiang zuu a zortermi phun hna i metabolome kha ningcang tein kan hlathlai, cun thawngpang chung i compound spectra cung ah hram bunh in mass spectrometric sining theory tampi kan tuah. A ttha bikmi ralvennak (OD) le aa chawkmi tinhmi (MT) theory hna i a biapimi chimchungnak hna hneksaknak caah phunglam. Thingkung metabolomics kong thawngpang tthencheunak cu OD theory he aa tlak, asinain MT theory nih herbivoms nih a chuahtermi metabolomics dynamics kong a chimchungmi he aa kalh. Micro in macro thlennak tahfung tiang, jasmonate signal cu OD a biapi bikmi a si tiah hngalh a si, cu lio ah ethylene signal nih MS/MS molecular network nih a langhtermi herbivore-specific lehnak kha a ttha tein a remh.
A phunphun in a ummi thilri hna cu thingram hna nih pawngkam he an i remh khawhnak ah aa telmi an si, a hlei in ral pawl kilvennak ah (1). Thingkung chung i hmuhmi a hleiin a phunphun in a ummi thilri thlennak nih kum hra chung a pawngkam riantuannak tampi kongah thukpi in hlathlainak a chuahter, cun thingram kilvennak theory cazin sau a ser, cu hna cu thingram le rungrul pehtlaihnak i thlennak le pawngkam tthanchonak an si. Hmuhtonnak hlathlainak nih a biapi mi lamhruainak a pek (2). Sihmanhsehlaw, hi thingram kilvennak theory hna nih hin hypothetical deductive reasoning phunglam lam kha an zulh lo, cu ah cun a biapi mi chimchungnak hna cu hlathlainak tlukruannak ah an um (3) i theory tthanchonak a hnu tthanchonak (4) thanchoternak caah hneksaknak in an hneksak hna. Technical rikhiahnak nih data laknak kha a hleiin metabolic phun hna ah a kham i a hleiin metabolites hna i a tlingmi hlathlainak kha a telh lo, cucaah theory tthanchonak caah a herhmi phun karlak tahchunhnak kha a kham (5). A tlingmi metabolomics data le thingram phun dangdang karlak ah metabolic hmunhma riantuan ning tahchunhnak caah a tlangpi tangka a um lo caah riantuannak scientific tthanchonak a kham.
Tandem mass spectrometry (MS/MS) metabolomics lei ah a thar in a chuakmi thil hna nih hin system clade phunkhat chung le karlak ah thlennak a ummi kha tling tein a langhter khawh, cun hi a harmi cawhmi hna karlak i a sining aa lawhnak kha tuaktan khawhnak ding caah tuaktannak lam he fonh khawh a si. Chemistry kong hngalh cia (5). Hlathlainak le tuaktannak ah a sangmi thiamnak fonhmi nih hin metabolic phunphun kong he pehtlai in ecological le evolutionary theories nih an tuahmi chimchungnak tampi caan sau hneksaknak caah a herhmi phunglam a ser. Shannon (6) nih 1948 ah a biapi bikmi capar ah a voikhatnak bik thawngpang theory a rak luhpi, thawngpang tuaktannak caah hrampi a rak bunh, cucu a hramthawk hmannak lengah rian tampi ah hman a si cang. Genomics ah, thawngpang theory cu a zulmi thawngpang (7) zat tuaktannak ah hlawhtling tein hman a si cang. Transcriptomics hlathlainak ah, thawngpang theory nih transcriptome chung i thlennak vialte kha a hlathlai (8). Hlan lio hlathlainak ah, thingram hna i tissue level i metabolic thiamnak langhternak caah metabolomics ah thawngpang theory i zatlang nunphung phunglam kha kan hman (9). Hika ah, MS/MS-based riantuan ning cu thawngpang theory i zatlang nunphung he kan fonh, cucu a tlangpi tangka ah a ummi thilri thlennak phunphun in a lang, cu nih cun thingram kilvennak theory i a biapi mi chimchungnak hna kha tahchunh awkah a si.
Thingkung kilvennak kong ruahnak phunglam hna cu a tlangpi in pakhat le pakhat aa telmi an si i phun hnih ah ṭhen khawh an si: kilvennak rianṭuannak cung ah hram bunh in thingram-specific metabolites i phawtzamhnak fianter aa zuammi hna, cu hna cu a ṭha bikmi kilvennak (OD) (10), aa chawkmi target (MT) (11) ) le muisam (12) theory an si, cu lioah a dang nih cun thilri hmuh khawhnak thlennak nih zeitindah a thlen timi fianternak an kawl thingram ṭhanchonak le carbon tibantuk a hleiin a ummi thilri pawl i khonnak: rawl tlukruannak ruahnak (13), ṭhanchonak tlukruannak ruahnak (14 ), le ṭhanchonak le thleidannak tlukruannak ruahnak (15). Theory phu hnih hna cu hlathlainak ah aa dangmi an si (4). Sihmanhsehlaw, riantuannak level ah i vennak riantuannak aa telmi theory pahnih nih thingram a sertu le a chuahter khomi venhimnak kong biaruahnak kha a uk: OD theory, cu nih cun thingram hna nih an man a fakmi chemical venhimnak ah an herh tikah lawng an hman tiah a ruah, tahchunhnak ah, an ei tikah. MT ruahnak nih cun lamhruainak metabolite thlennak axis a um lo, asinain metabolite cu aa thleng lo, cucaah a si khomi a ser. Phundang in chim ahcun, hi theory pahnih nih hin saram hna nih an tuk hna hnu ah a cangmi metabolic remhnak kongah aa kalhmi chimchungnak an tuah: defensive function (OD) le non-directed metabolic thlennak (MT) he metabolites pakhat lei kap in an i fonhnak karlak pehtlaihnak (11 ).
OD le MT ruahnak cheuhnak nih hin metabolome ah a chuakmi thlennak lawng si loin, hi metabolites pawl an i fonhnak nih a chuahpimi ecological le evolutionary thil sining zong aa tel, cu hna cu ecological pawngkam pakhatkhat ah hi metabolic thlennak hna i an i remh khawhnak man le hlawknak hna an si (16). Ruahnak pahnih nih hin a man a fak kho men asiloah a si kho men lo tiah a hleiin sermi thilri hna i an i vennak riantuannak kha an cohlan ko nain, OD le MT ruahnak cheuhnak a thleidangtu a biapi bikmi chimchungnak cu a chuakmi thlennak hna i an kalnak lam ah a um. OD theory chimchungnak nih hin atu tiangah hneksaknak lei ah a tam bik zohkhenhnak a hmuh. Hi hneksaknak hna ah hin greenhouse le thilnung dirhmun ah a ummi thilri phun dangdang hna i direct asiloah indirect in i vennak riantuannak kong hlathlainak le thingram tthanchonak dirhmun thlennak hna kong hlathlainak aa tel (17-19). Sihmanhsehlaw, atu tiang ahcun, zeibantuk thilnung hmanh an i thlennak kong vawleicung pumpi ah a tlingmi hlathlainak caah riantuan ningcang le zatlang relnak phunglam a um lo caah, theory pahnih karlak i a biapi bikmi aa dannak chimchungnak (cucu, thlennak lei thlennak lam) cu hneksak a hau rih. Hika ah, cu bantuk hlathlainak cu kan pek.
Thlaici-thleidannak metabolites hna i a biapi bikmi sining pakhat cu thingram pakhat lawng in, milu zat in aa lo ngaimi phun tiang an sining aa dang tukmi a si (20). Specialized metabolites ah zatlang thlennak tampi cu milu zat ah hmuh khawh a si, cu lioah a tthawngmi sining thleidannak cu phun zat ah a um tawn (20). Cucaah, thingram thlennak phunphun cu riantuannak phunphun ah a biapi bikmi a si, hmun dangdang ah i remh khawhnak a langhter, a hlei in rungrul le a tlangpi in a ei tawnmi saram hna nih an luhhnawh khawhnak aa dangmi hmun hna kha a langhter (21). Fraenkel (22) nih thingram-specific metabolites a umnak a ruang kong a ttialmi hramthawknak capar thawk in, rungrul phunphun he pehtlaihnak cu a biapi mi thimnak hneknak ah ruah a si, cun hi pehtlaihnak hna nih hin thlennawnnak lio ah thingram pawl a sersiam hna tiah zumh a si. Metabolism lam (23). Species metabolites phunphun ah phun dangdang aa dannak nih hin herbivorous strategy dohnak ah a ummi le a chuahter khomi thingram kilvennak he aa pehtlaimi pumsa lei tlukruannak zong a langhter khawh, zeicahtiah phun hnih hna cu pakhat le pakhat a ttha lomi pehtlaihnak an ngei tawn (24). Zeitik caan paoh ah ttha tein i vennak ngeih cu a ttha ko nain, i vennak he aa pehtlaimi a caan hmaan tein thlennak nih thingram hna kha a dang pumsa lei riantuannak ah a sunglawi mi thilri pawl hman khawhnak ah a fiangmi tthatnak a pek hna (19, 24), cun symbiosis a herhnak hrial khawh a si. A dang rawhralnak (25). Cun, rungrul saram hna nih an chuahtermi a hleiin a ummi thilri hna i an i remh ṭhannak nih hin milu ah rawhralnak a chuahter khawh (26), cun milu ah a hmunmi jasmonic acid (JA) signal ah a biapi tukmi thilnung thlennak hna kha direct in relnak a langhter khawh. JA signal sang le niam cu thingthei a eimi saram dohnak le saram phun pakhatkhat he i zuamnak karlak ah i thlennak a si (27). Cun, a hleiin sermi metabolite biosynthetic lam hna cu thlennawnnak chung ah rang tukin sunghnak le thlennak an tong lai, cu nih cun aa pehtlaimi phun hna lakah aa tluk lomi thlennak a chuahter lai (28). Hi polymorphisms pawl hi aa thlengmi thingthei ei ningcang (29) lehrulhnak ah rang tein dirh khawh an si, a sullam cu thingthei ei mibu aa thleng lengmangmi cu metabolic heterogeneity a chuahtertu a biapi bikmi thil a si tinak a si.
Hika ah, a tanglei buaibainak hna hi a bik in kan tawlrel. (I) Zeitindah thingram a eimi rungrul nih thingram metabolome kha a remh ṭhan? (Ii) Caan saupi i vennak theory chimchungnak hneksaknak caah tah khawhmi metabolic plasticity konglam a biapi bikmi hna cu zeidah an si? (Iii) Thlaici metabolome kha a tukmi caah aa dangmi lam in remh ṭhan ding maw, a si ahcun, thingram hormone nih a hleiin metabolic lehrulhnak remhnak ah zeibantuk rian dah a ṭuan, cun zeibantuk metabolites nih dah dohnak phun thleidannak ah a bawmh? (Iv) Ralvennak theory tampi nih an chimchungmi cu biological tissues level kip ah karhter khawh a si caah, chunglei tahchunhnak in saram phun karlak tahchunhnak tiang a chuahtermi metabolic lehrulhnak cu zeitluk in dah aa tlak tiah kan hal. Cu caah, kannih nih zuu nicotine hnah metabolome kha ningcang tein kan hlathlai, cucu ecological model thingram a si i a ttha ngaimi thlennak a ngeimi a si, cun a ttha ngaimi ramlak saram pahnih, Lepidoptera Datura (Ms) ( A fak tukmi, a tam deuh in ei) Solanaceae le Spodoptera (Spodoptera) ah, thinghnah coworm phunkhat an si “genus”, Solanaceae le a dang phun le chungkhar hna i an ngeitu thingram hna he Thingkung rawl. MS/MS metabolomics spectrum kha kan thleidan i OD le MT theory tahchunh awkah thawngpang theory statistical descriptors kan chuah. A biapimi metabolite pawl an sining langhter awkah a hleiin langhtermi map ser. Hlathlainak cu N. nasi le aa pehtlaimi zuu phun hna i an chuahkehnak milu tiang an karhter i thingram hormone signaling le OD induction karlak i aa dannak kha hlathlainak tuah chap a si.
