Numiemiewo takpɔkpɔ ŋuti nufiafia vovovowo naa nukpɔsusu ŋuti mɔfiame veviwo hena numiemiewo ƒe lãmenugbagbeviwo ƒe dɔwɔwɔ tɔxɛ ƒe ɖoɖowo me ɖeɖe, gake wogale woƒe nyagblɔɖi veviwo dom kpɔ. Le afisia la, míezã unbiased tandem mass spectrometry (MS/MS) analysis tsɔ ku nu me le ɖoɖo nu le metabolome si le atama ƒe ƒomevi siwo woɖe tso numiemie ɖekaɖekawo me yi amehawo kple ƒomevi siwo do ƒome kplikplikpli me, eye míewɔ dɔ tso mass spectrometric feature theories gbogbo aɖewo siwo wotu ɖe compound spectra dzi le nyatakakaawo me. Dɔwɔɖoɖo si woatsɔ ado nyagblɔɖi vevi siwo ku ɖe ametakpɔkpɔ nyuitɔ (OD) kple taɖodzinu si le ʋuʋum (MT) ƒe nufiafiawo kpɔ. Nyatakaka ƒe akpa si le numiemiewo ƒe metabolomics me sɔ kple OD nufiafia, gake etsi tre ɖe MT nufiafia ƒe nyagblɔɖi vevitɔ si ku ɖe metabolomics dynamics si gbeɖulawo hena vɛ ŋu. Tso micro va ɖo macro evolutionary scale dzi la, wode dzesi jasmonate ƒe dzesi be eyae nye nu vevitɔ si dea dzesi OD, esime ethylene ƒe dzesi na be woɖɔli nyuie le gbeɖuɖɔ ɖula tɔxɛ ƒe ŋuɖoɖo si ŋu MS/MS molecule network ŋlɔ ɖi la ŋu.
Nusiwo wotsɔ ɖɔa li nu tɔxɛ siwo ƒe wɔwɔme to vovoe nye nu vevitɔ siwo kpɔa gome le numiemiewo ƒe tɔtrɔ ɖe nutoa me ŋu me, vevietɔ le futɔwo taʋiʋli me (1). Alesi lãmenugbagbeviwo ƒe dɔwɔwɔ tɔxɛ si wokpɔna le numiemiewo me ƒe vovototo wɔnuku la ʋã numekuku deto ƒe bla nanewo le eƒe nu gbagbewo ƒe dɔwɔwɔ ƒe akpa geɖe ŋu, eye wòwɔ numiemiewo takpɔkpɔ ŋuti nufiafiawo ƒe xexlẽdzesi didi aɖe, siwo nye nɔnɔmetɔtrɔ kple lãwo ƒe agbenɔnɔ ƒe ŋgɔyiyi le numiemiewo kple nudzodzoewo dome kadodo me. Numekuku siwo wowɔ le nuteƒekpɔkpɔ me naa mɔfiame veviwo (2). Gake numiemiewo takpɔkpɔ ŋuti nufiafia siawo mezɔ ɖe nususugblɔ ƒe deductive reasoning ƒe ɖoɖowɔɖi ƒe mɔ dzi o, si me nyagblɔɖi veviwo nɔ numekuku ƒe ɖoƒe ɖeka (3) eye wodo wo kpɔ le dodokpɔ me be woado nukpɔsusu ƒe ŋgɔyiyi ƒe tsatsam si kplɔe ɖo ɖe ŋgɔ (4). Mɔ̃ɖaŋununya ƒe seɖoƒewo xea mɔ na nyatakakawo nuƒoƒoƒu ɖe lãmenugbagbeviwo ƒe dɔwɔwɔ ƒe hatsotso tɔxɛ aɖewo ko dzi eye womewɔa numekuku blibo le lãmenugbagbeviwo ƒe dɔwɔwɔ tɔxɛwo ŋu o, si wɔnɛ be woxea mɔ na hatsotsowo dome tsɔtsɔ sɔ kple wo nɔewo si le vevie na nukpɔsusu ƒe ŋgɔyiyi (5). Metobolomics ŋuti nyatakaka blibo kple ga ɖeka si woatsɔ asɔ kple metabolic space ƒe dɔwɔwɔ ƒe dɔwɔwɔ le numiemie ƒe hatsotso vovovowo dome ƒe anyimanɔmanɔ xea mɔ na agblea ƒe dzɔdzɔmeŋutinunya ƒe tsitsi.
Nu yeye siwo wowɔ le tandem mass spectrometry (MS/MS) metabolomics ƒe akpa dzi ateŋu ade dzesi metabolic ƒe tɔtrɔ siwo le system clade ƒomevi aɖe me kple wo dome bliboe, eye woateŋu atsɔe akpe ɖe akɔntabubu mɔnuwo ŋu atsɔ abu akɔnta le xɔtuɖoɖo ƒe sɔsɔ si le tsakatsaka sesẽ siawo dome ŋu. Dzɔdzɔmeŋusẽŋununya ŋuti sidzedze do ŋgɔ (5). Mɔ̃ɖaŋununya deŋgɔ siwo wotsɔ ƒo ƒui le numekuku kple kɔmpiuta zazã me naa ɖoɖo si hiã be woatsɔ ado nyagblɔɖi geɖe siwo nu gbagbewo kple wo nɔƒewo kple nɔnɔmetɔtrɔ ŋuti nufiafia siwo ku ɖe lãmenugbagbeviwo ƒe dɔwɔwɔ vovovowo ŋu gblɔ ɖi la kpɔ ɣeyiɣi didi. Shannon (6) to nyatakaka ŋuti nufiafia vɛ zi gbãtɔ le eƒe nyati vevitɔ me le ƒe 1948 me, eye wòɖo gɔmeɖoanyi na nyatakakawo me dzodzro le akɔntabubu me, si wozã le go geɖe me tsɔ wu alesi wozãe le gɔmedzedzea me. Le domenyiŋusẽfianu ŋuti nunya me la, wozã nyatakaka ŋuti nufiafia dzidzedzetɔe tsɔ bu nyatakaka siwo léa wo ɖe ɖoɖo nu ƒe agbɔsɔsɔme (7). Le transcriptomics numekuku me la, nyatakaka ŋuti nufiafia dzroa tɔtrɔ siwo katã va le transcriptome me (8). Le numekuku siwo míewɔ va yi me la, míezã akɔntabubu ƒe ɖoɖo si nye nyatakaka nufiafia ɖe lãmenugbagbeviwo ƒe dɔwɔwɔ ŋuti nunya ŋu tsɔ ɖɔ lãmenugbagbeviwo ƒe dɔwɔwɔ ŋuti nunya si le lãmenugbagbeviwo ƒe agbɔsɔsɔ me le numiemiewo me (9). Le afisia la, míetsɔ dɔwɔwɔ ƒe ɖoɖo si wotu ɖe MS/MS dzi la ƒo ƒu kple akɔntabubu ƒe ɖoɖo si nye nyatakaka ŋuti nufiafia, si ƒe dzesi enye lãmenugbagbeviwo ƒe tɔtrɔ le ga si wozãna ɖekae me, be míatsɔ numiemiewo takpɔkpɔ ŋuti nufiafia si nye metabolome si gbeɖulawo hena vɛ ƒe nyagblɔɖi veviwo asɔ kple wo nɔewo.
Zi geɖe la, numiemiewo takpɔkpɔ ƒe nukpɔsusu ƒe ɖoɖowo xɔa wo nɔewo eye woate ŋu ama wo ɖe hatsotso eve me: esiwo dzea agbagba be yewoaɖe numiemie tɔxɛwo ƒe nusiwo wotsɔna trɔa asi le nu ŋu ƒe mama me si wotu ɖe ametakpɔkpɔ ƒe dɔwɔwɔwo dzi, abe ametakpɔkpɔ nyuitɔ (OD) (10), taɖodzinu si le ʋuʋum (MT) (11) ) Kple dzedzeme (12) nufiafia, esime bubuwo dia mɔ̃ɖaŋununya me numeɖeɖewo le alesi tɔtrɔ siwo vaa numiemie ƒe nunɔamesiwo ƒe amesinɔnɔ me kpɔa ŋusẽ ɖe wo dzii ŋu tsitsi kple lãmenugbagbeviwo ƒe dɔwɔwɔ tɔxɛwo ƒe ƒuƒoƒo, abe carbon ene: nunyiame ƒe dadasɔ ƒe nukpɔsusu (13), tsitsi ƒe agbɔsɔsɔme ƒe nukpɔsusu (14 ), kple tsitsi kple vovototodedeameme ƒe dadasɔ ƒe nukpɔsusu (15). Nufiafia eveawo le numekuku ƒe ɖoƒe vovovowo (4). Gake nufiafia eve siwo ku ɖe ametakpɔdɔwo ŋu le dɔwɔwɔ ƒe ɖoɖo nu ɖua dzeɖoɖo si ku ɖe numiemiewo ƒe ametakpɔnu siwo wowɔna kple esiwo woate ŋu aʋã ŋu: OD nufiafia, si tsɔe be ne ehiã ko hafi numiemiewo dea ga woƒe atike xɔasiwo takpɔnu me, le kpɔɖeŋu me, ne woɖu wo Ne gbemelã aɖe dze wo dzi la, eyata le alesi wòate ŋu adzɔe be woadze wo dzi le etsɔme nu la, wodea atike si me ametakpɔnu ƒe dɔwɔwɔ le asi na (10); MT ƒe nukpɔsusua do susua ɖa be mɔfiame ƒe lãmenugbagbeviwo ƒe tɔtrɔ ƒe axis aɖeke meli o, gake metabolite la trɔna le vome, si wɔnɛ be wòte ŋu dzɔna be Woaxe mɔ na metabolite “ʋuʋu ƒe taɖodzinu” si dzea gbeɖuɖɔ ɖulawo dzi. Ne míagblɔe bubui la, nufiafia eve siawo gblɔa nya siwo tsi tre ɖe wo nɔewo ŋu ɖi le lãmenugbagbeviwo ƒe tɔtrɔ si dzɔna le gbeɖulawo ƒe amedzidzedze megbe ŋu: ƒomedodo si le lãmenugbagbeviwo ƒe lãmenugbagbeviwo ƒe ƒuƒoƒo ɖe mɔ ɖeka dzi kple ametakpɔkpɔ ƒe dɔwɔwɔ (OD) kple lãmenugbagbeviwo ƒe tɔtrɔ siwo womefia mɔe o (MT) dome (11 ).
OD kple MT ƒe nukpɔsusuwo meku ɖe tɔtrɔ siwo wodo ɖa le lãmenugbagbeviwo ƒe dɔwɔwɔ me ŋu ko o, ke boŋ nu gbagbewo kple wo nɔƒewo kple nɔnɔmetɔtrɔ me tsonu siwo dona tso nusiwo woɖɔna ɖo siawo ƒe ƒuƒoƒo me hã ɖe eme, abe asitɔtrɔ ƒe gazazãwo kple viɖe siwo dona tso lãmenugbagbeviwo ƒe tɔtrɔ siawo me le lãwo ƒe nɔnɔme tɔxɛ aɖe me ene (16). Togbɔ be nukpɔsusu eveawo siaa de dzesi ametakpɔdɔ si lãmenugbagbeviwo ƒe dɔwɔwɔ tɔxɛwo wɔna, si ate ŋu axɔ asi alo maxɔ asi o hã la, nyagblɔɖi vevitɔ si de vovototo OD kple MT ƒe nukpɔsusuwo dome la le lãmenugbagbeviwo ƒe tɔtrɔ siwo hena vɛ ƒe mɔfiame me. OD nufiafia ƒe nyagblɔɖie xɔ dodokpɔ ƒe susu wu vaseɖe fifia. Dodokpɔ siawo dometɔ aɖewoe nye nusɔsrɔ̃ tso lãmenugbagbevi vovovowo ƒe ametakpɔkpɔ ƒe dɔwɔwɔ tẽ alo esi mele tẽ o ŋu le atike tɔxɛwo ƒe lãkusiwo me le dzoxɔxɔxɔwo kple dzɔdzɔmenɔnɔmewo me, kpakple tɔtrɔ siwo vaa numiemiewo ƒe tsitsi ƒe afɔɖeɖe me (17-19). Gake vaseɖe fifia la, le dɔwɔwɔ ƒe ɖoɖo kple akɔntabubu ƒe ɖoɖo aɖeke si woatsɔ adzro nugbagbe ɖesiaɖe ƒe lãmenugbagbeviwo ƒe dɔwɔwɔ ƒe vovototo me le xexeame katã ƒe anyimanɔmanɔ ta la, vovototo vevitɔ si wogblɔ ɖi le nufiafia eveawo dome (si fia be lãmenugbagbeviwo ƒe tɔtrɔ ƒe mɔfiame) gakpɔtɔ le dodokpɔ me. Le afisia la, míetsɔ numekuku ma tɔgbe na.
Numiemie tɔxɛwo ƒe metabolite ƒe nɔnɔme vevitɔwo dometɔ ɖekae nye woƒe wɔwɔme ƒe vovototo si gbɔ eme le ɖoƒe ɖesiaɖe tso numiemie ɖeka dzi, amehawo dzi va ɖo ƒomevi siwo ɖi wo nɔewo dzi (20). Woate ŋu akpɔ agbɔsɔsɔme ƒe tɔtrɔ geɖe le metabolite tɔxɛwo me le amewo ƒe agbɔsɔsɔ ƒe dzidzenu nu, evɔ zi geɖe la, woléa nɔnɔme ƒe vovototo sesẽwo me ɖe asi le lã ƒomeviwo ƒe ɖoɖo nu (20). Eyata numiemiewo ƒe lãmenugbagbeviwo ƒe dɔwɔwɔ ƒe vovototoe nye dɔwɔwɔ ƒe vovototo ƒe akpa vevitɔ, si ɖea tɔtrɔ ɖe nɔnɔme vovovowo ŋu fiana, vevietɔ niche mawo siwo me nudzodzoe tɔxɛwo kple gbeɖula bɔbɔewo ƒe amedzidzedze ƒe mɔnukpɔkpɔ vovovowo le (21). Tso esime Fraenkel (22) ŋlɔ nyati yeye aɖe si ku ɖe susu siwo ta numiemie tɔxɛwo ƒe lãmenugbagbeviwo ƒe dɔwɔwɔ le wo me ŋu la, wobu kadodo kple nudzodzoe vovovowo be enye tiatia ƒe nyaƒoɖeamenu veviwo, eye woxɔe se be kadodo siawoe trɔ asi le numiemiewo ŋu le nɔnɔmetɔtrɔɣi. Lãmetsiŋusẽ ƒe mɔ (23). Vovototo siwo le lã ƒomeviwo dome le metabolite tɔxɛwo ƒe vovototo me hã ate ŋu aɖe ŋutilã ƒe dadasɔ si do ƒome kple numiemie ƒe ametakpɔkpɔ si nye constitutive kple inducible numiemiewo ta tso gbeɖuɖɔ ƒe mɔnuwo me hã afia, elabena zi geɖe la, ƒomevi eveawo do ƒome gbegblẽ kple wo nɔewo (24). Togbɔ be ate ŋu aɖe vi be woalé ametakpɔnu nyui me ɖe asi ɣesiaɣi hã la, lãmenugbagbeviwo ƒe tɔtrɔ le ɣeyiɣi nyuitɔ dzi si do ƒome kple ametakpɔkpɔ naa viɖe siwo dze ƒã le mɔɖeɖe na numiemiewo be woatsɔ nunɔamesi xɔasiwo ama ŋutilã ƒe gadede bubuwo me (19, 24), eye woaƒo asa na alesi wòhiã be woawɔ nu aduadu. Nusiwo gblẽ le ame ŋu le agbanɔamedzi me (25). Tsɔ kpe ɖe eŋu la, lãmenugbagbeviwo ƒe tɔtrɔ tɔxɛ siawo siwo nudzodzoewo ƒe gbeɖulawo hena vɛ ate ŋu ahe mama si gblẽa nu le ameha la me vɛ (26), eye ate ŋu aɖe dzɔdzɔmetɔtrɔ gã siwo va le jasmonic acid (JA) ƒe dzesi me, si ate ŋu alé ɖe te le ameha la me, ƒe nuxexlẽ tẽ afia. JA ƒe dzesi siwo kɔkɔ kple esiwo bɔbɔ nye asitsatsa le ametakpɔkpɔ ɖe gbeɖuɖɔ ɖulawo ŋu kple hoʋiʋli kple lã ƒomevi aɖewo koŋ dome (27). Tsɔ kpe ɖe eŋu la, metabolite biosynthetic mɔ tɔxɛwo abu kabakaba eye woatrɔ le nɔnɔmetɔtrɔɣi, si ana metabolite mama ɖe akpa vovovowo me le lã ƒomevi siwo do ƒome kplikplikpli dome ( 28 ). Woate ŋu aɖo nɔnɔme vovovo siawo anyi kabakaba le gbeɖulawo ƒe nɔnɔme siwo le tɔtrɔm ta (29), si fia be gbeɖulawo ƒe habɔbɔwo ƒe tɔtrɔ nye nu vevi aɖe si ʋãa lãmenugbagbeviwo ƒe tɔtrɔ.
