Afifide ho banbɔ ho nsusuwii ahorow de nsusuwii mu akwankyerɛ a ɛho hia ma de kyerɛkyerɛ sɛnea afifide mu nipadua mu nneɛma titiriw no yɛ adwuma no mu, nanso ɛda so ara wɔ hɔ a ɛsɛ sɛ wɔsɔ wɔn nkɔmhyɛ atitiriw no hwɛ. Ɛha yi, yɛde unbiased tandem mass spectrometry (MS / MS) nhwehwɛmu dii dwuma de hwehwɛɛ metabolome a tawa attenuated strains fi ankorankoro afifide mu kɔ nnipa dodow ne ahorow a ɛne wɔn ho wɔn ho di nsɛ mu wɔ nhyehyɛe kwan so, na yɛyɛɛ mass spectrometric feature theories dodow bi a egyina compound spectra so wɔ nsɛm no mu ho adwuma. Nhyehyɛe a wɔde bɛsɔ nkɔmhyɛ atitiriw a ɛfa ahobammɔ a eye sen biara (OD) ne botae a ɛkɔ (MT) nsusuwii ahorow ahwɛ. Amanneɛbɔ fã a ɛwɔ afifideɛ mu metabolomics mu no ne OD nsusuiɛ hyia, nanso ɛne nkɔmhyɛ titire a MT nsusuiɛ wɔ wɔ metabolomics dynamics a ɛnam mmoa a wɔdi nhabannuru so de ba no bɔ abira. Efi micro so kosi macro evolutionary scale so no, wɔkyerɛɛ jasmonate sɛnkyerɛnne no sɛ ade titiriw a ɛkyerɛ OD, bere a ethylene sɛnkyerɛnne no de fine-tuning ma herbivore-specific response a MS/MS molecular network no akyerɛw ho nkyerɛkyerɛmu no.
Nneɛma titiriw a ɛma nipadua no yɛ adwuma a ɛsono ne su ne nea ɛka ho titiriw wɔ afifide a ɛsakra ne nneɛma a atwa yɛn ho ahyia no mu, titiriw wɔ atamfo ho banbɔ mu (1). Nneɛma ahorow a ɛyɛ nwonwa a ɛwɔ nipadua mu nneɛma titiriw a wohu wɔ afifide mu no akanyan mfe du du pii a wɔde ayɛ nhwehwɛmu a emu dɔ wɔ ne nneɛma pii a ɛfa nneɛma a atwa yɛn ho ahyia ho no ho, na ayɛ afifide ahobammɔ ho nsusuwii ahorow a wɔahyehyɛ no tenten, a ɛyɛ adannandi ne abɔde a nkwa wom mu nkɔso a ɛba wɔ afifide ne nkoekoemmoa nkitahodi mu. Nhwehwɛmu a wɔde wɔn ho ahyɛ mu de akwankyerɛ a ɛho hia ma (2). Nanso, saa afifide ahobammɔ nsusuwii ahorow yi anni normative kwan a ɛne hypothetical deductive reasoning akyi, a na nkɔmhyɛ atitiriw no wɔ nhwehwɛmu gyinabea koro (3) na wɔsɔɔ hwɛe wɔ sɔhwɛ mu de maa nsusuwii nkɔso kyinhyia a edi hɔ no kɔɔ anim (4). Mfiridwuma mu anohyeto ahorow siw nsɛm a wɔboaboa ano no ano hye wɔ nipadua mu nneɛma a ɛsakra no akuw pɔtee bi mu na ɛyi nneɛma a ɛma nipadua no yɛ adwuma titiriw ho nhwehwɛmu a edi mũ fi mu, na ɛnam so siw akuw ahorow ntam ntotoho a ɛho hia ma nsusuwii mu nkɔso ano (5). Metabolomics data a edi mũ a enni hɔ ne sika biako a wɔde bɛtoto adwuma a wɔyɛ wɔ metabolic space a ɛwɔ afifide akuw ahorow ntam no ho no siw nyansahu mu onyin a ɛwɔ afuw no mu no kwan.
Nkɔsoɔ a ɛtwa toɔ wɔ tandem mass spectrometry (MS/MS) metabolomics mu no tumi kyerɛ nsakraeɛ a ɛba metabolic mu wɔ nhyehyɛeɛ clade bi a wɔde ama no mu ne wɔn ntam no yie, na wɔbɛtumi de akɔka akontabuo akwan ho de abu nsɛsoɔ a ɛwɔ saa afrafradeɛ a ɛyɛ den yi ntam. Nimdeɛ a wɔadi kan anya wɔ nnuruyɛ ho (5). Mfiridwuma a ɛkɔ anim a wɔaka abom wɔ nhwehwɛmu ne kɔmputa mu no ma wonya nhyehyɛe a ɛho hia a wɔde bɛsɔ nkɔmhyɛ pii a abɔde a nkwa wom ne adannandi ho nsusuwii ahorow a ɛfa nipadua mu nneɛma ahorow ho no ahwɛ bere tenten. Shannon (6) de nsɛm ho nkyerɛkyerɛ bae nea edi kan wɔ n’asɛm titiriw no mu wɔ 1948 mu, na ɔtoo fapem maa akontaabu mu nhwehwɛmu a ɛfa nsɛm ho, a wɔde adi dwuma wɔ nnwuma pii mu a ɛnyɛ nea wɔde dii dwuma mfiase no. Wɔ genomics mu no, wɔde nsɛm ho nsusuwii adi dwuma yiye de akyerɛ sɛnea nsɛm a ɛfa nnidiso nnidiso no te (7). Wɔ transcriptomics nhwehwɛmu mu no, nsɛm ho nsusuwii hwehwɛ nsakrae a ɛba wɔ transcriptome no mu nyinaa mu (8). Wɔ nhwehwɛmu a atwam mu no, yɛde akontaabu nhyehyɛe a ɛfa nsɛm ho nsusuwii ho no dii dwuma wɔ metabolomics mu de kyerɛkyerɛɛ nipadua mu nneɛma a ɛsakra ho nimdeɛ a ɛwɔ ntini dodow a ɛwɔ afifide mu no mu (9). Ɛha yi, yɛde adwumayɛ nhyehyɛe a egyina MS/MS so no ne akontaabu nhyehyɛe a ɛfa nsɛm ho nsusuwii ho, a ɛda nsow wɔ nipadua mu nneɛma a ɛsakra wɔ sika a wɔtaa de di dwuma mu no bom de toto nkɔmhyɛ atitiriw a ɛwɔ afifide ahobammɔ nsusuwii a ɛfa metabolome a mmoa a wodi nhabannuru de ba no ho.
Nsusuwii nhyehyɛe ahorow a ɛfa afifide ahobammɔ ho no taa yɛ nea ɛka wɔn ho wɔn ho nyinaa ho na wobetumi akyekyɛ mu abien: nea ɛbɔ mmɔden sɛ ɛbɛkyerɛkyerɛ afifide pɔtee bi mu nneɛma a ɛsakra no mu a egyina ahobammɔ dwumadi ahorow so, te sɛ ahobammɔ a eye sen biara (OD) (10), botae a ɛkɔ so (MT) (11) ) Ne sɛnea ɛte (12) nsusuwii, bere a afoforo hwehwɛ mfiridwuma mu nkyerɛkyerɛmu a ɛfa sɛnea nsakrae a ɛba wɔ nneɛma a ɛwɔ hɔ mu no ka afifide ho onyin ne metabolites titiriw a ɛboaboa ano, te sɛ carbon: aduannuru kari pɛ ho adwene (13), onyin dodow ho adwene (14 ), ne onyin ne nsonsonoe kari pɛ ho adwene (15). Nsusuwii ahorow abien no wɔ nhwehwɛmu gyinabea ahorow (4). Nanso, nsusuwii abien a ɛfa ahobammɔ dwumadi ahorow ho wɔ dwumadi gyinabea no di nkɔmmɔbɔ a ɛfa afifide nhyehyɛe ne nea wɔde kanyan ahobammɔ ho no so: OD nsusuwii, a ɛfa no sɛ afifide de wɔn sika hyɛ wɔn nnuru a wɔde bɔ wɔn ho ban a ne bo yɛ den no mu bere a ɛho hia nkutoo, sɛ nhwɛso no, bere a wɔnom Sɛ sare aboa bi tow hyɛ so a, enti, sɛnea ebetumi aba sɛ daakye ntua kyerɛ no, wɔde aduru a ɛwɔ ahobammɔ dwumadi bi ma (10); MT adwene no kyerɛ sɛ axis biara nni hɔ a ɛkyerɛ kwan sɛ metabolite sesa, nanso metabolite no sesa randomly, na ɛnam so ma ɛyɛ yiye Siw metabolic “motion target” a ɛtow hyɛ herbivores so no kwan. Ɔkwan foforo so no, saa nsusuwii abien yi ka nkɔmhyɛ ahorow a ɛne ne ho bɔ abira fa nipadua mu nneɛma a wɔsan yɛ no foforo a ɛba wɔ mmoa a wodi nhabannuru ntua akyi no ho: abusuabɔ a ɛda nipadua mu nneɛma a ɛboaboa ano wɔ ɔkwan biako so a ɛwɔ ahobammɔ dwumadi (OD) ne nipadua mu nsakrae a ɛnkyerɛ kwan (MT) ntam (11 ).
OD ne MT nsusuwii ahorow no nyɛ nsakrae a ɛba wɔ metabolome no mu nko, na mmom nea efi nneɛma a atwa yɛn ho ahyia ne adannandi mu ba wɔ saa nneɛma a ɛma nipadua no yɛ adwuma yi a wɔboaboa ano no mu ba, te sɛ ɛka a wɔbɔ wɔ nsakrae a ɛba wɔ nipadua mu nsakrae yi mu wɔ abɔde a nkwa wom tebea pɔtee bi mu no nso ho (16). Ɛwom sɛ nsusuwii abien no nyinaa hu dwuma a nipadua no mu nneɛma titiriw a ɛma nipadua no yɛ adwuma no bɔ ho ban, a ebia ne bo yɛ den anaasɛ ɛnyɛ nea ne bo yɛ den de, nanso nkɔmhyɛ titiriw a ɛma nsonsonoe da OD ne MT nsusuwii ahorow no ntam no gyina sɛnea nipadua no mu nsakrae a ɛde ba no kɔ no so. OD nsusuwii ho nkɔmhyɛ no na wɔde adwene asi so sen biara wɔ sɔhwɛ mu de besi nnɛ. Saa sɔhwɛ ahorow yi bi ne adesua a ɛfa ahobammɔ dwumadi ahorow a ɛwɔ ntini ahorow a ɛwɔ nnuru pɔtee bi mu wɔ greenhouses ne abɔde mu tebea horow mu, ne nsakrae a ɛba wɔ afifide nyin fã mu (17-19). Nanso, ɛde besi ha no, esiane sɛ wonni adwumayɛ nhyehyɛe ne akontaabu nhyehyɛe a wɔde bɛyɛ wiase nyinaa nhwehwɛmu a edi mũ wɔ abɔde biara mu nneɛma a ɛsakra ho nti, nsonsonoe titiriw a wɔahyɛ ho nkɔm wɔ nsusuwii abien no ntam (kyerɛ sɛ, ɔkwan a wɔfa so yɛ nsakrae a ɛba nipadua mu nneɛma mu) no da so ara wɔ hɔ a ɛsɛ sɛ wɔsɔ hwɛ. Ɛha yi, yɛde nhwehwɛmu a ɛte saa ma.
Afifide pɔtee bi mu nneɛma a ɛsakra no su titiriw biako ne wɔn nhyehyɛe ahorow a ɛtra so wɔ gyinabea ahorow nyinaa mu fi afifide biako, nnipa dodow so kosi mmoa ahorow a ɛte saa ara so (20). Wobetumi ahu nsakrae pii a ɛba dodow mu wɔ metabolites titiriw mu wɔ nnipa dodow nsenia mu, bere a wɔtaa kura nsonsonoe a emu yɛ den wɔ su mu wɔ mmoa ahorow no gyinabea (20). Enti, afifide mu nipadua mu nneɛma ahorow ne ade titiriw a ɛwɔ dwumadi ahorow mu, a ɛkyerɛ sɛnea wotumi sesa wɔn ho kɔ niches ahorow mu, titiriw saa niches a ɛsono ntua a nkoekoemmoa titiriw ne mmoa a wɔtaa di nhabannuru tumi ba no (21). Efi Fraenkel (22) asɛm a ɛyɛ nwonwa a ɛfa nea enti a afifide pɔtee bi mu nneɛma a ɛma nipadua no yɛ adwuma wɔ hɔ ho no, wɔabu nkitahodi a wɔne nkoekoemmoa ahorow di no sɛ nhyɛso a ɛho hia a wɔde paw nneɛma, na wogye di sɛ saa nkitahodi ahorow yi na ɛhyehyɛɛ afifide bere a na wɔreyɛ adannandi no. Nkwammoaa mu nkwaadɔm kwan (23). Nsonsonoe a ɛwɔ mmoa ahorow ntam wɔ metabolites soronko ahorow mu nso betumi ada nipadua mu kari pɛ a ɛbata constitutive ne inducible afifide ahobammɔ atia herbivorous akwan horow ho adi, efisɛ mpɛn pii no, ahorow abien no ne wɔn ho wɔn ho wɔ abusuabɔ bɔne ( 24 ). Ɛwom sɛ ebia mfaso bɛba so sɛ wɔbɛkɔ so akura ahobammɔ pa mu bere nyinaa de, nanso nsakrae a ɛba bere ano wɔ nipadua mu a ɛbata ahobammɔ ho no ma wonya mfaso a ɛda adi pefee wɔ afifide a ɛma afifide tumi kyekyɛ nneɛma a ɛsom bo ma nipadua mu sika afoforo (19, 24), na wɔkwati hia a ehia sɛ wɔyɛ biako. Nneɛma a wɔsɛe no a wɔde abɔ ho ban (25). Bio nso, saa nneɛma titiriw a wɔsan yɛ no foforo a nkoekoemmoa a wodi nhabannuru de ba no betumi ama wɔakyekyɛ nneɛma a ɛsɛe ade wɔ nnipa dodow no mu ( 26 ), na ebetumi ada abɔde mu nsakrae kɛse a ɛba wɔ jasmonic acid (JA) sɛnkyerɛnne mu, a ebia wɔbɛkɔ so akura mu wɔ nnipa dodow no mu no akenkan tẽẽ adi. JA nsɛnkyerɛnne a ɛkorɔn ne nea ɛba fam yɛ aguadi a ɛda ahobammɔ a wɔde tia mmoa a wodi nhabannuru ne akansi a wɔne mmoa ahorow pɔtee bi di ntam ( 27 ). Afei nso, metabolite biosynthetic akwan titiriw bɛhwere ntɛmntɛm na nsakrae bere a adannandi, na ɛde patchy metabolic kyekyɛ wɔ mmoa ahorow a ɛbɛn wɔn ho wɔn ho mu ( 28 ). Wobetumi de saa polymorphisms yi asi hɔ ntɛmntɛm de abua mmoa a wodi nhabannuru nhyehyɛe a ɛresakra ( 29 ), a ɛkyerɛ sɛ nhabannuru akuw a ɛsakrasakra no yɛ ade titiriw a ɛma nipadua mu nneɛma a ɛsakra no yɛ soronko.