Herbivorous tobacco hnah metabolome a tthatnak le a sining kong he pehtlai in a dihlak in map pakhat tlaih khawhnak ding caah, a hlan ah sermi hlathlainak le tuaktannak riantuan ning kha kan hman i thingram chuahmi chungin a ttha ngaimi data aa hngat lomi MS/MS spectra kha tling tein kan lak i kan thlen ( 9). Hi thleidan lomi lam (MS/MS tiah auhmi) nih a herh lomi compound spectra a ser khawh, cu cu hika ah langhtermi compound-level hlathlainak vialte caah hman khawh a si. Hi aa tthencheu mi thingram thlennak hna hi phunphun an si, thlennak za tampi in thong tampi tiang an i tel (hika ah 500-1000-s/MS/MS hrawng). Hika ah, thawngpang theory phunglam chung ah metabolic plasticity kha kan ruah, cun metabolome a phunphun le thiamnak kha metabolic frequency phawtzamhnak Shannon entropy cung ah hram bunh in kan tuak. A hlan ah hmanmi formula (8) hmangin, metabolome phunphun (Hj hmelchunhnak), metabolic profile thiamnak (δj hmelchunhnak) le metabolite pakhat lawng a simi metabolic thleidannak (Si hmelchunhnak) tahnak caah hman khawhmi hmelchunhnak phunphun kan tuak. Cun, kannih nih Relative Distance Plasticity Index (RDPI) kha kan hman i, ramlak saram hna i an metabolome inducibility kha tah khawh a si (Figure 1A) (30). Hi zatlang relnak phunglam chung ah, MS/MS spectrum cu a hrampi thawngpang unit ah kan chiah, cun MS/MS a tamnak kha frequency phawtzamhnak map ah kan thlen, cun cu chungin metabolome phunphun tuaktan awkah Shannon entropy kha kan hman. Metabolome thiamnak cu MS/MS spectrum pakhat i a tlangpi in a hlei in tah a si. Cucaah, herbivore induction hnu ah MS/MS phun cheukhat an karh ning cu spectral inducibility, RDPI le specialization ah aa thleng, cucu, δj index a karh, zeicahtiah specialized metabolites tam deuh an chuah i Si index sang an chuah. Hj phunphun index tlawmternak nih a chuakmi MS/MS zat a zorter, asiloah profile frequency phawtzamhnak cu aa khat lomi lei ah aa thleng, cu lioah a dihlak in a fiang lomi a zorter ti a langhter. Si index tuaktannak thawngin, zei MS/MS dah herbivore cheukhat nih an chuahter ti kha langhter khawh a si, cu he aa ralchanh in, zei MS/MS nih dah chuahternak kha a lehrulh lo, cucu MT le OD chimchungnak thleidan khawhnak caah a biapi bikmi hmelchunhnak a si.
(A) Herbivorous (H1 in Hx tiang) MS/MS data-inducibility (RDPI), diversity (Hj index), specialization (δj index) le metabolite specificity (Si index) caah hmanmi statistical descriptors. Specialization (δj) a karh ahcun, a tlangpi in, thingthei a eimi thilri phunphun an chuak lai ti a langhter, cu lioah phunphun (Hj) a zor ahcun metabolite chuahnak a zor asiloah phawtzamhnak map ah metabolite aa tluk lomi phawtzamhnak a langhter. Si man nih hin metabolite cu thil sining pakhatkhat (hika ah, thingthei a eimi) caah a hleiin a si maw asiloah a tluk tein a ummi a si maw ti kha a tuak. (B) Thawngpang theory axis hmangin venhimnak theory chimchungnak ruahnak hmanthlak. OD theory nih a chimchungmi cu herbivore tukdohnak nih hin ralvennak metabolites a karhter lai, cucaah δj a karhter lai. Cu lio caan te ah cun, Hj cu a zor, zeicahtiah profile cu metabolic thawngpang a zortermi lunghrinnak lei ah aa remh ṭhan. MT theory nih a chimchungmi cu, thingthei a eimi saram hna nih an tuknak nih hin metabolome ah lam dang thlennak a chuahter lai, cucaah Hj cu a karhmi metabolic thawngpang fiang lonak langhtertu ah a karhter lai i Si kha aa khat lomi phawtzamhnak a chuahter lai. Cun, aa cawhmi model, a tha bikmi MT zong kan chuahpi, cu ah cun a sang deuhmi i vennak man a ngeimi metabolite cheukhat cu a hleiin an karh lai (Si man a sang), cu lioah a dang nih cun random lehnak an langhter (Si man a niam).
Theihhngalhnak theory fianternak hmangin, OD theory kha kan fianter i, herbivore-induced special metabolite thlennak nih a chuahter lomi constitutive dirhmun ah (i) metabolic specificity (Si index) a karhter lai i metabolic specificity (δj index) a hruai lai. metabolic frequency phawtzamhnak kha leptin pum phawtzamhnak tam deuh ah thlennak. Metabolite pakhat tluk ah, ningcang tein Si phawtzamhnak a um lai tiah ruah a si, cuka ahcun metabolite nih a vennak man ning in Si man a karhter lai (Figure 1B). Hi lamthluan ah, MT theory kha kan fianter i, lungthawhnak nih (i) δj index a zortermi metabolites ah lam a kal lomi thlennak a chuahter lai, cun (ii) metabolic lunghrinnak a karh caah Hj index a karhter lai ti chimchungnak caah kan fianter. Asiloah randomness, cu cu Shannon entropy in a tlangpi in aa dangmi phun in tah khawh a si. Metabolic composition kongah, MT theory nih Si a tthencheunak kha a chimchung lai. Cheukhat thilri pawl cu thil sining pakhatkhat tangah an um i thil sining cheukhat cu thil sining pakhatkhat tangah an um lo, cun an i vennak man cu pawngkam cungah aa hngat ti kha ruah chih in, kannih nih aa cawhmi i vennak model zong kan chuahpi, cu ah cun δj le Hj cu Si lei ah pahnih in an i ṭhen. 1B).
Thawngpang theory fianternak axis cung ah a thar in fiantermi ralvennak theory chimchungnak hneksaknak caah, Nepenthes pallens (Figure 2A) hnah cung ah thiamsang (Ms) asiloah a tlangpi in (Sl) thingthei a eimi larvae kan cawm hna. MS/MS hlathlainak hmangin, rungrul rawl pek hnu ah lakmi thinghnah hnah chungin methanol chuahmi chungin a herh lomi MS/MS spectra (data file S1) 599 kan hmuh. MS/MS thlennak file chung i thawngpang chung i a ummi thilri remh ṭhannak hmuh khawhnak ding caah RDPI, Hj, le δj index hmannak nih lungthawh awk ngai a simi thil sining a langhter (Figure 2B). A tlangpi in a cangmi cu, thawngpang fianternak nih a langhter bantukin, rungrul nih thinghnah an ei peng tikah, caan a hung rauh deuh tikah, an pumsa thlennak vialte a karh: thingthei a ei hnu suimilam 72 ah, RDPI cu a karh ngaingai. A rawk lomi control he tahchunh tikah, Hj cu a zor ngaingai, cucu δj index in tahmi metabolic profile i a thiamnak a karh caah a si. Hi a langmi thil sining cu OD theory nih a chimchungmi he aa tlak, asinain MT theory nih a chimchungmi a biapi bikmi he aa kalh, mah nih cun metabolite level ah random (a kal lomi) thlennak hna cu i vennak caah hman a si tiah a zumh (Figure 1B). Hi saram pahnih hna i an kaa in chuahmi (OS) chuahtertu thilri le rawl ei ning cu aa dang ko nain, an rawl ei ning nih suimilam 24 le suimilam 72 chung rawl ṭuan caan chungah Hj le δj lei ah aa khatmi thlennak a chuahter. RDPI suimilam 72 chung lawng ah aa dannak a um. Ms rawl peknak nih a chuahtermi he tahchunh ahcun, Sl rawl peknak nih a chuahtermi a dihlak in metabolism cu a sang deuh.
(A) Hneksaknak suaisam: vok (S1) asiloah thiamsang (Ms) thingthei a ei tawnmi hna cu pitcher thingram hna i an hnah hna in an cawm hna, cu lio ah thingthei a ei tawnmi hna caah cun, Ms (W + OSMs) OS cu a tlarimi thinghnah dirhmun hma hna kha tawlrel awkah hman a si. S1 (W + OSSl) rungrul asiloah ti (W + W). Control (C) cu a rawk lomi thinghnah a si. (B) Inducibility (RDPI cu control chart he tahchunh), diversity (Hj index) le specialization (δj index) index cu special metabolite map (599 MS/MS; data file S1) caah tuakmi a si. Arfi hmelchunhnak nih direct herbivore rawl peknak le control phu karlak ah a biapi mi thleidannak a langhter (Student's t-test with paired t-test, *P<0.05 le ***P<0.001). ns, a biapi lo. (C) A tlangpi (a senmi kuang, amino acid, organic acid le chiti; data file S2) le a hleiin metabolite spectrum (a senmi kuang 443 MS/MS; data file S1) hna i an i zohchunhmi thingthei ei thlopnak hnu ah caan thlennak index. Color band nih 95% zumhnak karlak kha a chim duhmi a si. Arfi hmelchunhnak nih thlopnak le uknak karlak ah a biapi mi thleidannak a langhter [quadratic analysis of variance (ANOVA), cu hnu ah Tukey's honestly significant difference (HSD) cu post hoc multiple comparisons caah a si, *P<0.05, **P<0.01 Le *** P <0.001]. (D) Scatter plot le special metabolite profile (thlopbulnak phunphun he sample tuah tthanmi) thleidannak.