Le afisia la, míekpɔ kuxi siwo gbɔna gbɔ koŋ. (I) Aleke nudzodzoe si ɖua gbewo gbugbɔa numiemie ƒe metabolome ɖoa anyie? (Ii) Nukae nye nyatakaka ƒe akpa vevi siwo le metabolic plasticity me siwo woate ŋu abu agbɔsɔsɔme atsɔ ado ɣeyiɣi didi ƒe ametakpɔkpɔ ŋuti nufiafia ƒe nyagblɔɖiwo kpɔ? . . Be míawɔ esia la, míesrɔ̃ nu tso aŋgba ƒe aŋgba ƒe tɔtrɔ ŋu le ɖoɖo nu, si nye numiemie si nye nu gbagbewo ƒe kpɔɖeŋu numiemie si si lãmenugbagbeviwo ƒe dɔwɔwɔ tɔxɛ geɖe le, eye wòwɔa dɔ ɖe gbeɖuɖɔ ɖula eve siwo le afima, Lepidoptera Datura (Ms) ( Very aggressive, mainly eaten) ƒe lãviwo ŋu Le Solanaceae kple Spodoptera littoralis (Sl) dzi la, kotoku ƒe aŋgba me nudzodzoewo nye ƒomevi aɖe “ƒomevi”, kple numiemie siwo xɔa Solanaceae kple ƒome kple ƒome bubuwo ƒe amedzro bubuwo Numiemiewo ƒe nuɖuɖu. Míeɖe MS/MS metabolomics spectrum me eye míeɖe nyatakaka nufiafia ƒe akɔntabubu ŋuti nuŋlɔɖiwo ɖa be míatsɔ OD kple MT nufiafiawo asɔ kple wo nɔewo. Wɔ anyigbatata siwo ku ɖe nu tɔxɛwo ŋu be woaɖe alesi metabolite veviwo nye afia. Wokeke numekukua ɖe enu yi N. nasi kple atama ƒomevi siwo do ƒome kplikplikpli ƒe afimatɔwo gbɔ be woagaku vovototo si le numiemie ƒe lãmetsiŋusẽ ƒe dzesiwo kple OD ƒe ʋuʋu dome la me.
Be míate ŋu alé anyigbatata bliboa ku ɖe aŋgba ƒe metabolome ƒe plasticity kple wɔwɔme si le herbivorous atama me ŋu la, míezã numekuku kple akɔntabubu ƒe dɔwɔwɔ ƒe ɖoɖo si wowɔ va yi tsɔ ƒo nyatakaka siwo ƒe lolome deŋgɔ nu ƒu bliboe eye míeɖe wo ɖa le wo ɖokui si MS / MS spectra tso numiemie siwo woɖe tso numiemiewo me ( 9 ). Mɔnu sia si me vovototo mele o (si woyɔna be MS/MS) ate ŋu awɔ non-redundant compound spectra, si woate ŋu azã emegbe na compound-level numekuku siwo katã ŋu woƒo nu tsoe le afisia. Numiemie ƒe lãmenugbagbeviwo ƒe dɔwɔwɔ siawo siwo woɖe ɖa la nye ƒomevi vovovowo, siwo me lãmenugbagbeviwo ƒe dɔwɔwɔ alafa geɖe va ɖo akpe geɖe le (anɔ abe 500-1000-s/MS/MS ene le afisia). Le afisia la, míebua metabolic plasticity le nyatakaka nufiafia ƒe ɖoɖo nu, eye míebua metabolome ƒe vovototo kple dɔwɔwɔ ƒe agbɔsɔsɔme si wotu ɖe Shannon entropy si le metabolic frequency distribution dzi. Míezã mɔfiame si míewɔ va yi (8) tsɔ bu akɔnta le dzesiwo ƒe hatsotso aɖe si woate ŋu azã atsɔ adzidze metabolome vovovowo (Hj dzesi), metabolic profile specialization (δj indicator) kple metabolite specificity of a single metabolite (Si indicator) . Tsɔ kpe ɖe eŋu la, míewɔ Relative Distance Plasticity Index (RDPI) ŋudɔ tsɔ bu metabolome inducibility na gbeɖuɖɔ ɖulawo (Nɔnɔmetata 1A) (30). Le akɔntabubu ƒe ɖoɖo sia me la, míebua MS/MS ƒe ɣeyiɣi ƒe didime abe nyatakaka ƒe akpa vevitɔ ene, eye míewɔa dɔ tso MS/MS ƒe agbɔsɔsɔ si sɔ ŋu wòzua ɣeyiɣi ƒe mama ƒe nɔnɔmetata, eye emegbe míezãa Shannon entropy tsɔ bua metabolome ƒe vovototo tso eme. Wodzidzea metabolome ƒe etɔxɛnyenye to MS/MS spektrum ɖeka ƒe vovototodedeameme le mama dedie nu me. Eyata wotrɔa MS/MS ƒe hatsotso aɖewo ƒe agbɔsɔsɔ ƒe dzidziɖedzi le gbeɖulawo ƒe ʋuʋu megbe wòzua spectral inducibility, RDPI kple specialization, si fia be δj index ƒe dzidziɖedzi, elabena wowɔa metabolite tɔxɛ geɖe wu eye wowɔa Si index si lolo. Hj ƒe vovototodedeameme ƒe dzesi ƒe ɖiɖi ɖee fia be MS/MS siwo wowɔ ƒe xexlẽme dzi ɖe kpɔtɔ, alo profile frequency mama trɔna le mɔ si mewɔ ɖeka o nu, esime wòɖea eƒe kakaɖedzimanɔamesi bliboa dzi kpɔtɔna. To Si index ƒe akɔntabubu me la, ateŋu adzɔ be woate gbe ɖe MS/MS siwo gbeɖuɖɔ aɖewo ʋãna dzi, ke boŋ, MS/MS siwo mewɔa nu ɖe ​​induction la ŋu o, si nye dzesi vevi aɖe si dea vovototo MT kple OD ƒe nyagblɔɖi me.
(A) Akɔntabubu ƒe nuŋlɔɖi siwo wozãna na gbeɖulawo (H1 va ɖo Hx) MS/MS nyatakakawo ƒe ʋuʋu (RDPI), vovototodedeameme (Hj index), etɔxɛnyenye (δj index) kple metabolite specificity (Si index). Dzidziɖedzi le alesi gbegbe wowɔa nu tɔxɛe (δj) ɖee fia be, le mamã dedie nu la, woawɔ gbeɖuɖɔ ɖula tɔxɛ geɖe wu, gake vovototodedeameme (Hj) ƒe ɖiɖi ɖee fia be lãmenugbagbeviwo ƒe wɔwɔ dzi ɖe kpɔtɔ alo lãmenugbagbeviwo ƒe mama si mesɔ o le mama ƒe nɔnɔmetata me . Si ƒe asixɔxɔa daa nenye be metabolite la nye nɔnɔme aɖe koŋ tɔ (le afisia la, gbeɖula) alo le mɔ bubu nu la, wolée ɖe te le agbɔsɔsɔ ma ke me. (B) Nukpɔsusu ƒe nɔnɔmetata si ku ɖe ametakpɔkpɔ ŋuti nufiafia ƒe nyagblɔɖi ŋu to nyatakaka nufiafia ƒe axis zazã me. OD ƒe nufiafia gblɔe ɖi be gbeɖuɖɔ ɖulawo ƒe amedzidzedze ana ametakpɔnu ƒe lãmenugbagbeviwo nadzi ɖe edzi, si ana δj nadzi ɖe edzi. Le ɣeyiɣi ma ke me la, Hj dzi ɖena kpɔtɔna elabena wogbugbɔ ɖoa ​​ɖoɖo ɖe nɔnɔmea ŋu ɖo ta kakaɖedzimanɔamesi si dzi ɖe kpɔtɔ le lãmenugbagbeviwo ƒe dɔwɔwɔ ŋuti nyatakakawo ŋu. MT ƒe nufiafia gblɔe ɖi be gbeɖulawo ƒe amedzidzedze ahe tɔtrɔ siwo menye mɔfiame o vɛ le lãmenugbagbeviwo ƒe dɔwɔwɔ me, si ana Hj nadzi ɖe edzi abe nusi fia be lãmenugbagbeviwo ƒe dɔwɔwɔ ŋuti nyatakakawo ƒe kakaɖedzimanɔamesi dzi ɖe edzi eye wòana Si nama le vome. Míeɖo aɖaŋu hã be woawɔ kpɔɖeŋu si wotsaka, si nye MT nyuitɔ kekeake, si me woadzi metabolite aɖewo siwo ƒe ametakpɔkpɔ ƒe asixɔxɔwo lolo wu etɔxɛe (Si ƒe asixɔxɔ si lolo), gake bubuwo ya ɖea ŋuɖoɖo siwo wowɔna le vome fiana (Si ƒe asixɔxɔ si bɔbɔ wu).
Míezãa nyatakaka nufiafia ƒe numeɖelawo tsɔ ɖea OD nufiafia gɔme be míagblɔe ɖi be tɔtrɔ tɔxɛ siwo gbeɖulawo hena vɛ le metabolite nɔnɔme si me womeʋã o me ahe (i) metabolic specificity ƒe dzidziɖedzi (Si index) si ʋua metabolite specificity (δj index) The increase of) certain special metabolite groups with higher defense value, and (ii) the decrease of metabolome diversity (Hj) index le tɔtrɔ si va le lãmenugbagbeviwo ƒe dɔwɔwɔ ƒe mama zi geɖe me va zu leptin ƒe ŋutilã ƒe mama geɖe wu ta. Le metabolite ɖeka ƒe ɖoɖo nu la, wokpɔa mɔ na Si ƒe mama ɖe ɖoɖo nu, afisi metabolite la adzi Si ƒe asixɔxɔ ɖe edzi le eƒe ametakpɔnu ƒe asixɔxɔ nu (Nɔnɔmetata 1B). Le mɔ sia nu la, míeɖe MT ƒe nufiafia me be míagblɔe ɖi be dzidzɔkpɔkpɔ ahe (i) tɔtrɔ siwo menye mɔfiame o le metabolites me si ana δj index dzi naɖe akpɔtɔ, kple (ii) Hj index ƒe dzidziɖedzi le metabolite ƒe kakaɖedzimanɔamesi ƒe dzidziɖedzi ta. Alo randomness, si woate ŋu atsɔ Shannon entropy adzidze agbɔsɔsɔme le generalized diversity ƒe nɔnɔme me. Le lãmenugbagbeviwo ƒe wɔwɔme gome la, MT nufiafiaa agblɔ Si ƒe mama le vome ɖi. Esi míebu eŋu be metabolite aɖewo le nɔnɔme tɔxɛ aɖewo me le nɔnɔme tɔxɛ aɖewo me, eye nɔnɔme bubuwo mele nɔnɔme tɔxɛ aɖewo me o, eye woƒe ametakpɔkpɔ ƒe asixɔxɔ nɔ te ɖe nutoa dzi la, míedo ametakpɔnu ƒe kpɔɖeŋu si wotsaka hã ɖa, si me δj kple Hj woma ɖe eve me le Si Dzidziɖedzi le mɔwo katã nu, metabolite ƒe hatsotso aɖewo koe, siwo ƒe ametakpɔkpɔ ƒe asixɔxɔwo lolo wu, adzi Si ɖe edzi etɔxɛe, evɔ bubuwo ya akpɔ random mama (Nɔnɔmetata 1B).
Be míate ŋu ado ametakpɔkpɔ ŋuti nufiafia ƒe nyagblɔɖi si wogbugbɔ ɖe fia le nyatakaka ŋuti nufiafia ƒe nɔnɔmetata ƒe axadzi la, míenyi gbeɖuɖɔ ɖula bibi (Ms) alo generalist (Sl) ƒe lãviwo ɖe Nepenthes pallens ƒe aŋgbawo dzi (Nɔnɔmetata 2A). Míezã MS/MS numekuku tsɔ xɔ MS/MS spectra 599 siwo mehiã o (nyatakakawo ƒe faɛl S1) tso methanol si woɖe tso aŋgba ƒe lãkusi me si woƒo ƒu le ʋetsuviwo ƒe nuɖuɖu nana vɔ megbe. RDPI, Hj, kple δj ƒe xexlẽdzesiwo zazã atsɔ akpɔ nyatakaka siwo le MS/MS ƒe ɖoɖowɔɖi ƒe faɛlwo me ƒe gbugbɔgaɖoɖo le susu me ɖea nɔnɔme dodzidzɔnamewo fiana (Nɔnɔmetata 2B). Nusi le edzi yim bliboae nye be, abe alesi nyatakakafianu la ɖɔe ene la, ne ʋetsuviwo yi edzi le aŋgbawo ɖum la, alesi gbegbe lãmenugbagbeviwo ƒe lãmenugbagbeviwo ƒe tɔtrɔwo katã trɔnae dzina ɖe edzi le ɣeyiɣi aɖe megbe: gaƒoƒo 72 le gbeɖula la ƒe nuɖuɖu megbe la, RDPI dzina ɖe edzi ŋutɔ. Ne wotsɔe sɔ kple amesiwo dzi womegblẽ o la, Hj dzi ɖe kpɔtɔ ŋutɔ, si tso alesi lãmenugbagbeviwo ƒe dɔwɔwɔ ƒe nɔnɔme tɔxɛa dzi ɖe edzi gbɔ, si wotsɔ δj ƒe dzesi de dzesii. Nɔnɔme sia si dze abe ɖe wòdzɔ ene la sɔ kple OD nufiafia ƒe nyagblɔɖiwo, gake mewɔ ɖeka kple MT nufiafia ƒe nyagblɔɖi veviwo o, si xɔe se be wozãa tɔtrɔ siwo dzɔna le vome (siwo menye mɔfiame o) le metabolite ƒe agbɔsɔsɔ me abe ametakpɔnu ene (Nɔnɔmetata 1B). Togbɔ be gbeɖuɖɔ eve siawo ƒe nu me ʋuʋudedi (OS) elicitor ƒe agbɔsɔsɔ kple nuɖuɖu ƒe nɔnɔme to vovo hã la, woƒe nuɖuɖu tẽ na tɔtrɔ mawo tɔgbe va le Hj kple δj ƒe mɔ dzi le gaƒoƒo 24 kple gaƒoƒo 72 ƒe nuŋeɣiwo. Vovototo ɖeka koe dzɔ le gaƒoƒo 72 ƒe RDPI zazã me. Ne wotsɔe sɔ kple esi Ms nuɖuɖu hena vɛ la, lãmenugbagbeviwo ƒe dɔwɔwɔ bliboa si Sl nuɖuɖu hena vɛ la sɔ gbɔ wu.
(A) Dodokpɔ ƒe ɖoɖo: wotsɔa gbeɖuɖɔ ɖula siwo bɔ (S1) alo eŋutinunyala (Ms) gbeɖuɖɔ ɖulawo kple numiemie siwo wotsɔa tsi wɔe ƒe aŋgba siwo me dze mele o, gake le gbeɖuɖɔ si wowɔ abe ɖe wole gbe ɖuɖu ene gome la, wozãa OS si nye Ms (W + OSMs) tsɔ kpɔa abi ƒe aŋgba ƒe nɔnɔme siwo woɖo ɖi ƒe dodo gbɔ. S1 (W + OSSl) lãviwo alo tsi (W + W). Nusi dzi woɖuna (C) nye aŋgba si megblẽ o. (B) Inducibility (RDPI ne wotsɔe sɔ kple control chart), diversity (Hj index) kple specialization (δj index) index si wobu na metabolite map tɔxɛa (599 MS/MS; data file S1). Ɣletividzesiwo ɖe vovototo gã aɖewo fia le gbeɖuɖɔ ɖulawo ƒe nuɖuɖu tẽ kple ƒuƒoƒo si me wokpɔa wo gbɔ le dome (Student’s t-test with paired t-test, *P<0.05 and ***P<0.001). ns, mele vevie o. (C) Ɣeyiɣi ƒe vovototodedeameme ƒe dzesi si nye main (blue box, amino acid, organic acid kple sugar; data file S2) kple special metabolite spectrum (red box 443 MS/MS; data file S1) le gbeɖuɖɔ ƒe atikewɔwɔ si wowɔ abe ɖe wòle ene megbe. Amadede ƒe ƒuƒoƒoa fia kakaɖedzi ƒe dometsotso si nye 95%. Ɣletividzesi la fia vovototo gã si le atikewɔwɔ kple amesi dzi woɖu dome [quadratic analysis of variance (ANOVA), eye Tukey ƒe vovototo si ɖe dzesi le anukwareɖiɖi me (HSD) kplɔe ɖo na post hoc multiple comparisons, *P<0.05, **P<0.01 Eye *** P <0.001]. (D) Kakaɖedzi ƒe nɔnɔmetata tɔxɛwo kple metabolite ƒe nɔnɔmetata tɔxɛwo (kpɔɖeŋu siwo wogbugbɔ wɔ kple atikewɔwɔ vovovowo).