Ɛha yi, yedii ɔhaw ahorow a edidi so yi ho dwuma pɔtee. (I) Ɔkwan bɛn so na nkoekoemmoa a wodi nhabannuru no san hyehyɛ afifide no metabolome? (Ii) Dɛn ne nsɛm atitiriw a ɛwɔ metabolic plasticity mu a wobetumi akyerɛ dodow de asɔ nkɔmhyɛ ahorow a ɛfa bere tenten ahobammɔ nsusuwii ho ahwɛ? . (Iv) Esiane sɛ wobetumi atrɛw nkɔmhyɛ ahorow a ahobammɔ ho nsusuwii pii de ama no mu wɔ abɔde a nkwa wom ntini ahorow nyinaa mu nti, yebisaa sɛnea nipadua mu nneɛma a ɛsakra no ho mmuae a ɛde ba no fi emu ntotoho so kosi mmoa ahorow ntam ntotoho so? Nea ɛbɛyɛ na yɛatumi ayɛ eyi no, yɛasua ahaban mu nkwaadɔm a ɛwɔ tawa nicotine mu, a ɛyɛ abɔde a nkwa wom ho nhwɛso afifide a ɛwɔ nipadua mu nkwaadɔm titiriw a ɛdɔɔso, na etu mpɔn tia mmoa a wodi nhabannuru abien a ɛwɔ hɔ ankasa, Lepidoptera Datura (Ms) ( Very aggressive, mainly eaten) On Solanaceae and Spodoptera littoralis (Sl), cotton leaf worms are a kind of “genus”, ne afifide a ɛgye Solanaceae ne ahɔhoɔ foforɔ a ɛwɔ abusua ne mmusua foforɔ mu Afifideɛ aduane. Yɛyɛɛ MS/MS metabolomics spectrum no mu nhwehwɛmu na yeyii nsɛm ho nsusuwii akontaabu nkyerɛkyerɛmu ahorow de totoo OD ne MT nsusuwii ahorow ho. Yɛ specificity maps de kyerɛ sɛnea nneɛma atitiriw a ɛma nipadua no yɛ adwuma no te. Wɔtrɛw nhwehwɛmu no mu kɔɔ N. nasi ne tawa ahorow a ɛne wɔn bɔ abusua kɛse no mu de kɔɔ so yɛɛ covariance a ɛda afifide mu hormone sɛnkyerɛnne ne OD induction ntam no mu nhwehwɛmu.
Sɛnea ɛbɛyɛ a yɛbɛkyere asase mfonini a ɛfa plasticity ne nhyehyɛe a ahaban metabolome a herbivorous tawa ho no, yɛde nhwehwɛmu ne akontaabu adwumayɛ kwan a yɛadi kan ayɛ dii dwuma de boaboaa ne deconvolute a ɛkɔ akyiri ano na deconvolute high-resolution data independent MS / MS spectra fi afifide extracts ( 9 ). Saa kwan a wɔankyerɛ nsonsonoeɛ yi (a wɔfrɛ no MS/MS) yi tumi yɛ non-redundant compound spectra, a afei wɔbɛtumi de adi dwuma ama compound-level analyses a wɔaka ho asɛm wɔ ha nyinaa. Saa afifideɛ mu nneɛma a wɔayiyi afiri mu yi yɛ ahodoɔ, a ɛyɛ nneɛma ɔhaha kɔsi mpempem pii (bɛyɛ 500-1000-s/MS/MS wɔ ha). Ɛha yi, yɛsusu metabolic plasticity ho wɔ nsɛm ho nsusuiɛ nhyehyɛeɛ mu, na yɛbu metabolome no ahodoɔ ne adwumayɛ ho dodoɔ a egyina Shannon entropy a ɛwɔ metabolic frequency distribution no so. Yɛde nhyehyɛe (8) a wɔadi kan de adi dwuma no dii dwuma de buu nsɛnkyerɛnne ahorow bi a wobetumi de akyerɛ metabolome ahorow dodow (Hj kyerɛwtohɔ), metabolic profile specialization (δj indicator) ne metabolic specificity of a single metabolite (Si indicator) . Afei nso, yɛde Relative Distance Plasticity Index (RDPI) dii dwuma de kyerɛɛ dodow a metabolome inducibility a herbivores (Mfonini 1A) (30). Wɔ saa akontabuo nhyehyɛeɛ yi mu no, yɛfa MS/MS spectrum sɛ nsɛm titire kuo, na yɛyɛ MS/MS dodoɔ a ɛfa ho no ho adwuma kɔ frequency distribution map mu, na afei yɛde Shannon entropy di dwuma de bu metabolome ahodoɔ ho akontaa firi mu. Wɔde MS/MS spectrum biako pɔtee a ɛyɛ pɔtee no na ɛsusuw metabolome specialization no. Enti, MS/MS akuo binom dodoɔ a ɛkɔ soro wɔ herbivore induction akyi no dane kɔ spectral inducibility, RDPI ne specialization, kyerɛ sɛ, nkɔanim a ɛba δj index mu, ɛfiri sɛ wɔyɛ metabolites soronko pii na wɔyɛ Si index a ɛkorɔn. Hj ahodoɔ nkyerɛkyerɛmu no a wɔbrɛ aseɛ no kyerɛ sɛ MS/MS dodoɔ a wɔayɛ no so tew, anaasɛ profile frequency nkyekyɛmu no sesa wɔ ɔkwan a ɛnyɛ pɛ so, berɛ a ɛtew ne nyinaa mu adwenem naayɛ so. Ɛnam Si index akontabuo so no, ɛyɛ yie sɛ wɔbɛsi MS/MS a mmoa a wɔdi nhabannuru binom de ba no so dua, nea ɛne no bɔ abira no, MS/MS bɛn na ɛnyɛ ho biribi wɔ induction no ho, a ɛyɛ ade titire a ɛkyerɛ nsonsonoeɛ a ɛda MT ne OD nkɔmhyɛ mu.
(A) Akontaabu nkyerɛkyerɛmu a wɔde di dwuma ma nhabannuru (H1 kosi Hx) MS/MS data-inducibility (RDPI), ahorow ahorow (Hj index), specialization (δj index) ne metabolite pɔtee (Si index). Nkɔanim a ɛba wɔ sɛnea wɔyɛ soronko (δj) no kyerɛ sɛ, sɛ wɔkyekyem pɛpɛɛpɛ a, wɔbɛyɛ nhabannuru pɔtee a ɛma nipadua no yɛ adwuma pii, bere a nneɛma ahorow a ɛso tew (Hj) kyerɛ sɛ nneɛma a ɛma nipadua no yɛ adwuma no so tew anaasɛ nneɛma a ɛsakra no kyekyɛ a ɛnteɛ wɔ nkyekyɛmu asase mfonini no mu . Si bo no hwɛ sɛ ebia metabolite no yɛ pɔtee ma tebea bi (ha, herbivorous) anaasɛ nea ɛne no bɔ abira no, wɔakura mu wɔ dodow koro no ara mu. (B) Adwene mu mfonini a ɛkyerɛ defense theory nkɔmhyɛ a wɔde information theory axis di dwuma. OD nsusuwii kyerɛ sɛ herbivore ntua bɛma defense metabolites akɔ soro, na ɛnam so ama δj akɔ soro. Bere koro no ara mu no, Hj so tew efisɛ wɔsan hyehyɛ profile no kɔ nipadua no mu nsɛm a wontumi nsi pi a ɛso atew no so. MT nsusuwii no hyɛ nkɔm sɛ mmoa a wodi nhabannuru ntua no bɛma nsakrae a ɛnyɛ akwankyerɛ aba metabolome no mu, na ɛnam so ama Hj akɔ soro sɛ ade a ɛkyerɛ sɛ nipadua mu nsɛm a wontumi nsi pi wɔ nipadua mu no akɔ soro na ɛde Si a wɔkyekyɛ no kwa aba. Yɛsan nso de nhwɛsoɔ a wɔadi afra, MT a eye sen biara, a metabolites binom a ɛwɔ ahobammɔ gyinapɛn a ɛkorɔn no bɛkɔ soro titiriw (Si bo a ɛkorɔn), bere a afoforo da mmuae a wɔanhyɛ da (Si bo a ɛba fam) adi.
Sɛ yɛde nsɛm ho nsusuwii nkyerɛkyerɛmufo di dwuma a, yɛkyerɛ OD nsusuwii no ase de kyerɛ sɛ nsakrae titiriw a ɛba wɔ metabolite tebea a wɔanhyɛ da mu a herbivore de ba mu no bɛma (i) metabolic specificity (Si index) a ɛkɔ soro a ɛma metabonomic specificity (δj index) Nkɔanim a ɛba) metabolite akuw titiriw bi a ɛwɔ ahobammɔ bo a ɛkorɔn, ne (ii) metabolome ahorow ahorow (Hj) index a ɛkɔ soro esiane nsakrae a ɛba metabolic frequency kyekyɛ mu kɔ leptin nipadua kyekyɛ pii mu nti. Wɔ metabolite biako gyinabea no, wɔhwɛ kwan sɛ Si nkyekyɛmu a wɔahyehyɛ no nnidiso nnidiso, baabi a metabolite no bɛma Si bo akɔ soro sɛnea ne ahobammɔ bo te (Mfonini 1B). Wɔ saa kwan yi so no, yɛkyerɛkyerɛ MT nsusuwii no mu de hyɛ nkɔm sɛ anigyede bɛma (i) nsakrae a ɛnyɛ akwankyerɛ aba wɔ metabolites mu a ɛde δj index no so tew, ne (ii) Hj index no nkɔanim esiane metabolic a wontumi nsi pi a ɛkɔ soro nti. Anaasɛ randomness, a wobetumi de Shannon entropy akyerɛ dodow wɔ generalized diversity kwan so. Ɛdefa metabolic composition ho no, MT nsusuwii no bɛkyerɛ sɛnea Si bɛkyekyɛ random. Sɛ yɛsusu sɛ metabolite binom wɔ tebea pɔtee bi mu wɔ tebea pɔtee bi mu, na tebea afoforo nni tebea pɔtee bi ase, na wɔn ahobammɔ bo gyina nneɛma a atwa yɛn ho ahyia so a, yɛsan nso de ahobammɔ nhwɛso a wɔadi afra ho nyansahyɛ mae, a δj ne Hj kyekyɛ abien wɔ Si Nkɔanim wɔ akwan nyinaa so, metabolite akuw bi nkutoo, a wɔwɔ ahobammɔ bo a ɛkorɔn, bɛma Si akɔ soro titiriw, bere a afoforo benya random nkyekyɛmu (Mfonini 1B).
Sɛnea ɛbɛyɛ a yɛbɛsɔ defense theory prediction a wɔasan akyerɛkyerɛ mu no ahwɛ wɔ axis a ɛwɔ information theory descriptor no so no, yɛtetee expert (Ms) anaa generalist (Sl) herbivore larvae wɔ Nepenthes pallens nhaban so (Mfonini 2A). Yɛde MS/MS nhwehwɛmu dii dwuma, gyee MS/MS spectra 599 a ɛnyɛ nea ɛho nhia (data file S1) fii methanol a wɔayiyi afi nhaban ntini a wɔaboaboa ano wɔ ntontom aduan akyi. Sɛ yɛde RDPI, Hj, ne δj indexes di dwuma de yɛ nsɛm a ɛwɔ MS/MS nhyehyeɛ fael mu a wɔasan asiesie no ho mfonini a, ɛda nhwɛsoɔ a ɛyɛ anigyeɛ adi (Mfonini 2B). Nea ɛkɔ so wɔ ne nyinaa mu ne sɛ, sɛnea nsɛm ho nkyerɛkyerɛmufo no kyerɛkyerɛ mu no, bere a nwansena no kɔ so di nhaban no, sɛnea nipadua no mu nneɛma a ɛsakra no nyinaa yɛ foforo no kɔ soro bere tenten: nnɔnhwerew 72 akyi bere a aboa a odi nhabannuru no adi akyi no, RDPI no kɔ soro kɛse. Sɛ wɔde toto control a ɛnsɛee no ho a, Hj so tew kɛse, na ɛno fi sɛnea metabolic profile no yɛ soronko a ɛkɔ soro, a wɔde δj index no buu dodow no. Saa su a ɛda adi yi ne OD nsusuwii no nkɔmhyɛ ahorow no hyia, nanso ɛne MT nsusuwii no nkɔmhyɛ atitiriw no nhyia, a egye di sɛ wɔde nsakrae a ɛba kwa (a ɛnyɛ akwankyerɛ) a ɛba wɔ metabolite dodow mu no di dwuma sɛ ahobammɔ a wɔde kata wɔn ho (Mfonini 1B). Ɛwom sɛ ɛsono sɛnea mmoa abien a wodi nhabannuru yi anom secretion (OS) elicitor dodow ne wɔn aduan a wɔde ma no de, nanso wɔn aduan a wɔde ma tẽẽ no maa nsakrae a ɛte saa ara baa Hj ne δj akwankyerɛ mu wɔ nnɔnhwerew 24 ne nnɔnhwerew 72 otwa bere no mu. Nsonsonoe biako pɛ a ɛbae wɔ nnɔnhwerew 72 a wɔde yɛɛ RDPI no mu. Sɛ wɔde toto nea Ms aduan a wɔde ma no ho a, na nipadua mu nneɛma a ɛsakra a Sl aduan de ba no nyinaa dɔɔso.
(A) Nsɔhwɛ nhyehyɛe: wɔde mprako a wɔtaa di (S1) anaa animdefo (Ms) a wodi nhabannuru ma nhaban a wɔayi nkyene afi pitcher afifide mu, bere a wɔ simulated herbivory ho no, wɔde OS of Ms (W + OSMs) di dwuma de di puncture a standardized leaf positions wound ho dwuma. S1 (W + OSSl) nwansena anaa nsuo (W + W). Control (C) yɛ ahaban a ɛnsɛee. (B) Inducibility (RDPI a wɔde toto control chart ho), diversity (Hj index) ne specialization (δj index) index a wɔabu ho akontaa ama metabolite map soronko no (599 MS/MS; data file S1). Nsoromma kyerɛ nsonsonoe kɛse a ɛda mmoa a wodi nhabannuru a wɔde ma tẽẽ ne kuw a wɔhwɛ so no ntam (Student’s t-test with paired t-test, *P<0.05 and ***P<0.001). ns, ɛnyɛ nea ɛho hia. (C) Bere nhyehyɛe nkyerɛkyerɛmu a ɛkyerɛ titiriw (adaka bruu, amino acid, organic acid ne asikre; data fael S2) ne metabolite spectrum titiriw (adaka kɔkɔɔ 443 MS/MS; data fael S1) wɔ nhabannuru a wɔde di dwuma sɛnea wɔayɛ no akyi. Kɔla band no kyerɛ 95% ahotoso ntamgyinafo. Nsoromma no kyerɛ nsonsonoe kɛse a ɛda ayaresa ne nea wɔde di dwuma no ntam [quadratic analysis of variance (ANOVA), a Tukey nokwaredi mu nsonsonoe a ɛho hia (HSD) di akyi ma post hoc ntotoho pii, *P<0.05, **P<0.01 Na *** P <0.001]. (D) Specialization of scatter plots ne soronko metabolite profiles (nhwɛsode a wɔasan ayɛ a wɔde ayaresa ahorow ayɛ).