Metabolome level ah herbivore nih a chuahtermi remhthannak cu pumpak metabolites level thlennak ah a lang maw langhter lo ti hlathlai awkah, Nepenthes pallens hnah ah a hlan ah hlathlainak tuahmi herbivore doh khawhnak a ngeimi metabolites kha kan zoh hmasa. Phenolic amides cu hydroxycinnamamide-polyamine conjugates an si i rungrul hna nih an ei lio ah an i pum i rungrul hna an riantuan ning a zorter ti a si (32). Aa pehtlaimi MS/MS i a hramthawknak pawl kha kan kawl hna i an i fonhmi kinetic curve pawl kha kan suai hna (Figure S1). Khuaruahhar awk a um lo, phenol chuahmi hna cu ramlak saram dohnak ah direct in aa tel lomi, chlorogenic acid (CGA) le rutin tibantuk hna cu ramsa ei hnu ah an zor. Cu he aa dang in, thingthei a eimi saram nih phenol amides kha a tthawng tukmi ah an ser khawh. Thlaici a eimi saram pahnih hna i pehzulh tein rawl peknak nih phenolamides hna i an lungthawhnak spectrum aa khatmi a chuahter, cun hi pattern cu phenolamides hna i de novo sernak caah a hleiin a fiang. Cu thil cangmi cu 17-hydroxygeranyl nonanediol diterpene glycosides (17-HGL-DTGs) lamthluan hlathlainak tuah tikah hmuh khawh a si lai, mah nih hin acyclic diterpenes tampi a chuahter i a ttha ngaimi anti-herbivore riantuannak (33) a chuahter, cu chung ah Ms Feeding with Sl nih aa lo ngaimi langhternak a chuahter (Figure S1).
Direct herbivore rawl peknak hneksaknak ah a chuak khomi thatlonak cu herbivore hna nih an hnah ei zat le rawl an ei caan aa dannak a si, cu nih cun hma le herbivore nih a chuahtermi herbivore-specific effects kha hloh a harter. A chuakmi thinghnah thlennak lehrulhnak i herbivore phun thleidannak kha ttha deuh in tawlrel khawhnak ding caah, Ms le Sl larvae rawl peknak kha kan lakmi OS (OSM le OSS1) kha aa khatmi thinghnah dirhmun i a tlarimi puncture W ah hman colh in kan i zohchunh. Hi tuahsernak cu W + OS thlopnak tiah auhmi a si, cun tissue tlaunak rate le zat aa dannak a chuahter loin herbivore nih a chuahtermi lehnak aa thawk caan kha a dikthliar in a khiah i induction kha a tlukruanter (Figure 2A) (34). MS/MS hlathlainak le tuaktannak pipeline hmangin, 443 MS/MS spectra (data file S1) kan hmuh, cu hna cu direct feed hneksaknak in a hlan ah an rak i fonhmi spectra he an i tluk. Hi MS/MS data set i thawngpang theory hlathlainak nih a langhter mi cu, thinghnah-specialized metabolomes pawl kha thinghnah a eimi saram pawl i zohchunhnak in reprogram tuahnak nih OS-specific inducements a langhter (Figure 2C). A bik in, OSS1 thlopnak he tahchunh ahcun, OSM nih suimilam 4 ah metabolome thiamnak a ṭhanchoter. Hngalh awk ngai a simi cu, direct herbivore rawl peknak hneksaknak data set he tahchunh ahcun, Hj le δj kha coordinate ah hmangin two-dimensional space ah hmuh khawhmi metabolic kinetics le caan saupi chung herbivore thlopnak lemsoinak lehrulhnak ah metabolome specialization lamhruainak cu aa khatmi in a karh (Figure 2D). Cu caan te ah cun, amino acid, organic acid le sugars (data file S2) hna kha kan tuak i, metabolome thiamnak ah aa tinhmi karhternak hi lemsoimi thingthei a eimi saram hna lehrulhnak ah central carbon metabolism remh ṭhannak ruangah a si maw ti hlathlainak tuah awkah kan tuak ( Figure S2). Hi thil sining hi ttha deuh in fianternak caah, a hlan ah kan i ruahmi phenolamide le 17-HGL-DTG lamthluan hna i an i thlennak kha kan zohfel chin. OS-specific induction of herbivores cu phenolamide metabolism chung ah aa dangmi remhthannak pattern ah aa thleng (Figure S3). Coumarin le caffeoyl tthencheu aa telmi phenolic amides cu OSS1 nih a ttha deuh in a chuahter, cu lio ah OSMs nih ferulyl conjugates a chuahter. 17-HGL-DTG lamthluan caah, a tanglei malonylation le dimalonylation thilchuak hna nih aa dangmi OS chuahternak kha hmuh a si (Figure S3).
A hnu ah, OS-induced transcriptome plasticity kha time-course microarray data set hmangin kan hlathlai, mah nih hin OSMs hmannak kha herbivors ah rosette thingram hnah hna thlopnak ah a zohchunh. Sampling kinetics cu hi metabolomics hlathlainak ah hmanmi kinetics he aa tlak (35). Caan karlak ah metabolic plasticity a karh chin lengmangmi metabolome remhthannak he tahchunh tikah, Ms nih a chuahtermi thinghnah hna ah caan tawite chung ṭialmi puahnak kan hmuh, cuka ahcun transcriptome inducibility (RDPI) le specialization (δj) cu 1 ah an um. transcriptome thiamnak (Figure S4). Metabolic gene chungkhar (P450, glycosyltransferase, le BAHD acyltransferase tibantuk) nih a cunglei langhtermi hlanlio thiamnak sang model zulh in, a hrampi metabolism in a chuakmi sining tthencheunak in a hleiin metabolite pawl fonhnak ah an i tel. Hlathlainak ah, phenylalanine lam kha hlathlainak tuah a si. Hlathlainak nih a fehtermi cu phenolamide metabolism ah a hrampi genes pawl cu a hnuh lomi thingram hna he tahchunh tikah herbivores ah OS-induced a sang ngai, cun an langhternak pattern ah aa naih ngai. Hi lamthluan cunglei ah a ummi MYB8 le a sining genes PAL1, PAL2, C4H le 4CL nih ṭialnak hramthawknak a langhter. Phenolamide a donghnak fonhnak ah rian a ttuanmi acyltransferases, AT1, DH29, le CV86 tibantuk hna nih caan saupi a raumi upregulation pattern an langhter (Figure S4). A cunglei hmuhmi nih a langhter mi cu transcriptome thiamnak hramthawknak le a hnu ah metabolomics thiamnak tthanchonak cu aa pehtlaimi lam an si, cucu a tthawngmi ralvennak lehrulhnak a thawktu synchronous regulatory system ruangah a si kho men.
Thlaici hormone signaling ah remhthannak nih thingram hna an pumsa sining thlennak caah thingram a ei mi konglam a fonhmi uknak tthen pakhat bantuk in rian a ttuan. Herbivore zohchunhnak hnu ah, a biapi mi thingram hormone phun hna i an i fonhmi thlennak kha kan tah i an karlak i caan karlak i langhternak kha kan hmuh [Pearson pehtlaihnak tlukruannak (PCC)> 0.4] (Figure 3A). Ruahning bantuk in, biosynthesis he aa pehtlaimi thingram hormone pawl cu thingram hormone co-expression network chung ah an i pehtlai. Cun, thlennak phunphun nih a chuahtermi thingram hormone pawl langhter awkah hi network ah hin metabolic specificity (Si index) cu map tuah a si. Herbivorous specific response a biapi bikmi hmun pahnih an suai: pakhat cu JA bu ah a um, cuka ahcun JA (a nunnak ah a cawlcangmi JA-Ile) le a dang JA chuahkehnak nih Si score sang bik an langhter; a dang pakhat cu ethylene (ET) a si. Gibberellin nih cun thingthei a eimi saram thleidannak ah tlawmpal lawng a karhter, cu lio ah thingram hormone dang, cytokinin, auxin, le abscisic acid tibantuk hna nih cun thingthei eimi saram caah a tlawm deuhmi induction thleidannak an ngei. W + W lawng hmannak he tahchunh ahcun, OS hmannak (W + OS) hmangin JA chuahmi thilri hna i a sangbik man karhternak cu a hrampi in JAs hna i a fekmi a hleiin langhtertu ah thlen khawh a si. Ruah lo piin, OSM le OSS1 cu aa dangmi elicitor content he JA le JA-Ile aa khatmi khonnak a chuahter ti a si. OSS1 he aa dang in, OSM cu OSMs nih a bik in le fakpi in a chuahter, cu lio ah OSS1 nih cun basal hma hna lehrulhnak kha a karhter lo (Figure 3B).
(A) Herbivore-induced plant hormone khonnak kinetics zohchunhnak PCC tuaktannak cung ah hram bunh in co-expression network hlathlainak. Node nih thingram hormone pakhat lawng a aiawh, cun node ngan le hme nih thlopnak karlak ah thingram hormone he aa pehtlaimi Si index a aiawh. (B) rong dangdang nih a langhtermi thlopnak phunphun ruangah thinghnah ah JA, JA-Ile le ET an i khonnak: apricot, W + OSM; sen, W + OSSl; sen, W + W; gray, C (control) . Arfi hmelchunhnak nih thlopnak le uknak karlak ah a biapi mi thleidannak a langhter (two-way ANOVA hnu ah Tukey HSD post hoc multiple comparison, *** P <0.001). JA biosynthesis le a tlamtlin lomi hmuh khawhnak spectrum (irAOC le irCOI1) ah (C)697 MS/MS (data file S1) le ETR1 ah a tlamtling lomi ET signal he (D)585 MS/MS (data file S1) kong thawngpang theory hlathlainak. Arfi hmelchunhnak nih W+OS thlopnak le a rawk lomi uknak karlak ah a biapi mi thleidannak a langhter (two-way ANOVA hnu ah Tukey HSD post hoc multiple comparison, *P<0.05, **P<0.01 le ***P<0.001). (E) Specialization dohnak aa ṭhekmi graph pawl. Ron pawl nih hin thlennawnnak phun dangdang a aiawh; hmelchunhnak nih thlopnak phun dangdang a aiawh: tlangthum, W + OSS1; rectangle, W + OSM; C kulh
A hnu ah, JA le ET biosynthesis (irAOC le irACO) le hmuhning (irCOI1 le sETR1) ah a biapi bikmi step ah a derthawmmi Nepenthes (irCOI1 le sETR1) timi thlennawnnak phun kha kan hman i, hi thingram hormone pahnih hna i an thlennak kha kan hlathlai. A hlan i hneksaknak hna he aa tlak in, a lawngmi (EV) thingram hna ah herbivore-OS luhternak le OSM nih a chuahtermi Hj index a dihlak in a zornak kha kan fehter, cu lioah δj index cu a karh. OSS1 nih a chuahtermi lehnak nakin lehnak cu a lang deuh. Hj le δj kha coordinate ah hmanmi line pahnih graph nih a bik in deregulation a langhter (Figure 3E). A fiang bikmi thil sining cu JA signal a ngei lomi phun hna ah, thingthei a eimi hna nih an chuahtermi metabolome phunphun le thiamnak thlennak cu a dih deng cang (Figure 3C). Cu he aa dang in, sETR1 thingram hna ah dai tein ET hmuh khawhnak nih, JA signaling nakin herbivorous metabolism thlennak ah a dihlak in a chuahpimi thil cu a niam deuh ko nain, OSM le OSS1 lungthawhnak karlak ah Hj le δj indices i dannak kha a zorter (Figure 3D le Figure S5). . Mah nih hin JA signal thlennak riantuannak hrampi lengah, ET signal thlennak nih hin ramlak saram hna i an phun-thleidannak metabolic lehrulhnak zong a ttha tein remh khawhnak rian a ttuan ti a langhter. Hi ttha tein remh khawhnak riantuannak he aa tlak in, sETR1 thingram hna ah a dihlak in metabolome chuahter khawhnak ah thlennak a um lo. Khatlei kam in, sETR1 thingram hna he tahchunh tikah, irACO thingram hna nih hin thingthei a eimi saram hna nih an chuahtermi thlennak a tlangpi in aa khatmi an chuahter, asinain OSM le OSS1 zuamcawhnak karlak ah Hj le δj mark aa dang tukmi an langhter (Figure S5).