Be míaku nu me akpɔe ɖa be gbeɖuɖɔ ɖulawo ƒe tɔtrɔ si wowɔna le metabolome ƒe ɖoɖo nu la dzena le tɔtrɔ siwo vaa lãmenugbagbeviwo ƒe dɔwɔwɔ ɖekaɖekawo ƒe agbɔsɔsɔ me hã la, míetsɔ susu ɖo nusiwo woɖɔna ɖo siwo ŋu wosrɔ̃ nu tsoe va yi le Nepenthes pallens ƒe aŋgbawo me siwo ŋu kpeɖodzi le be gbeɖulawo te ŋu nɔa te ɖe enu la ŋu. Phenolic amides nye hydroxycinnamamide-polyamine conjugates siwo ƒoa ƒu le nudzodzoewo ƒe gbeɖuɖɔ ƒe dɔwɔwɔ me eye wonya be eɖea nudzodzoewo ƒe dɔwɔwɔ dzi kpɔtɔna (32). Míedi MS/MS siwo sɔ la ƒe ŋgɔdonyawo eye míewɔ woƒe ʋuʋudedi ƒe ʋuʋudedi siwo woƒo ƒu ƒe nɔnɔmetata (Nɔnɔmetata S1). Mewɔ nuku o be phenol-nu siwo mekpɔa gome le ametakpɔkpɔ tso gbeɖuɖɔ ɖulawo abe chlorogenic acid (CGA) kple rutin ene me tẽ o la dzi ɖena kpɔtɔna le gbeɖuɖɔ vɔ megbe. To vovo na ema la, gbeɖuɖɔ ɖulawo ate ŋu ana phenol amides nasẽ ŋutɔ. Gbeɖuɖɔ ɖula eveawo ƒe nuɖuɖu atraɖii na phenolamides ƒe dzidzɔkpɔkpɔ ƒe ɣeyiɣi ɖeka kloe, eye nɔnɔme sia dze ƒã vevietɔ le phenolamides ƒe wɔwɔme de novo me. Woakpɔ nudzɔdzɔ ma ke ne wole 17-hydroxygeranyl nonanediol diterpene glycosides (17-HGL-DTGs) mɔ dzi dzrom, si wɔa acyclic diterpenes gbogbo aɖewo siwo wɔa dɔ nyuie siwo tsi tre ɖe gbeɖuɖɔ ɖulawo ŋu (33), siwo dometɔ Ms Feeding with Sl he nyagbɔgblɔ ƒe nɔnɔme ma tɔgbe vɛ (Nɔnɔmetata S1)).
Nusi ate ŋu agblẽ le gbeɖuɖɔ ɖulawo ƒe nuɖuɖu nana dodokpɔwo ŋu tẽe nye vovototo si le aŋgba ɖuɖu ƒe agbɔsɔsɔ kple gbeɖuɖɔ ɖulawo ƒe nuɖuɖu ɣeyiɣi me, si na wòsesẽna be woaɖe gbeɖuɖɔ ɖulawo ƒe ŋusẽkpɔɖeamedzi tɔxɛ siwo abi kple gbeɖulawo hena vɛ la ɖa. Be míate ŋu akpɔ gbeɖuɖɔ ƒomevi tɔxɛ si le aŋgba ƒe lãmenugbagbeviwo ƒe dɔwɔwɔ ƒe ŋuɖoɖo si wodo ɖa la gbɔ nyuie wu la, míewɔ Ms kple Sl lãviwo ƒe nuɖuɖu ƒe kpɔɖeŋu to OS si woƒo ƒu yeyee (OSM kple OSS1) zazã enumake ɖe aŋgba ƒe nɔnɔme siwo mewɔa tɔtrɔ o ƒe ʋuʋudedi W si woɖo ɖi la ŋu. Woyɔa mɔnu sia be W + OS dɔdamɔnu, eye wòɖoa dzidzenu si woatsɔ awɔ dɔe to ɣeyiɣi si tututu gbeɖuɖɔ ɖula la hea ŋuɖoɖoa ƒe gɔmedzedze me evɔ mehea vovototo siwo le lãmenugbagbeviwo ƒe bu ƒe agbɔsɔsɔ alo agbɔsɔsɔ me ƒe ŋusẽkpɔɖeamedzi tɔtɔame vɛ o (Nɔnɔmetata 2A) (34). Míezã MS/MS numekuku kple akɔntabubu ƒe mɔ̃a tsɔ xɔ MS/MS spektrum 443 (nyatakakawo ƒe faɛl S1), siwo sɔ kple spektrum siwo woƒo ƒu va yi tso nuɖuɖu tẽ dodokpɔwo me. Nyatakaka ŋuti nufiafia me dzodzro le MS/MS nyatakaka ƒuƒoƒo sia me ɖee fia be aŋgba tɔxɛ ƒe metabolomewo ƒe ɖoɖowɔwɔ gbugbɔgawɔ to gbeɖulawo ƒe nɔnɔmetata wɔwɔ me ɖe OS-koŋ ƒe dzideƒonamewo fia (Nɔnɔmetata 2C). Vevietɔ, ne wotsɔe sɔ kple OSS1 dɔdamɔnu la, OSM na metabolome ƒe etɔxɛnyenye dzi ɖe edzi le gaƒoƒo 4 me. Edze be míade dzesii be ne wotsɔe sɔ kple gbeɖuɖɔ ɖulawo ƒe nuɖuɖu tẽ dodokpɔ ŋuti nyatakakawo ƒe ƒuƒoƒo la, lãmenugbagbeviwo ƒe dɔwɔwɔ ƒe ʋuʋu si wokpɔ le susu me le akpa eve ƒe teƒe si wozã Hj kple δj abe kɔordinate ene kple metabolome etɔxɛ ƒe mɔfiame le gbeɖuɖɔ ƒe atikewɔwɔ si wowɔ abe ɖe wowɔe ene le ɣeyiɣi aɖe megbe ƒe ŋuɖoɖo me Dzidziɖedzi si sɔ (Nɔnɔmetata 2D). Le ɣeyiɣi ma ke me la, míebu agbɔsɔsɔme si le amino acids, organic acids kple sugars me (nyatakakawo ƒe faɛl S2) be míaku nu me akpɔe ɖa be metabolome ŋuti nunya ƒe dzidziɖedzi sia si woɖo taɖodzinu na la tso central carbon metabolism ƒe gbugbɔgaɖoɖo le gbeɖuɖɔ ɖula siwo wowɔ abe ɖe wole eŋu ene gbɔ hã ( Nɔnɔmetata S2 ). Be míaɖe nɔnɔme sia me nyuie wu la, míegalé ŋku ɖe phenolamide kple 17-HGL-DTG mɔ siwo ŋu míeƒo nu tsoe va yi ƒe lãmenugbagbeviwo ƒe ƒuƒoƒo ƒe ʋuʋu ŋu. Wotrɔa gbeɖulawo ƒe OS-koŋ ƒe ʋuʋu wòzua vovototodedeameme ƒe tɔtrɔ ƒe ɖoɖo le phenolamide ƒe lãmenugbagbeviwo ƒe dɔwɔwɔ me (Nɔnɔmetata S3). Phenolic amides siwo me coumarin kple caffeoyl ƒe akpawo le lae OSS1 lɔ̃na wu, gake OSMwo ya ʋãa ferulyl conjugates ƒe ʋuʋu tɔxɛ aɖe. Le 17-HGL-DTG mɔa gome la, wokpɔ vovototo OS ƒe ʋuʋu to malonylation si le anyime kple dimalonylation ƒe nuwɔna me (Nɔnɔmetata S3).
Eyome, míesrɔ̃ nu tso OS-si na transcriptome plasticity ŋu to time-course microarray data set zazã me, si wɔa OSMs zazã tsɔ daa dɔ na rosette numiemie ƒe aŋgbawo ƒe aŋgbawo le gbeɖulawo me. Sampling kinetics koŋ sɔ kple kinetics si wozã le metabolomics numekuku sia me ( 35 ). Ne míetsɔe sɔ kple metabolome ƒe gbugbɔgaɖoɖo si me metabolic plasticity dzina ɖe edzi vevietɔ le ɣeyiɣi aɖe megbe la, míekpɔa ɣeyiɣi kpui aɖe ƒe nuŋɔŋlɔ ƒe gbagbã le aŋgba siwo Ms hena vɛ me, afisi transcriptome inducibility (RDPI) kple specialization (δj) le 1 Gaƒoƒo ƒe dzidziɖedzi gã aɖe, kple vovototo (Hj) le ɣeyiɣi sia me, BMP1 ƒe ɖeɖefia dzi ɖe kpɔtɔ ŋutɔ, eye ɖiɖiɖeme kplɔe ɖo ƒe transcriptome ƒe etɔxɛnyenye (Nɔnɔmetata S4). Domenyiŋusẽfianu ƒe domenyiŋusẽfianu ƒe ƒomewo (abe P450, glycosyltransferase, kple BAHD acyltransferase ene) kpɔa gome le lãmenugbagbeviwo ƒe tɔtrɔ tɔxɛwo nuƒoƒoƒu tso xɔtuɖoɖo ƒe akpa siwo wokpɔ tso lãmenugbagbeviwo ƒe dɔwɔwɔ gbãtɔ me me, le etɔxɛnyenye ƒe kpɔɖeŋu gbãtɔ si míeyɔ va yi la dzi. Abe nudzɔdzɔ ŋuti numekuku ene la, woda phenylalanine ƒe mɔa kpɔ. Kukuɖenuŋua ɖo kpe edzi be domenyiŋusẽfianu vevi siwo le phenolamide ƒe asitɔtrɔ me la nye esiwo OS hena vɛ ŋutɔ le gbeɖuɖɔɖulawo me ne wotsɔe sɔ kple numiemie siwo womehe o, eye wowɔ ɖeka kplikplikpli le woƒe ɖeɖefia ƒe ɖoɖowo me. Nuŋɔŋlɔ ƒe akpa si nye MYB8 kple domenyiŋusẽfianu siwo nye PAL1, PAL2, C4H kple 4CL le mɔ sia ƒe dzigbe gome ɖe nuŋɔŋlɔ ƒe gɔmedzedze kaba fia. Acyltransferases siwo wɔa akpa aɖe le phenolamide ƒe ƒuƒoƒo mamlɛtɔ me, abe AT1, DH29, kple CV86 ene, ɖea dziyiyi ƒe ɖoɖo si nɔa anyi didi fiana (Nɔnɔmetata S4). Nusiwo míede dzesii le etame ɖee fia be transcriptome ƒe etɔxɛnyenye ƒe gɔmedzedze kaba kple metabolomics ƒe etɔxɛnyenye ƒe ŋgɔyiyi emegbe nye mɔnu si wotsɔ kpe ɖe eŋu, si ate ŋu atso synchronous regulatory system si dzea ametakpɔkpɔ ƒe ŋuɖoɖo sẽŋu aɖe gɔme gbɔ.
Numiemiewo ƒe lãmetsiŋusẽ ƒe dzesiwo ƒe tɔtrɔ yeyea wɔa dɔ abe ɖoɖowɔha si ƒoa gbeɖuɖɔ ŋuti nyatakakawo nu ƒu be woagbugbɔ awɔ ɖoɖo ɖe numiemiewo ƒe ŋutilã ƒe dɔwɔwɔ ŋu ene. Le gbeɖulawo ƒe nɔnɔmetata wɔwɔ vɔ megbe la, míedzidze numiemie ƒe lãmetsiŋusẽ ƒe hatsotso veviwo ƒe ƒuƒoƒo ƒe ŋusẽkpɔɖeamedzi eye míekpɔ ɣeyiɣi ƒe ɖekawɔwɔ ƒe ɖeɖefia le wo dome le susu me [Pearson correlation coefficient (PCC)> 0.4] (Nɔnɔmetata 3A). Abe alesi wokpɔ mɔe ene la, numiemie ƒe lãmetsi siwo do ƒome kple nugbagbewo ƒe wɔwɔme do ƒome le numiemie ƒe lãmetsiŋusẽ ƒe co-expression network la me. Tsɔ kpe ɖe eŋu la, wowɔa metabolic specificity (Si index) ƒe nɔnɔmetata ɖe network sia dzi be wòahe susu ayi numiemie ƒe lãmetsi siwo dɔdamɔnu vovovowo hena vɛ dzi. Woɖe gbeɖulawo ƒe ŋuɖoɖo tɔxɛ ƒe akpa vevi eve: ɖeka le JA ƒuƒoƒoa me, afisi JA (eƒe nugbagbeŋusẽ ƒe nɔnɔme JA-Ile) kple JA me nu bubu siwo woɖe tso eme la ɖea Si ƒe dzesi gãtɔ fiana le; eveliae nye ethylene (ET). Gibberellin ɖe dzidziɖedzi si sɔ koe fia le gbeɖuɖɔ ɖulawo ƒe vovototodedeameme me, gake numiemie ƒe lãmetsi bubuwo abe cytokinin, auxin, kple abscisic acid ene ya ƒe induction ƒe vovototodedeameme mesɔ gbɔ na gbeɖulawo o. Ne wotsɔe sɔ kple W + W ɖeɖeko zazã la, woate ŋu atrɔ JA ƒe nusiwo woɖe tso eme ƒe asixɔxɔ deŋgɔ ƒe dzidziɖedzi to OS dɔwɔwɔ (W + OS) dzi koŋ wòazu JAwo ƒe dzesi tɔxɛ sesẽ aɖe. Le mɔ si womele mɔ kpɔm na o nu la, wonya be OSM kple OSS1 siwo me elicitor ƒe agbɔsɔsɔme to vovo le la hea JA kple JA-Ile ƒe ƒuƒoƒo ɖekae vɛ. To vovo na OSS1 la, OSMwoe ʋãa OSM koŋ eye wòsẽna, gake OSS1 ya medoa abi siwo le gɔmedzedzea me ƒe ŋuɖoɖo ɖe ŋgɔ o (Nɔnɔmetata 3B).
(A) Co-expression network numekuku si wotu ɖe PCC akɔntabubu dzi le herbivore-si wotsɔ ƒo numiemie ƒe lãmetsiwo ƒe ƒuƒoƒo kinetics simulation. Node la tsi tre ɖi na numiemie ƒe lãmetsi ɖeka, eye node la ƒe lolome tsi tre ɖi na Si index si le numiemie ƒe lãmetsiŋusẽ ŋu koŋ le atikewɔwɔwo dome. (B) JA, JA-Ile kple ET ƒe ƒuƒoƒo ɖe aŋgbawo me si tso atikewɔwɔ vovovo siwo woɖe fia to amadede vovovowo me gbɔ: apricot, W + OSM; blɔ, W + OSSl, kple blɔ; ameyibɔ, W + W; grey, C (dziɖuɖu) . Ɣletividzesiwo ɖe vovototo gã aɖewo fia le atikewɔwɔ kple edzikpɔkpɔ dome (mɔ eve dzi ANOVA si kplɔe ɖo kple Tukey HSD post hoc multiple comparison, *** P <0.001). Nyatakaka ŋuti nufiafia me dzodzro le (C)697 MS/MS (nyatakakawo ƒe faɛl S1) me le JA ƒe nugbagbewo ƒe wɔwɔme kple nukpɔkpɔ ƒe vovototo si gblẽ (irAOC kple irCOI1) kple (D)585 MS/MS (nyatakakawo ƒe faɛl S1) le ETR1 me kple ET ƒe dzesi si gblẽ le gbeɖuɖɔ ɖulawo ƒe atikewɔwɔ eve siwo wowɔ abe ɖe wodze numiemie ƒe fliwo kple ʋu ƒuƒlu (EV) dzi ɖuɖu ƒe numiemiewo vɛ. Ɣletividzesiwo ɖe vovototo gã aɖewo fia le W+OS dɔdamɔnu kple esi megblẽ o dome (mɔ eve dzi ANOVA si kplɔe ɖo kple Tukey HSD post hoc multiple comparison, *P<0.05, **P<0.01 kple ***P<0.001). (E) Tsitretsitsi ɖe dɔ tɔxɛ wɔwɔ ŋu ƒe nɔnɔmetata siwo kaka. Amadedeawo tsi tre ɖi na ƒomevi vovovo siwo ƒe domenyiŋusẽfianu wogbugbɔ trɔ; dzesiawo tsi tre ɖi na atikewɔwɔmɔnu vovovowo: dzogoe etɔ̃, W + OSS1; dzogoe ene me, W + OSM; ƒo xlã C
Eyome, míezãa Nepenthes ƒe ƒomevi si ŋu wotrɔ asi le le domenyiŋusẽfianu me (irCOI1 kple sETR1) le JA kple ET biosynthesis (irAOC kple irACO) kple nukpɔsusu (irCOI1 kple sETR1) ƒe afɔɖeɖe veviwo me tsɔ daa numiemie ƒe lãmetsi eve siawo ƒe asitɔtrɔ le gbeɖuɖɔ ɖulawo ŋu kpɔna The relative contribution of reprogramming. Le ɖekawɔwɔ me kple dodokpɔ siwo míewɔ va yi la, míeɖo kpe gbeɖuɖɔ-OS ƒe ʋuʋu le numiemie ƒuƒlu siwo tsɔa nu (EV) me dzi (Nɔnɔmetata 3, C vaseɖe D) kple Hj ƒe dzesi ƒe ɖiɖi bliboa si OSM hena vɛ, esime δj ƒe dzesi dzi ɖe edzi. Ŋuɖoɖoa dzena wu ŋuɖoɖo si OSS1 ʋãna. Fli eve ƒe nɔnɔmetata si zã Hj kple δj abe kɔordinate ene la ɖe deregulation tɔxɛa fia (Nɔnɔmetata 3E). Nusi dze ƒã wu enye be le ƒomevi siwo me JA dzesi mele o me la, metabolome ƒe vovototo kple tɔtrɔ tɔxɛ siwo gbeɖuɖɔ ɖulawo hena vɛ la nu yi keŋkeŋ kloe (Nɔnɔmetata 3C). To vovo na esia la, ET ƒe nukpɔsusu si me ɖoɖoezizi le le sETR1 numiemiewo me, togbɔ be ŋusẽ si wòkpɔna ɖe tɔtrɔ siwo vaa gbeɖuɖɔ ɖulawo ƒe lãmenugbagbeviwo me dzi le bliboa me le ʋɛ ŋutɔ wu JA ƒe dzesiwo hã la, eɖea vovototo si le Hj kple δj ƒe dzesiwo me le OSM kple OSS1 ƒe dzodzo dome dzi kpɔtɔna (Nɔnɔmetata 3D kple Nɔnɔmetata S5). . Esia ɖee fia be tsɔ kpe ɖe JA ƒe dzesiwo ƒe tɔtrɔ ƒe dɔ vevitɔ ŋu la, ET ƒe dzesiwo ƒe tɔtrɔ hã wɔa dɔ abe gbeɖuɖɔ ɖulawo ƒe lãmenugbagbeviwo ƒe dɔwɔwɔ ƒe ŋuɖoɖo si le lã ƒomevi aɖewo koŋ ŋu la ɖɔɖɔɖo nyuie ene. Le ɖekawɔwɔ me kple asitɔtrɔ nyuie ƒe dɔwɔwɔ sia la, tɔtrɔ aɖeke meva le metabolome inducibility bliboa me le sETR1 numiemiewo me o. Le go bubu me la, ne wotsɔe sɔ kple sETR1 numiemiewo la, irACO numiemiewo he lãmenugbagbeviwo ƒe tɔtrɔ siwo gbeɖuɖɔ ɖulawo hena vɛ ƒe amplitude bliboa sɔ kple wo nɔewo vɛ, gake woɖe Hj kple δj ƒe xexlẽdzesi siwo to vovo ŋutɔ fia le OSM kple OSS1 kuxiwo dome (Nɔnɔmetata S5).
