He aratohu ariā nui ngā ariā parepare tipu rerekē hei whakamārama i ngā tauira o te pākia o te tipu, engari me whakamātautau tonu ā rātou matapae matua. I konei, i whakamahia e mātou te tātaritanga ine papatipu tandem kore whakaaro (MS/MS) hei tūhura i te metabolome o ngā momo tupeka kua ngoikore mai i ngā tipu takitahi ki ngā taupori me ngā momo whanaunga tata, ā, i tukatukahia he maha o ngā ariā āhuatanga ine papatipu i runga i ngā ira matū i roto i te mōhiohio. Anga hei whakamātautau i ngā matapae matua o te parepare pai (OD) me ngā ariā whāinga neke (MT). He rite te wāhanga mōhiohio o te metabolomics tipu ki te ariā OD, engari e whakahē ana i te matapae matua o te ariā MT mō ngā hihiri metabolomics i puta mai i ngā kararehe kai otaota. Mai i te tauine whanaketanga moroiti ki te tauine whanaketanga nui, i tautuhia te tohu jasmonate hei whakatau matua o te OD, i te mea i whakaratohia e te tohu ethylene te whakatikatika pai o te urupare motuhake ki te kararehe kai otaota i whakamāramahia e te whatunga ngota ngota MS/MS.
Ko ngā matū motuhake me ngā hanganga rerekē ngā tino kaiuru ki te urutau tipu ki te taiao, inā koa hei tiaki i ngā hoariri (1). Nā te kanorau whakamiharo o te pākia motuhake i kitea i roto i ngā tipu kua whakaoho ake i ngā tekau tau o te rangahau hohonu mō ōna āhuatanga maha o ngā mahi taiao, ā, kua hanga he rārangi roa o ngā ariā tiaki tipu, arā, ko te whanaketanga whanaketanga me te whanaketanga taiao o ngā taunekeneke tipu-ngārara. He aratohu nui ngā rangahau wheako (2). Heoi, kāore ēnei ariā tiaki tipu i whai i te ara paerewa o te whakaaro whakaaro whakaaro, i reira ko ngā matapae matua i te taumata kotahi o te tātari (3) ā, i whakamatautauria mā te whakamātautau hei whakatairanga i te huringa whanaketanga ariā e whai ake nei (4). Ko ngā herenga hangarau e whakawhāiti ana i te kohikohinga raraunga ki ngā kāwai pākia motuhake, ā, ka ārai i te tātari whānui o ngā matū motuhake, nā reira ka ārai i ngā whakataurite i waenga i ngā kāwai he mea nui mō te whanaketanga ariā (5). Ko te kore o ngā raraunga pākia whānui me te moni kotahi hei whakataurite i te reremahi tukatuka o te wāhi pākia i waenga i ngā rōpū tipu rerekē e aukati ana i te pakeketanga pūtaiao o te mara.
Ka taea e ngā whanaketanga hou o te mara o te tandem mass spectrometry (MS/MS) metabolomics te whakaahua whānui i ngā huringa pākia i roto, i waenga hoki i ngā momo o tētahi pūnaha clade, ā, ka taea te whakakotahi me ngā tikanga rorohiko hei tatau i te ōritetanga hanganga i waenga i ēnei ranunga uaua. Te mōhiotanga o mua ki te matū (5). Ko te whakakotahitanga o ngā hangarau matatau i roto i te tātari me te rorohiko e whakarato ana i tētahi anga e tika ana mō te whakamātautau roa o ngā matapae maha i mahia e ngā ariā taiao me te whanaketanga o te kanorau pākia. I whakaurua e Shannon (6) te ariā mōhiohio mō te wā tuatahi i roto i tana tuhinga matua i te tau 1948, e whakatakoto ana i te turanga mō te tātari pāngarau o ngā mōhiohio, kua whakamahia i roto i ngā mara maha atu i tua atu i tōna tono taketake. I roto i te genomics, kua angitu te tono ariā mōhiohio ki te ine i ngā mōhiohio tiaki raupapa (7). I roto i te rangahau transcriptomics, ka tātarihia e te ariā mōhiohio ngā huringa whānui i roto i te transcriptome (8). I roto i ngā rangahau o mua, i whakamahia e mātou te anga tatauranga o te ariā mōhiohio ki te metabolomics hei whakaahua i te tohungatanga pākia o te taumata kiko i roto i ngā tipu (9). I konei, ka whakakotahi mātou i te tukanga mahi e hangai ana ki te MS/MS me te anga tatauranga o te ariā mōhiohio, e tohuhia ana e te kanorau pākia i roto i te moni noa, hei whakatairite i ngā matapae matua o te ariā tiaki tipu o te metabolome i whakaohokia e ngā kararehe kai otaota.