Sɛnea ɛbɛyɛ na yɛahwehwɛ sɛ ebia nhabannuru a wodi a ɛma wɔsan yɛ no foforo wɔ metabolome gyinabea no da adi wɔ nsakrae a ɛba ankorankoro nipadua mu nneɛma a ɛsakra no dodow mu no, yedii kan de yɛn adwene sii nneɛma a ɛsakra a wɔadi kan asua ho ade wɔ Nepenthes pallens nhaban a wɔada no adi sɛ nhabannuru a wodi no ko tia no so. Phenolic amides yɛ hydroxycinnamamide-polyamine conjugates a ɛboaboa ano bere a nkoekoemmoa di nhabannuru no na wonim sɛ ɛtew nkoekoemmoa adwumayɛ so ( 32 ). Yɛhwehwɛɛ MS/MS a ɛne no hyia no animdefo na yɛyɛɛ wɔn cumulative kinetic curves no ho mfonini (Mfonini S1). Ɛnyɛ nwonwa sɛ wɔtew phenol a efi mu ba a ɛnyɛ nea ɛka ho tẽẽ wɔ ahobammɔ a wɔde ko tia mmoa a wodi nhabannuru te sɛ chlorogenic acid (CGA) ne rutin no so bere a wɔadi nhabannuru akyi. Nea ɛne eyi bɔ abira no, mmoa a wodi nhabannuru betumi ama phenol amides ayɛ nea tumi wom kɛse. Ɛkame ayɛ sɛ mmoa abien a wodi nhabannuru no aduan a wɔkɔɔ so de maa wɔn no maa phenolamides a ɛkanyan wɔn no yɛɛ pɛ, na na saa nhyehyɛe yi da adi titiriw wɔ phenolamides a wɔyɛ de novo no mu. Wɔbɛhunu adeyɛ korɔ no ara berɛ a wɔrehwehwɛ 17-hydroxygeranyl nonanediol diterpene glycosides (17-HGL-DTGs) kwan no mu, a ɛma acyclic diterpenes dodoɔ bi a ɛwɔ anti-herbivore dwumadie a ɛtu mpɔn (33), a Ms Feeding with Sl na ɛkanyan nkyerɛkyerɛmu a ɛte saa ara (Mfonini S1)).
Mfomso a ebetumi aba wɔ sɔhwɛ ahorow a wɔde ma mmoa a wodi nhabannuru tẽẽ so ne nsonsonoe a ɛwɔ nhaban a wodi ne bere a mmoa a wodi nhabannuru nya mu, na ɛma ɛyɛ den sɛ wobeyi nkɛntɛnso a akuru ne mmoa a wodi nhabannuru de ba mmoa a wodi nhabannuru pɔtee bi so no afi hɔ. Sɛnea ɛbɛyɛ a yebedi herbivore species specificity a ɛwɔ induced leaf metabolic response no ho dwuma yiye no, yɛyɛɛ Ms ne Sl larvae aduan a wɔde ma ho mfonini denam OS a wɔaboaboa ano foforo (OSM ne OSS1) a yɛde dii dwuma ntɛm ara wɔ standard puncture W a ɛwɔ ahaban gyinabea a ɛkɔ so daa no so. Wɔfrɛ saa adeyɛ yi W + OS ayaresa, na ɛma induction no yɛ gyinapɛn denam bere pɔtee a wɔde fi ase ma mmuae a herbivore no de ba a ɛmma nsonsonoe a ɛwɔ ntini a ɛyera no dodow anaa dodow mu no nsunsuanso a ɛyɛ ahodwiriw mma (Mfonini 2A) (34). Yɛde MS/MS nhwehwɛmu ne akontabuo nsuo afiri no dii dwuma, gyee MS/MS spectra 443 (data file S1), a ɛne spectra a wɔadi kan aboaboa ano afiri direct feed experiments mu no hyia. Nsɛm ho nsusuwii nhwehwɛmu a wɔyɛe wɔ MS/MS data nhyehyɛe yi mu no kyerɛe sɛ nhaban-specialized metabolomes a wɔsan yɛe denam herbivores a wɔyɛ no sɛnea wɔyɛ no so no kyerɛɛ OS-specific inducements (Mfonini 2C). Titiriw no, sɛ wɔde toto OSS1 ayaresa ho a, OSM maa metabolome specialization nyaa nkɔso wɔ nnɔnhwerew 4 mu. Ɛfata sɛ yɛhyɛ no nsow sɛ sɛ wɔde toto tẽẽ herbivore feeding experimental data set ho a, metabolic kinetics a wɔde wɔn adwene sii so wɔ afã abien ahunmu a wɔde Hj ne δj dii dwuma sɛ coordinates ne directionality a metabolome specialization no de bua simulated herbivore treatment wɔ bere mu Nkɔanim a ɛkɔ so daa (Mfonini 2D). Bere koro no ara mu no, yɛkyerɛɛ dodow a ɛwɔ amino acids, organic acids ne asikre mu (data file S2) de hwehwɛɛ sɛ ebia saa nkɔanim a wɔde asi wɔn ani so yi wɔ metabolome nimdeɛ mu no fi central carbon metabolism a wɔsan hyehyɛ no foforo de bua mmoa a wodi nhabannuru a wɔayɛ no sɛnea ɛte no ( Mfonini S2 ). Sɛnea ɛbɛyɛ na yɛakyerɛkyerɛ saa nhyehyɛe yi mu yiye no, yɛkɔɔ so hwɛɛ metabolic accumulation kinetics a ɛwɔ phenolamide ne 17-HGL-DTG akwan a yɛadi kan aka ho asɛm no mu. OS-specific induction a herbivores no dane kɔ differential rearrangement pattern mu wɔ phenolamide metabolism mu (Mfonini S3). Phenolic amides a coumarin ne caffeoyl afã horow wom no, OSS1 na ɛpɛ, bere a OSM ahorow no kanyan ferulyl conjugates a wɔde hyɛ mu pɔtee. Wɔ 17-HGL-DTG kwan no ho no, wohuu nsonsonoe OS induction denam downstream malonylation ne dimalonylation nneɛma so (Mfonini S3).
Afei, yɛde bere-kɔ so microarray data set a ɛyɛ sɛnea wɔde OSM ahorow di dwuma de sa rosette afifide nhaban nhaban wɔ mmoa a wodi nhabannuru mu no suaa transcriptome plasticity a OS de ba no ho ade. Sampling kinetics no titiriw ne kinetics a wɔde dii dwuma wɔ saa metabolomics nhwehwɛmu yi mu no hyia ( 35 ). Sɛ yɛde toto metabolome reconfiguration a metabolic plasticity kɔ soro titiriw bere tenten a, yehu bere tiaa mu transcription bursts wɔ nhaban a Ms de ba mu, faako a transcriptome inducibility (RDPI) ne specialization (δj) wɔ 1 Na nkɔanim kɛse wɔ nnɔnhwerew mu, na ahorow ahorow (Hj) wɔ saa bere yi mu, na BMP1 a wɔda no adi no so tew kɛse, na ɛno akyi no, ahomegye no bae a ɛfa transcriptome titiriw ho (Mfonini S4). Nkwammoaa mu nkwaadɔm mmusua (te sɛ P450, glycosyltransferase, ne BAHD acyltransferase) de wɔn ho hyɛ ɔkwan a wɔfa so boaboa nneɛma titiriw a ɛma nipadua no yɛ adwuma ano fi nhyehyɛe mu akuw a wonya fi nipadua mu nkwaadɔm titiriw mu no mu, di mfitiase nhwɛso a ɛkorɔn a yɛadi kan aka ho asɛm no akyi. Sɛ́ asɛm bi a wɔayɛ ho nhwehwɛmu no, wɔyɛɛ phenylalanine kwan no mu nhwehwɛmu. Nhwehwɛmu no sii so dua sɛ awosu mu nkwaadɔm titiriw a ɛwɔ phenolamide metabolism mu no yɛ nea OS de ba kɛse wɔ mmoa a wodi nhabannuru mu sɛ wɔde toto afifide a wɔantwetwe wɔn ho a, na ɛne wɔn nkyerɛkyerɛmu nhyehyɛe hyia kɛse. Transcription factor MYB8 ne structure genes PAL1, PAL2, C4H ne 4CL a ɛwɔ saa kwan yi atifi no kyerɛɛ sɛ wɔhyɛɛ nkyerɛwee ase ntɛm. Acyltransferases a ɛdi dwuma wɔ phenolamide nhyiamu a ɛtwa toɔ mu, te sɛ AT1, DH29, ne CV86, da upregulation nhyehyɛeɛ a ɛkyɛ adi (Mfonini S4). Atifi hɔ nsɛm a yɛahu no kyerɛ sɛ transcriptome specialization a wofi ase ntɛm ne metabolomics specialization a wɔma ɛkɔ anim akyiri yi no yɛ coupled mode, a ebia efi synchronous regulatory system a efi ase yɛ defense mmuae a tumi wom.
Nneɛma a wɔsan hyehyɛ wɔ afifide mu hormone sɛnkyerɛnne mu no yɛ adwuma sɛ mmara kwan so ade a ɛka nsɛm a ɛfa nhabannuru ho bom de san yɛ afifide nipadua ho nhyehyɛe. Wɔ herbivore simulation akyi no, yɛsusuu cumulative dynamics of key plant hormone categories na yɛyɛɛ bere mu co-expression a ɛda wɔn ntam no ho mfonini [Pearson correlation coefficient (PCC)> 0.4] (Mfonini 3A). Sɛnea wɔhwɛ kwan no, afifide mu hormone ahorow a ɛfa biosynthesis ho no wɔ abusuabɔ wɔ afifide mu hormone co-expression network no mu. Bio nso, wɔayɛ metabolic specificity (Si index) ho mfonini akɔ saa network yi so de asi afifide mu hormone ahorow a ayaresa ahorow de ba no so dua. Wɔatwe mmeae atitiriw abien a herbivorous mmuae pɔtee bi wɔ: biako wɔ JA akuw no mu, baabi a JA (ne abɔde a nkwa wom a ɛyɛ adwuma JA-Ile) ne JA a efi mu ba afoforo kyerɛ Si nkontaa a ɛkorɔn sen biara; ɔfoforo no ne ethylene (ET). Gibberellin daa nkɔanim a ɛkɔ fam pɛ adi wɔ mmoa a wodi nhabannuru pɔtee mu, bere a afifide mu nkwaadɔm afoforo te sɛ cytokinin, auxin, ne abscisic acid, nni induction pɔtee a ɛba fam ma mmoa a wodi nhabannuru. Sɛ wɔde toto W + W nkutoo a wɔde di dwuma ho a, JA derivatives no bo a ɛkorɔn a wɔma ɛyɛ kɛse denam OS application (W + OS) so no, wobetumi adan no titiriw ayɛ no JAs ho sɛnkyerɛnne pɔtee a emu yɛ den. Nea wɔnhwɛ kwan no, wonim sɛ OSM ne OSS1 a ɛsono elicitor mu no ma JA ne JA-Ile boaboa ano saa ara. Nea ɛne OSS1 bɔ abira no, OSM yɛ nea OSM ahorow na ɛkanyan no pɔtee na ɛyɛ den, bere a OSS1 ntumi mma akuru a ɛwɔ ase no mmuae nyɛ kɛse (Mfonini 3B).
(A) Co-expression network nhwehwɛmu a egyina PCC akontaabu a ɛfa herbivore-a ɛma afifide hormone boaboa ano kinetics simulation so. Node no gyina hɔ ma afifide mu hormone biako, na node no kɛse gyina hɔ ma Si index pɔtee a ɛfa afifide hormone ho wɔ ayaresa ahorow ntam. (B) JA, JA-Ile ne ET a ɛboaboa ano wɔ nhaban mu a ɛnam ayaresa ahorow a wɔde kɔla ahorow kyerɛ so: apricot, W + OSM; bruu, W + OSSl, a ɛyɛ bruu; tuntum, W + W; grey, C (ahyɛde) . Nsoromma kyerɛ nsonsonoe kɛse a ɛda ayaresa ne nea wɔde di dwuma ntam (akwan abien ANOVA a Tukey HSD post hoc multiple comparison di akyi, *** P <0.001). Amanneɛbɔ nsusuiɛ nhwehwɛmu a ɛfa (C)697 MS/MS (data file S1) a ɛwɔ JA biosynthesis ne impaired perception spectrum (irAOC ne irCOI1) ne (D)585 MS/MS (data file S1) wɔ ETR1 a ɛwɔ ET sɛnkyerɛnne a asɛe Ayaresa mmienu a wɔayɛ no sɛdeɛ ɛteɛ wɔ herbivore ayaresa mu no kanyan afifideɛ nhama ne empty vehicle (EV) control plants. Nsoromma kyerɛ nsonsonoe kɛse a ɛda W + OS ayaresa ne nea ɛnsɛee no ntam (akwan abien ANOVA a Tukey HSD post hoc ntotoho pii di akyi, *P <0.05, **P <0.01 ne ***P <0.001). (E) Graf ahorow a apete a ɛkyerɛ ɔsɔretia a ɛbɔ apete wɔ specialization ho. Kɔla ahorow no gyina hɔ ma ahorow ahorow a wɔayɛ wɔn awosu mu nsakrae; nsɛnkyerɛnnede no gyina hɔ ma akwan horow a wɔfa so sa yare: ahinanan, W + OSS1; ahinanan, W + OSM; kurukuruwa C
Afei, yɛde Nepenthes a wɔayɛ no mmerɛw (irCOI1 ne sETR1) a wɔayɛ no awosu mu nsakrae di dwuma wɔ anammɔn atitiriw a ɛwɔ JA ne ET biosynthesis (irAOC ne irACO) ne nkate (irCOI1 ne sETR1) mu de hwehwɛ afifide mu nkwaadɔm abien yi mu nneɛma a ɛsakra wɔ nhabannuru a wodi nhabannuru so The relative contribution of reprogramming. Nea ɛne sɔhwɛ ahorow a atwam hyia no, yesii so dua sɛ wɔde herbivore-OS ba afifide a empty carrier (EV) mu (Mfonini 3, C kosi D) ne Hj index a ɛso tew nyinaa a OSM de bae no so dua, bere a δj index no kɔɔ soro. Mmuae no da adi kɛse sen mmuae a OSS1 kanyan no. Ntrɛwmu abien graph a wɔde Hj ne δj di dwuma sɛ coordinates kyerɛ deregulation pɔtee no (Mfonini 3E). Nea ɛda adi kɛse ne sɛ wɔ nnomaa ahorow a wonni JA sɛnkyerɛnne mu no, ɛkame ayɛ sɛ metabolome ahorow ne nsakrae titiriw a mmoa a wodi nhabannuru de ba no fi hɔ koraa (Mfonini 3C). Nea ɛne eyi bɔ abira no, ET adwene a ɛyɛ komm wɔ sETR1 afifide mu, ɛwom sɛ nkɛntɛnso a ɛwɔ nsakrae a ɛba wɔ nhabannuru a wodi mu no nyinaa sua koraa sen JA sɛnkyerɛnne de, nanso brɛ nsonsonoe a ɛwɔ Hj ne δj indices mu wɔ OSM ne OSS1 excitations ntam no ase (Mfonini 3D ne Mfonini S5). . Eyi kyerɛ sɛ wɔ adwuma titiriw a JA sɛnkyerɛnne kwan so nsakrae yɛ akyi no, ET sɛnkyerɛnne kwan so nsakrae nso yɛ adwuma sɛ nhabannuru a wodi nhabannuru no nipadua mu nneɛma a ɛsakra wɔ mmoa ahorow pɔtee bi mu no ho nhyehyɛe pa. Nea ɛne saa adwuma a wɔde siesie nneɛma yiye yi hyia no, nsakrae biara amma metabolome inducibility nyinaa mu wɔ sETR1 afifide mu. Ɔkwan foforo so no, sɛ wɔde toto sETR1 afifide ho a, irACO afifide no maa nsakrae a ɛba nipadua mu nneɛma mu a ɛnam mmoa a wodi nhabannuru so de ba no nyinaa yɛɛ kɛse, nanso ɛdaa Hj ne δj nkontaahyɛde ahorow a ɛsono kɛse adi wɔ OSM ne OSS1 nsɛnnennen ntam (Mfonini S5).