Be míate ŋu ade dzesi metabolite tɔxɛ siwo wɔ akpa vevi aɖe le gbeɖulawo ƒe ŋuɖoɖo ɖe lã ƒomevi aɖewo koŋ ŋu eye míatrɔ asi le woƒe wɔwɔme ŋu nyuie to ET dzesiwo dzi la, míezã structural MS/MS mɔnu si woto vɛ va yi. Mɔnu sia ɖoa ŋu ɖe bi-clustering mɔnu ŋu be woagaƒo nya ta tso metabolic ƒomea ŋu tso MS/MS kakɛwo [normalized dot product (NDP)] kple similarity score si wotu ɖe neutral loss (NL) dzi. MS/MS nyatakaka siwo wotu to ET ƒe domenyiŋusẽfianu ƒe fliwo me dzodzro me la na MS/MS 585 (nyatakakawo ƒe faɛl S1), si wokpɔ gbɔ to wo ƒoƒo ƒu ɖe MS/MS modules vevi adre (M) me (Nɔnɔmetata 4A) . Module siawo dometɔ aɖewo yɔ fũ kple metabolite tɔxɛ siwo woɖe dzesi va yi: le kpɔɖeŋu me, M1, M2, M3, M4 kple M7 yɔ fũ kple phenol derivatives vovovowo (M1), flavonoid glycosides (M2), acyl sugars (M3 Kple M4), kple 17-HGL-DTG (M7). Tsɔ kpe ɖe eŋu la, wobua akɔnta le lãmenugbagbeviwo ƒe dɔwɔwɔ ŋuti nyatakaka tɔxɛ (Si index) si le metabolite ɖeka si le module ɖesiaɖe me ŋu, eye woate ŋu akpɔ eƒe Si mama le susu me. Kpuie ko la, MS/MS spectra siwo ɖea gbeɖuɖɔ kple domenyiŋusẽfianu ƒe vovototo gã fiana la ƒe dzesi enye Si ƒe asixɔxɔ gãwo, eye kurtosis ƒe akɔntabubuwo ɖe lãfu ƒe mama fia le ɖusime asike ƒe dzogoe dzi. Wokpɔ colloid mama mawo dometɔ ɖeka le M1 me, afisi phenol amide ɖe Si ƒe akpa si lolo wu fia le (Nɔnɔmetata 4B). Herbivorous inducible 17-HGL-DTG si míeyɔ va yi le M7 me ɖe Si ƒe dzesi si sɔ fia, si fia be vovototo ƒe ɖoɖowɔwɔ si sɔ le OS ƒomevi eveawo dome. To vovo na ema la, metabolite tɔxɛ siwo wowɔ le ɖoɖo nu ƒe akpa gãtɔ, abe rutin, CGA, kple acyl sukliwo ene, le Si ƒe dzesi siwo bɔbɔ wu dome. Be woateŋu aku nu me nyuie wu le xɔtuɖoɖo ƒe sesẽ kple Si ƒe mama le metabolite tɔxɛwo dome la, wowɔ molecule network na module ɖesiaɖe (Nɔnɔmetata 4B). OD nufiafia ƒe nyagblɔɖi vevi aɖe (si woƒo nu tsoe kpuie le nɔnɔmetata 1B me) enye be ele be metabolite tɔxɛwo ƒe gbugbɔgaɖoɖo le gbeɖuɖu vɔ megbe nahe tɔtrɔ siwo le mɔ ɖeka dzi vɛ le metabolites siwo ƒe ametakpɔkpɔ ƒe asixɔxɔ gã le me, vevietɔ to woƒe tɔxɛnyenye dzi ɖeɖe kpɔtɔ me (si to vovo na mama le vome) Mɔnu) Defensive metabolite si wogblɔ ɖi to MT nufiafia me. Phenol-nu siwo ƒo ƒu ɖe M1 me ƒe akpa gãtɔ do ƒome kple nudzodzoewo ƒe dɔwɔwɔ ƒe ɖiɖi le dɔwɔwɔ gome ( 32 ). Ne míetsɔ Si ƒe asixɔxɔ siwo le M1 ƒe lãmenugbagbeviwo me sɔ kple wo nɔewo le aŋgba siwo wodo ɖa kple aŋgba siwo le EV dzi ɖum ƒe aŋgba siwo le wo me dome le gaƒoƒo 24 me la, míekpɔe be lãmenugbagbeviwo ƒe tɔtrɔ ƒe vovototodedeameme ƒe vovototodedeameme le nudzodzoe siwo ɖua gbe megbe la le dzidzim ɖe edzi ŋutɔ (Nɔnɔmetata 4C). Wokpɔ Si ƒe asixɔxɔ ƒe dzidziɖedzi tɔxɛa le phenolamides siwo ʋlia wo ɖokui ta me ko, gake womekpɔ Si ƒe asixɔxɔ ƒe dzidziɖedzi aɖeke le phenol bubuwo kple metabolite siwo womenya o siwo nɔa anyi ɖekae le module sia me o. Esia nye kpɔɖeŋu tɔxɛ aɖe, si do ƒome kple OD ƒe nufiafia. Nusiwo koŋ wogblɔ ɖi tso lãmenugbagbeviwo ƒe tɔtrɔ siwo gbeɖuɖɔ ɖulawo hena vɛ ŋu la sɔ. Be míadoe akpɔ nenye be OS-koŋ ƒe ET ye na phenolamide spectrum ƒe tɔxɛ sia he vɛ la, míewɔ metabolite Si index ƒe nɔnɔmetata eye míena vovototo ƒe ɖeɖefia ƒe asixɔxɔ le OSM kple OSS1 dome le EV kple sETR1 domenyiŋusẽfianuwo me (Nɔnɔmetata 4D ). Le sETR1 me la, vovototo si phenamide hena vɛ le OSM kple OSS1 dome dzi ɖe kpɔtɔ ŋutɔ. Wozã bi-clustering mɔnu hã ɖe MS/MS nyatakaka siwo woƒo ƒu le ƒomevi siwo me JA mesɔ gbɔ o me be woatsɔ aƒo nya ta le MS/MS modules vevi siwo ku ɖe JA-regulated metabolic specialization ŋu (Nɔnɔmetata S6).
(A) 585 MS/MS ƒe ƒuƒoƒo me tsonu siwo wotu ɖe kakɛ si woama (NDP ƒe sɔsɔ) kple akpaɖekedzimademade si woama (NL ƒe sɔsɔ) dzi na be module (M) la sɔ kple ƒuƒoƒo si wonya, alo to Metabolite ƒe wɔwɔme si womenya o alo esi wometrɔ nyuie o me. Le module ɖesiaɖe xa la, woɖe metabolite (MS/MS) specific (Si) mama fia. (B) Modular molecular network: Nodes tsi tre ɖi na MS/MS kple goawo, NDP (dzẽ) kple NL (blɔ) MS/MS xexlẽdzesiwo (cut-off,> 0.6). Wode ama metabolite specificity index (Si) si woɖo ɖe ɖoɖo nu la ɖe module la dzi (miame) eye wowɔ nɔnɔmetata na molecule network (ɖusime). (C) Module M1 si EV numiemie le constitutive (dziɖuɖu) kple induced nɔnɔme (simulated herbivore) le gaƒoƒo 24 me: molecule network diagram (Si asixɔxɔ nye node ƒe lolome, defensive phenolamide woɖe dzesi le blɔ me). (D) M1 molecule network diagram si nye spectrum line sETR1 kple EV kple ET ƒe nukpɔsusu si gblẽ: phenolic compound si wotsɔ green circle node tsi tre ɖi na, kple vovototo gã (P value) si le W + OSM kple W + OSS1 dɔdamɔnuawo dome abe Node ƒe lolome ene. CP, N-kafeoyl-tyrosine, si nye N-kafeoyl-tyrosine; CS, N-kafeoyl-spermidine, si nye N-kafeoyl-spermidine; FP, si nye N-ferulic acid si nye ester-uric acid; FS, N-ferulyl-spermidine, kple eƒe lãmesẽnyawo gbɔ kpɔkpɔ; CoP, N’, N “-Kumarolyl-tyrosine; CFS, N’, N”-caffeoyl, feruloyl-spermidine;Lycium barbarum le amegãxi Vi me, Nick.O-AS, O-acyl sukli.
Míegakeke numekukua ɖe enu tso Nepenthes ƒe domenyiŋusẽfianu ɖeka si gbɔdzɔ gbɔ va ɖo dzɔdzɔmetɔwo ƒe hatsotsowo me, afisi woƒo nu tso tɔtrɔ sesẽ siwo le lã ƒomeviwo me le gbeɖulawo ƒe JA ƒe agbɔsɔsɔ kple metabolite ƒe agbɔsɔsɔ tɔxɛwo me va yi le dzɔdzɔmetɔwo ƒe hatsotsowo me ( 26 ). Zã nyatakaka sia nàtsɔ aƒo nu tso dɔlékui 43 ŋu. Numiemie ƒomevi 123 siwo tso N. pallens me ye le dɔlékui siawo me. Woɖe numiemie siawo tso nuku siwo woƒo ƒu le teƒe vovovowo le Utah, Nevada, Arizona, kple California (Nɔnɔmetata S7), míebu metabolome ƒe vovototo (si woyɔna le afisia be amewo ƒe xexlẽme ƒe agbɔsɔsɔ) β vovototo) kple etɔxɛnyenye si OSM hena vɛ. Le ɖekawɔwɔ me kple numekuku siwo wowɔ va yi la, míekpɔ tɔtrɔ geɖe le lãmenugbagbeviwo ƒe dɔwɔwɔ me le Hj kple δj axes dzi, si ɖee fia be vovototo gã aɖe le dɔlékuiwo ƒe lãmenugbagbeviwo ƒe dɔwɔwɔ ɖe gbeɖuɖɔ ɖulawo ŋu ƒe plasticity me (Nɔnɔmetata S7). Habɔbɔ sia ɖo ŋku nusiwo wokpɔ va yi ku ɖe JAs ƒe tɔtrɔ ƒe ŋusẽ si gbeɖuɖɔ ɖulawo hena vɛ ŋu, eye wòlé asixɔxɔ gã aɖe me ɖe asi le ameha ɖeka me (26, 36). To JA kple JA-Ile zazã tsɔ do Hj kple δj dome le dzidzenu ƒe ƒomedodo bliboa kpɔ me la, míekpɔe be ƒomedodo nyui ɖedzesi aɖe le JA kple metabolome β ƒe vovototo kple etɔxɛnyenye ƒe dzesi dome (Nɔnɔmetata S7). Esia fia be gbeɖuɖɔ ɖulawo ƒe vovototodedeameme le JAs ƒe ʋuʋu si wokpɔ le amewo ƒe xexlẽme ƒe ɖoɖo nu ate ŋu atso lãmenugbagbeviwo ƒe tɔtrɔ vevi siwo tso tiatia tso nudzodzoe gbeɖulawo dome gbɔ.
Numekuku siwo wowɔ va yi ɖee fia be atama ƒomeviwo to vovo ŋutɔ le ƒomevi kple ŋuɖoɖo ɖe lãmenugbagbeviwo ƒe dɔwɔwɔ ƒe ametakpɔnu siwo wodona kple esiwo wowɔna ŋu le mɔ aɖe nu. Woxɔe se be nudzodzoewo ƒe agbɔsɔsɔ ƒe nyaƒoɖeamenu, numiemiewo ƒe agbenɔnɔ, kple ametakpɔnu wɔwɔ ƒe gazazãwoe kpɔa tɔtrɔ siawo siwo vaa gbeɖuɖɔ ɖulawo ƒe dzesiwo ƒe tɔtrɔ kple ametakpɔkpɔ ŋutetewo dzi le teƒe si lã ƒomevi aɖe tsina le. Míesrɔ̃ nu tso aŋgba ƒe metabolome ƒe tɔtrɔ si Nicotiana ƒomevi ade siwo tso Dziehe Amerika kple Anyiehe Amerika ƒe gbeɖuɖɔ ɖulawo hena vɛ ƒe ɖekawɔwɔ ŋu. Lã ƒomevi siawo do ƒome kplikplikpli kple Nepenthes Dziehe Amerika, si nye Nicolas Bociflo. La, N. nikotinis, Nikotiana n. gbe si gbɔdzɔ, Nicotiana tabacum, atama si le fli me, atama (Nicotiana spegazzinii) kple atama ƒe aŋgba ƒe atama (Nicotiana obtusifolia) (Nɔnɔmetata 5A) (37). Numiemie siawo dometɔ ade, siwo dome N. please ƒomevi si wonya nyuie hã le, nye ƒe sia ƒe ƒe numiemie siwo le petunia clade me, eye obtusifolia N. nye nɔvinyɔnu clade Trigonophyllae ƒe numiemie siwo nɔa anyi ɖaa (38). Emegbe la, wowɔ W + W, W + OSM kple W + OSS1 induction ɖe lã ƒomevi adre siawo dzi be woasrɔ̃ nu tso nudzodzoewo ƒe nuɖuɖu ƒe lãmenugbagbeviwo ƒe tɔtrɔ le lã ƒomeviwo ƒe ɖoɖo nu ŋu.
(A) Bootstrap phylogenetic tree si wotu ɖe alesi wòate ŋu adzɔe wu [na nuklia glutamine ƒe wɔwɔme (38)] kple Nicotiana ƒomevi adre siwo do ƒome kplikplikpli (amadede vovovowo) ƒe anyigbamama dzi (37). (B) Kakaɖedzi ƒe nɔnɔmetata si me vovototo tɔxɛwo le na Nicotiana ƒomevi adre ƒe lãmenugbagbeviwo ƒe dɔwɔwɔ ƒe nɔnɔmewo (939 MS/MS; data file S1). Le lã ƒomeviwo ƒe ɖoɖo nu la, metabolome ƒe vovototodedeameme do ƒome gbegblẽ kple alesi gbegbe wowɔ etɔxɛe. Woɖe ƒomedodo si le lã ƒomeviwo ƒe ɖoɖo nu le lãmenugbagbeviwo ƒe dɔwɔwɔ ƒe vovototo kple etɔxɛnyenye kpakple JA ƒe ƒuƒoƒo dome fia le nɔnɔmetata 2. S9. Amadede, ƒomevi vovovowo; dzogoe etɔ̃, W + OSS1; dzogoe ene me, W + OSM; (C) Woɖo Nicotiana JA kple JA-Ile ƒe ŋusẽkpɔɖeamedziwo ɖe OS ƒe dzidzɔkpɔkpɔ ƒe amplitude nu (mɔ eve ANOVA kple Tukey HSD le tsɔtsɔ sɔ kple wo nɔewo megbe, * P <0.05, ** P <0.01 kple * ** Na W + OS kple W + W tsɔtsɔ sɔ kple wo nɔewo, P <0.001). Aɖaka me nɔnɔmetata si me woɖe (D) vovototodedeameme kple (E) lã ƒomevi ɖesiaɖe ƒe etɔxɛnyenye fia le gbeɖuɖɔ kple methyl JA (MeJA) ƒe nɔnɔmetata wɔwɔ vɔ megbe. Ɣletividzesi la fia vovototo gã si le W + OS kple W + W alo lanolin kpe ɖe W (Lan + W) alo Lan kpe ɖe MeJA (Lan + MeJa) kple Lan dziɖuɖu (mɔ eve dzi numekuku le vovototo ŋu, si kplɔe ɖo kple Tukey ƒe HSD post hoc multiple comparison , *P<0.05, **P<0.01 kple ***P<0.001).