Ko ngā anga ariā mō te parepare tipu he mea whakauru tahi tetahi ki tetahi, ā, ka taea te wehewehe kia rua ngā wāhanga: ko ērā e ngana ana ki te whakamārama i te tohatoha o ngā matūkai motuhake ki te tipu i runga i ngā mahi parepare, pērā i te parepare tino pai (OD) (10), te whāinga neke (MT) (11) ) Me te ariā āhua (12), ko ētahi e rapu ana i ngā whakamārama ā-pūkaha mō te pānga o ngā huringa o te wātea rauemi ki te tipu tipu me te kohikohinga o ngā matūkai motuhake, pērā i te waro: te ariā taurite matūkai (13), te ariā tere tipu (14 ), me te ariā taurite tipu me te rerekētanga (15). Kei ngā taumata tātari rerekē ngā huinga ariā e rua (4). Heoi, e rua ngā ariā e pā ana ki ngā mahi parepare i te taumata mahi e whakahaere ana i te kōrero mō ngā parepare tipu pumau me te ārai ka taea te whakaoho: te ariā OD, e whakaaro ana ka haumi ngā tipu ki ā rātou parepare matū utu nui ina hiahiatia anake, hei tauira, ina horomia. Ina whakaeke mai he kararehe tarutaru, nō reira, i runga i te āheinga o te whakaeke a meake nei, ka tohaina te matū me te mahi parepare (10); E kī ana te ariā MT kāore he tuaka o te huringa ahunga o te metabolite, engari ka huri matapōkere te metabolite, ā, ka puta te tūponotanga ki te aukati i te "whāinga nekehanga" pākia o ngā kararehe kai otaota e whakaeke ana. Arā, he rerekē ngā matapae a ēnei ariā e rua mō te whakahounga pākia e puta ana i muri i te whakaekenga o ngā kararehe kai otaota: te whanaungatanga i waenga i te kohikohinga kotahi-ara o ngā metabolite me te mahi tiaki (OD) me ngā huringa pākia kore-ara (MT) (11).
Ko ngā whakapae OD me MT ehara i te mea ko ngā huringa i puta i roto i te metabolome anake, engari ko ngā hua ā-taiao me te whanaketanga o te kohikohinga o ēnei metabolites, pērā i ngā utu urutau me ngā painga o ēnei huringa pākia i roto i tētahi taiao ā-taiao motuhake (16). Ahakoa e mōhio ana ngā whakapae e rua ki te mahi tiaki o ngā metabolites motuhake, he utu nui pea, he kore utu rānei, ko te matapae matua e wehewehe ana i ngā whakapae OD me MT kei roto i te ahunga o ngā huringa pākia i puta. Ko te matapae o te ariā OD te mea i arohia nuitia tae noa ki tēnei wā. Kei roto i ēnei whakamātautau te ako i ngā mahi tiaki tika, ā-kore rānei o ngā kiko rerekē o ngā matū motuhake i roto i ngā whare kati kōtuhi me ngā āhuatanga taiao, tae atu ki ngā huringa i te wāhanga whanaketanga tipu (17-19). Heoi, tae noa ki tēnei wā, nā te korenga o tētahi rerenga mahi me tētahi anga tatauranga mō te tātari whānui ā-ao mō te kanorau pākia o tētahi rauropi, ko te matapae rerekētanga matua i waenga i ngā ariā e rua (arā, te ahunga o ngā huringa pākia) me whakamātautau tonu. I konei, ka whakaratohia e mātou tētahi tātaritanga pēnei.