Thlaici a eimi saram hna i an phun-thleidannak lehrulhnak ah a biapi mi bawmhnak a ngeimi a hleiin sermi thilri pawl hngalh khawhnak ding le ET signal hmangin an chuah khawhnak ding caah, a hlan ah sermi structural MS/MS phunglam kha kan hman. Hi lam nih hin MS/MS tthencheunak [normalized dot product (NDP)] le neutral loss (NL) cung ah hram bunhmi aa lawhnak mark chungin metabolic chungkhar kha thlen ṭhan awkah bi-clustering lam kha aa hngat. ET transgenic lines hlathlainak in sermi MS/MS data set nih 585 MS/MS (data file S1) a chuahter, cucu MS/MS modules (M) pasarih ah fonh in a tawlrel (Figure 4A). Hi module cheukhat cu a hlan ah langhtermi special metabolites he a khat: tahchunhnak ah, M1, M2, M3, M4 le M7 cu phenol derivatives phunphun (M1), flavonoid glycosides (M2), acyl sugars (M3 le M4), le 17-HGL-DTG (M7) in an rum. Cun, module pakhat cio ah metabolite pakhat cio i metabolic konglam (Si index) kha tuak a si, cun a Si phawtzamhnak kha a fiangmi in hmuh khawh a si. A tawinak in, MS/MS spectra nih a sangmi herbivory le genotype thleidannak cu a sangmi Si man in langhter a si, cun kurtosis statistics nih orhlei hngawng kam ah vun tthencheunak a langhter. Cu bantuk a tlawmmi colloid phawtzamhnak pakhat cu M1 ah hmuh a si, cu ah cun phenol amide nih Si tthennak a sang bik a langhter (Figure 4B). A hlan ah kan rak chimmi herbivorous inducible 17-HGL-DTG cu M7 ah a tlawm deuhmi Si score a langhter, OS phun hnih karlak ah a tlawm deuhmi thleidannak uknak a langhter. Cu he aa dang in, rutin, CGA, le acyl sugars tibantuk a hlei in chuahmi thilri tam deuh cu Si mark a niam bik ah an i tel. A hleiin metabolites karlak ah a sining a harmi le Si phawtzamhnak kha ttha deuh in hlathlai khawhnak ding caah, module pakhat cio caah molecular network pakhat cio ser a si (Figure 4B). OD theory (Figure 1B ah a tawinak in langhtermi) i a biapi mi chimchungnak cu herbivory hnu ah special metabolites pawl i remhthannak nih hin a sangmi venhimnak man a ngeimi metabolites ah lam khatlei thlennak a chuahpi awk a si, a hlei in an thleidannak (random distribution he aa ralchanh in) Mode) MT theory nih a chimchungmi venhimnak metabolite. M1 ah aa pummi phenol chuahmi tam deuh cu rungrul riantuan ning a zornak he riantuannak in aa pehtlaimi an si (32). Suimilam 24 chung ah EV control thingram hna i a chuakmi thinghnah le aa telmi thinghnah karlak ah M1 metabolites chung i Si man kha tahchunh tikah, herbivory rungrul hnu ah metabolites tampi i metabolic specificity cu a karh chin lengmangmi a si ti kha kan hmuh (Figure 4C). Si man a karhnak cu aa vennak phenolamides lawng ah hmuh a si, asinain hi module chung i a ummi phenol dang le theih lomi metabolites ah Si man a karhnak a um lo. Mah cu a hleiin sermi model a si, cucu OD theory he aa pehtlaimi a si. Thlaici a eimi saram hna nih an chuahtermi thilri thlennak kong chimchungnak bik cu aa khat. Phenolamide spectrum i a hleiin a ummi hi OS-specific ET nih a chuahter maw ti hneksaknak caah, metabolite Si index kha kan suai i EV le sETR1 genotypes ah OSM le OSS1 karlak ah aa dangmi langhternak man kan chuahter (Figure 4D ). sETR1 ah, OSM le OSS1 karlak ah phenamide nih a chuahtermi thleidannak cu tampi a zor. Bi-clustering phunglam cu JA-regulated metabolic specialization he aa pehtlaimi a biapi bikmi MS/MS modules pawl hngalh khawhnak ding caah JA a tlawmmi phun hna ah lakmi MS/MS data zongah hman a si (Figure S6).
(A) 585 MS/MS tthencheunak (NDP aa lawhnak) le aa hrawmmi neutral sunghnak (NL aa lawhnak) cung ah hram bunh in fonhmi hmuhmi nih module (M) cu theihmi compound chungkhar he aa tlakmi a siter, asiloah theih lomi asiloah a ttha lomi Metabolite fonhmi nih a chuahpi. Module pakhat cio pawngah, metabolite (MS/MS) a hleiin (Si) phawtzamhnak kha langhter a si. (B) Modular molecular network: Nodes nih MS/MS le edges, NDP (sen) le NL (sen) MS/MS mark (cut-off,> 0.6) an aiawh. A tthencheu mi metabolite thleidannak index (Si) cu module (orhlei) cung ah hram bunh in rong in thlen a si i molecular network (orhlei) ah map tuah a si. (C) EV thingram M1 cu suimilam 24 chung ah a ummi (control) le a chuahtermi dirhmun (simulated herbivore) ah a um: molecular network diagram (Si man cu node ngan a si, a vennak phenolamide cu a sen in langhter a si). (D) EV le ET hmuh khawhnak a tlaumi spectrum line sETR1 i M1 molecular network diagram: a senmi kuang node nih a langhtermi phenolic compound, le Node ngan le W + OSM le W + OSS1 thlopnak karlak ah a biapi mi thleidannak (P man). 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 in wolfberry Son; Nick. O-AS, O-acyl sugar.
A derthawmmi Nepenthes genotype pakhat lawng in a chuakmi milu tiang hlathlainak cu kan kauhter chin, cuka ahcun thingthei a ei mi JA tlukruannak le a hleiin metabolite tlukruannak ah a tthawngmi intraspecific thlennak cu a ummi milu ah a rak langhter cang (26). Hi data set hi germplasm 43 khumh awkah hmang. Hi germplasms hna hi N. pallens chungin thingram phun 123 an si. Hi thingram hna hi Utah, Nevada, Arizona, le California (Figure S7) ah a ummi hmun dangdang ah lakmi thlaici chungin lakmi an si, metabolome phunphun (hika ah milu zat tiah auhmi) β phunphun) le OSM nih a chuahtermi thiamnak kha kan tuak. Hlan lio hlathlainak he aa tlak in, Hj le δj axes ah metabolic thlennak phunphun kan hmuh, cucu germplasms nih herbivores cungah an metabolic lehrulhnak ah a biapi mi thleidannak an ngei ti a langhter (Figure S7). Hi bu nih hin thingthei a eimi saram hna nih an chuahtermi JAs thlennak aa thleng lengmangmi kong he pehtlai in a hlan i hmuhtonnak hna kha a kan hngalhter ṭhan, cun milu pakhat ah man sang tukmi a ngeih peng (26, 36). Hj le δj karlak ah a dihlak in pehtlaihnak hneksaknak caah JA le JA-Ile hmannak thawngin, JA le metabolome β phunphun le thiamnak index karlak ah a biapi mi pehtlaihnak tha a um ti kan hmuh (Figure S7). Mah nih hin milu zat ah hmuhmi JAs chuahternak ah herbivour-induced heterogeneity cu rungrul herbivores chungin thimnak nih a chuahtermi a biapi mi metabolic polymorphisms ruangah a si kho men tiah a langhter.
Hlanlio hlathlainak nih a langhter ning ahcun zuu phun hna cu an phun le an i hngatchannak ah an i dang tuk i a chuahtertu le a dirhmi metabolic defenses cungah an i hngatchan. Anti-herbivore signal kuatnak le ralvennak thilti khawhnak ah thlennak hna hi rungrul tamnak, thingram nunnak caan, le phunkhat a ṭhannak hmun ah ralvennak chuah man nih a uk tiah zumh a si. North America le South America ah a ummi Nicotiana phun paruk hna i thinghnah a eimi saram hna nih an chuahtermi thinghnah metabolome remhnak aa khatmi kha kan hlathlai. Hi phun hna hi Nepenthes North America he aa pehtlaimi an si, cu hna cu Nicolas Bociflo an si. La, N. nicotinis, Nicotiana n. a dermi ramlak, Nicotiana tabacum, linear tobacco, tobacco (Nicotiana spegazzinii) le tobacco leaf tobacco (Nicotiana obtusifolia) (Figure 5A) (37). Hi phun hna lak ah paruk, a ttha tein langhtermi phun N. zaangfahnak tein, petunia phun chung i kum fatin a nungmi thingram an si, cun obtusifolia N. cu unau phun Trigonophyllae (38) chung i kum tampi a nungmi an si. Cu hnu ah, W + W, W + OSM le W + OSS1 induction cu hi phun sarih hna cungah rungrul rawl peknak phun-level metabolic remhthannak hlathlainak caah tuah a si.
(A) A ttha bikmi a si khomi [nuclear glutamine sernak caah (38)] le aa naihniammi Nicotiana phun sarih (rong aa dangmi) (37) hmunhma tthencheunak cung ah hram bunhmi bootstrap phylogenetic thingkung. (B) Nicotiana phun sarih (939 MS/MS; data file S1) hna i an pumsa thlennak sining caah a hleiin aa dangmi tthencheunak hmanthlak. Saram phun tlukruannak ah, metabolome phunphun cu thiamnak sang he aa pehtlaimi a si lo. Metabolic phunphun le thiamnak le JA khonnak karlak i phun-level pehtlaihnak hlathlainak cu hmanthlak 2 ah hmuh khawh a si. S9. Ron, phun dangdang; tlangthum, W + OSS1; rectangle, W + OSM; (C) Nicotiana JA le JA-Ile dynamics cu OS excitation amplitude (two-way ANOVA le Tukey HSD post-multiple comparison, * P <0.05, ** P <0.01 le * ** W + OS le W + W tahchunhnak caah, P <0.001) ningin an i tthen. Herbivorous le methyl JA (MeJA) i zohchunh hnu ah (D) phunphun le (E) phunkhat cio i an thiamnak box plot. Arfi hmelchunhnak nih W + OS le W + W asiloah lanolin plus W (Lan + W) asiloah Lan plus MeJA (Lan + MeJa) le Lan control (two-way analysis of variance, followed by Tukey's HSD post hoc multiple comparison , *P<0.05, **P<0.01 le ***P<0.01) karlak ah a biapi mi thleidannak a langhter.