Míezã dual cluster mɔnu tsɔ de dzesi module 9 siwo me 939 MS/MS (data file S1) le. MS/MS ƒe wɔwɔme si wogbugbɔ ɖo to atikewɔwɔ vovovowo me to vovo ŋutɔ le module vovovowo dome le lã ƒomeviwo dome (Nɔnɔmetata S8). Hj (si woyɔna le afisia be lã ƒomevi vovovowo ƒe γ-vovototo) kple δj kpɔkpɔ le susu me ɖee fia be lã ƒomevi vovovowo ƒoa ƒu ɖe hatsotso vovovowo me ŋutɔ le lãmenugbagbeviwo ƒe dɔwɔwɔ ƒe teƒea, afisi zi geɖe la, lã ƒomeviwo ƒe mama dzena le wu dzidzɔkpɔkpɔ. Negbe N. linear kple N. obliquus koe la, woɖea induction ƒe ŋusẽkpɔɖeamedzi vovovowo fiana (Nɔnɔmetata 5B). To vovo na ema la, lã ƒomeviwo abe N. purpurea kple N. obtusifolia ƒe lãmenugbagbeviwo ƒe dɔwɔwɔ ƒe ŋuɖoɖo si medzena gaglãa le atikewɔwɔ me o, gake lãmenugbagbeviwo ƒe dɔwɔwɔ le vovovo wu. Lã ƒomevi tɔxɛwo ƒe mama le lãmenugbagbeviwo ƒe dɔwɔwɔ ƒe ŋuɖoɖo si wodo ɖa me na ƒomedodo manyomanyo ɖedzesi aɖe le etɔxɛnyenye kple gamma ƒe vovototo dome (PCC = -0.46, P = 4.9×10-8). Tɔtrɔ siwo OS hena vɛ le JAs ƒe agbɔsɔsɔ me do ƒome nyuie kple metabolome ƒe etɔxɛnyenye, eye wòdo ƒome gbegblẽ kple metabolic gamma ƒe vovototo si lã ƒomevi ɖesiaɖe ɖena fiana (Nɔnɔmetata 5B kple Nɔnɔmetata S9). Enyo be míade dzesii be lã ƒomevi siwo woyɔna le nuƒoƒo me be “dzesi ƒe ŋuɖoɖo” ƒomevi siwo le Nɔnɔmetata 5C me, abe Nepenthes nematodes, Nepenthes nepenthes, Nepenthes acute, kple Nepenthes attenuated ene, he dzesi ɖedzesiwo vɛ le miniti 30 me. OS-koŋ ƒe JA kple JA-Ile ƒe dɔvɔ̃ siwo do nyitsɔ laa, esime dɔlékui bubu siwo woyɔna be “signal unresponsive”, abe Nepenthes mills, Nepenthes powdery kple N. obtusifolia koe ɖea JA-Ile Edge induction fiana OS tɔxɛ aɖeke manɔmee (Nɔnɔmetata 5C). Le lãmenugbagbeviwo ƒe dɔwɔwɔ ƒe ɖoɖo nu, abe alesi míegblɔe le etame ene la, na Nepenthes si gbɔdzɔ la, nusiwo wɔa nu ɖe ​​dzesi ŋu ɖe OS ƒe vovototodedeameme fia eye wodzi δj ɖe edzi ŋutɔ, esime woɖe Hj dzi kpɔtɔ. Womekpɔ OS-koŋ ƒe gɔmedzedze ƒe ŋusẽkpɔɖeamedzi sia le lã ƒomevi siwo woda ɖe hatsotso si nye dzesi non-reactive ƒomeviwo me o (Nɔnɔmetata 5, D kple E). Woma OS-koŋ ƒe metabolitewo zi geɖe wu le dzesi-ŋuɖoɖo ƒomeviwo dome, eye dzesi ƒe ƒuƒoƒo siawo ƒoa ƒu kple lã ƒomevi siwo ƒe dzesi ƒe ŋuɖoɖo gbɔdzɔ, gake ƒomevi siwo ƒe dzesi ƒe ŋuɖoɖo gbɔdzɔ la meɖea wo nɔewo dzi nɔnɔ fiana boo o (Nɔnɔmetata S8 ). Nusi do tso eme sia ɖee fia be OS-koŋ ƒe JAwo ƒe ʋuʋu kple OS-koŋ ƒe tɔtrɔ si le anyime metabolome la do ƒome le lã ƒomeviwo ƒe ɖoɖo nu.
Eyome, míezã lanolin-mi si me methyl JA (MeJA) le tsɔ da dɔ na numiemiewo be míaku nu me akpɔe ɖa be woxe mɔ ɖe kadodomɔnu siawo nu le JA si wotsɔ exogenous JA, si anɔ numiemiewo ƒe cytoplasm me, ƒe anyinɔnɔ ta hã. Deesterification kabakaba nye JA. Míekpɔ nɔnɔme ma ke si nye tɔtrɔ vivivi tso lã ƒomevi siwo ɖoa dzesi dzesiwo gbɔ yi lã ƒomevi siwo mewɔa nu ɖe ​​dzesiwo ŋu o si tso JA ƒe nana atraɖii gbɔ (Nɔnɔmetata 5, D kple E). Kpuie ko la, MeJA dɔdamɔnu gbugbɔ wɔ ɖoɖo ɖe linear nematodes, N. obliquus, N. aquaticus, N. pallens, kple N. mikimotoi ƒe metabolomes ŋu vevie, si na δj dzi ɖe edzi ŋutɔ eye Hj dzi ɖe kpɔtɔ. N. purpurea ɖe δj ƒe dzidziɖedzi koe fia, gake menye Hj o. N. obtusifolia, si wokpɔ va yi be eƒoa JA ƒe agbɔsɔsɔ si bɔbɔ ŋutɔ nu ƒu la hã mewɔa nu nyuie ɖe MeJA ƒe atikewɔwɔ ŋu le metabolome ƒe tɔtrɔ yeye gome o. Nusiwo do tso eme siawo ɖee fia be JA wɔwɔ alo dzesiwo ƒe tɔtrɔ le ŋutilã me seɖoƒe li na le lã ƒomevi siwo mewɔa nu ɖe ​​dzesiwo ŋu o me. Be míado nukpɔsusu sia kpɔ la, míesrɔ̃ nu tso lã ƒomevi ene (N. pallens, N. mills, N. pink kple N. microphylla) siwo W + W, W + OSMs kple W + OSS1 Transcriptome (39) hena vɛ ŋu. Le ɖekawɔwɔ me kple metabolome ƒe tɔtrɔ ƒe ɖoɖo la, lã ƒomeviawo ma nyuie le transcriptome ƒe teƒea, siwo dome N. attenuated ɖe RDPI si OS hena vɛ si lolo wu fia, gake N. gracilis ya bɔbɔ wu (Nɔnɔmetata 6A). Gake wokpɔe be transcriptome ƒe vovototo si N. oblonga hena vɛ lae nye esi bɔbɔ wu le lã ƒomevi eneawo dome, si to vovo na N. oblonga ƒe metabolic vovototo si lolo wu si woɖe fia va yi le lã ƒomevi adre me. Numekuku siwo wowɔ va yi ɖee fia be domenyiŋusẽfianu siwo do ƒome kple ametakpɔkpɔ ƒe dzesi gbãtɔwo, siwo dome JA dzesiwo hã le, ɖea ametakpɔkpɔ ƒe ŋuɖoɖo gbãtɔ siwo gbeɖuɖɔ ɖulawo ƒe elicitors hena vɛ le Nicotiana ƒomeviwo me ƒe vovototodedeameme me (39). Ne wotsɔ JA ƒe dzesimɔwo sɔ kple wo nɔewo le lã ƒomevi ene siawo dome la, wokpɔ nɔnɔme dodzidzɔname aɖe (Nɔnɔmetata 6B). Domenyiŋusẽfianu akpa gãtɔ siwo le mɔ sia me, abe AOC, OPR3, ACX kple COI1 ene, ɖe induction ƒe agbɔsɔsɔ gã aɖe fia le lã ƒomevi ene siawo me. Ke hã, domenyiŋusẽfianu vevi aɖe, JAR4, trɔa JA wòzua eƒe nugbagbeŋusẽ ƒe nɔnɔme si nye JA-Ile ƒe nuŋɔŋlɔ siwo woƒo ƒu, eye eƒe nuŋɔŋlɔ ƒe agbɔsɔsɔme le sue ŋutɔ, vevietɔ le N. mills, Nepenthes pieris kple N. microphylla me. Tsɔ kpe ɖe eŋu la, domenyiŋusẽfianu bubu si nye AOS ƒe nuŋɔŋlɔ koe womekpɔ le N. bifidum me o. Tɔtrɔ siawo le domenyiŋusẽfianu ƒe ɖeɖefia me ateŋu anye nusi gbɔ nɔnɔme siwo gbɔ eme siwo JA ƒe wɔwɔme sue si le dzesi ƒe anergic ƒomeviwo me kple N. gracilis ƒe ʋuʋu hena vɛ.
(A) Nyatakaka ŋuti nufiafia me dzodzro le atama ƒomevi ene siwo do ƒome kplikplikpli siwo wotsɔ wɔ kpɔɖeŋui le aɖabaƒoƒo 30 le gbeɖuɖɔ ɖuɖu vɔ megbe ƒe nuŋɔŋlɔ ƒe ŋuɖoɖo gbãtɔwo ƒe ɖoɖowɔwɔ yeyee ŋu. Wobua RDPI to aŋgba siwo gbeɖuɖɔɖula OS hena vɛ tsɔtsɔ sɔ kple abidziɖuɖu me. Amadedeawo fia lã ƒomevi vovovowo, eye dzesiawo fia atikewɔwɔmɔnu vovovowo. (B) Domenyiŋusẽfianu ƒe ɖeɖefia le JA ƒe dzesimɔwo me me dzodzro le lã ƒomevi ene dome. Woɖe JA mɔ si wowɔ bɔbɔe la fia le aɖaka ƒe nɔnɔmetata la xa. Amadede vovovowo fia alesi wowɔa dɔ vovovowoe. Ɣletividzesi la ɖee fia be vovototo gã aɖe le W + OS dɔdamɔnu kple W + W dzi ɖuɖu dome (na Student ƒe t-dodokpɔ hena vovototo eveveve, *P<0.05, **P<0.01 kple ***P<0.001). OPDA, si nye 12-oxophytodienoic acid, si nye aɖi si me wodea amii; OPC-8: 0,3-oxo-2 (2 '(Z)-pentenyl)-siklopentane-1-oktanoic acid.
Le akpa mamlɛtɔ me la, míesrɔ̃ nu tso alesi nudzodzoe ƒomevi aɖewo koŋ ƒe numiemie ƒomevi vovovowo ƒe metabolome ƒe tɔtrɔ ate ŋu anɔ te ɖe gbeɖulawo nu ŋu. Numekuku siwo wowɔ va yi te gbe ɖe Nicotiana ƒomea dzi. Woƒe tsitretsitsi ɖe Ms kple lãviwo ŋu to vovo ŋutɔ (40). Le afisia la, míesrɔ̃ nu tso kadodo si le kpɔɖeŋu sia kple woƒe metabolic plasticity dome ŋu. Esi míezã atama ƒomevi ene siwo le etame, eye míedo ƒomedodo si le metabolome si gbeɖulawo hena vɛ ƒe vovototo kple etɔxɛnyenye kple numiemiewo ƒe tsitretsitsi ɖe Ms kple Sl ŋu dome kpɔ la, míekpɔe be tsitretsitsi, vovototodedeameme kple etɔxɛnyenye na generalist Sl All do ƒome nyuie, esime ƒomedodo si le tsitretsitsi ɖe nyɔnu bibiwo ŋu kple etɔxɛnyenye dome gbɔdzɔ, eye ƒomedodo si le vovototodedeameme dome la meɖe dzesi o (Nɔnɔmetata S10). Le S1 ƒe tsitretsitsi gome la, N. chinensis si gbɔdzɔ kple N. gracilis siaa, siwo woɖe fia va yi be woɖea JA ƒe dzesi ƒe tɔtrɔ ƒe agbɔsɔsɔme kple metabolome ƒe plasticity siaa fiana la, ƒe ŋuɖoɖo to vovo kura ɖe gbeɖulawo ƒe ʋuʋu ŋu, eye woɖe tsitretsitsi gã ma tɔgbe hã fia. Gbɔdɔdɔ.
Le ƒe blaade siwo va yi me la, numiemiewo takpɔkpɔ ŋuti nufiafia na nukpɔsusu ƒe ɖoɖo aɖe, si dzi numekulawo nɔ te ɖo gblɔ numiemiewo ƒe lãmenugbagbeviwo ƒe dɔwɔwɔ tɔxɛwo ƒe nɔnɔmetɔtrɔ kple dɔwɔwɔ ƒe agbɔsɔsɔ gã aɖe ɖi. Nufiafia siawo ƒe akpa gãtɔ mewɔna ɖe nutsotso sesẽwo ƒe ɖoɖo si sɔ dzi o (41). Wodoa nyagblɔɖi veviwo (3) ɖa le numekuku ƒe ɖoɖo ma ke me. Ne nyagblɔɖi veviwo dodokpɔ ɖe mɔ be woaku nukpɔsusu tɔxɛwo me la, esia ana be The field is moving forward. Woado alɔ wò, gake nàgbe ame bubuwo (42). Ke boŋ nufiafia yeyea gblɔa nya ɖi le numekuku ƒe ɖoƒe vovovowo eye wòtsɔa nusiwo ŋu wobu nyuie siwo ɖɔa nu yeye aɖe kpena ɖe eŋu (42). Gake woate ŋu aɖe nufiafia eve siwo wodo ɖa le dɔwɔwɔ ƒe ɖoɖo nu, MT kple OD, me bɔbɔe be wonye lãmenugbagbeviwo ƒe tɔtrɔ tɔxɛ siwo gbeɖuɖɔ ɖulawo hena vɛ ƒe nyagblɔɖi veviwo: OD ƒe nufiafia xɔe se be tɔtrɔ siwo vaa lãmenugbagbeviwo ƒe dɔwɔwɔ ƒe “teƒe” tɔxɛwo me la nyea mɔfiame ŋutɔ . MT ƒe nufiafia xɔe se be tɔtrɔ siawo manye esiwo metso mɔ dzi o eye woanɔ teƒe si le vome le lãmenugbagbeviwo ƒe dɔwɔwɔ ƒe teƒea, eye wodina be lãmenugbagbeviwo ƒe dɔwɔwɔ ƒe asixɔxɔ gã aɖe nanɔ wo si. Wodo OD kple MT ƒe nyagblɔɖiwo me dzodzro va yi kpɔ to a priori “ametakpɔnu” ƒe atikewo ƒe hatsotso kpui aɖe zazã me. Dodokpɔ siawo siwo wotu ɖe metabolite dzi xea mɔ na ŋutete be woaku nu me le metabolome ƒe tɔtrɔ ƒe kekeme kple mɔzɔzɔ ŋu le gbeɖuɣi, eye meɖea mɔ be dodokpɔ le akɔntabubu ƒe ɖoɖo si mewɔa tɔtrɔ o me nabia nyagblɔɖi vevi siwo woate ŋu abu abe bliboe ene Agbɔsɔsɔme na tɔtrɔ siwo va le numiemie ƒe metabolome me. Le afisia la, míezã mɔ̃ɖaŋununya yeye si le metabolomics me si wotu ɖe akɔntabubu MS dzi eye míewɔ deconvolution MS numekuku le nyatakaka nufiafia ƒe nuŋlɔɖiwo ƒe ga bliboa me tsɔ do vovototo si le eve siwo wodo ɖa le xexeame katã ƒe metabolomics ƒe ɖoɖo nu la kpɔ. Nufiafia sia ƒe nyagblɔɖi vevitɔ. Wozã nyatakaka ŋuti nufiafia le go geɖe me, vevietɔ le nugbagbe ƒomevi vovovowo kple nunyiame ƒe sisi ŋuti numekuku me (43). Gake le alesi míenya nu la, esiae nye dɔ gbãtɔ si wozã tsɔ ɖɔ numiemiewo ƒe lãmenugbagbeviwo ƒe dɔwɔwɔ ŋuti nyatakaka teƒe eye wokpɔ nu gbagbewo ƒe lãmenugbagbeviwo ƒe kuxi siwo ku ɖe ɣeyiɣi kpui aɖe ƒe lãmenugbagbeviwo ƒe tɔtrɔ le nutome ƒe dzesiwo ŋu gbɔ kpɔkpɔ me. Vevietɔ la, ŋutete si le mɔnu sia ŋu la le eƒe ŋutete be wòatsɔ nɔnɔme siwo le numiemie ƒomeviwo me kple wo dome asɔ kple wo nɔewo atsɔ alé ŋku ɖe alesi gbeɖulawo trɔ tso lã ƒomevi vovovowo me va zu lã ƒomevi vovovowo dome nɔnɔmetɔtrɔ gã ƒe nɔnɔmewo le nɔnɔmetɔtrɔ ƒe ɖoƒe vovovowo me. Nuwo ƒe lãmenugbagbeviwo ƒe dɔwɔwɔ.