Ko tētahi o ngā āhuatanga tino nui o ngā matū matū motuhake ki ngā tipu ko tō rātou kanorau hanganga tino nui i ngā taumata katoa mai i ngā tipu takitahi, ngā taupori ki ngā momo rite (20). He maha ngā huringa tauanga i roto i ngā matū matū motuhake ka kitea i te tauine taupori, ko ngā rerekētanga kounga kaha ka mau tonu i te taumata momo (20). Nō reira, ko te kanorau pākia o ngā tipu te āhuatanga matua o te kanorau mahi, e whakaata ana i te urutau ki ngā momo āputa rerekē, inā koa ko ngā āputa me ngā momo whakaekenga rerekē e ngā pepeke motuhake me ngā kararehe kai otaota noa (21). Mai i te tuhinga hou a Fraenkel (22) mō ngā take mō te noho o ngā matū matū motuhake ki ngā tipu, kua kiia ngā taunekeneke me ngā momo pepeke he pēhanga kōwhiringa nui, ā, e whakaponohia ana ko ēnei taunekeneke te mea i hanga i ngā tipu i te wā whanaketanga. Ara pākia (23). Ko ngā rerekētanga i waenga i ngā momo i roto i te kanorau o ngā matū matū motuhake ka whakaata pea i te taurite ā-tinana e pā ana ki te parepare tipu pumau me te ārai tipu ka taea te whakaoho i ngā rautaki kararehe kai otaota, i te mea he maha ngā momo e rua e hono kino ana tetahi ki tetahi (24). Ahakoa he pai te pupuri i te parepare pai i ngā wā katoa, ko ngā huringa pākia wa e pā ana ki te parepare ka whakarato i ngā painga mārama i roto i te tuku i ngā tipu ki te tohatoha rauemi nui ki ētahi atu haumitanga ā-tinana (19, 24), me te karo i te hiahia mō te hononga tahi. Te kino ā-tauira (25). Hei tāpiri, ko ēnei whakaritenga anō o ngā matū motuhake i puta mai i ngā pepeke kai otaota ka arahi pea ki te tohatoha kino i roto i te taupori (26), ā, ka whakaata pea i ngā pānui tika o ngā huringa taiao nui i roto i te tohu waikawa jasmonic (JA), ka mau tonu pea i roto i te taupori. Ko ngā tohu JA teitei me te iti he tauwhitinga i waenga i te parepare ki ngā kai otaota me te whakataetae ki ngā momo motuhake (27). Hei tāpiri, ka tere te ngaro me te hurihanga o ngā ara biosynthetic metabolite motuhake i te wā whanaketanga, ka hua ake he tohatoha pākia koretake i waenga i ngā momo whanaunga tata (28). Ka taea te whakatū tere i ēnei polymorphisms hei whakautu ki te hurihanga o ngā tauira kai otaota (29), ko te tikanga ko te rerekētanga o ngā hapori kai otaota he mea matua e akiaki ana i te rerekētanga pākia.
I konei, i whakatauhia e mātou ngā raruraru e whai ake nei. (I) Me pēhea te whakarite anō a te pepeke kai otaota i te metabolome tipu? (Ii) He aha ngā wāhanga mōhiohio matua o te kirihou pākia ka taea te ine hei whakamātautau i ngā matapae o te ariā parepare mō te wā roa? (Iii) Me whakarite anō rānei te metabolome tipu i tētahi ara ahurei ki te kaiwhakaeke, mēnā koinā, he aha te mahi a te homoni tipu ki te whakarite i tētahi urupare pākia motuhake, ā, ko ēhea ngā metabolite e whai wāhi ana ki te motuhaketanga momo o te parepare? (Iv) I te mea ka taea te whakawhānui i ngā matapae i hangaia e te maha o ngā ariā parepare puta noa i ngā taumata katoa o ngā kiko koiora, i pātai mātou me pēhea te ōrite o te urupare pākia i puta mai i te whakataurite ā-roto ki te whakataurite i waenga i ngā momo? Hei whakatutuki i tēnei, kua rangahauhia e mātou te metabolome rau o te nikotīni tupeka, he tipu tauira rauropi he nui te pākia o te tinana, ā, he whai hua ki ngā torongū o ngā kararehe kaiwhenua taketake e rua, ko Lepidoptera Datura (Ms) (He tino pukuriri, he mea kainga te nuinga). Mō Solanaceae me Spodoptera littoralis (Sl), he momo "momo" ngā kutukutu rau miro, me ngā tipu manaaki o Solanaceae me ētahi atu manaaki o ētahi atu momo me ngā whānau. Kai tipu. I tātarihia e mātou te whānuitanga metabolomics MS/MS, ā, i tangohia ngā whakaahuatanga tatauranga ariā mōhiohio hei whakataurite i ngā ariā OD me MT. Waihangahia he mahere motuhake hei whakaatu i te tuakiri o ngā metabolites matua. I whakawhānuihia te tātaritanga ki te taupori taketake o N. nasi me ngā momo tupeka e pā tata ana ki a rātou hei tātari atu i te rerekētanga i waenga i te tohu homoni tipu me te whakaoho OD.