Sɛnea ɛbɛyɛ a yebehu metabolites titiriw a ɛwɔ mmoa a ɛho hia wɔ mmoa ahorow pɔtee bi a wodi nhabannuru ho mmuae mu na yɛasiesie wɔn a wɔyɛ no yiye denam ET nsɛnkyerɛnne so no, yɛde structural MS/MS kwan a wɔadi kan ayɛ no dii dwuma. Saa kwan yi gyina bi-clustering kwan no so de san kyerɛ metabolic abusua no fi MS/MS asinasin [normalized dot product (NDP)] ne nsɛdi nkontaahyɛde a egyina neutral loss (NL) so. MS/MS data set a wɔnam ET transgenic lines nhwehwɛmu so yɛɛ no ​​maa MS/MS 585 (data file S1), a wɔsiesiee no denam wɔn a wɔboaboaa ano yɛɛ no ​​MS/MS module atitiriw ason (M) (Mfonini 4A) . Saa module yi bi yɛ nea metabolites soronko a wɔadi kan ayɛ no ahyɛ mu ma: sɛ nhwɛso no, M1, M2, M3, M4 ne M7 wɔ phenol a efi mu ba ahorow (M1), flavonoid glycosides (M2), acyl asikre (M3 Ne M4), ne 17-HGL-DTG (M7) pii. Bio nso, wɔbu metabolic specific information (Si index) a ɛwɔ metabolite baako a ɛwɔ module biara mu no ho akontaa, na wɔtumi hu ne Si kyekyɛ wɔ nkate mu. Ne tiawa mu no, MS/MS spectra a ɛda herbivory ne genotype specificity a ɛkorɔn adi no yɛ nea Si gyinapɛn a ɛkorɔn na ɛda adi, na kurtosis akontaabu kyerɛ sɛnea aboa nhoma kyekyɛ wɔ dua nifa so. Wohuu colloid kyekyɛ a ɛte saa a ɛyɛ mmerɛw no biako wɔ M1 mu, a phenol amide daa Si fã a ɛkorɔn sen biara adi (Mfonini 4B). Herbivorous inducible 17-HGL-DTG a yɛadi kan aka ho asɛm wɔ M7 mu no daa Si nkontabuo a ɛkɔ fam adi, a ɛkyerɛ sɛ nsonsonoeɛ nhyehyɛeɛ a ɛkɔ fam wɔ OS ahodoɔ mmienu no ntam. Nea ɛne eyi bɔ abira no, nneɛma titiriw a wɔde yɛ nneɛma a ɛma nipadua no yɛ adwuma, te sɛ rutin, CGA, ne acyl asikre dodow no ara ka Si nkontaahyɛde a ɛba fam sen biara no ho. Sɛnea ɛbɛyɛ a wɔbɛhwehwɛ nhyehyɛe mu nsɛnnennen ne Si kyekyɛ wɔ metabolites soronko ntam yiye no, wɔyɛɛ molecule network maa module biara (Mfonini 4B). OD nsusuwii no nkɔmhyɛ a ɛho hia (a wɔabɔ no mua wɔ Mfonini 1B mu) ne sɛ ɛsɛ sɛ metabolites soronko a wɔsan hyehyɛ wɔ herbivory akyi no ma nsakrae a ɛkɔ ɔkwan biako so ba metabolites a ɛwɔ ahobammɔ bo a ɛkorɔn mu, titiriw denam wɔn pɔtee a wɔma ɛkɔ soro (a ɛne random distribution bɔ abira) Mode) Defensive metabolite a MT nsusuwii no hyɛ ho nkɔm. Phenol a efi mu ba a wɔaboaboa ano wɔ M1 mu no mu dodow no ara ne nkoekoemmoa adwumayɛ a ɛso tew no wɔ abusuabɔ wɔ dwumadi mu ( 32 ). Bere a yɛde Si gyinapɛn ahorow a ɛwɔ M1 metabolites mu toto nhaban a wɔahyɛ no den ne nhaban a ɛyɛ EV control afifide no ntam wɔ nnɔnhwerew 24 mu no, yehui sɛ metabolic pɔtee a ɛwɔ metabolites pii mu wɔ nkoekoemmoa a wodi nhabannuru akyi no wɔ nkɔanim kɛse (Mfonini 4C). Wɔhunuu nkɔanim pɔtee a ɛbaa Si boɔ mu wɔ phenolamides a ɛbɔ wɔn ho ban nkutoo mu, nanso wɔanhunu nkɔanim biara wɔ Si boɔ mu wɔ phenols foforɔ ne metabolites a wonnim a ɛbom tra saa module yi mu. Eyi yɛ nhwɛso titiriw, a ɛne OD nsusuwii no wɔ abusuabɔ. Nkɔmhyɛ atitiriw a ɛfa nsakrae a ɛbɛba wɔ nipadua mu nneɛma a mmoa a wodi nhabannuru de ba ho no yɛ nea ɛkɔ so daa. Sɛnea ɛbɛyɛ a yɛbɛsɔ ahwɛ sɛ ebia saa particularity a ɛwɔ phenolamide spectrum no mu no, OS-specific ET na ɛde bae no, yɛyɛɛ metabolite Si index no ho mfonini na yɛde nsonsonoe a ɛda OSM ne OSS1 ntam wɔ EV ne sETR1 genotypes no mu bae (Mfonini 4D ). Wɔ sETR1 mu no, nsonsonoe a phenamide de ba OSM ne OSS1 ntam no so tew kɛse. Wɔde bi-clustering kwan no nso dii dwuma wɔ MS/MS data a wɔaboaboa ano wɔ nnomaa a JA nnɔɔso mu de asusuw MS/MS module atitiriw a ɛfa JA-regulated metabolic specialization ho (Mfonini S6).
(A) Nneɛma a ɛfiri MS/MS 585 a wɔaboaboa ano a egyina asinasin a wɔkyɛ (NDP nsɛsoɔ) ne adehwereɛ a wɔkyɛ (NL nsɛsoɔ) so no ma module (M) no ne aduru abusua a wonim no hyia, anaasɛ ɛnam Metabolite dodoɔ a wonnim anaasɛ ɛnnyɛ yie so. Wɔ module biara nkyɛn no, wɔakyerɛ metabolite (MS/MS) pɔtee (Si) nkyekyɛmu. (B) Modular molecular network: Nodes gyina hɔ ma MS/MS ne anoano, NDP (kɔkɔɔ) ne NL (blue) MS/MS nkontabuo (cut-off,> 0.6). Graded metabolite specificity index (Si) no de kɔla gyinaa module no so (benkum so) na wɔde yɛɛ map kɔɔ molecule network (nifa so). (C) Module M1 a EV afifide a ɛwɔ constitutive (control) ne induced tebea mu (simulated herbivore) wɔ nnɔnhwerew 24 mu: molecular network diagram (Si bo yɛ node kɛse, wɔde bruu asi defensive phenolamide so dua). (D) M1 molecular network diagram a ɛkyerɛ spectrum line sETR1 a EV ne ET nkateɛ asɛe: phenolic compound a wɔde green circle node gyina hɔ ma, ne nsonsonoeɛ kɛseɛ (P value) a ɛda W + OSM ne W + OSS1 ayaresa ntam sɛ Node kɛseɛ. CP, N-caffeoyl-tyrosine a wɔde yɛ aduru; CS, N-caffeoyl-spermidine, ne nea wɔde yɛ aduru; FP, N-ferulic acid a ɛyɛ ester-uric acid, a ɛyɛ aduru a ɛma obi nya ahoɔden; FS, N-ferulyl-spermidine, ne nea ɛwɔ nipadua no mu; CoP, N’, N “-Kumarolyl-tyrosine; CFS, N’, N”-caffeoyl, feruloyl-spermidine, Lycium barbarum a ɛwɔ ɔkraman Ba ​​mu, Nick.O-AS, O-acyl asikre.
Yɛkɔɔ so trɛw nhwehwɛmu no mu fii Nepenthes genotype biako a wɔayɛ no mmerɛw so kɔɔ abɔde mu nnipa dodow so, faako a wɔadi kan aka nsakrae a emu yɛ den a ɛba wɔ nhabannuru JA dodow ne metabolite dodow pɔtee mu ho asɛm wɔ abɔde mu nnipa dodow mu ( 26 ). Fa saa data set yi kata germplasms 43 so. Saa mmoawa yi yɛ afifide ahorow 123 a efi N. pallens mu. Wɔfaa saa afifideɛ yi firii aba a wɔboaboaa ano wɔ kurom hɔ mmeaeɛ ahodoɔ wɔ Utah, Nevada, Arizona, ne California (Mfonini S7), yɛbuu metabolome ahodoɔ (wɔfrɛ no ha sɛ nnipa dodoɔ) β ahodoɔ) ne specialization a OSM de baeɛ no ho akontaa. Nea ɛne nhwehwɛmu a atwam hyia no, yehuu nsakrae ahorow pii a ɛba nipadua mu nneɛma mu wɔ Hj ne δj axes no so, a ɛkyerɛ sɛ germplasms wɔ nsonsonoe kɛse wɔ plasticity a ɛwɔ wɔn nipadua mu nneɛma mu mmuae a wɔde ma wɔ herbivores ho no mu (Mfonini S7). Saa ahyehyɛde yi kae nea wɔadi kan ahu wɔ JAs nsakrae a ɛyɛ nnam a mmoa a wodi nhabannuru de ba no ho, na akura bo a ɛkorɔn yiye mu wɔ nnipa biako mu (26, 36). Ɛdenam JA ne JA-Ile a yɛde dii dwuma de sɔɔ level correlation a ɛda Hj ne δj ntam no hwɛe so no, yehui sɛ abusuabɔ pa a ɛho hia wɔ JA ne metabolome β ahorow ne specialization index ntam (Mfonini S7). Eyi kyerɛ sɛ heterogeneity a herbivore-induced wɔ JAs induction a wɔahu wɔ population level no betumi afi metabolic polymorphisms atitiriw a ɛnam paw a wɔpaw fii nkoekoemmoa herbivores mu ba.
Nhwehwɛmu ahorow a atwam ada no adi sɛ ɛsono tawa ahorow no kɛse wɔ ne su ne sɛnea egyina nipadua no mu nneɛma a ɛbɔ wɔn ho ban a wɔde ba ne nea wɔde hyɛ mu so kakra no mu. Wogye di sɛ nsakrae yi a ɛba wɔ anti-herbivore signal transduction ne ahobammɔ tumi mu no nam nkoekoemmoa dodow nhyɛso, afifide asetra kyinhyia, ne ahobammɔ ho ka a wɔbɔ wɔ niche a mmoa ahorow bi nyin no so na ɛhwɛ so. Yɛyɛɛ sɛnea nhaban mu metabolome a wɔsan yɛ no foforo a Nicotiana ahorow asia a wodi nhabannuru a wofi Amerika Atifi fam ne Amerika Kesee Fam de ma no yɛ pɛpɛɛpɛ no ho nhwehwɛmu. Saa mmoa ahorow yi ne Nepenthes Amerika Atifi fam, a ɔne Nicolas Bociflo, wɔ abusuabɔ kɛse. La, N. nicotinis, Nicotiana n., ne nea ɛwɔ nipadua no mu. sare a ayɛ mmerɛw, Nicotiana tabacum, tawa a ɛyɛ linear, tawa (Nicotiana spegazzinii) ne tawa ahaban tawa (Nicotiana obtusifolia) (Mfonini 5A) (37). Saa mmoa yi mu asia, a N. please a wɔakyerɛ wɔn su yiye ka ho, yɛ afe afe afifide a ɛwɔ petunia clade mu, na obtusifolia N. yɛ afe afe afifide a ɛwɔ onuawa clade Trigonophyllae (38). Akyiri yi, wɔyɛɛ W + W, W + OSM ne W + OSS1 induction wɔ saa mmoa ahorow nson yi so de suaa sɛnea nkoekoemmoa aduan a wɔsan de ma wɔn nipadua mu nneɛma a wɔsan yɛ wɔ mmoa ahorow no mu no ho ade.