Principal component analysis (PCA) trɔa nyatakaka gbogbo aɖewo ƒe ƒuƒoƒo wòzua didime dzi ɖeɖe kpɔtɔ ƒe teƒe ale be woate ŋu aɖe nyatakakaawo ƒe nɔnɔme vevitɔ me, eyata zi geɖe la, wozãnɛ abe numekukumɔnu ene tsɔ ɖea nyatakakaawo me, abe deconvolution metabolome ene. Ke hã, didime dzi ɖeɖe kpɔtɔ ana nyatakaka siwo le nyatakakaawo me ƒe akpa aɖe abu, eye PCA mate ŋu ana nyatakaka agbɔsɔsɔme si ku ɖe nɔnɔme siwo sɔ etɔxɛe na lãwo ƒe agbenɔnɔ ŋuti nufiafia ŋu o, abe: alesi gbeɖulawo gbugbɔ ɖoa ​​nu vovovowo ɖe ɖoɖo nu le agble tɔxɛwo me (le kpɔɖeŋu me, kesinɔnuwo, mama) Kple agbɔsɔsɔ) metabolites? Nusiwo wotsɔna trɔa asi le nu ŋu kawoe nye nusiwo gblɔa nɔnɔme si me gbeɖuɖɔ ɖula aɖe le? Le nu tɔxɛwo, vovototodedeameme kple nusiwo woateŋu awɔ ƒe nukpɔsusu nu la, nyatakaka siwo le aŋgba tɔxɛ ƒe lãmenugbagbeviwo ƒe nɔnɔme me gblẽna, eye wokpɔe be gbeɖulawo ƒe nuɖuɖu ateŋu ana lãmenugbagbeviwo ƒe dɔwɔwɔ tɔxɛ nawɔ dɔ. Le mɔ si míenɔ mɔ kpɔm na o nu la, míede dzesii be, abe alesi woɖe eme le nyatakaka ŋuti nufiafia ƒe dzesi siwo wozã me ene la, lãmenugbagbeviwo ƒe dɔwɔwɔ ƒe nɔnɔme si dona tso eme la sɔ kple wo nɔewo ŋutɔ le gbeɖuɖɔ eve (si ɖua nu le zã me ƒe generalist Sl) kple Solanaceae ŋuti nunyala bibi Aƒenɔ ƒe amedzidzedze megbe. Togbɔ be woƒe nuɖuɖu ƒe nuwɔna kple woƒe susu tsɔtsɔ ɖe nu ŋu to vovo kura hã. Fatty acid-amino acid conjugate (FAC) ƒe gɔmedzela le OS me (31). To gbeɖuɖɔ OS zazã atsɔ ada abi siwo wodo ɖe lãme na ame siwo woɖo ɖe ɖoɖo nu me la, gbeɖuɖɔ ɖulawo ƒe atikewɔwɔ si wowɔ abe ɖe wole dzɔdzɔm ene hã ɖe nɔnɔme ma tɔgbe fia. Mɔnu sia si woɖo ɖe ɖoɖo nu si wotsɔ srɔ̃a alesi numiemiewo wɔa nui ɖe gbeɖuɖɔ ɖulawo ƒe amedzidzedze ŋu la ɖea nusiwo tɔtɔa ame siwo tɔtrɔ siwo vaa gbeɖuɖɔ ɖulawo ƒe nuɖuɖu ƒe nɔnɔme me, siwo hea nusiwo gblẽa nu le wo ŋu le mɔ vovovowo nu le ɣeyiɣi vovovowo me hena vɛ la ɖa (34). FAC, si wonya be eya koŋue hea OSM vɛ la ɖea JAS kple numiemiewo ƒe lãmetsiŋusẽ bubuwo ƒe dɔwɔwɔ dzi kpɔtɔna le OSS1 me, gake OSS1 ya ɖea edzi zi gbɔ zi alafa geɖe (31). Gake OSS1 na JA ƒe ƒuƒoƒo ƒe agbɔsɔsɔ ma tɔgbe ne wotsɔe sɔ kple OSM. Woɖee fia va yi be JA ƒe ŋuɖoɖo le Nepenthes si gbɔdzɔ me la sea veve ɖe OSM nu ŋutɔ, afisi FAC ate ŋu alé eƒe dɔwɔwɔ me ɖe asi ne wotsɔ tsi ƒoe 1:1000 gɔ̃ hã ( 44 ). Eyata ne wotsɔe sɔ kple OSM la, togbɔ be FAC si le OSS1 me le sue ŋutɔ hã la, esɔ gbɔ be wòahe JA ƒe dɔvɔ̃ si sɔ vɛ. Numekuku siwo wowɔ va yi ɖee fia be woate ŋu azã protein siwo le abe porin ene (45) kple oligosaccharides (46) abe molecule ƒe kpeɖodzinuwo ene atsɔ aʋã numiemiewo ƒe ametakpɔkpɔ ƒe ŋuɖoɖowo le OSS1 me. Gake womenya haɖe nenye be elicitor siawo siwo le OSS1 me gbɔe JA ƒe ƒuƒoƒo si wokpɔ le numekuku si wowɔ fifia me tso o.
Togbɔ be numekuku ʋɛ aɖewo koe li siwo ƒo nu tso vovototo si le lãmenugbagbeviwo ƒe asibidɛ siwo dona tso gbeɖuɖɔ vovovowo alo exogenous JA alo SA (salicylic acid) zazã me (47) hã la, ame aɖeke meɖe fu na gbeɖuɖɔ ƒomevi aɖewo koŋ ƒe tɔtɔ le numiemiewo ƒe gbewo ƒe kadodo me kple eƒe ŋusẽkpɔɖeamedzi ɖe nusiwo koŋ ŋu The overall impact of metabolism is systematically studied. ame ŋutɔ ƒe nyatakakawo. Kukuɖenuŋu sia gaɖo kpe edzi be lãmetsiŋusẽ ƒe kadodo si le ememe kple numiemie ƒe lãmetsiŋusẽ bubu siwo menye JAs o la trɔa asi le lãmenugbagbeviwo ƒe tɔtrɔ si gbeɖuɖɔ ɖulawo hena vɛ ƒe tɔtrɔ tɔxɛ ŋu. Vevietɔ míekpɔe be ET si OSM hena vɛ la lolo ŋutɔ wu esi OSS1 hena vɛ. Mɔnu sia sɔ kple FAC ƒe nyatakaka geɖe wu le OSM me, si nye nɔnɔme si hiã eye wòsɔ gbɔ na ET ƒe gbagbã (48). Le numiemiewo kple gbeɖulawo dome kadodo gome la, ET ƒe dzesidɔ si wòwɔna ɖe numiemie tɔxɛwo ƒe lãmenugbagbeviwo ƒe dɔwɔwɔ dzi gakpɔtɔ nyea ɣeaɖewoɣi eye wòɖoa taɖodzinu ɖe ​​ƒuƒoƒo ɖeka si wotsɔ ƒo ƒui ŋu. Tsɔ kpe ɖe eŋu la, numekuku akpa gãtɔ zã ET alo eƒe ŋgɔdonuwo alo mɔxenu vovovowo zazã le gota tsɔ srɔ̃ nu tso ET ƒe ɖoɖowɔwɔ ŋu, siwo dometɔ aɖewoe nye atike siawo zazã le egodo ahe nugbegblẽ le ame ŋu geɖe siwo menye tẽ o vɛ. Le míaƒe sidzedze nu la, numekuku sia tsi tre ɖi na numekuku gã gbãtɔ si wowɔ ɖe ɖoɖo nu le akpa si ET wɔna le ET zazã me be woawɔ numiemie siwo trɔ domenyiŋusẽfianu siwo gblẽ eye woakpɔ wo adze sii be woawɔ ɖoɖo ɖe numiemiewo ƒe metabolome ƒe ŋusẽkpɔɖeamedzi ŋu. Herbivore-specific ET induction ate ŋu atrɔ asi le metabolome ƒe ŋuɖoɖo ŋu mlɔeba. Nusi ɖe dzesi wue nye domenyiŋusẽfianu ƒe asitɔtrɔ le ET biosynthesis (ACO) kple perception (ETR1) domenyiŋusẽfianuwo ŋu si ɖe phenolamides ƒe de novo nuƒoƒoƒu si le gbeɖuɖɔ ɖulawo koŋ ŋu fia. Woɖee fia va yi be ET ate ŋu atrɔ asi le nicotine ƒe ƒuƒoƒo si JA hena vɛ ŋu nyuie to putrescine N-methyltransferase ƒe ɖoɖowɔwɔ me ( 49 ). Gake le mɔ̃ɖaŋununya ƒe nukpɔsusu nu la, womenya alesi ET trɔa asi le phenamide ƒe dɔwɔwɔ ŋu nyuie o. Tsɔ kpe ɖe ET ƒe dzesiwo ƒe tɔtrɔ ƒe dɔwɔwɔ ŋu la, woate ŋu atrɔ asi le lãmenugbagbeviwo ƒe dɔwɔwɔ ŋu hã ayi S-adenosyl-1-methionine gbɔ be wòaɖɔ gadede polyaminophenol amides me ɖo. S-adenosyl-1-methionine nye ET kple Common domedome na polyamine biosynthetic mɔ. Ele be woagasrɔ̃ nu tso mɔnu si dzi ET ƒe dzesi tona ɖɔa phenolamide ƒe agbɔsɔsɔ ɖo ŋu.
Ɣeyiɣi didi aɖee nye sia le lãmenugbagbeviwo ƒe dɔwɔwɔ tɔxɛ siwo ƒe wɔwɔme womenya o ƒe agbɔsɔsɔ ta la, ŋkuléle ɖe lãmenugbagbeviwo ƒe hatsotso tɔxɛ aɖewo ŋu vevie mete ŋu bu ɣeyiɣi aɖe ƒe tɔtrɔ siwo vaa lãmenugbagbeviwo ƒe dɔwɔwɔ vovovowo ŋu le nugbagbewo ƒe kadodo megbe o. Fifia la, wotu ɖe nyatakaka ŋuti nufiafia me dzodzro dzi la, nu vevitɔ si do tso MS/MS spectrum xɔxɔ si wotu ɖe metabolite siwo mekpɔa ame ŋkume o dzi me enye be gbeɖula siwo ɖua gbeɖulawo alo wowɔa woƒe kpɔɖeŋua yi edzi le aŋgba ƒe lãmenugbagbeviwo ƒe dɔwɔwɔ ƒe vovototo bliboa dzi ɖem kpɔtɔ esime wòle eƒe etɔxɛnyenye ƒe seƒe dzim ɖe edzi. Metabolome ƒe vovototodedeameme ƒe dzidziɖedzi sia si gbeɖuɖɔ ɖulawo hena vɛ hena ɣeyiɣi kpui aɖe la do ƒome kple transcriptome ƒe vovototodedeameme ƒe dzidziɖedzi si wɔa dɔ aduadu. Nusi kpena ɖe metabolome tɔxɛ sia si lolo wu ŋu wu (si ƒe Si ƒe asixɔxɔ lolo wu) enye metabolite tɔxɛ si si gbeɖuɖɔ ƒe dɔwɔwɔ si woɖe dzesi va yi le. Kpɔɖeŋu sia sɔ kple OD nufiafia ƒe nyagblɔɖi, gake MT ƒe nyagblɔɖi si ku ɖe metabolome gbugbɔgawɔ ƒe ɖoɖo si wowɔna le vome ŋu mewɔ ɖeka o. Ke hã, nyatakaka sia sɔ kple kpɔɖeŋu si wotsaka ƒe nyagblɔɖi hã (MT nyuitɔ kekeake; Nɔnɔmetata 1B), elabena metabolite bubu siwo womede dzesii o siwo si ametakpɔkpɔ ƒe dɔwɔwɔwo womenya o ate ŋu akplɔ Si ƒe mama si wowɔ le vome la yome kokoko.
Kpɔɖeŋu ɖedzesi aɖe si numekuku sia galé ɖe te enye be tso nɔnɔmetɔtrɔ ƒe ɖoɖo sue aɖe (numiemie ɖeka kple atama ƒe xexlẽme) dzi va ɖo nɔnɔmetɔtrɔ ƒe dzidzenu si lolo wu (atama ƒomevi siwo do ƒome kplikplikpli) dzi la, nɔnɔmetɔtrɔ ƒe habɔbɔ ƒe ɖoɖo vovovowo le “ametakpɔnu nyuitɔ kekeake” me Vovototo gãwo le gbeɖulawo ƒe ŋutetewo me. Moore kple exɔlɔ̃wo. (20) kple Kessler kple Kalske (1) do susua ɖa le wo ɖokui si be woatrɔ nugbagbe ƒomevi vovovowo ƒe dɔwɔwɔ ƒe ɖoɖo etɔ̃ siwo Whittaker (50) de vovototo wo dome gbã la woazu atike vovovowo ƒe ɣeyiɣi ƒe tɔtrɔ siwo woɖo kple esiwo wòhe vɛ; agbalẽŋlɔla siawo dometɔ aɖeke meƒo nu tso eŋu kpuie Dɔwɔwɔ siwo wowɔna tsɔ ƒoa lãmenugbagbeviwo ƒe dɔwɔwɔ ŋuti nyatakakawo nu ƒu le agbɔsɔsɔ gã me hã megblɔ alesi woabu akɔnta le lãmenugbagbeviwo ƒe dɔwɔwɔ vovovowo ŋu tso nyatakaka siawo me o. Le numekuku sia me la, tɔtrɔ suesuesue siwo woawɔ le Whittaker ƒe dɔwɔwɔ ƒe hatsotsowo me abu α-metabolic diversity be enye MS/MS spectra ƒe vovototo le numiemie aɖe me, eye β-metabolic diversity abe intraspecific intraspecific metabolism of a group of populations Space, kple γ-metabolic diversity anye kekeɖenudɔ na numekuku le lã ƒomevi mawo tɔgbe ŋu.
JA ƒe dzesi le vevie na gbeɖulawo ƒe lãmenugbagbeviwo ƒe dɔwɔwɔ ƒe ŋuɖoɖo vovovowo. Gake agbɔsɔsɔme dodokpɔ sesẽ aɖeke mele akpa si JA ƒe nugbagbewo ƒe wɔwɔme ƒe ɖoɖowɔwɔ le lãmenugbagbevi ƒomeviwo me wɔna le metabolome vovovowo me ŋu o, eye nenye be JA ƒe dzesi nye teƒe si wozãna le mɔ gbadza nu na nuteɖeamedzi ƒe lãmenugbagbeviwo ƒe tɔtrɔ le nɔnɔmetɔtrɔ gã si kɔ wu me la, womenya kokoko o. Míede dzesii be Nepenthes gbeɖula ƒe gbeɖuɖɔ ƒe nɔnɔme hea metabolome ƒe etɔxɛnyenye vɛ eye metabolome ƒe etɔxɛnyenye ƒe tɔtrɔ le Nicotiana ƒomeviwo ƒe xexlẽme me kple le Nicotiana ƒomevi siwo do ƒome kplikplikpli dome la do ƒome nyuie le ɖoɖo nu kple JA ƒe dzesiwo. Tsɔ kpe ɖe eŋu la, ne JA ƒe dzesia gblẽ la, woate fli ɖe lãmenugbagbeviwo ƒe dɔwɔwɔ ƒe vovototo si gbeɖuɖɔ ɖula ɖeka ƒe domenyiŋusẽfianu hena vɛ la me (Nɔnɔmetata 3, C kple E). Esi wònye be Nepenthes ƒe xexlẽme si gbɔdzɔ le dzɔdzɔme nu ƒe lãmenugbagbeviwo ƒe dɔwɔwɔ ƒe tɔtrɔwo ƒe akpa gãtɔ nyea agbɔsɔsɔme ta la, tɔtrɔ siwo vaa lãmenugbagbeviwo ƒe β vovovowo kple woƒe vovototodedeameme me le numekuku sia me ate ŋu atso nusiwo me lãmenugbagbeviwo ƒe dɔwɔwɔ sɔ gbɔ ɖo ƒe hatsotsowo ƒe dzodzo sesẽ gbɔ koŋ. Nusiwo wotsɔ ƒo ƒui ƒe hatsotso siawo ɖua metabolome ƒe nɔnɔmetata ƒe akpa aɖe dzi eye wòhea kadodo nyui aɖe vɛ kple JA ƒe dzesiwo.