Dual cluster phunglam hmangin, 939 MS/MS (data file S1) chungin module 9 kan hmuh. Thlopnak phunphun in remh ṭhanmi MS/MS fonhmi cu phun dangdang karlak ah aa dang tuk (Figure S8). Hj (hika ah phun-level γ-diversity tiah auhmi) le δj kha mitthlam ah cuanter tikah phun dangdang hna cu metabolic hmun ah phu dangdang ah an i fonh ti kha a langhter, cuka ahcun phun-level tthencheunak cu lungthawhnak nakin a lang deuh tawn. N. linear le N. obliquus ti lo ahcun, annih nih a kau ngaimi aa thleng lengmangmi thlennak thlennak an langhter (Figure 5B). Cu he aa dang in, N. purpurea le N. obtusifolia tibantuk phun hna nih cun thlopnak ah a fiang lomi metabolic lehrulhnak an ngei, asinain metabolome cu a phunphun deuh. A chuahtermi thlennak lehnak i phun-thleidannak phawtzamhnak nih thiamnak le gamma phunphun karlak ah a biapi mi a ttha lomi pehtlaihnak a chuahter (PCC = -0.46, P = 4.9×10-8). OS-induced thlennak cu JAs levels ah metabolome thiamnak he a ttha lei in aa pehtlai, cun phunkhat cio nih an langhtermi metabolic gamma phunphun he a chia lei in aa pehtlai (Figure 5B le Figure S9). Figure 5C ah “signal response” tiah auhmi phun hna, Nepenthes nematodes, Nepenthes nepenthes, Nepenthes acute, le Nepenthes attenuated tibantuk hna nih minutes 30 ah hmelchunhnak nganpi an chuahter ti kha hngalh awk tlak a si. Nai ah OS-specific JA le JA-Ile zawtnak a chuah lio ah, a dang bacteria “signal unresponsive” tiah auhmi, Nepenthes mills, Nepenthes powdery le N. obtusifolia tibantuk nih OS specificity zeihmanh um loin JA-Ile Edge induction lawng an langhter (Figure 5C). A cunglei ah kan chim cang bantuk in, metabolic level ah, a dermi Nepenthes caah, signal-a lehrulhmi thilri hna nih OS thleidannak an langhter i δj kha tampi in an karhter, cu lioah Hj kha an zorter. Hi OS-specific priming effect cu signal non-reactive species tiah tthenmi phun ah hmuh a si lo (Figure 5, D le E). OS-specific metabolites cu signal-a lehrulhmi phun hna karlak ah an i hrawm deuh, cun hi signal phu hna cu signal a lehmi a der deuhmi phun hna he an i fonh, cu lioah signal a lehmi a der deuhmi phun hna nih cun pakhat le pakhat i hngatchannak tlawm deuh an langhter (Figure S8 ). Hi hmuhmi nih hin OS-specific induction of JAs le OS-specific reconfiguration of downstream metabolome cu species level ah an i pehtlaih ti a langhter.
A hnu ah, thingram hna thlopnak caah methyl JA (MeJA) aa telmi lanolin paste kan hman i, hi pehtlaihnak lam hna hi thingram hna cytoplasm ah a ummi exogenous JA nih a hmanmi JA hmuh khawhnak nih a kham maw ti hlathlainak caah kan hman. Rapid desterification cu JA a si. JA pehzulh tein pekchanhnak nih a chuahtermi signal-a lehrulhmi phun in signal-a lehrulh lomi phun ah tlawmpal in aa thlengmi sining aa khatmi kan hmuh (Figure 5, D le E). A tawinak in, MeJA thlopnak nih linear nematodes, N. obliquus, N. aquaticus, N. pallens, le N. mikimotoi hna i an metabolomes kha fakpi in a remh hna, cu nih cun δj a karhter ngaingai i Hj a zorter. N. purpurea nih δj lawng a karhter, asinain Hj cu a langhter lo. N. obtusifolia, a hlan ah JAs tlawm tukmi a khonmi a si tiah langhter a si cang, MeJA thlopnak zongah metabolome remhthannak lei ah a ttha lo ngai in a lehrulh. Hi hmuhmi nih hin JA chuahternak asiloah signal thlennak cu signal a lehrulh lomi saram phun hna ah pumsa lei in kham a si ti a langhter. Hi ruahnak hneksaknak caah, W + W, W + OSMs le W + OSS1 Transcriptome (39) nih a chuahtermi phun li (N. pallens, N. mills, N. pink le N. microphylla) kha kan hlathlai. Metabolome remhnak phunglam he aa tlak in, phun pawl cu transcriptome hmun ah ttha tein an i tthen, cu hna lakah N. attenuated nih OS-induced RDPI a sang bik a langhter, N. gracilis nih a niam bik a langhter (Figure 6A). Sihmanhsehlaw, N. oblonga nih a chuahtermi transcriptome phunphun cu phun pali lakah a niam bik a si ti hmuh a si, N. oblonga i a sang bikmi metabolomic phunphun a hlan ah phun sarih ah langhtermi he aa ralchanh. Hlanlio hlathlainak nih a langhter ning ahcun, JA hmelchunhnak telh in, hlanlio i vennak hmelchunhnak he aa pehtlaimi genes phu pakhat nih Nicotiana phun ah herbivore he aa pehtlaimi elicitors nih a chuahtermi hlanlio venhimnak lehrulhnak a hleiin a fianter (39). Hi phun li karlak ah JA thawngthanhnak lam tahchunhnak nih a hngalh a nuammi sining pakhat a langhter (Figure 6B). Hi lamthluan chung i a ummi gene tam deuh, AOC, OPR3, ACX le COI1 tibantuk hna nih hi phun li ah hin a sang deuhmi induction an langhter. Sihmanhsehlaw, a biapimi gene, JAR4 nih JA cu a nunnak ah a cawlcangmi JA-Ile khonmi ttialmi phun ah a thlen, cun a ttialmi tlukruannak cu a niam tuk, a hlei in N. mills, Nepenthes pieris le N. microphylla ah. Cun, N. bifidum ah AOS gene dang pakhat i a ttialmi lawng hmuh a si lo. Gene langhternak ah thlennak hna hi signal anergic phun ah JA chuahnak a tlawmmi le N. gracilis chuahternak nih a chuahtermi a fakmi phenotypes caah tuanvo a lakmi a si kho.
(A) Herbivory induction hnu minutes 30 ah sample lakmi aa pehtlaimi zuu phun li hna i a hmasa bik ṭialmi lehnak reprogramming kong thawngpang theory hlathlainak. RDPI cu herbivore OS nih a chuahtermi thinghnah le hma khamnak tahchunhnak in tuak a si. Rong nih phun dangdang a langhter, cun hmelchunhnak nih thlopnak phun dangdang a langhter. (B) Saram phun li lakah JA thawngthanhnak lam ah gene langhternak hlathlainak. A fawitermi JA lam cu box plot pawngah langhter a si. Ron aa dangmi nih thil tuahning aa dangmi a langhter. Arfi hmelchunhnak nih W + OS thlopnak le W + W uknak karlak ah a biapi mi thleidannak a um ti a langhter (Pairwise thleidannak caah Student t-test caah, *P<0.05, **P<0.01 le ***P<0.001). OPDA, 12-oxophytodienoic acid; OPC-8: 0,3-oxo-2(2′(Z)-pentenyl)-cyclopentane-1-octanoic acid.
A donghnak ah, rungrul phun dangdang hna i an metabolome remhthannak nih thingthei a eimi saram hna kha zeitindah a tei khawh hna ti kha kan hlathlai. Hlan lio hlathlainak nih Nicotiana phun kha a biapi ah a chiah. Ms le larvae an doh khawhnak cu aa dang tuk (40). Hika ah, hi model le an metabolic plasticity karlak pehtlaihnak kha kan hlathlai. A cunglei zuu phun li hmangin, le herbivores nih a chuahtermi metabolome phunphun le thiamnak karlak pehtlaihnak le thingram hna nih Ms le Sl an doh khawhnak karlak pehtlaihnak hneksaknak in, generalist Sl All doh khawhnak, phunphun le thiamnak cu a ṭhami pehtlaihnak kan hmuh, cu lioah thiamsang nu pawl dohnak le thiamnak karlak pehtlaihnak cu a der, cun thleidannak he pehtlaihnak cu a biapi lo (Figure 10). S1 doh khawhnak he pehtlai in, a dertermi N. chinensis le N. gracilis pahnih, a hlan ah JA signal thlennak tlukruannak le metabolome plasticity pahnih an langhter cangmi hna nih, herbivore luhternak ah aa dang tukmi lehrulhnak an ngei, cun annih zong nih aa khatmi doh khawhnak sang an langhter. Nu le pa sualnak.
A luancia kum sawmruk chung ah, thingram kilvennak theory nih theory phunglam a kan pek, cu cungah hram bunh in hlathlaitu hna nih thingram a hleiin a ummi thilri hna i an thlennak le an riantuannak tampi an rak chimchung. Hi theory tam deuh nih hin a fekmi ruahnak (41) timi a tlangpi phunglam kha an zulh lo. Hlathlainak tlukruannak ah a biapimi chimchungnak (3) an chuahpi. A biapimi chimchungnak hna hneksaknak nih a bik in ruahnak cheuhnak hna kha hlathlainak tuah khawhnak a pek tikah, mah nih hin The field is moving forward a tuah lai. Thazaang pe, asinain midang kha hlawt hna (42). Cu nakin, theory thar nih hlathlainak tthen dangdang ah chimchungnak a tuah i fianternak ruahnak cheuhnak tthen thar a chap (42). Sihmanhsehlaw, riantuannak tlukruannak ah chuahpimi theory pahnih, MT le OD, cu fawi tein a biapi mi chimchungnak ah a biapi mi chimchungnak ah fianter khawh a si: OD theory nih cun a hlei in thlennak “hmun” ah thlennak cu lamhruainak sangpi a ngeimi a si tiah a zumh. MT theory nih cun hi thlennak hna hi lam a kal lomi le metabolic hmun ah a ummi an si lai tiah a zumh, cun a sangmi ralvennak man metabolites an ngei tawn. OD le MT chimchungnak hna i a hlan i hlathlainak hna cu a priori “vennak” compound tlawmte hmangin hneksaknak an tuah cang. Hi metabolite-centric hneksaknak hna nih hin herbivory lio ah metabolome remhthannak a kauhnak le a kalning hlathlainak tuah khawhnak kha a kham, cun a dihlak in ruah khawhmi a biapi mi chimchungnak a herhmi aa khatmi zatlang cazin chung ah hneksaknak tuahnak nawl a pe lo Thlaici metabolome thlennak kha tuaktan. Hika ah, tuaktannak MS cung ah hram bunhmi metabolomics ah thiamnak thar kan hman i vawleicung metabolomics level ah ruahnak cheuhnak pahnih karlak i thleidannak hneksaknak caah thawngpang theory fiantertu hna i a tlangpi tangka ah deconvolution MS hlathlainak kan tuah. Hi theory i a biapi bikmi chimchungnak. Thawngpang theory cu rian tampi ah hman a si cang, a hlei in biodiversity le rawl tthatnak kong hlathlainak ah hman a si cang (43). Sihmanhsehlaw, kan theih khawh tawk ahcun, hi cu thingram hna i an thilri thlennak konglam langhternak le pawngkam hmelchunhnak hna lehrulhnak ah caan tawite chung thilri thlennak he aa pehtlaimi ecological buaibainak hna tawlrelnak caah hmanmi application hmasa bik a si. A bik in, hi lamthluan thilti khawhnak cu thingram phun dangdang chung le karlak i a ummi thil sining hna kha tahchunh khawhnak ah a um i, zeitindah thingram a ei mi hna cu phun dangdang in thlennak tlukruannak tlukruannak ah phun dangdang karlak macroevolutionary thil sining hna ah an i thlen timi zohfel khawhnak ah a um. Metabolism.