(A) Bootstrap phylogenetic dua a egyina nea ebetumi aba kɛse [ma nuklea glutamine a wɔyɛ (38)] ne asasesin a Nicotiana ahorow ason a ɛne wɔn ho wɔn ho di nkitaho kɛse (kɔla ahorow) (37) so. (B) Apete mfonini a ɛkyerɛ ahorow ahorow soronko ma Nicotiana ahorow ason (939 MS/MS; data fael S1) nipadua mu nneɛma a ɛsakra no. Wɔ mmoa ahorow no gyinabea no, metabolome ahorow ahorow ne sɛnea wɔyɛ soronko no wɔ abusuabɔ bɔne. Nhwehwɛmu a ɛfa abusuabɔ a ɛda metabolic ahorow ne specialization ne JA a wɔboaboa ano ntam wɔ species-level correlation ntam no wɔakyerɛ wɔ Mfonini 2. S9. Kɔla, ahorow ahorow; ahinanan, W + OSS1; ahinanan, W + OSM; (C) Wɔde Nicotiana JA ne JA-Ile dynamics no toto ho sɛnea OS excitation amplitude te (akwan abien ANOVA ne Tukey HSD akyi ntotoho, * P <0.05, ** P <0.01 ne * ** Sɛ wɔde toto W + OS ne W + W ho a, P <0.001). Adaka mu mfonini a ɛkyerɛ (D) ahorow ahorow ne (E) soronkoyɛ a ɛfa mmoa ahorow biara ho bere a wɔayɛ nhabannuru ne methyl JA (MeJA) ho mfonini akyi. Nsoromma no kyerɛ nsonsonoe kɛse a ɛda W + OS ne W + W anaa lanolin a wɔde W (Lan + W) aka ho anaa Lan a wɔde MeJA aka ho (Lan + MeJa) ne Lan control (akwan abien nhwehwɛmu a ɛfa nsonsonoe ho, a Tukey HSD post hoc multiple comparison di akyi , *P<0.05, **P<0.01 ne ***P<0.001) ntam.
Yɛde dual cluster kwan no dii dwuma no, yɛhunuu module 9 a ɛwɔ 939 MS/MS (data file S1). MS/MS a wɔde ayaresa ahodoɔ asan asiesie no ahodoɔ no gu ahodoɔ kɛseɛ wɔ module ahodoɔ mu wɔ mmoa ahodoɔ ntam (Mfonini S8). Sɛ yɛyɛ Hj (a wɔfrɛ no ha sɛ γ-ahorow ahorow a ɛwɔ mmoa ahorow mu) ne δj ho mfonini wɔ yɛn adwenem a, ɛda adi sɛ mmoa ahorow ahorow boaboa wɔn ho ano yɛ akuw ahorow koraa wɔ nipadua mu nneɛma a ɛsakra no mu, baabi a mpɛn pii no mpaapaemu a ɛwɔ mmoa ahorow mu no da adi kɛse sen anigye. Sɛ yɛyi N. linear ne N. obliquus a, wɔda induction nsunsuansoɔ a ɛyɛ nnam pii adi (Mfonini 5B). Nea ɛne eyi bɔ abira no, mmoa ahorow te sɛ N. purpurea ne N. obtusifolia nni nipadua mu nneɛma a ɛsakra a ɛda adi kɛse wɔ ayaresa ho, nanso nipadua mu nkwaadɔm no gu ahorow kɛse. Nkyekyɛmu pɔtee a ɛfa nipadua mu nneɛma a ɛma nipadua no yɛ adwuma ho no maa abusuabɔ bɔne a ɛho hia baa specialization ne gamma ahorow ahorow ntam (PCC = -0.46, P = 4.9×10-8). Nsakraeɛ a OS de ba JAs dodoɔ mu no ne metabolome specialization wɔ abusuabɔ pa, na ɛne metabolic gamma ahodoɔ a ɛda adi wɔ mmoa ahodoɔ biara mu no wɔ abusuabɔ bɔne (Mfonini 5B ne Mfonini S9). Ɛfata sɛ yɛhyɛ no nsow sɛ mmoa ahorow a wɔka ho asɛm wɔ kasa mu sɛ “sɛnkyerɛnne kwan so mmuae” ahorow wɔ Mfonini 5C mu, te sɛ Nepenthes nematodes, Nepenthes nepenthes, Nepenthes acute, ne Nepenthes attenuated no de nsɛnkyerɛnne a ɛho hia bae wɔ simma 30 mu. Nnansa yi OS-specific JA ne JA-Ile a ɛpae gui, bere a mmoawa afoforo a wɔfrɛ wɔn “signal unresponsive”, te sɛ Nepenthes mills, Nepenthes powdery ne N. obtusifolia nkutoo kyerɛ JA-Ile Edge induction a OS pɔtee biara nni mu (Mfonini 5C). Wɔ metabolic level, sɛnea yɛaka wɔ atifi hɔ no, wɔ attenuated Nepenthes ho no, signal-responsive nneɛma no daa OS pɔtee adi na ɛmaa δj kɔɔ soro kɛse, bere a ɛtew Hj so. Wɔanhu saa OS-specific priming effect yi wɔ species a wɔakyekyɛ mu sɛ signal non-reactive species (Mfonini 5, D ne E). Wɔtaa kyɛ OS-specific metabolites wɔ signal-responsive species ntam, na saa signal clusters yi ne species a wɔn signal responses yɛ mmerɛw no bom, bere a species a wɔn signal responses yɛ mmerɛw no nkyerɛ sɛ wɔmfa wɔn ho nhyɛ wɔn ho wɔn ho so kɛse (Mfonini S8 ). Saa aba yi kyerɛ sɛ OS-specific induction of JAs ne OS-specific reconfiguration of the downstream metabolome no ka bom wɔ species level.
Afei, yɛde lanolin paste a methyl JA (MeJA) wom dii dwuma de saa afifide ho adwuma de hwehwɛɛ sɛ ebia saa coupling modes yi anohyeto wɔ JA a ɛwɔ hɔ a exogenous JA de di dwuma, a ɛbɛyɛ afifide cytoplasm mu no. Deesterification a ɛba ntɛmntɛm ne JA. Yehuu saa su koro no ara a ɛfa nsakrae a ɛba nkakrankakra fi nnomaa a wɔyɛ nsɛnkyerɛnne so kɔ nnomaa a wɔnyɛ sɛnkyerɛnne kwan so a ɛnam JA a wɔkɔ so de ma no so ba no ho (Mfonini 5, D ne E). Ne tiawa mu no, MeJA ayaresa san yɛɛ linear nematodes, N. obliquus, N. aquaticus, N. pallens, ne N. mikimotoi metabolomes ho nhyehyɛe denneennen, na ɛde δj kɔɔ soro kɛse na Hj so tew bae. N. purpurea daa nkɔanim adi wɔ δj mu nkutoo, nanso ɛnyɛ Hj. N. obtusifolia a wɔadi kan ada no adi sɛ ɛboaboa JA dodow a ɛba fam koraa ano no nso nyɛ adwuma yiye wɔ MeJA ayaresa ho wɔ metabolome a wɔsan hyehyɛ no mu. Saa nsunsuansoɔ yi kyerɛ sɛ JA a wɔyɛ anaa nsɛnkyerɛnneɛ a wɔde kɔ nipadua mu no anohyetoɔ wɔ nipadua mu wɔ mmoa ahodoɔ a wɔnyɛ nsɛnkyerɛnneɛ ho mmuaeɛ mu. Sɛ yɛbɛsɔ saa adwene yi ahwɛ a, yɛyɛɛ mmoa ahorow anan (N. pallens, N. mills, N. pink ne N. microphylla) a W + W, W + OSMs ne W + OSS1 Transcriptome (39) de ba no ho nhwehwɛmu. Nea ɛne metabolome remodeling nhyehyɛe no hyia no, wɔatew mmoa ahorow no mu yiye wɔ transcriptome space no mu, a emu bi mu no N. attenuated daa RDPI a OS de bae a ɛkorɔn sen biara adi, bere a N. gracilis nyaa nea ɛba fam sen biara (Mfonini 6A). Nanso, wohui sɛ transcriptome ahorow a N. oblonga de ba no na ɛba fam sen biara wɔ mmoa ahorow anan no mu, na ɛne N. oblonga metabolic ahorow ahorow a ɛkorɔn sen biara a wɔadi kan ada no adi wɔ mmoa ahorow ason mu no bɔ abira. Nhwehwɛmu a atwam no ada no adi sɛ awosu mu nkwaadɔm bi a ɛfa ahobammɔ nsɛnkyerɛnne a edi kan ho, a JA nsɛnkyerɛnne ka ho, kyerɛkyerɛ sɛnea ahobammɔ ho mmuae a edi kan a elicitors a ɛfa nhabannuru ho de ba wɔ Nicotiana ahorow mu no mu ( 39 ). Sɛ wɔde JA sɛnkyerɛnne akwan a ɛwɔ saa mmoa ahorow anan yi ntam toto ho a, ɛdaa nhwɛso bi a ɛyɛ anigye adi (Mfonini 6B). Awosu mu nkwaadɔm dodow no ara a ɛwɔ saa kwan yi so, te sɛ AOC, OPR3, ACX ne COI1, daa induction a ɛkɔ soro kakra adi wɔ saa mmoa ahorow anan yi mu. Nanso, awosu mu abɔde titiriw bi, JAR4, dan JA ma ɛyɛ nea ɛyɛ adwuma wɔ abɔde mu a ɛyɛ JA-Ile nkyerɛwee a wɔaboaboa ano, na ne nkyerɛwee dodow sua koraa, titiriw wɔ N. mills, Nepenthes pieris ne N. microphylla mu. Bio nso, wɔanhu awosu mu abɔde foforo AOS no nkyerɛwee nkutoo wɔ N. bifidum mu. Ebia nsakrae yi a ɛba wɔ awosu mu nkwaadɔm a wɔda no adi mu no na ɛde phenotypes a ɛtra so a JA a ɛba fam wɔ signal anergic ahorow mu ne N. gracilis a ɛba no de ba no ba.
(A) Amanneɛbɔ ho nsusuwii mu nhwehwɛmu a wɔyɛe wɔ tawa ahorow anan a ɛne wɔn ho wɔn ho di nsɛ a wɔfaa wɔn nhwɛsode wɔ simma 30 akyi wɔ nhabannuru a wɔde ma akyi no ho mmuae a wɔde kyerɛw nsɛm a edi kan a wɔsan yɛ ho nhyehyɛe ho. Wɔnam nhaban a herbivore OS de ba no a wɔde toto nea wɔde siw kuru no ho no so na ebu RDPI. Kɔla ahorow no kyerɛ mmoa ahorow, na sɛnkyerɛnnede ahorow no kyerɛ akwan horow a wɔfa so sa yare. (B) Nhwehwɛmu a wɔyɛe wɔ awosu mu nkwaadɔm a wɔda no adi wɔ JA sɛnkyerɛnne akwan so wɔ mmoa ahorow anan mu. Wɔakyerɛ JA kwan a wɔayɛ no mmerɛw no wɔ adaka mu mfonini no nkyɛn. Kɔla ahorow kyerɛ akwan horow a wɔfa so yɛ adwuma. Nsoromma no kyerɛ sɛ nsonsonoe kɛse da W + OS ayaresa ne W + W a wɔde di dwuma no ntam (ma Osuani no t-sɔhwɛ a ɛfa nsonsonoe a ɛwɔ abien abien mu no, *P<0.05, **P<0.01 ne ***P<0.001). OPDA, 12-oxophytodienoic acid a ɛwɔ nipadua no mu; OPC-8: 0,3-oxo-2 (2 '(Z)-pentenyl)-cyclopentane-1-octanoic acid.
Wɔ ɔfã a etwa to no mu no, yesuaa sɛnea nkoekoemmoa ahorow pɔtee bi a wɔsan yɛ no foforo wɔ afifide ahorow mu metabolome mu no betumi ayɛ nea ɛko tia mmoa a wodi nhabannuru ho ade. Nhwehwɛmu a atwam no sii Nicotiana abusua no so dua. Ɛsono sɛnea wɔko tia Ms ne larvae kɛse (40). Ɛha yi, yɛsuaa abusuabɔ a ɛda saa nhwɛso yi ne wɔn metabolic plasticity ntam. Yɛde tawa ahorow anan a ɛwɔ atifi hɔ no dii dwuma, na yɛsɔɔ abusuabɔ a ɛda metabolome a egu ahorow ne nea ɛyɛ soronko a afifide a wodi nhabannuru de ba ne sɛnea afifide tumi ko tia Ms ne Sl ntam hwɛe no, yehui sɛ sɛnea wɔko tia, sɛnea wɔko tia Ms ne Sl a ɛyɛ soronko no nyinaa wɔ abusuabɔ pa, bere a abusuabɔ a ɛda mmea a wɔyɛ abenfo a wɔko tia ne specialization ntam no yɛ mmerɛw, na abusuabɔ a ɛda ahorow ahorow ntam no nyɛ nea ɛho hia (Mfonini S10). Ɛdefa S1 a wɔko tia ho no, N. chinensis a wɔayɛ mmerɛw ne N. gracilis a wɔadi kan akyerɛ sɛ ɛda JA sɛnkyerɛnne kwan so nsakrae dodow ne metabolome plasticity nyinaa adi no nyinaa nyaa mmuae soronko kɛse wɔ herbivore induction ho, na wɔsan nso daa ahoɔden a ɛkorɔn a ɛte saa ara adi. Ɔbarima ne ɔbea nna.
Wɔ mfe aduosia a atwam no mu no, afifide ahobammɔ ho nsusuwii ama nsusuwii nhyehyɛe bi ama, a nhwehwɛmufo agyina so ahyɛ afifide mu nneɛma titiriw a ɛma nipadua no yɛ adwuma no adannandi ne dwumadi dodow bi a ɛdɔɔso ho nkɔm. Saa nsusuwii ahorow yi mu dodow no ara nni ɔkwan a wɔtaa fa so de nsusuwii a emu yɛ den akyi ( 41 ). Wɔde nkɔmhyɛ atitiriw (3) ho nyansahyɛ ma wɔ nhwehwɛmu gyinabea koro no ara mu. Sɛ nkɔmhyɛ atitiriw a wɔsɔ hwɛ no ma kwan ma wɔhwehwɛ nsusuwii pɔtee bi mu a, eyi bɛma The field is moving forward. Wɔboa wo, nanso pow afoforo (42). Mmom no, nsusuwii foforo no hyɛ nkɔm wɔ nhwehwɛmu gyinabea ahorow mu na ɛde nkyerɛkyerɛmu nsusuwii foforo ka ho (42). Nanso, ɛnyɛ den sɛ wɔbɛkyerɛkyerɛ nsusuwii abien a wɔahyɛ ho nyansa wɔ dwumadi mu, MT ne OD, mu sɛ nkɔmhyɛ ahorow a ɛho hia a ɛfa nsakrae titiriw a ɛba nipadua mu a mmoa a wodi nhabannuru de ba ho: OD nsusuwii gye di sɛ nsakrae a ɛba nipadua mu nneɛma a ɛsakra “ahunmu” titiriw mu no yɛ akwankyerɛ kɛse . MT nsusuwii no gye di sɛ nsakrae yi bɛyɛ nea ɛnyɛ akwankyerɛ na ɛbɛba randomly wɔ metabolic space, na ɛtaa nya defense value metabolites a ɛkorɔn. Wɔde a priori “defense” compounds teateaa bi asɔ OD ne MT nkɔmhyɛ ahorow mu nhwehwɛmu ahorow a atwam no ahwɛ. Saa sɔhwɛ ahorow a ɛfa metabolite ho yi siw tumi a wɔde bɛhwehwɛ baabi a metabolome san hyehyɛ no kɛse ne ɔkwan a ɛkɔ so wɔ bere a wɔredi nhabannuru no mu, na ɛmma kwan mma sɔhwɛ a wɔyɛ wɔ akontaabu nhyehyɛe a ɛkɔ so daa mu no nhwehwɛ nkɔmhyɛ atitiriw a wobetumi asusuw ho sɛ ne nyinaa Kyerɛ nsakrae dodow a ɛba afifide mu metabolome mu. Ɛha yi, yɛde mfiridwuma foforo a ɛwɔ metabolomics mu a egyina kɔmputa MS so dii dwuma na yɛyɛɛ deconvolution MS nhwehwɛmu wɔ sika a wɔde di dwuma wɔ nsɛm ho nsusuwii nkyerɛkyerɛmufo mu nyinaa mu de sɔɔ nsonsonoe a ɛda abien a wɔahyɛ ho nyansa wɔ wiase nyinaa metabolomics gyinabea no hwɛe. Nkɔmhyɛ titiriw a ɛwɔ saa nsusuwii yi mu. Wɔde nsɛm ho nsusuwii adi dwuma wɔ mmeae pii, titiriw wɔ abɔde a nkwa wom ne aduannuru a ɛsen ho nhwehwɛmu mu (43). Nanso, sɛnea yenim no, eyi ne dwumadie a ɛdi kan a wɔde di dwuma de kyerɛkyerɛ afifideɛ mu nipadua mu nkwaadɔm ho nsɛm atenaeɛ mu na wɔdi nneɛma a atwa yɛn ho ahyia ho haw a ɛfa bere tiaa mu nipadua mu nsakraeɛ a ɛba de bua nneɛma a atwa yɛn ho ahyia ho nsɛnkyerɛnneɛ ho dwuma. Titiriw no, tumi a saa kwan yi tumi de toto afifide ahorow mu ne wɔn ntam ho de hwehwɛ sɛnea mmoa a wodi nhabannuru adan afi mmoa ahorow ahorow mu akɔ mmoa ahorow ntam adannandi kɛse nhyehyɛe ahorow mu wɔ adannandi gyinabea ahorow mu. Nkwammoaa mu nneɛma a ɛsakra.