Esi wònye be atama ƒomevi si do ƒome kplikplikpli ƒe nugbagbeŋusẽ ƒe dɔwɔwɔ to vovo kura ta la, wodea dzesi nusiwo woɖɔna ɖo koŋ le nɔnɔme ƒe akpa dzi, eyata ewɔa numekuku wu. Nyatakaka ŋuti nufiafia ƒe dɔwɔwɔ le lãmenugbagbeviwo ƒe dɔwɔwɔ ƒe nɔnɔme si wolé ŋu ɖee fia be gbeɖuɖɔ ƒe ʋuʋu nana asitsatsa si le lãmenugbagbeviwo ƒe gamma ƒe vovototo kple etɔxɛ dome la nu sẽna ɖe edzi. JA ƒe dzesi wɔa akpa vevi aɖe le asitsatsa sia me. Metabolome ƒe etɔxɛnyenye ƒe dzidziɖedzi sɔ kple OD ƒe nyagblɔɖi vevitɔ eye wòdo ƒome nyuie kple JA ƒe dzesi, gake JA ƒe dzesi ya do ƒome gbegblẽ kple metabolic gamma ƒe vovototo. Kpɔɖeŋu siawo ɖee fia be numiemiewo ƒe OD ŋutete koŋue nɔa te ɖe JA ƒe plasticity dzi, eɖanye le microevolutionary scale alo le evolutionary scale si lolo wu dzi o. Exogenous JA zazã dodokpɔ siwo ƒo asa na JA biosynthesis ƒe gbɔdzɔgbɔdzɔwo gaɖee fia be woate ŋu ade vovototo atama ƒomevi siwo do ƒome kplikplikpli me ɖe dzesi-ŋuɖoɖo kple dzesi-ŋuɖoɖo o ƒomeviwo me, abe alesi woƒe JA ƒe nɔnɔme kple metabolome plasticity si gbeɖulawo hena vɛ ene. Dzesi ƒomevi siwo mewɔa dɔ o mate ŋu awɔ nu ɖe ​​wo ŋu o le woƒe ŋutete be woawɔ JA si le lãmenugbagbeviwo me ta eye le esia ta woƒe ŋutilã ƒe seɖoƒewo le wo ŋu. Esia ate ŋu atso domenyiŋusẽfianu vevi aɖewo ƒe tɔtrɔ le JA ƒe dzesimɔ (AOS kple JAR4 le N. crescens me) ƒe... Nusi do tso eme sia te gbe ɖe edzi be tɔtrɔ siwo vaa lãmetsiŋusẽ ƒe nukpɔsusu kple alesi wowɔa nui le ememe gbɔ koŋue na lã ƒomeviwo dome nɔnɔmetɔtrɔ gã siawo.
Tsɔ kpe ɖe kadodo si le numiemiewo kple gbeɖulawo dome ŋu la, lãmenugbagbeviwo ƒe lãmenugbagbeviwo ƒe vovototodedeameme didi do ƒome kple nukpɔsusu ƒe ŋgɔyiyi veviwo katã le numekuku le nugbagbewo ƒe tɔtrɔ ɖe nutome ŋu kple nɔnɔme sesẽ siwo ƒe nɔnɔmewo trɔna ŋu. Esi nyatakaka agbɔsɔsɔme si egbegbe MS dɔwɔnuwo xɔna le dzidzim ɖe edzi ta la, nukpɔsusuwo dodokpɔ le lãmenugbagbeviwo ƒe dɔwɔwɔ ƒe vovototo ŋu ate ŋu aƒo ame ɖekaɖekawo/ƒomeviwo ƒe lãmenugbagbeviwo ƒe vovototowo ta azɔ eye woawɔ xexeame katã ƒe numekuku atsɔ aɖe nɔnɔme siwo womele mɔ kpɔm na o afia. Le numekuku gãwo wɔwɔ me la, kpɔɖeŋunyagbɔgblɔ vevi aɖee nye susu si nye be woasusu anyigbatata siwo ŋu gɔmesese le siwo woate ŋu azã atsɔ aku nyatakakawo me. Eyata nu vevi aɖe si do tso MS/MS ƒe lãmenugbagbeviwo ƒe dɔwɔwɔ si me ameŋkumemakpɔmakpɔ le kple nyatakaka ŋuti nufiafia ƒe ƒuƒoƒo si li fifia me enye be ena metrik bɔbɔe aɖe si woate ŋu azã atsɔ awɔ anyigbatatawo atsɔ adzro lãmenugbagbeviwo ƒe tɔtrɔ ƒe vovototowo me le hatsotso vovovowo ƒe dzidzenu vovovowo dzi. Enye mɔnu sia ƒe nudidi vevitɔ. Nusɔsrɔ̃ tso micro/macro evolution kple nutoa me ƒe lãwo ƒe agbenɔnɔ ŋu.
Le macro-evolutionary level la, nu vevitɔ si le Ehrlich kple Raven (51) ƒe numiemie kple nudzodzoewo ƒe co-evolution nufiafia me enye be woagblɔe ɖi be tɔtrɔ si le lã ƒomeviwo dome metabolic diversity me gbɔe numiemiewo ƒe dzidzime vovovowo tso. Gake le ƒe blaatɔ̃ siwo va yi tso esime wota agbalẽ vevi sia me la, ƒã hafi wodo nukpɔsusu sia kpɔ (52). Esia tso dzidzime ƒe nɔnɔme siwo le lãmenugbagbeviwo ƒe dɔwɔwɔ ƒe nɔnɔme siwo sɔ le numiemie siwo le didiƒe ʋĩ ƒe dzidzimewo me gbɔ koŋ. Woate ŋu azã nusi mebɔ o la atsɔ aɖo seke na taɖodzinu ƒe numekukumɔnu. MS/MS ƒe dɔwɔwɔ ƒe ɖoɖo si li fifia si ŋu nyatakaka ŋuti nufiafia wɔ dɔ tsoe la bua MS/MS ƒe wɔwɔme ƒe sɔsɔ si le metabolite siwo womenya o ƒe agbɔsɔsɔme (metsɔ metabolite tiatia do ŋgɔ o) eye wòtrɔa MS/MS siawo wozua MS/MS ƒe hatsotso aɖe, ale le dɔnyala ƒe lãmenugbagbeviwo ƒe dɔwɔwɔ me Wotsɔa macro-evolutionary model siawo sɔna kple wo nɔewo le classification scale me. Akɔntabubu ƒe dzesi bɔbɔewo. Dɔwɔwɔa sɔ kple dzidzimeŋutinunya me dzodzro, si ate ŋu azã ɖoɖo si nu wowɔa nu ɖe ​​ɖoɖo nu le atsɔ akpɔ alesi gbegbe ame vovovowo alo nɔnɔmewo trɔnae ƒe agbɔsɔsɔme evɔ womagblɔe ɖi do ŋgɔ o.
Le nugbagbeŋusẽ ƒe ɖoɖo nu la, Firn kple Jones (53) ƒe dodokpɔ ƒe nukpɔsusu ɖee fia be woléa lãmenugbagbeviwo ƒe dɔwɔwɔ ƒe vovototo me ɖe asi le ɖoɖo vovovowo me be woana nu xoxowo be woawɔ nugbagbeŋusẽ ƒe dɔwɔna siwo me lãmenugbagbeviwo ƒe dɔwɔwɔ siwo medo ƒome kple wo nɔewo tsã o alo esiwo wotsɔ ɖo wo teƒe la ŋudɔ. Nyatakaka ŋuti nufiafia mɔnuwo naa ɖoɖo si me woate ŋu adzidze nɔnɔmetɔtrɔ ƒe tɔtrɔ siawo siwo ku ɖe metabolite tɔxɛwo ŋu siwo dzɔna le metabolite ƒe etɔxɛɣi me be wonye nɔnɔmetɔtrɔ ƒe dodokpɔ ƒe ɖoɖo si wodo ɖa ƒe akpa aɖe: tɔtrɔ si wowɔna le nugbagbewo me tso nusiwo le tẽ si le sue dzi yi esiwo le tẽ ŋutɔ gbɔ Nusiwo woxe mɔ na le nuto aɖe me.
Le wo katã me la, le molecule biology ƒe gɔmedzedze la, woto numiemiewo takpɔkpɔ ŋuti nufiafia veviwo vɛ, eye ame geɖe bua mɔnu siwo wotu ɖe deductive hypothesis dzi be wonye mɔ ɖeka kolia si dzi woato awɔ ŋgɔyiyi le dzɔdzɔmeŋutinunya me. Esia tso mɔ̃ɖaŋununya ƒe seɖoƒe siwo le metabolome bliboa dzidzedze me gbɔ koŋ. Togbɔ be mɔnu siwo wotu ɖe nukpɔsusuwo dzi ɖea vi ŋutɔ le nusiwo gbɔ wòtso ƒe mɔnu bubuwo tiatia me hã la, woƒe ŋutete be woado míaƒe gɔmesese ɖe ŋgɔ le nugbagbeŋusẽ ƒe kadodowo ŋu la se ɖe afi aɖe wu akɔntabubumɔnu siwo li fifia le egbegbe dzɔdzɔmeŋutinunya si me wozãa nyatakakawo le vevie me. Eyata nufiafia siwo womate ŋu agblɔ ɖi o la gbɔ nyatakaka siwo li ƒe ŋutete ŋu sasasã, eyata womate ŋu aɖe nususugblɔ ƒe mɔfiame/dodokpɔ ƒe tsatsam si nye ŋgɔyiyi le numekuku ƒe akpaa dzi la ɖa o (4). Míekpɔe do ŋgɔ be akɔntabubu ƒe dɔwɔwɔ ƒe ɖoɖo si nye metabolomics si woto vɛ le afisia ate ŋu agbugbɔ ɖetsɔleme aɖo nyitsɔ laa (alesi) kple mamlɛawo (nukatae) nya siwo ku ɖe metabolic vovovowo ŋu, eye wòawɔ akpa aɖe le ɣeyiɣi yeye si me wofia mɔe le nukpɔsusu nu ƒe nyatakakawo ŋuti dzɔdzɔmeŋutinunya me. Ɣeyiɣia gadzro nufiafia vevi siwo ʋã dzidzime siwo va yi me ake.
Wowɔa gbeɖulawo ƒe nuɖuɖu tẽ to instar larva alo Sl larva evelia nyinyi ɖe Pale-colored pitcher plant aŋgba ɖeka dzi le rose seƒoƒo ƒe numiemie ɖeka dzi, eye numiemie 10 gbugbɔgawɔ le numiemie ɖesiaɖe me. Wotsɔa nulénu siwo wotsɔ blaa nudzodzoewo ƒe lãviwo, eye woƒoa aŋgba ƒe lãkusi susɔeawo nu ƒu le gaƒoƒo 24 kple 72 le dɔlékuiwo xɔxɔ vɔ megbe eye wotsɔa wo dea tsikpe me kabakaba, eye woɖea nusiwo woɖɔna ɖo.
Srɔ̃ gbeɖuɖɔ ƒe atikewɔwɔ le mɔ si sɔ ŋutɔ nu. Mɔnuae nye be woazã avɔ ƒe nɔnɔme ƒe tasiaɖamfɔwo atsɔ ato ŋù siwo le fli etɔ̃ me le numiemiea ƒe aŋgba etɔ̃ siwo keke bliboe ƒe titina ƒe akpa ɖesiaɖe le avɔ ƒe seƒoƒotsihea ƒe tsitsi ƒe afɔɖeɖea me, eye woatsɔ 1:5 Diluted Ms. Alo azã asibidɛ siwo ŋu asigɛ le atsɔ ade S1 OS abi si woɖe la me. Ƒo aŋgba aɖe eye nàtrɔ asi le eŋu abe alesi míegblɔe ɖe etame ene. Zã mɔnu si ŋu míeƒo nu tsoe va yi nàtsɔ aɖe lãmenugbagbeviwo ƒe dɔwɔwɔ gbãtɔwo kple numiemiewo ƒe lãmetsiwo (54).
Le exogenous JA zazã gome la, wotsɔa lanolin paste 20μl si me MeJA 150μg (Lan + MeJA) le, kple lanolin 20μl kpe ɖe abi ƒe atikewɔwɔ (Lan + W) ŋu, alo zãa lanolin dzadzɛ 20μl abe alesi woatsɔ akpɔ wo dzii ene wɔa dɔ na rose-seƒoƒo ƒe numiemie ade siwo le ƒomevi ɖesiaɖe me ƒe aŋgba etɔ̃awo. Woŋe aŋgbaawo le gaƒoƒo 72 megbe le atikewɔwɔ vɔ megbe, wotsɔ wo de tsikpe me enumake le nitrogen si nye tsi me, eye wodzra wo ɖo ɖe -80°C vaseɖe esime woazã wo.
Wode dzesi JA kple ET ƒe domenyiŋusẽfianu ƒe hatsotso ene, siwo nye irAOC (36), irCOI1 (55), irACO kple sETR1 (48), le míaƒe numekuku ƒuƒoƒoa me. irAOC ɖee fia vevie be JA kple JA-Ile ƒe agbɔsɔsɔ dzi ɖe kpɔtɔ, evɔ irCOI1 ya mesea veve ɖe JAwo nu o. Ne wotsɔe sɔ kple EV la, JA-Ile ƒe ƒuƒoƒo dzi ɖe edzi. Nenema ke irACO aɖe ET ƒe wɔwɔ dzi akpɔtɔ, eye ne wotsɔe sɔ kple EV la, sETR1, si mesea veve ɖe ET nu o la, ana ET ƒe wɔwɔ nadzi ɖe edzi.
Wozãa fotoacoustic laser spectrometer (Sensor Sense ETD-300 ɣeyiɣi ŋutɔŋutɔ me ET sensor) tsɔ wɔa ET dzidzedze le mɔ si mewɔa nuvevi ame o nu. Le atikewɔwɔ vɔ megbe teti la, wolã aŋgbaawo ƒe afã hetsɔ wo de ahuhɔ̃e me nugoe si ƒe kpekpeme nye mililita 4 si wotu nu ɖo me, eye woɖe mɔ na ta la be wòaƒo ƒu le gaƒoƒo 5 me. Le dzidzedzea me la, wotsɔ ya dzadzɛ si ƒe sisi le lita 2/gaƒoƒo me klɔa nugoe ɖesiaɖe hena miniti 8, si to nusi Sensor Sense na do ŋgɔ be wòaɖe CO2 kple tsi ɖa.
Wota microarray ƒe nyatakakawo gbã le (35) me eye wodzra wo ɖo ɖe Dukɔa ƒe Nu gbagbewo Ŋuti Nyatakaka Dɔwɔƒe (NCBI) ƒe Gene Expression Comprehensive Database (accession number GSE30287). Woɖe nyatakaka siwo sɔ kple aŋgba siwo W + OSMs ƒe atikewɔwɔ kple atike si me gblẽ o he vɛ hena numekuku sia. Raw ƒe sesẽme nye log2. Hafi woawɔ akɔntabubuwo me dzodzro la, wotrɔ gɔmedzedzea eye wowɔe wòsɔ ɖe eƒe alafa memamã 75 lia nu to R kɔmpiuta dɔwɔɖoɖoa zazã me.
Woxɔ Nicotiana ƒomeviwo ƒe RNA ƒe ɖoɖowɔwɔ (RNA-seq) nyatakaka gbãtɔ tso NCBI Short Reading Archives (SRA), dɔa ƒe xexlẽdzesi nye PRJNA301787, si ŋu Zhou et al. (39) eye nàyi edzi abe alesi woɖe eme le (56) me ene. Wotia nyatakaka xoxo siwo ŋu W + W, W + OSM kple W + OSS1 wɔ dɔ tsoe siwo sɔ kple Nicotiana ƒomeviwo hena numekuku le numekuku sia me, eye wowɔ dɔ tso wo ŋu le mɔ si gbɔna nu: Gbã la, wotrɔ RNA-seq xexlẽ xoxoawo ɖe FASTQ nɔnɔme me. HISAT2 trɔa FASTQ wòzua SAM, eye SAMtools trɔa SAM faɛlwo wozua BAM faɛl siwo woɖo ɖe ɖoɖo nu. Wozãa StringTie tsɔ bua domenyiŋusẽfianu ƒe ɖeɖefia, eye eƒe ɖeɖefia mɔnu enye be kakɛwo nɔa gɔmeɖoanyi kakɛ akpe ɖeka ɖesiaɖe me le nuŋɔŋlɔ kakɛ miliɔn ɖeka siwo woɖo ɖe ɖoɖo nu me.