Principal component analysis (PCA) nih cun multivariate data set kha dimensionality reduction space ah a thlen i cucaah data i a tlangpi in a kalning kha a fianter khawh, cucaah deconvolution metabolome tibantuk data set kha hlathlainak tuahnak ah hman a si tawn. Sihmanhsehlaw, a tlawmternak nih data set chung i a ummi thawngpang cheukhat a sung lai, cun PCA nih ecological theory he aa pehtlaimi sining hna kong he pehtlai in zatlang thawngpang a pek kho lai lo, cu hna cu: zeitindah herbivores nih a hleiin rian (tahchunhnak ah, rumnak, phawtzamhnak) le tamnak) metabolites ah a phunphun in an sersiam? Zei metabolites nih dah herbivore pakhatkhat a chuahtertu dirhmun a chimchungtu an si? A hleiin, a phunphun le a chuahter khomi hmuhning in, thinghnah-a hleiin metabolite profile chung i a ummi thawngpang cu a rawk, cun thingthei a eimi saram einak nih a hleiin metabolism a chuahter khawh ti kha hmuh a si. Ruah lo piin, kan hmuhmi cu, hmanmi thawngpang theory hmelchunhnak ah langhtermi bantuk in, a chuakmi thilri thlennak dirhmun cu saram pahnih (zan ah rawl a eimi generalist Sl) le Solanaceae thiamsang Ms. nih an tuk hna hnu ah aa tluk ngaingai. An rawl ei ning le an lungthin peknak cu aa dang ngai ko nain. OS (31) ah thau acid-amino acid fonhmi (FAC) thawktu. A tlarimi hliamhma thlopnak caah herbivore OS hmannak thawngin, herbivore thlopnak lemsoinak zong nih aa lo ngaimi sining a langhter. Thlaici nih ramlak saram nih an tuk hna tikah an lehrulh ning zohchunh awk ah a tlarimi phunglam nih hin saram rawl ei ning thlennak nih a chuahtermi buaibainak thil hna kha a hloh, cu nih cun caan dangdang ah rawhralnak a phunphun a chuahter (34). OSM a chuahtertu bik tiah theihmi FAC nih OSS1 ah JAS le a dang thingram hormone lehrulhnak a zorter, OSS1 nih cun a let za tampi a zorter (31). Sihmanhsehlaw, OSS1 nih OSM he tahchunh tikah JA khonnak tlukruannak a chuahter. A hlan ah a derthawmmi Nepenthes ah JA lehrulhnak cu OSM cungah a ttha tuk ti kha langhter a si cang, cuka ahcun FAC nih ti he 1:1000 in cawh hmanh ah a cawlcanghnak a kilven khawh (44). Cucaah, OSM he tahchunh ahcun, OSS1 chung i FAC cu a niam tuk ko nain, JA zawtnak a chuahter khomi a si. Hlan lio hlathlainak nih porin-bantuk protein (45) le oligosaccharides (46) cu OSS1 ah thingram kilvennak lehrulhnak chuahter awkah molecule hmelchunhnak ah hman khawh a si ti a langhter. Sihmanhsehlaw, OSS1 chung i hi elicitors pawl hi atu lio hlathlainak ah hmuhmi JA khonnak caah tuanvo a ngeimi an si maw ti cu a fiang rih lo.
Herbivors phun dangdang asiloah exogenous JA asiloah SA (salicylic acid) (47) hmannak nih a chuahtermi aa dangmi metabolic kutdonghrolh kong a langhtermi hlathlainak tlawmte lawng an um ko nain, ahohmanh nih herbivors phun-specific perturbation ah thingram hrampi network le a thlennak kha a buaiter lo. pumpak konglam. Hi hlathlainak nih hin JAs ti lo cu thingram hormone dang hna he chunglei hormone network pehtlaihnak nih herbivores nih a chuahtermi metabolic remhnak a hleiin a ser ti kha a fehter chin. A bik in, OSM nih a chuahtermi ET cu OSS1 nih a chuahtermi nakin a tam deuh ti kha kan hmuh. Hi mode hi OSM chung i FAC tam deuh he aa tlak, cucu ET puahnak chuahternak caah a herhmi le a za tawk dirhmun a si (48). Thingkung le thingthei a eimi saram hna karlak pehtlaihnak kongkau ah, ET nih thingram-specific metabolite dynamics cung ah a langhtermi riantuannak cu a tlawmpal lawng a si rih i compound phu pakhat lawng a target. Cun, hlathlainak tam deuh nih ET asiloah a hlan i a chuakmi thilri asiloah khamnak phunphun kha ET uknak hlathlainak caah lenglei in hmannak an hman, cu hna lakah hi lenglei chemical hmannak nih a hleiin a ttha lomi thil tampi a chuahter lai. Kan theih khawh tawk ahcun, hi hlathlainak nih hin ET hmannak ah ET nih a ttuanvo a tthatnak le a tlamtling lomi transgenic thingram pawl chuah khawhnak ding caah thingram metabolome dynamics tawlrel khawhnak ding caah a hmasa bik a nganmi ningcang tein zohfelnak a aiawh. Herbivore-specific ET induction nih a donghnak ah metabolome lehrulhnak kha a thlen khawh. A biapi bikmi cu ET biosynthesis (ACO) le hmuh khawhnak (ETR1) genes hna i thlennawnnak thlennak a si i, cu nih cun herbivore-specific de novo phenolamides khonnak a langhter. ET nih putrescine N-methyltransferase (49) a uk khawhnak thawngin JA-induced nicotine khonnak kha a ttha tein a remh khawh ti kha a hlan ah langhter a si cang. Sihmanhsehlaw, seh lei zoh tikah, ET nih zeitindah phenamide chuahternak kha a ttha tein a remh timi cu a fiang lo. ET i signal thlennak riantuannak lengah, polyaminophenol amides ah tangka chiahnak kha tawlrel awkah S-adenosyl-1-methionine ah thlen khawh a si. S-adenosyl-1-methionine cu ET le polyamine biosynthetic lamthluan ah a tlangpi in a ummi a si. ET signal nih phenolamide dat a uk khawhnak lam cu hlathlainak tuah a herh rih.
Caan saupi chung, a sining theih lomi a hleiin a ummi thilri phunphun an tam tuk caah, a hleiin thilri thlennak phun hna ah fakpi in lungthin peknak nih biological pehtlaihnak hnu ah thilri thlennak phunphun caan thlennak kha fek tein a tuaktan kho lo. Atu lio ah, thawngpang theory hlathlainak cung ah hram bunh in, thleidannak a ngei lomi metabolites cung ah hram bunh in MS/MS spectrum hmuhnak nih a chuahpimi a biapi bikmi cu, herbivors nih herbivors a eimi asiloah a zohchunhmi nih leaf metabolome a dihlak in metabolic phunphun a zorter i a thiamnak a karhter. Herbivores nih a chuahtermi metabolome thleidannak ah caan tawite karhnak cu transcriptome thleidannak ah aa bawmmi karhternak he aa pehtlai. Hi a ngan deuhmi metabolome thleidannak (Si man sang deuh a ngeimi) ah a bawmtu bikmi sining cu a hlan ah langhtermi thingthei a eimi riantuannak a ngeimi a hleiin metabolite a si. Hi model cu OD theory chimchungnak he aa tlak, asinain metabolome reprogramming randomness he aa pehtlaimi MT chimchungnak cu aa tlak lo. Sihmanhsehlaw, hi data cu cawhmi model (a tha bikmi MT; Figure 1B) chimchungnak he zong aa tlak, zeicahtiah a dang sining langhter lomi metabolites pawl cu theih lomi venhimnak riantuannak he random Si phawtzamhnak kha an zulh rih men lai.
Hi hlathlainak nih a tlaihmi hngalh awk tlakmi sining pakhat cu micro-evolution tlukruannak (thingram pakhat le zuu phun) in a ngan deuhmi thlennawnnak tlukruannak (a naih tein aa pehtlaimi zuu phun) tiang, thlennawnnak dirhmun aa dangmi hna cu “a ṭha bikmi kilvennak” ah an um. Thlaici a eimi saram hna an thilti khawhnak ah a biapi tukmi thleidannak a um. Moore le a dangdang. (20). hi caṭialtu hna nih hin a tawinak in an chim lo. Metabolome data nganpi laknak caah tuah ningcang hna zong nih hi data chungin metabolic phunphun tuak ning a langhter lo. Hi hlathlainak ah, Whittaker riantuannak thleidannak ah remhnak hmete nih α-metabolic phunphun kha thingram pakhatkhat ah MS/MS spectra phunphun a si tiah a ruah lai, cun β-metabolic phunphun cu milu phu pakhat i a hrampi intraspecific metabolism ah a ruah lai.
JA signal cu thingthei a eimi saram hna i an thlennak phunphun caah a herhmi a si. Sihmanhsehlaw, JA biosynthesis i intraspecific uknak nih metabolome phunphun ah a bawmhnak kongah a fekmi zatlang hneksaknak a um lo, cun JA signal cu a sang deuhmi macroevolutionary tahfung ah lungretheihnak nih a chuahtermi metabolic phunphun caah a tlangpi hmun a si maw ti cu a fiang rih lo. Nepenthes thingthei a ei mi hna nih hin metabolome thiamnak a chuahter i Nicotiana phun hna le aa pehtlaimi Nicotiana phun hna lakah metabolome thiamnak aa dangmi cu JA signaling he aa pehtlaimi a si ti kha kan hmuh. Cun, JA signal a rawh tikah, genotype pakhat lawng a simi herbivore nih a chuahtermi metabolic specificity cu a rawk lai (Figure 3, C le E). A sining tein a dermi Nepenthes milu hna i an metabolic spectrum thlennak cu a tam deuh cu zat lei a si caah, hi hlathlainak ah metabolic β phunphun le a hleiin thlennak cu metabolite a tammi compound phun hna i an lungthawhnak a tthawngmi ruangah a si kho men. Hi thilri phun hna nih hin metabolome profile cheukhat kha an uk i JA signals he pehtlaihnak tha a chuahter.