Principal component analysis (PCA) dane multivariate data set kɔ dimensionality reduction space mu sɛdeɛ ɛbɛyɛ a wɔbɛtumi akyerɛkyerɛ data no su titire mu, enti wɔtaa de di dwuma sɛ nhwehwɛmu kwan a wɔfa so hwehwɛ data set no mu, te sɛ deconvolution metabolome. Nanso, dimensionality reduction bɛhwere nsɛm a ɛwɔ data set no mu no fã bi, na PCA ntumi mfa nsɛm dodow a ɛfa su ahorow a ɛfa abɔde a nkwa wom ho nsusuwii ho titiriw mma, te sɛ: sɛnea herbivores san hyehyɛ ahorow ahorow wɔ mfuw titiriw mu (sɛ nhwɛso no, ahonyade, nkyekyɛmu) Ne dodow) metabolites? Nneɛma a ɛma nipadua no yɛ adwuma bɛn na ɛkyerɛ tebea a wɔde ma aboa bi a odi nhabannuru? Sɛ yɛhwɛ nneɛma pɔtee, ahodoɔ ne inducibility a, nsɛm a ɛwɔ ahaban pɔtee metabolite profile no mu no porɔw, na wɔhunu sɛ herbivores adidi tumi ma metabolism pɔtee bi yɛ adwuma. Nea yɛnhwɛ kwan no, yehui sɛ, sɛnea wɔaka ho asɛm wɔ nsɛm ho nsusuwii nsɛnkyerɛnne a wɔde dii dwuma no mu no, nipadua mu nneɛma a ɛsakra tebea a efi mu ba no wɔ ntua kɛse wɔ ntua a mmoa abien a wodi nhabannuru (oduruyɛfo a odi nhabannuru anadwo) ne Solanaceae ho ɔbenfo Ms. Ɛwom sɛ ɛsono wɔn aduan a wɔde ma ne sɛnea wɔde wɔn adwene si so kɛse de. Fatty acid-amino acid conjugate (FAC) a wɔde fi ase wɔ OS mu (31). Ɛdenam OS a wɔde sa nhabannuru a wɔde saa akuru a wɔahyɛ da ayɛ so no, nnuru a wɔde di nhabannuru a wɔayɛ no sɛnea ɛte no nso daa su a ɛte saa ara adi. Saa ɔkwan a wɔahyɛ da ayɛ a wɔfa so yɛ sɛnea afifide yɛ wɔn ade wɔ ntua a wɔde di nhabannuru ho no yi nneɛma a ɛyɛ basaa a nsakrae a ɛba mmoa a wodi nhabannuru adidi mu, a ɛde ɔsɛe a ɛsonosonoe ba wɔ mmere ahorow mu no fi hɔ (34). FAC a wonim sɛ ɛno na ɛde OSM ba titiriw no brɛ JAS ne afifide mu nkwaadɔm afoforo a ɛma nipadua no yɛ adwuma no ase wɔ OSS1 mu, bere a OSS1 brɛ ase mpɛn ɔhaha pii ( 31 ). Nanso, OSS1 maa JA boaboaa ano dodow a ɛte saa ara bae bere a wɔde totoo OSM ho no. Wɔadi kan ada no adi sɛ JA mmuae a ɛwɔ Nepenthes a ayɛ mmerɛw mu no yɛ nea ɛyɛ mmerɛw yiye wɔ OSM ho, baabi a FAC betumi akura ne dwumadi mu mpo sɛ wɔde nsu afra mu 1:1000 a ( 44 ). Enti, sɛ wɔde toto OSM ho a, ɛwom sɛ FAC a ɛwɔ OSS1 mu no sua koraa de, nanso ɛdɔɔso sɛ ɛbɛma JA a ɛpae a ɛdɔɔso. Nhwehwɛmu ahorow a atwam ada no adi sɛ wobetumi de protein ahorow a ɛte sɛ porin (45) ne oligosaccharides (46) adi dwuma sɛ molecule nsɛnkyerɛnne de akanyan afifide ahobammɔ mmuae wɔ OSS1 mu. Nanso, ɛda so ara yɛ nea wontumi nhu sɛ ebia saa elicitors a ɛwɔ OSS1 mu yi na ɛma JA a wɔahu wɔ mprempren nhwehwɛmu no mu no boaboa ano anaa.
Ɛwom sɛ nhwehwɛmu kakraa bi na ɛwɔ hɔ a ɛkyerɛkyerɛ nsonsonoe a ɛwɔ nipadua mu nsateaa nkyerɛwee a efi mmoa ahorow a wodi nhabannuru anaa exogenous JA anaa SA (salicylic acid) a wɔde di dwuma mu ba no mu (47), nanso obiara nhaw nhabannuru ahorow pɔtee bi a ɛhaw adwene wɔ afifide sare ntam ne nkɛntɛnso a enya wɔ pɔtee no so The overall impact of metabolism is systematically studied. ankorankoro ho nsɛm. Saa nhwehwɛmu yi san sii so dua sɛ abusuabɔ a ɛda nipadua no mu nkwaadɔm ntam nkitahodi ne afifide mu nkwaadɔm afoforo a ɛnyɛ JAs ntam no na ɛkyerɛ sɛnea nipadua mu nkwaadɔm mu nsakrae a mmoa a wodi nhabannuru de ba no yɛ pɔtee. Titiriw no, yehui sɛ ET a OSM de ba no sõ kɛse sen nea OSS1 de ba no. Saa mode yi ne FAC content pii a ɛwɔ OSM mu no hyia, a ɛyɛ tebea a ɛho hia na ɛdɔɔso a wɔde kanyan ET burst ( 48 ). Wɔ nkitahodi a ɛda afifide ne wɔn a wodi nhabannuru ntam no mu no, ET sɛnkyerɛnne dwumadi wɔ afifide pɔtee bi mu nneɛma a ɛsakra no so no da so ara yɛ nea ɛba bere ne bere mu na ɛde n’ani si kuw biako pɛ a wɔde afrafra so. Bio nso, nhwehwɛmu dodow no ara de ET a wɔde di dwuma wɔ akyi anaa nea edii n’anim anaa nnuru ahorow a esiw ano adi dwuma de asua ET no nhyehyɛe ho ade, a nea ɛka ho ne nnuru a wɔde di dwuma wɔ akyi yi bɛma wɔanya nsunsuanso bɔne pii a ɛnyɛ pɔtee. Sɛnea yenim no, nhwehwɛmu yi gyina hɔ ma nhyehyɛe mu nhwehwɛmu kɛse a edi kan a ɛfa dwuma a ET di wɔ ET a wɔde di dwuma de yɛ na wohu afifide a wɔadan wɔn awosu mu nkwaadɔm a asɛe de ayɛ afifide mu metabolome nkɔso ho nhyehyɛe. Herbivore-specific ET induction betumi awiei koraa no sesa metabolome mmuae. Nea ɛho hia sen biara ne transgenic manipulation a ɛfa ET biosynthesis (ACO) ne perception (ETR1) awosu mu nkwaadɔm a ɛdaa herbivore pɔtee bi de novo a ɛboaboa phenolamides ano adi. Wɔadi kan ada no adi sɛ ET betumi asiesie nicotine a JA de ba no yiye denam putrescine N-methyltransferase a ɛhwɛ so no so ( 49 ). Nanso, sɛ yɛhwɛ mfiridwuma a, ɛnyɛ nea ɛda adi pefee sɛnea ET siesie phenamide a wɔde ba no yiye. Wɔ ET sɛnkyerɛnne kwan so nsakrae dwumadi akyi no, wobetumi nso de metabolic flux no akɔ S-adenosyl-1-methionine mu de ahwɛ sika a wɔde hyɛ polyaminophenol amides mu no so. S-adenosyl-1-methionine yɛ ET ne Common ntamgyinafo a polyamine biosynthetic kwan. Ɛsɛ sɛ wosua ɔkwan a ET sɛnkyerɛnne fa so hwɛ phenolamide dodow so.
Bere tenten ni, esiane sɛ nneɛma titiriw a ɛma nipadua no yɛ adwuma a wonnim no dɔɔso nti, adwene a wɔde sii nipadua no mu nneɛma akuw pɔtee bi so kɛse no ntumi nsusuw bere mu nsakrae a ɛba nipadua mu nneɛma ahorow mu wɔ abɔde a nkwa wom nkitahodi akyi no ho yiye. Mprempren, a egyina nsɛm ho nsusuwii nhwehwɛmu so no, nea efi MS/MS spectrum a wonya a egyina metabolites a enni animhwɛ so ba titiriw ne sɛ mmoa a wodi nhabannuru a wodi anaa wɔyɛ wɔn a wodi nhabannuru ho mfonini no kɔ so tew nhaban metabolome no nipadua mu nneɛma ahorow nyinaa so bere a ɛma ne titiriw no kɔ soro no. Saa bere tiaa mu nkɔanim a ɛkɔ soro wɔ metabolome pɔtee a mmoa a wodi nhabannuru de ba yi ne nkɔanim a ɛkɔ soro wɔ adwumayɛ mu wɔ transcriptome pɔtee mu no wɔ abusuabɔ. Ade a ɛboa kɛse ma saa metabolome specialization kɛse yi (a ɛwɔ Si bo a ɛkorɔn) ne metabolite titiriw a ɛwɔ herbivorous dwumadi a kan no na ɛyɛ ne su. Saa nhwɛso yi ne OD nsusuwii no nkɔmhyɛ hyia, nanso MT ho nkɔmhyɛ a ɛfa metabolome reprogramming a ɛba kwa no ho no nhyia. Nanso, saa nsɛm yi nso ne nkɔmhyɛ a ɛfa nhwɛso a wɔadi afra (MT a eye sen biara; Mfonini 1B) no hyia, efisɛ ebia metabolites afoforo a wonnim a wonnim wɔn ahobammɔ dwumadi ahorow da so ara di Si kyekyɛ a wɔanhyɛ da akyi.
Nhwɛso a ɛda nsow a nhwehwɛmu yi kyere bio ne sɛ efi adannandi nketenkete (afifide biako ne tawa dodow) so kosi adannandi kwan kɛse so (tawa ahorow a ɛne wɔn ho wɔn ho wɔ abusuabɔ kɛse) so no, adannandi nhyehyɛe ahorow a ɛsonosonoe wɔ “ahobammɔ a eye sen biara” mu Nsonsonoe kɛse wɔ mmoa a wodi nhabannuru tumi mu. Moore ne afoforo. (20) ne Kessler ne Kalske (1) de wɔn ho hyɛɛ nyansa sɛ wɔbɛdan abɔdeɛ a nkwa wom ahodoɔ a ɛyɛ adwuma mmiɛnsa a mfitiaseɛ no Whittaker (50) kyerɛɛ nsonsonoeɛ no mu no ayɛ no nsakraeɛ a ɛba wɔ nnuru ahodoɔ mu a ɛyɛ ne berɛ mu nsakraeɛ a ɛde ba; saa akyerɛwfo yi mu biara anbɔ mua Akwan a wɔfa so boaboa metabolome ho nsɛm ano kɛse nso nkyerɛ sɛnea wobebu nipadua mu nneɛma ahorow ho akontaa afi saa nsɛm yi mu. Wɔ saa nhwehwɛmu yi mu no, nsakraeɛ nketenkete a wɔbɛyɛ wɔ Whittaker dwumadie nkyekyɛmu mu no bɛbu α-metabolic ahodoɔ sɛ MS/MS spectra ahodoɔ a ɛwɔ afifideɛ bi a wɔde ama mu, ne β-metabolic ahodoɔ sɛ mfitiaseɛ intraspecific metabolism a ɛwɔ nnipa dodoɔ kuw bi mu Ahunmu, ne γ-metabolic ahodoɔ bɛyɛ ntrɛmu a ɛfa nhwehwɛmu a wɔyɛ wɔ afifideɛ a ɛte saa ara ho.
JA sɛnkyerɛnne no ho hia ma herbivore metabolic mmuae ahorow pii. Nanso, dodow mu sɔhwɛ a emu yɛ den nni hɔ a ɛkyerɛ mmoa a JA biosynthesis nhyehyɛe a ɛwɔ nkwaboaa mu no de ma wɔ metabolome ahorow mu, na sɛ́ ebia JA sɛnkyerɛnne yɛ beae titiriw a adwennwen de ba nipadua mu nneɛma ahorow a ɛsonosonoe wɔ macroevolutionary nsenia a ɛkorɔn so no da so ara yɛ nea wontumi nhu. Yɛhunuu sɛ Nepenthes herbivorous su a ɛyɛ herbivorous no ma metabolome specialization na nsakraeɛ a ɛba metabolome specialization mu wɔ Nicotiana ahodoɔ dodoɔ mu ne Nicotiana ahodoɔ a ɛbɛn wɔn ho no mu no ne JA signaling wɔ abusuabɔ pa wɔ nhyehyɛeɛ kwan so. Bio nso, sɛ JA sɛnkyerɛnne no sɛe a, wɔbɛtwa metabolic specificity a genotype herbivore biako de ba no mu (Mfonini 3, C ne E). Esiane sɛ nsakrae a ɛba Nepenthes nnipa dodow a wɔayɛ mmerɛw wɔ abɔde mu no mu dodow no ara yɛ dodow nti, nsakrae a ɛba nipadua mu β ahorow ne nea ɛyɛ pɔtee wɔ nhwehwɛmu yi mu no betumi afi nnuru akuw a ɛwɔ nipadua mu nneɛma a ɛdɔɔso mu a ɛkanyan denneennen no kɛse. Saa afrafra akuw yi di metabolome profile no fã bi so na ɛde abusuabɔ pa a ɛne JA nsɛnkyerɛnne ba.