Acclaim chromatographic column (150 mm x 2.1 mm; particle size 2.2μm) si wozã le numekukua me kple 4 mm x 4 mm dzɔla sɔti la nye nu ɖeka ma ke. Wozãa vovototo eve siwo gbɔna le Dionex UltiMate 3000 Ultra High Performance Liquid Chromatography (UHPLC) ɖoɖoa me: miniti 0 va ɖo 0.5, isocratic 90% A [tsi si me ion mele o, 0.1% (v/v) acetonitrile kple 0.05% formic acid], 10% B (Acetonitrile kple 0.05% formic acid); 0.5 va ɖo 23.5 miniti, gradient phase nye 10% A kple 90% B, le wo nɔewo yome; 23.5 va ɖo 25 miniti, isocratic 10% A kple 90% B. Sisi ƒe agbɔsɔsɔme nye 400μl/min. Le MS ƒe numekukuwo katã gome la, do sɔti me tsi si woɖena ɖa la ɖe quadrupole kple time-of-flight (qTOF) analyzer si me electrospray dzɔtsoƒe si le dɔ wɔm le positive ionization mode me (capillary voltage, 4500 V; capillary outlet 130 V ; Drying temperature 200°C; drying airflow 10 litres/min) me.
Wɔ MS / MS kakɛwo me dzodzro (si woyɔna le afisia be MS / MS) si mehiã o alo womate ŋu ade vovototo wo dome o tso nyatakakaawo gbɔ be nàkpɔ xɔtuɖoɖo ŋuti nyatakaka tso lãmenugbagbeviwo ƒe dɔwɔwɔ ƒe nɔnɔme bliboa si woate ŋu ade dzesii ŋu. MS/MS mɔnu si me vovototo aɖeke mele o ƒe nukpɔsusu nɔ te ɖe nyateƒe si wònye be fesre si ɖea agbɔsɔsɔme ɖe vovo ŋutɔ le quadrupole la ŋu [eyata bu mass-to-charge ratio (m/z) ƒe dzesiwo katã be wonye kakɛwo]. Le susu sia ta la, esi wònye be Impact II dɔwɔnua mete ŋu wɔ CE ƒe ʋuʋu o ta la, wowɔ numekuku geɖe siwo le wo ɖokui si tsɔ zã dzidziɖedzi si tso ƒoƒo ƒe vovototodedeameme ƒe ƒoƒo ƒe ŋusẽ (CE) ƒe asixɔxɔwo me. Kpuie ko la, gbã la, dzro kpɔɖeŋua me to UHPLC-electrospray ionization/qTOF-MS dzi to mass spectrometry mode ɖeka zazã me (mamã ƒe nɔnɔme sue siwo dona tso in-source fragmentation me), scanning tso m/z 50 va ɖo 1500 le gbugbɔgagblɔ ƒe tsɔtsɔme si nye 5 Hz. Zã nitrogen abe gas si doa go wo nɔewo ene hena MS/MS numekuku, eye nàwɔ dzidzenu siwo le wo ɖokui si le ʋuʋudedi ƒe ʋuʋudedi vovovo ene siwo gbɔna siwo tso ƒoƒo me: 20, 30, 40, kple 50 eV. Le dzidzedze ƒe ɖoɖo bliboa me la, quadrupole la ƒe mass isolation fesre gãtɔ kekeake, tso m/z 50 va ɖo 1500. Ne woɖo ŋgɔgbe ŋutilã m/z kple isolation width dodokpɔ ɖe 200 la, mass range la wɔa dɔ le eɖokui si to dɔwɔnua ƒe dɔwɔwɔ ƒe kɔmpiuta dɔwɔɖoɖo kple 0 Da dzi. Scan for mass fragments abe alesi wòle le mass mode ɖeka me ene. Zã sodium formate (50 ml isopropanol, 200 μl formic acid kple 1 ml 1M NaOH tsi me tsi) hena agbɔsɔsɔme dzidzedze. Wozãa Bruker ƒe dzidzedzekpɔkpɔ ƒe mɔnu si sɔ pɛpɛpɛ ŋutɔ tsɔ doa nyatakaka faɛl la kpɔna ne wowɔ ɣeyiɣi ƒe didime si woɖo ɖi le mama dedie nu vɔ. Zã Data Analysis v4.0 kɔmpiuta dɔwɔɖoɖo (Brook Dalton, Bremen, Germany) ƒe dɔdɔ ƒe dɔwɔwɔ nàtsɔ atrɔ nyatakaka xoxoawo ƒe faɛlwo ɖe NetCDF ƒe nɔnɔme me. Wodzra MS/MS nyatakakawo ƒe ƒuƒoƒoa ɖo ɖe metabolomics nyatakakadzraɖoƒe si le ʋuʋu ɖi MetaboLights (www.ebi.ac.uk) me kple xexlẽdzesi si nye accession. MTBLS1471 ƒe xexlẽme.
MS/MS kpekpe ate ŋu ava eme to kadodo me dzodzro le MS1 kple MS/MS ƒe nyonyome ƒe dzesiwo dome na ƒoƒo ƒe ŋusẽ si le sue kple esi lolo kpakple se yeye siwo wowɔ. Wozãa R ŋɔŋlɔdzesi la tsɔ wɔa kadodo me dzodzro le ŋgɔdonya ƒe mama na nusi wowɔ ŋu, eye wozãa C# ŋɔŋlɔdzesi (https://github.com/MPI-DL/indiscriminant-MS-MS-assembly-pipeline) tsɔ wɔa seawo dzi.
Be míaɖe alakpa vodada nyui siwo tso megbe toɣliɖeɖe kple alakpa kadodo si dona tso m/z ƒe nɔnɔme aɖewo kpɔkpɔ le kpɔɖeŋu ʋee aɖewo ko me dzi akpɔtɔ la, míezãa “filled peak” dɔwɔwɔ si le R package XCMS (hena megbe toɣliɖeɖe ɖɔɖɔɖo) Should be used to replace the “NA” (undetected peak) intensity. Ne wozã fill peak function la, “0′′ intensity values ​​geɖewo gakpɔtɔ le data set la me siwo akpɔ ŋusẽ ɖe correlation calculation dzi.Ekema, míetsɔa data processing me tsonu siwo míekpɔ ne wozã filled peak function kple ne wome filled peak function zã o la sɔna kple wo nɔewo, eye míebua fill toɣliɖeɖe ƒe asixɔxɔ si wotu ɖe average ɖɔɖɔɖo ƒe asixɔxɔ si wobu dzi, eye emegbe míetsɔa 0 intensity value siawo ɖɔlia background value si wobu ɖeko wobua nɔnɔme siwo ƒe sesẽme wu megbenya ƒe teƒe etɔ̃ eye wobua wo be wonye “ta vavãwo.” Le PCC ƒe akɔntabubuwo gome la, m/z dzesi siwo le sample precursor (MS1) kple fragment data sets siwo ƒe taƒoƒo vavãtɔ enyi ya teti koe wobuna.
Ne ŋgɔdonya ƒe nyonyome ƒe nɔnɔme ƒe sesẽme le kpɔɖeŋu bliboa me do ƒome vevie kple nyonyome ƒe nɔnɔme ma ke si wotsɔ de ƒoƒo ƒe ŋusẽ sue alo gã me ƒe sesẽme si dzi ɖe kpɔtɔ, eye womede dzesi nɔnɔmea be isotope ƒe taƒoƒo to CAMERA dzi o la, woate ŋu aɖe egɔme ɖe edzi. Emegbe, to akɔntabubu le ŋgɔdonya-nuwɔna eve siwo katã ate ŋu anɔ anyi le s 3 me (fesre si wobu be woalé ɣeyiɣi si woatsɔ alé nu ɖe ​​te ƒe kɔkɔƒe) me la, wowɔa ƒomedodo ŋuti numekuku. Ne m/z ƒe asixɔxɔ bɔbɔ wu ŋgɔdonya ƒe asixɔxɔ eye MS/MS ƒe mama dzɔ le kpɔɖeŋu teƒe ɖeka ma ke le nyatakakawo ƒe ƒuƒoƒoa me kple ŋgɔdonya si me woɖee tsoe ko hafi wobua eŋu be enye kakɛ.
Míenɔ te ɖe se bɔbɔe eve siawo dzi ɖea kakɛ siwo wogblɔ siwo ƒe m/z ƒe asixɔxɔwo lolo wu m/z si le ŋgɔdonya si wode dzesii la ɖa, eye míenɔa te ɖe kpɔɖeŋu ƒe nɔƒe si ŋgɔdonya la dze le kple kakɛ si wogblɔ dzi. Ate ŋu adzɔ hã be woatia nyonyome ƒe nɔnɔme siwo dona tso kakɛ geɖe siwo le dzɔtsoƒe me siwo wowɔ le MS1 nɔnɔme me abe ŋgɔdolawo ene, si ana woawɔ MS/MS ƒe ƒuƒoƒo siwo mehiã o. Be míaɖe nyatakaka ƒe tsɔtsɔke sia dzi akpɔtɔ la, ne NDP ƒe sɔsɔme si le spektraawo me wu 0.6, eye wole chromatogram “pcgroup” si CAMERA ŋlɔ ɖi la me la, míaƒo wo nu ƒu ɖekae. Mlɔeba la, míeƒoa CE ƒe emetsonu ene siwo katã do ƒome kple ŋgɔdonya kple kakɛwo nu ƒu ɖe deconvoluted composite spectrum mamlɛtɔ me to tiatia ƒe sesẽme ƒe taƒoƒo si lolo wu le candidate peaks katã dome kple m/z ƒe asixɔxɔ ɖeka le ƒoƒo ƒe ŋusẽ vovovowo me. Wotu dɔwɔwɔ ƒe afɔɖeɖe siwo kplɔe ɖo ɖe nukpɔsusu si nye composite spectrum dzi eye wobua CE nɔnɔme vovovo siwo hiã be woatsɔ adzi kakɛ ƒe kakaɖedzi ɖe edzi, elabena kakɛ aɖewo koe woate ŋu ade dzesii le gododo ƒe ŋusẽ tɔxɛ aɖe te.
Wozã RDPI (30) tsɔ bu akɔnta le lãmenugbagbeviwo ƒe dɔwɔwɔ ƒe nɔnɔme ƒe inducibility ŋu. Wokpɔa metabolic spectrum diversity (Hj index) tso MS/MS ŋgɔdolawo ƒe agbɔsɔsɔ me to Shannon entropy si nye MS/MS frequency distribution zazã me to equation si gbɔna si Martínez et al. (8) . Hj = −∑i = 1mPijlog2(Pij) afisi Pij sɔ kple i-th MS/MS ƒe xexlẽme si sɔ le j-th kpɔɖeŋua me (j = 1, 2,..., m) (i = 1, 2, ..., m) t).
Woɖea metabolic specificity (Si index) gɔme be enye MS/MS si wona ƒe nyagbɔgblɔ ƒe dzedzeme le ƒomedodo me kple zi gbɔ zi geɖe le kpɔɖeŋu siwo ŋu wole nu ƒom tsoe dome. Wobu MS/MS ƒe vovototodedeameme be Si = 1t (∑j = 1tPijPilog2PijPi) .
Zã mɔfiame si gbɔna nàtsɔ adzidze metabolome-koŋ ƒe δj ƒe dzesi le j kpɔɖeŋu ɖesiaɖe me, kple MS/MS tɔxɛ ƒe mamã dedie δj = ∑i = 1mPijSi
Woɖoa MS/MS spectra ɖe ɖoɖo nu le eve eve me, eye wotua akɔnta le alesi woɖi wo nɔewoe ŋu ɖe dzesi eveawo dzi. Gbã la, to NDP si woɖo ɖi (si woyɔna hã be cosine correlation mɔnu) zazã me, zã sɔsɔŋusẽ si gbɔna tsɔ de dzesi akpa ƒe sɔsɔ le spektrum NDP = (∑iS1 & S2WS1, iWS2, i) 2∑iWS1, i2∑iWS2, i2 afisi S1 kple S2 Le Le ɖekawɔwɔ me la, na spektrum 1 kple spektrum 2, kpakple WS1, i kple WS2, i tsi tre ɖi na kpekpeme si wotu ɖe taƒoƒo ƒe sesẽme dzi be vovototo si le i-th common peak dome le spectra eveawo dome mede 0.01 Da o. Wobu kpekpeme la ale: W = [sesẽme si de ŋgɔ wu] m [nɔnɔme] n, m = 0.5, n = 2, abe alesi MassBank doe ɖa ene.
Wowɔ dzesidemɔnu evelia ŋudɔ, si bia be woaku NL si woama le MS/MS dome la me. Be míawɔ esia la, míezã NL ƒe xexlẽdzesi 52 siwo míedoa goe enuenu le MS ƒe mama ƒe ɖoɖoa me le tandem me, kple NL si le tẽ wu (nyatakakawo ƒe faɛl S1) si ŋu woŋlɔ nu ɖo va yi na MS/MS spectrum si le evelia ƒe metabolite siwo le Nepenthes ƒomevi si gbɔdzɔ la me ( 9, 26). Wɔ vektor eve si nye 1 kple 0 na MS/MS ɖesiaɖe, si sɔ kple NL aɖewo ƒe fifia kple esi meli o. Wonɔ te ɖe Euclidean ƒe didime ƒe sɔsɔ dzi bu NL ƒe sɔsɔ ƒe xexlẽdzesi na NL vektor eve ɖesiaɖe.
Be míawɔ ƒuƒoƒo eve ƒe ƒuƒoƒo la, míezã R package DiffCoEx, si wotu ɖe Weighted Gene Co-expression Analysis (WGCNA) ƒe kekeɖenudɔwɔwɔ dzi. Míezã NDP kple NL dzesidede matrikiwo le MS/MS spektrumwo me, míezã DiffCoEx tsɔ bu akɔnta le ƒomedodo matriki si wotsɔ sɔ kple wo nɔewo ŋu. Wowɔa binary clustering to “cutreeDynamic” parameter ɖoɖo ɖe mɔnu = “hybrid”, cutHeight = 0.9999, deepSplit = T, kple minClusterSize = 10. Woɖe DiffCoEx ƒe R dzɔtsoƒe code tso file 1 bubu me to Tesson et al. (57); Woateŋu akpɔ R WGCNA kɔmpiuta dɔwɔɖoɖo si hiã le https://horvath.genetics.ucla.edu/html/CoexpressionNetwork/Rpackages/WGCNA.
Be míate ŋu awɔ MS/MS molecule network numekuku la, míebu akɔnta le spectral connectivity eve eve ŋu si wotu ɖe NDP kple NL similarity types dzi, eye míezã Cytoscape software tsɔ kpɔ network topology le susu me to organic layout zazã me le CyFilescape yFiles layout algorithm extension application me.
Zã R version 3.0.1 nàtsɔ awɔ akɔntabubu me dzodzro le nyatakakaawo ŋu. Wozã mɔ eve ƒe vovototo me dzodzro (ANOVA) tsɔ da akɔntabubu ƒe vevienyenye kpɔ, eye Tukey ƒe vovototo si ɖe dzesi le anukwareɖiɖi me (HSD) le dodokpɔ si wowɔ le ɣeyiɣi aɖe megbe kplɔe ɖo. Be woate ŋu adzro vovototo si le gbeɖuɖɔ ƒe atikewɔwɔ kple esi dzi woɖu dome la, wozã Student ƒe t dodokpɔ tsɔ da kpɔɖeŋu ƒe hatsotso eve siwo ƒe vovototo ɖekae la ƒe asike eve mama kpɔ.
Ne èdi kpeɖeŋutɔ nyati sia la, taflatse kpɔ http://advances.sciencemag.org/cgi/content/full/6/24/eaaz0381/DC1
Esia nye nyati si woateŋu akpɔ le mɔ gbadza nu si woma le Creative Commons Attribution-Non-Commercial License ƒe sewo nu, si ɖe mɔ ɖe eŋudɔwɔwɔ, mama kple egbugbɔgawɔ le mɔnu ɖesiaɖe me, nenye be zazã mamlɛtɔ menye asitsatsa ƒe viɖe ta o eye gɔmeɖoanyia nye be dɔ gbãtɔa sɔ. Ɖeɖe fiã.
De dzesii: Ðeko míebia tso asiwò be nàtsɔ wò e-mail adrɛs ana ale be amesi nèkafu na axaa nanya be yedi be yeakpɔ e-mail la eye be menye nyatakaka gbegblẽe wònye o. Mímalé e-mail adrɛs aɖeke ɖe mɔ̃ dzi o.
Wozãa biabia sia tsɔ doa amedzro kpɔ nenye be amedzroe nènye eye woxea mɔ na nyatakaka gbegblẽwo ɖoɖo ɖe amewo le wo ɖokui si.
Nyatakaka ŋuti nufiafia naa xexeame katã ƒe ga si woatsɔ asɔ kple metabolome tɔxɛwo kple dodokpɔ ƒe ametakpɔkpɔ ŋuti nufiafiawo ƒe nyagblɔɖi.
Nyatakaka ŋuti nufiafia naa xexeame katã ƒe ga si woatsɔ asɔ kple metabolome tɔxɛwo kple dodokpɔ ƒe ametakpɔkpɔ ŋuti nufiafiawo ƒe nyagblɔɖi.
©2021 Amerikatɔwo ƒe Dzɔdzɔmeŋutinunya ƒe Ŋgɔyiyi Habɔbɔ. gomenɔamesiwo katã le esi. AAAS nye HINARI, AGORA, OARE, CHORUS, CLOCKSS, CrossRef kple COUNTER ƒe hadɔwɔla. Dzɔdzɔmeŋutinunya ƒe Ŋgɔyiyi ISSN 2375-2548.