Zeicatiah amah he aa pehtlaimi zuu phun hna i an biochemical mechanism cu aa dang tuk caah, metabolites pawl cu a hleiin sining lei ah an langhter hna caah, hlathlainak lei ah a si deuh. Information theory nih a tlaihmi metabolic profile a tawlrel ning nih herbivorous induction nih metabolic gamma phunphun le thiamnak karlak i chawlehnak a zualter ti a langhter. JA signal nih hi chawlehnak ah a biapi tukmi hmunhma a lak. Metabolome thiamnak a karhmi cu OD chimchungnak he aa tlak i JA signal he a ṭhami pehtlaihnak a ngei, cu lioah JA signal cu metabolic gamma phunphun he a ṭhalomi pehtlaihnak a ngei. Hi model hna nih hin thingram hna i OD thilti khawhnak cu JA i a tthawnnak nih a khiah bik ti a langhter, microevolutionary tahfung ah siseh, a ngan deuhmi thlennak tahfung ah siseh. JA biosynthesis tlamtlin lonak a hrialmi exogenous JA hmannak hneksaknak nih a naihmi zuu phun pawl cu signal-a lehrulhmi le signal-a lehrulh lomi phun ah thleidan khawh an si ti kha a langhter chin, an JA phun le herbivores nih a chuahtermi metabolome plasticity bantuk in. Signal a leh lomi phun hna cu an chunglei JA an chuah khawh lo caah an leh kho lo i cucaah pumsa lei rikhiahnak an ngei. Mah cu JA thawngthanhnak lam (N. crescens ah AOS le JAR4) chung i a biapimi gene cheukhat thlennak ruangah a si kho. Hi hmuhmi nih hin hi phun dangdang thlennak sining hna hi chunglei hormone hmuhning le lehrulhnak thlennak nih a hruai hna ti a langhter.
Thingkung le thingthei a eimi saram hna karlak pehtlaihnak lengah, thilri thlennak phunphun kawlnak cu pawngkam he aa tlakmi thilnung thlennak le a harmi phenotypic sining thlennak hlathlainak ah a biapi mi ruahnak tthanchonak vialte he aa pehtlai. Tuchan MS thilri hna nih an hmuhmi data a karh chin lengmang caah, metabolic phunphun kongah ruahnak cheuhnak hneksaknak nih pumpak/phung metabolite thleidannak kha a lonh khawh cang i ruahlopi in a cangmi thil pawl langhter awkah vawleicung pumpi hlathlainak a tuah khawh cang. A nganmi hlathlainak tuahnak ah, a biapi mi tahchunhnak cu data hlathlainak caah hman khawhmi sullam a ngeimi map sernak ruahnak a si. Cucaah, a tu lio ah thleidannak a ngei lomi MS/MS metabolomics le thawngpang theory fonhmi nih a chuahpimi a biapi mi cu, taxonomic tahfung dangdang ah metabolic phunphun zoh khawhnak ding caah map sernak ah hman khawhmi a sawhsawh mi tahfung a pek. Hi lamthluan nih a herhmi a hrampi a si. Micro/macro thlennak le zatlang nunphung kong hlathlainak.
Macro-evolutionary level ah, Ehrlich le Raven (51) nih an sermi thingram-rungrul co-evolution theory hrampi cu phun dangdang thlennak aa dangmi nih thingram phun dangdang a chuahtertu a si tiah chimchung ding a si. Sihmanhsehlaw, hi rian biapi chuah hnu kum sawmnga chungah, hi ruahnak cheuhnak hi hneksak a si bal lo (52). Hihi a ruang bik cu a hlatnak thingram phun hna karlak ah aa tlukmi thlennak sining hna i phylogenetic sining hna ruangah a si. Rarity cu target hlathlainak lamthluan hna kha fek tein dirh khawhnak caah hman khawh a si. Atu lio MS/MS riantuan ning cu thawngpang theory nih a tawlrelmi nih theih lomi thilri hna i MS/MS sining aa lawhnak kha a tah (a hlan ah thilri thimnak um loin) i mah MS/MS pawl kha MS/MS phunkhat ah a thlen hna, cucaah thiamsang thilri thlennak ah Hi macro-evolutionary model pawl cu thleidannak tahfung ah an tahchunh hna. A sawhsawh mi zatlang relnak hmelchunhnak. Mah thil kalning cu phylogenetic hlathlainak he aa lo, mah nih cun a hlankan in chimchungnak um loin a phunphun in thlennak asiloah sining thlennak rate kha tuaktan awkah a zulmi tlukruannak kha a hman khawh.
Biochemical level ah, Firn le Jones (53) nih an tuahmi thleidannak ruahnak nih a langhter mi cu, a hlan ah aa pehtlaimi asiloah aa thlengmi metabolites hna i an nunnak cawlcanghnak tuah khawhnak ding caah thilri pawl pek khawhnak ding caah, metabolic phunphun cu level dangdang ah a um peng. Theihhngalhnak theory lamthluan nih hin metabolite-specific thlennak thlennak hna cu metabolite thleidannak chung ah a cangmi hna hi thlennawnnak thleidannak kalning ah aa telmi ah tah khawh a si: a tlawmmi in a sangmi thlennak ah biologically active adaptation Pawngkam pakhatkhat i a khammi metabolites hna.
A dihlak in, molecular biology a hramthawk lio ah, a biapi mi thingram kilvennak theory pawl an rak ser, cun deductive hypothesis-driven methods cu scientific thanchonak lam pakhat lawng a si tiah mi tampi nih an rak ruah. Mah cu a bik in metabolome dihlak tahnak ah thiamnak lei rikhiahnak ruangah a si. Ruahnak cheuhnak nih a hruaimi lam hna cu a dang a ruang a chuahtertu thil sining hna thimnak ah a hleiin a ttha ko nain, biochemical network kong kan hngalhthiamnak a thanchoter khawhnak cu atu chan data-intensive science ah a ummi tuaktannak lam hna nakin a tlawm deuh. Cucaah, chimchung khawh lomi theory pawl cu hmuh khawhmi data leng ah an um caah, hlathlainak lei ah tthanchonak a ummi ruahnak cheuhnak formula/hneksaknak tthanchonak cu hloh khawh a si lo (4). Hika ah langhtermi metabolomics tuaktannak riantuannak nih naite (zeitindah) le a donghnak (zeicah) metabolic phunphun kong ah lungthawhnak a chuahter ṭhan khawh lai ti kha kan hmuhchung, cun theory lei in lamhruaimi data science chan thar ah a bawmh khawh lai. Cu chan nih cun a hlan chan hna thazaang a petu a biapi mi ruahnak hna kha a zohfel tthan hna.
Herbivore rawl peknak cu a pahnihnak instar larva asiloah Sl larva kha pangpar a parmi thingram pakhat i a senmi pitcher thingram hnah pakhat cung ah cinnak in tuah a si, thingram pakhat ah thingram 10 aa khat. Rungrul larvae pawl cu clamps in an hren hna, cun a tangmi hnah tissue cu zawtnak an ngeih hnu suimilam 24 le 72 ah an lak hna i rang tein an hnim hna, cun metabolites pawl cu an chuah hna.
Herbivorous thlopnak kha aa tlak ngaimi lam in i zohchunh. A tuah ning cu puan suaimi kedan hmangin puan hri a tthang lio ah thingram a kau ngaimi thinghnah pathum hna i a laifang ah hling tlar pathum kha cawh, cun 1:5 Diluted Ms kha thuh colh. Asiloah kutdonghrolh hmangin S1 OS kha hma chungah khumh. A cunglei langhtermi bantuk in thinghnah pakhat kha lak law thlen. A hlan ah langhtermi lam kha hmang law a hrampi thilri le thingram hormone pawl kha chuah (54).
Exogenous JA hmannak caah, phunkhat cio i pangpar par paruk hna i an hnah pathum cu 150μg MeJA (Lan + MeJA) aa telmi lanolin paste 20μl le hma thlopnak (Lan + W) 20μl he thlopbul an si, asiloah 20μl lanolin thianghlim kha uk khawhnak caah hman an si. Thlaici pawl cu thlop hnu suimilam 72 ah an lak hna, ti nitrogen ah an lumter hna, cun hman hlan tiang -80°C ah an chiah hna.
JA le ET transgenic phun li, irAOC (36), irCOI1 (55), irACO le sETR1 (48) hna cu kan hlathlainak phu ah hmuh an si. irAOC nih JA le JA-Ile tlawmternak fakpi in a langhter, irCOI1 cu JAs cungah a tuar kho lo. EV he tahchunh ahcun, JA-Ile khonnak a karh. Cu bantuk cun, irACO nih ET chuahternak a zorter lai, cun EV he tahchunh ahcun, ET a tuar kho lomi sETR1 nih ET chuahternak a karhter lai.
Photoacoustic laser spectrometer (Sensor Sense ETD-300 real-time ET sensor) cu ET tahnak kha a luhhnawh lomi in tuah awkah hman a si. Thlopnak an tuah hnu tlawmpal ah, thinghnah cheukhat cu an tan hna i 4-ml kharmi thlalang thawl chungah an chiah hna, cun a lu cung hmun cu suimilam 5 chungah a khon khawh nakhnga an tuah. Tahnak chungah, vial pakhat cio cu minutes 8 chung thli thiang 2 liters/hour in an thlet hna, cu thli cu a hlan ah Sensor Sense nih CO2 le ti chuah khawhnak ding caah a pekmi catalyst chungin a rak kal cang.
Microarray data cu a hramthawk ah (35) ah chuah a si i National Center for Biotechnology Information (NCBI) Gene Expression Comprehensive Database (accession number GSE30287) ah chiah a si. W + OSMs thlopnak le a rawk lomi control nih a chuahtermi thinghnah he aa pehtlaimi data cu hi hlathlainak caah chuah a si. A rawkmi thazaang cu log2 a si. Statistical hlathlainak tuah hlan ah, R software package hmangin a hrampi cu thlen le a 75th percentile ah a tlarimi ah ser a si.
Nicotiana phun hna i a hramthawk RNA zulhphung (RNA-seq) data cu NCBI Short Reading Archives (SRA) in lakmi a si, project nambar cu PRJNA301787 a si, cucu Zhou et al nih an thanh. (39) le (56) ah langhtermi ningin kalpi. Nicotiana phun he aa tlakmi W + W, W + OSM le W + OSS1 nih an tawlrelmi raw data cu hi hlathlainak ah hlathlainak caah thim an si, cun a tanglei bantuk in an tuah: Pakhatnak ah, raw RNA-seq relmi cu FASTQ format ah an thlen. HISAT2 nih FASTQ cu SAM ah a thlen, cun SAMtools nih SAM file pawl kha tthencheu mi BAM file ah a thlen. StringTie cu gene langhternak tuaktannak caah hman a si, cun a langhternak lam cu a zulmi ṭialmi tthencheunak million pakhat ah base tthencheunak thongkhat ah tthencheunak a um.
Hlathlainak ah hmanmi Acclaim chromatographic column (150 mm x 2.1 mm; particle size 2.2μm) le 4 mm x 4 mm ralvengtu column cu thilri pakhat in an ser. A tanglei binary gradient cu Dionex UltiMate 3000 Ultra High Performance Liquid Chromatography (UHPLC) system ah hman a si: 0 in 0.5 minutes, isocratic 90% A [deionized ti, 0.1% (v/v) acetonitrile le 0.05% formic acid] B, 10% le acetonitrile 0.05% formic acid); 0.5 in 23.5 minutes karlak ah, a ttumchuknak tthen cu 10% A le 90% B, ciocio; 23.5 in 25 minutes, isocratic 10% A le 90% B. A luan ning cu 400μl/min a si. MS hlathlainak vialte caah, column eluent kha quadrupole le time-of-flight (qTOF) hlathlainak ah thun, cu ah cun electrospray source cu positive ionization mode (capillary voltage, 4500 V; capillary outlet 130 V ; Drying temperature 200°C; drying temperature 10 liters/min) in rian a ttuan.