Esiane sɛ ɛsono sɛnea tawa ahorow a ɛne no wɔ abusuabɔ kɛse no mu nneɛma a nkwa wom yɛ adwuma no yɛ soronko koraa nti, wɔkyerɛ nneɛma a ɛsakra no pɔtee wɔ su afã no mu, enti ɛyɛ nhwehwɛmu kɛse. Information theory’s processing of captured metabolic profile da no adi sɛ herbivorous induction ma aguadi a ɛda metabolic gamma ahorow ne specialization ntam no yɛ kɛse. JA sɛnkyerɛnne no di dwuma titiriw wɔ saa aguadi yi mu. Nkɔanim a ɛba wɔ metabolome specialization mu no ne OD nkɔmhyɛ titiriw no hyia na ɛne JA sɛnkyerɛnne wɔ abusuabɔ pa, bere a JA sɛnkyerɛnne no ne metabolic gamma ahorow wɔ abusuabɔ bɔne. Saa nhwɛsoɔ yi kyerɛ sɛ afifideɛ OD tumi no gyina JA plasticity so titire, sɛ ɛyɛ microevolutionary scale anaasɛ evolutionary scale kɛseɛ. Exogenous JA application experiments a ɛkwati JA biosynthesis sintɔ ahorow no da no adi bio sɛ ​​wobetumi akyerɛ tawa ahorow a ɛne wɔn ho wɔn ho di nkitaho kɛse no mu nsonsonoe mu ayɛ no signal-responsive ne signal-non-responsive species, te sɛ wɔn mode of JA ne metabolome plasticity a herbivores de ba no ara pɛ. Nnua a wɔnyɛ ho biribi wɔ sɛnkyerɛnne kwan so no ntumi nyɛ ho hwee esiane sɛ wontumi nnya JA a ɛwɔ nipadua no mu nti ma enti wɔhyɛ nipadua mu anohyeto ahorow ase. Ebia eyi fi nkwaadɔm mu nsakrae a ɛba awosu mu nkwaadɔm atitiriw bi mu wɔ JA sɛnkyerɛnne kwan so (AOS ne JAR4 wɔ N. crescens mu) of. Nea afi mu aba yi si so dua sɛ ebia nsakrae a ɛba wɔ nipadua mu nkwaadɔm a ɛwɔ nipadua no mu ho adwene ne sɛnea wɔyɛ ho biribi mu titiriw na ɛde saa nkwaboaa ahorow ntam adannandi kɛse yi ba.
Wɔ nkitahodi a ɛda afifide ne mmoa a wodi nhabannuru ntam akyi no, nhwehwɛmu a wɔyɛ wɔ nipadua mu nneɛma ahorow mu no ne nsusuwii mu nkɔso a ɛho hia nyinaa a aba wɔ nhwehwɛmu a wɔyɛ wɔ abɔde a nkwa wom a wɔsakra wɔn ho ma nneɛma a atwa yɛn ho ahyia ne su ahorow a ɛyɛ den a ɛba wɔ nipadua mu no dannan mu no wɔ abusuabɔ. Esiane sɛ nsɛm dodow a nnɛyi MS nnwinnade nya no kɔ soro nti, mprempren nsusuwii hunu sɔhwɛ a ɛfa nipadua mu nneɛma ahorow ho no betumi aboro ankorankoro/akuw ahorow mu nneɛma a ɛsakra mu nsonsonoe so na wɔayɛ wiase nyinaa nhwehwɛmu de ada nhwɛso ahorow a wɔnhwɛ kwan adi. Wɔ nhwehwɛmu kɛseɛ mu no, kasakoa a ɛho hia ne adwene a ɛne sɛ wɔbɛsusu asase mfonini a nteaseɛ wom a wɔbɛtumi de ahwehwɛ nsɛm mu. Enti, ade titiriw a efi mprempren MS/MS metabolomics a enni animhwɛ ne nsɛm ho nsusuwii a wɔaka abom no mu aba ne sɛ ɛma metric a ɛnyɛ den a wobetumi de ayɛ asase mfonini de ahwehwɛ metabolic ahorow no mu wɔ taxonomic nsenia ahorow so. Ɛyɛ ɔkwan yi ahwehwɛde titiriw. Micro/macro evolution ne mpɔtam hɔ abɔde a nkwa wom ho adesua.
Wɔ macro-evolutionary level no, afifide ne nkoekoemmoa co-evolution theory a Ehrlich ne Raven (51) de mae no mu titiriw ne sɛ wɔbɛka sɛ nsakrae a ɛba interspecies metabolic diversity mu no na ɛde afifide abusua ahorow a egu ahorow ba. Nanso, wɔ mfe aduonum a atwam fi bere a wotintim saa nhoma titiriw yi mu no, wɔntaa nsɔ saa nsusuwii hunu yi nhwɛ (52). Eyi fi abusua mu su ahorow a ɛwɔ nipadua mu nneɛma a ɛsakra no su a wɔde toto ho wɔ afifide abusua ahorow a ɛwɔ akyirikyiri no mu titiriw. Wobetumi de rarity no adi dwuma de anchor target analysis akwan. Mprempren MS/MS adwumayɛ nhyehyɛe a wɔde nsɛm ho nsusuwii di ho dwuma no kyerɛ MS/MS nhyehyɛe mu nsɛdi a ɛwɔ metabolites a wonnim (a wonni kan paw metabolite) na ɛdannan saa MS/MS ahorow yi ma ɛbɛyɛ MS/MS kuw, ma enti wɔ adwumayɛfo nipadua mu nneɛma a wɔde yɛ adwuma mu no Wɔde saa macro-evolutionary models yi toto ho wɔ classification scale mu. Akontaabu ho nsɛnkyerɛnne a ɛnyɛ den. Adeyɛ no te sɛ phylogenetic analysis, a ebetumi de ntoatoaso a ɛne ne ho hyia adi dwuma de akyerɛ sɛnea nneɛma ahorow anaa suban ahorow no dannan no ntɛmntɛm a wonni kan nhyɛ nkɔm.
Wɔ biochemical level no, screening hypothesis a Firn ne Jones (53) yɛeɛ no kyerɛ sɛ wɔkura metabolic ahodoɔ mu wɔ level ahodoɔ mu de ma raw materials a wɔde bɛyɛ biological dwumadie a kane no na ɛnyɛ abusuabɔ anaasɛ wɔde asi ananmu metabolites. Amanneɛbɔ ho nsusuwii akwan ma nhyehyɛe bi a wobetumi akyerɛ saa adannandi nsakrae a ɛfa metabolite pɔtee bi a ɛba bere a wɔreyɛ metabolite titiriw yi dodow sɛ adannandi nhwehwɛmu nhyehyɛe a wɔahyɛ ho nyansa no fã: abɔde a nkwa wom mu nsakrae a ɛyɛ adwuma fi pɔtee a ɛba fam kɔ ​​pɔtee a ɛkorɔn mu Nkwammoaa a wɔbara wɔ nneɛma a atwa yɛn ho ahyia bi mu.
Ne nyinaa mu no, wɔ molecule abɔde a nkwa wom ho adesua mfiase no, wɔyɛɛ afifide ahobammɔ ho nsusuwii ahorow a ɛho hia, na wobu akwan a wɔde deductive hypothesis di dwuma kɛse sɛ ɛno nkutoo ne ɔkwan a wɔfa so nya nyansahu mu nkɔso. Eyi fi mfiridwuma mu anohyeto ahorow a ɛwɔ metabolome no nyinaa susuw mu titiriw. Ɛwom sɛ akwan a wɔfa so de nsusuwii hunu di dwuma no ho wɔ mfaso titiriw wɔ akwan afoforo a ɛde ba a wɔpaw mu de, nanso tumi a wotumi ma yɛn ntease a yɛwɔ wɔ biochemical networks ho no nya nkɔso no sua sen akontaabu akwan a ɛwɔ hɔ mprempren wɔ nnɛyi nyansahu a egye data pii mu no. Enti, nsusuwii ahorow a wontumi nhyɛ ho nkɔm no boro nsɛm a ɛwɔ hɔ no so koraa, enti wontumi nyi nsusuwii hunu nhyehyɛe/sɔhwɛ kyinhyia a ɛfa nkɔso a ɛba wɔ nhwehwɛmu mu no mfi hɔ (4). Yɛhwɛ kwan sɛ kɔmputa so adwumayɛ nhyehyɛe a ɛfa metabolomics ho a wɔde aba ha no betumi asan akanyan anigye wɔ nnansa yi (sɛnea) ne nea etwa to (dɛn nti) nsɛm a ɛfa metabolic ahorow ahorow ho no ho, na aboa ma bere foforo a ɛfa data nyansahu a wɔde nsusuwii kyerɛ kwan no aba. Bere no san hwehwɛɛ nsusuwii ahorow a ɛho hia a ɛkanyan awo ntoatoaso a atwam no mu.
Wɔyɛ nhabannuru aduan tẽẽ denam instar larva anaa Sl larva a wɔtete no wɔ Pale-colored pitcher plant ahaban biako a ɛwɔ rose nhwiren afifide biako so, a afifide biara yɛ nsɛso 10. Wɔde krampɔn kyekyeree nkoekoemmoa nwansena no, na wɔboaboaa nhaban mu ntini a aka no ano wɔ nnɔnhwerew 24 ne 72 akyi wɔ ɔyare no akyi na wɔde yɛɛ nwini ntɛmntɛm, na woyii nneɛma a ɛma nipadua no yɛ adwuma no fii mu.
Yɛ ayaresa a wɔde di nhabannuru ho mfonini wɔ ɔkwan a ɛne ne ho di nsɛ kɛse so. Ɔkwan no ne sɛ wɔde ntama nsusuwii ntwahonan bedi dwuma de atow nsɔe a ɛtoatoa so abiɛsa wɔ afifide no nhaban abiɛsa a atrɛw koraa no mfinimfini fã biara bere a ntama nhwiren no nyin no, na wɔde 1:5 Diluted Ms. Anaasɛ wɔde nsateaa a wɔde nsateaa ahyɛ mu de S1 OS bɛhyɛ kuru a wɔato mu no mu ntɛm ara. Twa ahaban bi na yɛ ho adwuma sɛnea yɛaka ho asɛm wɔ atifi hɔ no. Fa ɔkwan a yɛadi kan aka ho asɛm no yiyi nneɛma atitiriw a ɛma nipadua no yɛ adwuma ne afifide mu nkwaadɔm (54).
Wɔ exogenous JA dwumadie mu no, wɔde lanolin paste 20μl a MeJA 150μg (Lan + MeJA) wom, ne lanolin 20μl a wɔde ka kuru aduru (Lan + W) ho, anaasɛ wɔde lanolin kronkron 20μl di dwuma sɛ nea wɔde siw ano. Wɔtwaa nhaban no wɔ nnɔnhwerew 72 akyi wɔ aduruyɛ akyi, na wɔde guu nitrogen nsu mu ntɛm ara, na wɔde siee -80°C kosii sɛ wɔde bedi dwuma.
Wɔahu JA ne ET transgenic lines anan, a ɛne irAOC (36), irCOI1 (55), irACO ne sETR1 (48), wɔ yɛn nhwehwɛmu kuw no mu. irAOC daa no adi denneennen sɛ JA ne JA-Ile dodow so atew, bere a na irCOI1 nyɛ nea ɛyɛ mmerɛw wɔ JA ahorow ho. Sɛ wɔde toto EV ho a, JA-Ile a ɛboaboa ano no kɔɔ soro. Saa ara nso na irACO bɛtew ET a wɔyɛ so, na sɛ wɔde toto EV ho a, sETR1 a ɛnte nka wɔ ET ho no bɛma ET a wɔyɛ no akɔ soro.
Wɔde photoacoustic laser spectrometer (Sensor Sense ETD-300 bere ankasa ET sensor) di dwuma de yɛ ET susuw a ɛnyɛ nea wɔde hyɛ mu. Bere a wɔsaa no yare no akyi pɛɛ no, wotwitwaa nhaban no fã de guu ahwehwɛ toa a wɔatoto mu a ne kɛse yɛ mililita 4 mu, na wɔmaa ti no boaboaa ano wɔ nnɔnhwerew 5 ntam. Wɔ susudua no mu no, wɔde mframa kronkron a ɛyɛ lita 2/dɔnhwerew biara hohoroo ahina biara mu simma 8, a na adi kan afa ade a ɛma nneɛma yɛ kɛse a Sensor Sense de mae mu de yi CO2 ne nsu fii mu.
Wɔdii kan tintim microarray data no wɔ (35) mu na wɔde siee National Center for Biotechnology Information (NCBI) Gene Expression Comprehensive Database (accession number GSE30287). Wɔyii nsɛm a ɛne nhaban a W + OSMs ayaresa ne nea ɛnsɛee no de bae no hyia maa nhwehwɛmu yi. Raw intensity no yɛ log2. Ansa na wɔbɛyɛ akontabuo nhwehwɛmu no, wɔdanee mfitiaseɛ no na wɔde yɛɛ no ​​sɛdeɛ ɛteɛ no kɔɔ ne ɔha mu nkyekyɛmu a ɛtɔ so 75 denam R software package no so.
Wɔgyee mfitiaseɛ RNA sequencing (RNA-seq) data a ɛfa Nicotiana ahodoɔ ho no firii NCBI Short Reading Archives (SRA), adwuma no nɔma ne PRJNA301787, a Zhou ne ne mfɛfoɔ na wɔbɔɔ ho amanneɛ. (39) na kɔ so sɛnea wɔaka ho asɛm wɔ (56) mu no. Wɔpaw raw data a W + W, W + OSM ne W + OSS1 a ɛne Nicotiana ahorow hyia no dii ho dwuma no sɛ wɔmfa nyɛ nhwehwɛmu wɔ nhwehwɛmu yi mu, na wɔyɛɛ ho adwuma wɔ ɔkwan a edidi so yi so: Nea edi kan no, wɔdanee raw RNA-seq akenkan no kɔɔ FASTQ format mu. HISAT2 dane FASTQ kɔ SAM, na SAMtools dane SAM fael kɔ BAM fael a wɔahyehyɛ mu. Wɔde StringTie bu awosu mu nkwaadɔm a wɔda no adi ho akontaa, na ɔkwan a wɔfa so da no adi ne sɛ asinasin wɔ nnyinaso asinasin apem biara mu wɔ nkyerɛwee asinasin ɔpepem a wɔahyehyɛ no nnidiso nnidiso biara mu.
Acclaim chromatographic column (150 mm x 2.1 mm; particle size 2.2μm) a wɔde dii dwuma wɔ nhwehwɛmu no mu ne 4 mm x 4 mm guard column no yɛ ade koro no ara. Wɔde binary gradient a edidi so yi di dwuma wɔ Dionex UltiMate 3000 Ultra High Performance Liquid Chromatography (UHPLC) nhyehyɛe no mu: simma 0 kosi 0.5, isocratic 90% A [nsu a wɔayi ion afi mu, 0.1% (v/v) acetonitrile ne 0.05% formic acid], 10% B (Acetonitrile ne 0.05% formic acid); Simma 0.5 kosi 23.5, gradient phase no yɛ 10% A ne 90% B, sɛnea ɛte biara; 23.5 kosi 25 simma, isocratic 10% A ne 90% B. Nsuo a ɛsen no yɛ 400μl/min. Wɔ MS nhwehwɛmu nyinaa mu no, fa column eluent no gu quadrupole ne time-of-flight (qTOF) analyzer a wɔde electrospray source a ɛyɛ adwuma wɔ positive ionization mode mu (capillary voltage, 4500 V; capillary outlet 130 V ; Drying temperature 200°C; drying airflow 10 liters/min) mu.
Yɛ MS / MS asinasin nhwehwɛmu (a wɔbɛfrɛ no MS / MS wɔ ha) a ɛho nhia anaasɛ wontumi nyi nsonsonoe mfi data no mu na ama woanya nhyehyɛe ho nsɛm a ɛfa metabolic profile a wotumi hu nyinaa ho. Adwene a ɛfa MS/MS kwan a wɔmfa nyiyim nni dwuma ho no gyina nokwasɛm a ɛyɛ sɛ quadrupole no wɔ mass isolation window kɛse paa [enti, susuw mass-to-charge ratio (m/z) nsɛnkyerɛnne nyinaa sɛ asinasin]. Esiane eyi nti, esiane sɛ Impact II adwinnade no antumi anyɛ CE tilt nti, wɔde collision-induced dissociation collision energy (CE) values ​​a ɛkɔ soro dii dwuma yɛɛ nhwehwɛmu ahorow pii a ɛde ne ho. Ne tiawa mu no, di kan hwehwɛ nhwɛsode no mu denam UHPLC-electrospray ionization/qTOF-MS so denam single mass spectrometry mode (mpaapaemu tebea a ɛba fam a ɛnam in-source fragmentation so de ba), scanning fi m/z 50 kosi 1500 wɔ repetition frequency a ɛyɛ 5 Hz. Fa nitrogen di dwuma sɛ mframa a ɛbɔ mu ma MS/MS nhwehwɛmu, na yɛ susudua a ɛde ne ho wɔ mpaapaemu ahoɔden ahorow anan a edidi so yi a ɛnam nhyiam so ba no so: 20, 30, 40, ne 50 eV. Wɔ susudua no nyinaa mu no, quadrupole no na ɛwɔ mass isolation window kɛseɛ paa, ɛfiri m/z 50 kɔsi 1500. Sɛ wɔde anim nipadua m/z ne isolation width experiment no si 200 a, mass range no yɛ adwuma ankasa denam adwinnadeɛ no dwumadie software ne 0 Da so. Scan ma mass fragments sɛnea ɛte wɔ single mass mode mu. Fa sodium formate (50 ml isopropanol, 200 μl formic acid ne 1 ml 1M NaOH nsuo mu aduru) di dwuma ma mass calibration. Sɛ wɔde Bruker’s high-precision calibration algorithm di dwuma a, wɔde calibration data fael no bere a wɔatu mmirika awie spectrum no wɔ bere bi a wɔde ama mu akyi. Fa Data Analysis v4.0 softwea (Brook Dalton, Bremen, Germany) no export dwumadie no di dwuma de dane raw data fael no kɔ NetCDF format mu. Wɔde MS/MS data nhyehyɛe no asie wɔ metabolomics database a wɔabue MetaboLights (www.ebi.ac.uk) a ɛwɔ accession nɔma no mu. MTBLS1471 na ɛwɔ hɔ.
MS/MS nhyiamu no betumi abam denam nkitahodi nhwehwɛmu a ɛda MS1 ne MS/MS su pa nsɛnkyerɛnne ntam ama ahoɔden a ɛba fam ne nea ɛkɔ soro a ɛbɔ ne mmara foforo a wɔde adi dwuma. Wɔde R nkyerɛwde no di dwuma de hu nkitahodi nhwehwɛmu a ɛfa nea edi kan no nkyekyɛmu ho, na wɔde C# nkyerɛwde (https://github.com/MPI-DL/indiscriminant-MS-MS-assembly-pipeline) di dwuma de di mmara no ho dwuma.
Sɛnea ɛbɛyɛ a yɛbɛtew mfomso a ɛyɛ papa a ɛnam akyi dede ne atoro abusuabɔ a ɛnam m/z nneɛma bi a yehu wɔ nhwɛsode kakraa bi pɛ mu de ba no so no, yɛde “filled peak” adwuma a ɛwɔ R package XCMS (ma akyi dede nteɛso) no di dwuma Ɛsɛ sɛ wɔde si “NA” (undetected peak) ahoɔden ananmu. Sɛ wɔde fill peak function no di dwuma a, “0′′ intensity values ​​pii da so ara wɔ data set no mu a ɛbɛka correlation akontabuo no Afei, yɛde data processing aba a yɛnyaeɛ berɛ a wɔde filled peak function no dii dwuma ne berɛ a filled peak function no nni dwuma no toto ho, na yɛbu background noise value no ho akontaa gyina average corrected estimated values ​​no so, na afei yɛde saa 0 intensity values ​​yi si ananmu wobuu nneɛma a emu yɛ den boro akyi bo mmɔho abiɛsa nkutoo na wobuu no sɛ “nokware bepɔw.” Wɔ PCC akontabuo mu no, m/z nsɛnkyerɛnneɛ a ɛwɔ sample precursor (MS1) ne fragment data sets a anyɛ yie koraa no ɛwɔ nokware peaks awotwe nko ara na wɔsusu ho.
Sɛ ahoɔden a ɛwɔ precursor quality feature no mu wɔ sample no nyinaa mu no ne ahoɔden a ɛso atew a ɛwɔ quality feature koro no ara a wɔde hyɛ collision ahoɔden a ɛba fam anaa kɛse ase no wɔ abusuabɔ kɛse, na wɔamfa CAMERA ankyerɛw feature no sɛ isotope peak a, wobetumi akyerɛkyerɛ mu bio. Afei, ɛdenam akontaabu a ɛfa precursor-product pairs a ebetumi aba nyinaa ho wɔ s 3 mu (bere a wɔasusuw sɛ wɔde sie mfɛnsere ma peak retention) so no, wɔyɛ nkitahodi nhwehwɛmu no. Sɛ m/z boɔ no sua sene precursor boɔ no na MS/MS mpaapaemu ba wɔ nhwɛsoɔ beaeɛ korɔ no ara wɔ data set no mu sɛ precursor a wɔnya firi mu no nko ara na wɔbu no sɛ ɛyɛ asinasin.
Yegyina saa mmara mmienu a ɛnyɛ den yi so yi asinasin a wɔakyerɛ a m/z boɔ boro m/z a ɛwɔ anim a wɔakyerɛ no mu, na egyina nhwɛsoɔ gyinabea a animdifoɔ no pue ne asinasin a wɔakyerɛ no so. Ɛsan nso yɛ yie sɛ wɔbɛpaw su pa a ɛnam in-source fragments pii a wɔayɛ wɔ MS1 mode mu no ayɛ no sɛ candidate precursors, na ɛnam so ama MS/MS compounds a ɛho nhia aba. Sɛnea ɛbɛyɛ a yɛbɛtew saa data redundancy yi so no, sɛ NDP nsɛso a ɛwɔ spectra ahorow no mu boro 0.6, na wɔyɛ chromatogram “pcgroup” a CAMERA de nkyerɛkyerɛmu ma no dea a, yɛbɛka abom. Awiei koraa no, yɛka CE aba anan a ɛbata precursor ne asinasin no nyinaa bom kɔ deconvoluted composite spectrum a etwa to no mu denam ahoɔden a ɛkorɔn sen biara a yɛpaw wɔ candidate peaks nyinaa mu a m/z bo yɛ pɛ wɔ collision ahoɔden ahorow mu no so. Anamɔn a ɛdi akyire a wɔfa so yɛ adwuma no gyina adwene a ɛfa composite spectrum ho so na ɛsusu CE tebea ahodoɔ a ɛhia na ama ayɛ yie sɛ ɛbɛpaapae, ɛfiri sɛ wɔtumi hunu asinasin bi wɔ ahoɔden pɔtee bi a ɛbɔ mu nkutoo ase.
Wɔde RDPI (30) dii dwuma de buu inducibility a ɛwɔ metabolic profile no ho akontaa. Metabolic spectrum diversity (Hj index) no fi MS/MS precursors dodow a wɔde Shannon entropy a ɛwɔ MS/MS frequency distribution a wɔde equation a edidi so yi a Martínez ne ne mfɛfo kyerɛkyerɛɛ mu no di dwuma no mu. (8) no. Hj = −∑i = 1mPijlog2(Pij) a Pij ne mpɛn dodow a MS/MS a ɛto so i wɔ j-t nhwɛsode no mu no hyia (j = 1, 2,..., m) (i = 1, 2, ..., m) t).
Wɔkyerɛ metabolic specificity (Si index) ase sɛ MS/MS a wɔde ama no nkyerɛkyerɛmu a ɛfa mpɛn dodow a ɛda nhwɛsode ahorow a wɔresusuw ho no ntam ho. Wɔbu MS/MS pɔteeyɛ ho akontaa sɛ Si = 1t (∑j = 1tPijPilog2PijPi) .
Fa fomula a edidi so yi susuw metabolome-specific δj index a ɛwɔ j sample biara mu, ne MS/MS specificity δj = ∑i = 1mPijSi
Wɔde MS/MS spectra no hyia abien abien, na wogyina nkontabuo mmienu no so bu nsɛsoɔ no ho akontaa. Nea edi kan no, denam standard NDP (a wɔsan frɛ no cosine correlation kwan) so no, fa nsɛso a edidi so yi di dwuma de nkontaa afã nsɛso a ɛda spectra NDP = (∑iS1 & S2WS1, iWS2, i) 2∑iWS1, i2∑iWS2, i2 a S1 ne S2 a S1 ne S2 hyia no, ma spectrum 1 ne spectrum 2, ne WS1, i ne WS2, i gyina hɔ ma mu duru a egyina peak intensity so sɛ nsonsonoe a ɛwɔ i-th common peak a ɛda spectra abien no ntam no nnu 0.01 Da. Wɔabu mu duru no sɛnea edidi so yi: W = [peak intensity] m [quality] n, m = 0.5, n = 2, sɛnea MassBank kyerɛ no.
Wɔde ɔkwan a ɛto so abien a wɔfa so nya nkontaahyɛde dii dwuma, a na ɛfa NL a wɔkyɛ wɔ MS/MS ntam no mu nhwehwɛmu ho. Nea ɛbɛyɛ na yɛatumi ayɛ eyi no, yɛde NL lists 52 a wɔtaa hyia wɔ MS mpaapaemu nhyehyɛe no mu no dii dwuma wɔ tandem mu, ne NL a ɛyɛ pɔtee kɛse (data file S1) a wɔadi kan akyerɛw ho asɛm ama MS/MS spectrum a ɛwɔ metabolites a ɛto so abien a ɛwɔ Nepenthes ahorow a wɔayɛ mmerɛw no mu ( 9, 26). Yɛ binary vector a ɛyɛ 1 ne 0 ma MS/MS biara, a ɛne mprempren ne nea enni hɔ a ɛwɔ NL bi mu no hyia. Wogyina Euclidean akyirikyiri nsɛsoɔ so bu NL nsɛsoɔ nkontabuo ma NL vectors mmienu biara.
Sɛ yɛbɛyɛ dual clustering a, yɛde R package DiffCoEx a egyina Weighted Gene Co-expression Analysis (WGCNA) ntrɛwmu so dii dwuma. Yɛde NDP ne NL scoring matrices a ɛwɔ MS/MS spectra mu dii dwuma no, yɛde DiffCoEx dii dwuma de buu ntotoho nkitahodi matrix no ho akontaa. Wɔyɛ binary clustering denam “cutreeDynamic” parameter a wɔde besi ɔkwan = “hybrid”, cutHeight = 0.9999, deepSplit = T, ne minClusterSize = 10. Wɔtwee DiffCoEx R fibea koodu no fii fael foforo 1 a Tesson ne ne mfɛfo yɛe no so. (57) . Wobetumi ahu R WGCNA softwea a wɔhwehwɛ no wɔ https://horvath.genetics.ucla.edu/html/CoexpressionNetwork/Rpackages/WGCNA.
Sɛnea ɛbɛyɛ a yɛbɛyɛ MS/MS molecular network nhwehwɛmu no, yɛbuu paired spectral connectivity a egyina NDP ne NL nsɛsoɔ ahodoɔ so, na yɛde Cytoscape software yɛɛ network topology ho mfonini wɔ yɛn adwene mu de organic layout dii dwuma wɔ CyFilescape yFiles layout algorithm extension application no mu.
Fa R version 3.0.1 yɛ akontabuo nhwehwɛmu wɔ data no so. Wɔde akwan abien nhwehwɛmu a ɛfa nsonsonoe ho (ANOVA) na ɛhwɛɛ akontaabu mu ntease, na wɔde Tukey nokwaredi mu nsonsonoe a ɛho hia (HSD) akyi sɔhwɛ dii akyi. Sɛnea ɛbɛyɛ a wɔbɛhwehwɛ nsonsonoe a ɛda aduru a wɔde di nhabannuru ne nea wɔde di dwuma no ntam no, wɔde Student’s t sɔhwɛ yɛɛ nhwɛsode akuw abien a ɛwɔ nsonsonoe koro no mu nkyekyɛmu a ɛwɔ dua abien no mu.
Sɛ wopɛ nneɛma foforo a wode bɛka saa asɛm yi ho a, yɛsrɛ wo hwɛ http://advances.sciencemag.org/cgi/content/full/6/24/eaaz0381/DC1
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Amanneɛbɔ ho nsusuwii ma amansan sika a wɔde bɛtoto metabolome soronko ne sɔhwɛ ahobammɔ nsusuwii ahorow ho nkɔmhyɛ ho.
Amanneɛbɔ ho nsusuwii ma amansan sika a wɔde bɛtoto metabolome soronko ne sɔhwɛ ahobammɔ nsusuwii ahorow ho nkɔmhyɛ ho.
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