MS / MS tthencheunak hlathlainak (a hnu ah MS / MS tiah auh a si lai) kha data he aa pehtlaimi asiloah thleidan khawh lomi tuah law a dihlak in hmuh khawhmi thilri thlennak sining kong he pehtlai in sining kong theihternak hmuh khawh a si. Thleidannak a ngei lomi MS/MS phunglam ruahnak cu quadrupole nih a ngan tukmi thilri thleidannak window a ngeih timi cungah aa hngat [cucaah, thilri-le-charge ratio (m/z) signal vialte kha tthencheu bantuk in ruat hna]. Hi ruang ah, Impact II thilri nih CE tilt a ser khawh lo caah, a karhmi i pahnak-induced dissociation collision energy (CE) man hmangin aa hngatmi hlathlainak zeimawzat an tuah. A tawinak in, UHPLC-electrospray ionization/qTOF-MS hmangin sample kha hmasa bik ah zohfel hmasa law, single mass spectrometry mode (in-source fragmentation nih a chuahtermi tlawmte tthencheunak dirhmun) hmangin, m/z 50 in 1500 tiang 5 Hz tluk in tthencheunak frequency in zohfelnak tuah. MS/MS hlathlainak caah nitrogen kha i pahnak gas ah hmang law, a tanglei i pahnak nih a chuahtermi tthencheunak voltage phun li ah aa hngatmi tahnak tuah: 20, 30, 40, le 50 eV. Tahnak kalning chung vialte ah, quadrupole nih a ngan bikmi rit thleidannak window a ngei, m/z 50 in 1500 tiang. Hmailei pum m/z le thleidannak kauhnak hneksaknak cu 200 ah chiah a si tikah, rit tthennak cu thilri riantuannak software le 0 Da nih amah tein a rian a ttuanter. Mass pakhat lawng mode bantuk in mass tthencheunak kha zohfel. Mass calibration caah sodium formate (50 ml isopropanol, 200 μl formic acid le 1 ml 1M NaOH aqueous solution) kha hmang. Bruker's high-precision tahfung algorithm hmangin, caan khiahmi chung ah a tlangpi in spectrum a kal hnu ah data file cu tahfung tuah a si. Raw data file pawl kha NetCDF format ah thlen awkah Data Analysis v4.0 software (Brook Dalton, Bremen, Germany) i chuahnak riantuannak kha hmang. MS/MS data set cu a ongmi metabolomics database MetaboLights (www.ebi.ac.uk) ah luh khawhnak nambar he chiah a si cang. MTBLS1471.
MS/MS pumhnak cu MS1 le MS/MS quality signals karlak pehtlaihnak hlathlainak in a niam le a sangmi i pahnak thazaang le phung thar hmanmi hna caah tuah khawh a si. R script cu thilri ah a hlan i a ummi thilri phawtzamhnak pehtlaihnak hlathlainak tuah khawhnak caah hman a si, cun C# script (https://github.com/MPI-DL/indiscriminant-MS-MS-assembly-pipeline) cu phunglam hna tuah khawhnak caah hman a si.
Hnulei aw-an ruangah a chuakmi a hmaan lomi a ttha mi palhnak le sample tlawmte lawng ah m/z sining cheukhat hmuhchuah ruangah a chuakmi a hmaan lomi pehtlaihnak zorternak caah, R package XCMS (hnulei aw-an remhnak caah) i “filled peak” riantuannak kha kan hman. Fill peak function hman tikah, pehtlaihnak tuaktannak a hnorsuan dingmi “0′′ intensity values ​​data set chungah tampi an um rih. Cun, filled peak function hman tikah le filled peak function hman lo tikah hmuhmi data processing results kha kan tahchunh, cun a tlangpi in remhmi cung ah hram bunh in hnulei aw-an man kha kan tuak, cun mah man hna kha 0 in kan thlen hna tuakmi hnulei man. Kannih zong nih a tthawngmi hnulei man let thum a lonhmi sining lawng kan ruah i “a hmaanmi sangbik” tiah kan ruah hna. PCC tuaktannak caah, sample precursor (MS1) le a tlawmbik a dikmi sangbik pariat a ngeimi fragment data set hna i m/z signals lawng kha ruah chih an si.
Sample dihlak chung i a hlan i a tthatnak sining a tthawnnak cu a tlawmmi asiloah a sangmi i pahnak thazaang a tongmi a tthatnak sining aa khatmi a tthawnnak a zormi he aa pehtlaimi a si ahcun, cun sining cu CAMERA nih isotope peak tiah min a pek lo ahcun, a dang tein fianter khawh a si. Cun, 3 s chung ah a si khomi precursor-product pahnih vialte tuaktannak in (a sangbik tlaih khawhnak caah tuaktanmi tlaih khawhnak caan), pehtlaihnak hlathlainak tuah a si. m/z man cu a hlan man nakin a niam deuh i MS/MS tthencheunak cu data set chung i a chuahkehnak precursor he aa khatmi sample hmun ah a chuah lawngah, tthennak ah ruah a si.
Hi phunglam sawhsawh pahnih cungah hram bunh in, hngalhmi a hramthawknak m/z nakin a ngan deuhmi m/z man a ngeimi a hleiin langhtermi tthencheunak hna kha kan chuah hna, cun a hlankan in a langmi sample dirhmun le a langhtermi tthencheunak cungah hram bunh in. Cun MS1 ​​mode in chuahmi in-source tthencheunak tampi nih a chuahpimi tthatnak sining hna kha candidate precursors ah thim khawh a si, cucaah a herh lomi MS/MS compound pawl a chuahter. Hi data tlawmternak caah, spectra hna i NDP aa lawhnak nih 0.6 a lonh ahcun, cun CAMERA nih a langhtermi chromatogram “pcgroup” ah an i tel ahcun, kan fonh hna lai. A donghnak ah, aa dangmi i pahnak thazaang ah m/z man aa khatmi candidate peak vialte lakah a sang bikmi tthawnnak peak thim in a donghnak deconvoluted composite spectrum ah precursor le fragments he aa pehtlaimi CE hmuhmi pali vialte kha kan fonh hna. A hnu i tuahsernak step pawl cu composite spectrum timi ruahnak cungah hram bunh in an si i tthencheunak a chuah khawhnak ding caah a herhmi CE thil sining phunphun kha an lak, zeicahtiah tthencheunak cheukhat cu a hleiin i pahnak thazaang tangah lawng hmuh khawh an si.
RDPI (30) cu metabolic profile a chuahter khawhnak tuaktannak caah hman a si. Metabolic spectrum diversity (Hj index) cu Martínez le a hawile nih an langhtermi a tanglei equation hmangin MS/MS frequency distribution Shannon entropy hmangin MS/MS precursors tamnak in lakmi a si. (8). Hj = −∑i = 1mPijlog2(Pij) cuka ah Pij cu j-th sample (j = 1, 2,..., m) (i = 1, 2, …, m) t) chung i i-th MS/MS i aa pehtlaimi frequency he aa tlak.
Metabolic specificity (Si index) cu ruahmi sample karlak i a cangmi he pehtlai in pekmi MS/MS langhternak aa khatmi tiah fianter a si. MS/MS thleidannak cu Si = 1t (∑j = 1tPijPilog2PijPi) tiah tuak a si
j sample pakhat cio i metabolome-specific δj index tah awkah a tanglei formula kha hmang law, MS/MS specificity δj = ∑i = 1mPijSi a tlangpi kha hmang
MS/MS spectra cu pahnih in an i tluk, cun aa lawhnak cu mark pahnih cungah hram bunh in tuak a si. Pakhatnak ah, standard NDP (cosine pehtlaihnak lam tiah auhmi) hmangin, a tanglei equation hmangin spectra NDP = (∑iS1 & S2WS1, iWS2, i) 2∑iWS1, i2∑iWS2, i2 ah S1 le S2 he aa tlak in, spectrum 1 le 2 caah, WS1 le WS, i2 spectra pahnih karlak i i-th common peak i dannak cu 0.01 Da nak tlawm deuh a si timi peak intensity cung ah hram bunh in a ritmi kha a aiawh. A ritmi cu a tanglei bantuk in tuak a si: W = [a sangbik thazaang] m [tthatnak] n, m = 0.5, n = 2, MassBank nih ruahnak a pekmi bantuk in.
A pahnihnak mark peknak lam cu hman a si, cu ah cun MS/MS karlak ah aa hrawmmi NL hlathlainak aa tel. Cu caah, MS tthencheunak kalning ah ton tawnmi NL cazin 52 kha kan hman, cun a derthawmmi Nepenthes phun ( 9, 26) i a changtu thilri hna i MS/MS spectrum caah a hlan ah an rak tialmi a bik in NL (data file S1) kha kan hman. MS/MS pakhat cio caah 1 le 0 in binary vector ser, NL cheukhat a tu lio le a um lomi he aa tlak. Euclidean hlatnak aa lawhnak cungah hram bunh in, NL aa lawhnak mark cu binary NL vector pahnih cio caah tuak a si.
Dual clustering tuah awkah, R package DiffCoEx kan hman, cucu Weighted Gene Co-expression Analysis (WGCNA) karhternak cungah hram aa bunh. MS/MS spectra i NDP le NL mark peknak matrices hmangin, tahchunhnak pehtlaihnak matrix tuaktannak caah DiffCoEx kan hman. Binary clustering cu “cutreeDynamic” parameter kha method = “hybrid”, cutHeight = 0.9999, deepSplit = T, le minClusterSize = 10 ah chiah in tuah a si. DiffCoEx i R source code cu Tesson et al nih a dang file 1 in an lak. (57); A herhmi R WGCNA software package cu https://horvath.genetics.ucla.edu/html/CoexpressionNetwork/Rpackages/WGCNA ah hmuh khawh a si.
MS/MS molecular network hlathlainak tuah khawhnak ding caah, NDP le NL aa lawhnak phun cung ah hram bunh in aa pehtlaimi spectral pehtlaihnak kan tuak, cun CyFilescape yFiles layout algorithm extension application ah organic layout hmangin network topology hmuh khawhnak ding caah Cytoscape software kan hman.
Data cung ah zatlang hlathlainak tuah awkah R version 3.0.1 kha hmang. Statistical significance cu two-way analysis of variance (ANOVA) hmangin tuaktan a si, cu hnu ah Tukey's honestly significant difference (HSD) post-hoc test nih a zulh. Herbivorous thlopnak le control karlak i dannak hlathlainak tuah khawhnak ding caah, aa khatmi thleidannak a ngeimi sample phu hnih hna i a tthen hnih tthencheunak kha Student's t test hmangin hlathlainak tuah a si.
Hi capar caah bawmtu thilri caah, http://advances.sciencemag.org/cgi/content/full/6/24/eaaz0381/DC1 zoh
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Thawngpang theory nih special metabolomes tahchunhnak le hneksaknak kilvennak theory chimchungnak caah vawleicung pumpi tangka a pek.
Thawngpang theory nih special metabolomes tahchunhnak le hneksaknak kilvennak theory chimchungnak caah vawleicung pumpi tangka a pek.
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