Hei hopu i tētahi mahere whānui mō te kirihou me te hanganga o te metabolome rau o te tupeka kai otaota, i whakamahia e mātou tētahi tukanga tātari me te tatau kua whakawhanakehia i mua hei kohikohi me te wetewete i ngā ira MS/MS motuhake raraunga taumira teitei mai i ngā tangohanga tipu (9). Ka taea e tēnei tikanga kore-whakarerekētanga (e kiia nei ko te MS/MS) te hanga i ngā ira matū kore-taumaha, ka taea te whakamahi mō ngā tātari taumata-pūhui katoa kua whakaahuatia i konei. He momo rerekē ēnei ira matū tipu wetewetehia, kei roto ko ngā rau ki ngā mano o ngā ira matū (tata ki te 500-1000-s/MS/MS i konei). I konei, ka whakaarohia e mātou te kirihou matū i roto i te anga o te ariā mōhiohio, ā, ka ine i te kanorau me te ngaiotanga o te metabolome i runga i te entropy Shannon o te tohatoha auau matū. Mā te whakamahi i te tātai kua whakatinanahia i mua (8), i tatauhia e mātou he huinga tohu ka taea te whakamahi hei ine i te kanorau metabolome (tohu Hj), te motuhaketanga kōtaha matū (tohu δj) me te motuhaketanga matū o tētahi ira matū kotahi (tohu Si). Hei tāpiritanga, i whakamahia e mātou te Relative Distance Plasticity Index (RDPI) hei ine i te āheinga o ngā kararehe kai otaota ki te whakaoho i te metabolome (Pikitia 1A) (30). I roto i tēnei anga tatauranga, ka tirohia e mātou te whānuitanga MS/MS hei waeine mōhiohio taketake, ā, ka tukatukahia te nui o te MS/MS ki roto i tētahi mahere tohatoha auau, kātahi ka whakamahia te Shannon entropy hei whakatau tata i te kanorau metabolome mai i a ia. Ka inehia te motuhaketanga metabolome e te toharite motuhaketanga o tētahi whānuitanga MS/MS kotahi. Nō reira, ko te pikinga o te nui o ētahi akomanga MS/MS i muri i te whakaoho i te kararehe kai otaota ka hurihia hei āheinga spectral, RDPI me te motuhaketanga, arā, ko te pikinga o te taupū δj, nā te mea ka puta he metabolites motuhake ake, ā, ka puta he taupū Si teitei. Ko te whakahekenga o te taupū kanorau Hj e whakaatu ana kua whakaitihia te maha o ngā MS/MS kua puta, kua rerekē rānei te tohatoha auau kōtaha ki tētahi ahunga iti ake te ōrite, me te whakaiti i tōna koretake whānui. Mā te tātaitanga o te taupū Si, ka taea te whakamārama ko ēhea MS/MS i whakaohokia e ētahi kararehe kai otaota, engari, ko ēhea MS/MS kāore e aro ki te whakaoho, he tohu matua tēnei hei wehewehe i te matapae MT me te OD.
(A) Ngā whakaahuatanga tatauranga e whakamahia ana mō te whakaawe raraunga MS/MS kai otaota (H1 ki Hx) (RDPI), te kanorau (taupū Hj), te motuhaketanga (taupū δj) me te motuhaketanga o te matūkai (taupū Si). Ko te pikinga ake o te tohu motuhaketanga (δj) e tohu ana, i te toharite, ka nui ake ngā matūkai kai otaota ka puta, ko te hekenga o te kanorau (Hj) e tohu ana i te hekenga o te whakaputanga o ngā matūkai, te tohatoha koretake rānei o ngā matūkai i roto i te mahere tohatoha. Ka aromatawai te uara Si mēnā he motuhake te matūkai ki tētahi āhuatanga kua hoatu (i konei, kai otaota) ka puritia rānei i te taumata kotahi. (B) Te hoahoa ariā o te matapae ariā parepare mā te whakamahi i te tuaka ariā mōhiohio. E matapae ana te ariā OD ka whakanui ake te whakaekenga a ngā kai otaota i ngā matūkai parepare, ā, ka piki ake te δj. I taua wā anō, ka heke te Hj nā te mea kua whakaritea anōtia te kōtaha ki te whakaiti i te koretake o ngā mōhiohio pākia. E matapaetia ana e te ariā MT ka puta he huringa kore-aronga i roto i te metabolome i te whakaekenga a ngā kararehe kai otaota, ā, ka piki ake te Hj hei tohu mō te pikinga o te koretake o ngā mōhiohio pākia, ā, ka puta he tohatoha matapōkere o te Si. I whakaarohia hoki e mātou he tauira whakauru, ko te MT pai rawa atu, e tino piki ake ai ētahi metabolites me ngā uara parepare teitei ake (he uara Si teitei), ko ētahi e whakaatu ana i ngā urupare matapōkere (he uara Si iti iho).
Mā te whakamahi i ngā whakaahuatanga ariā mōhiohio, ka whakamāramahia e mātou te ariā OD hei matapae ko ngā huringa metabolite motuhake i puta mai i ngā kararehe kai otaota i roto i te āhua pumau kāore i puta mai ka arahi ki (i) te pikinga ake o te motuhaketanga metabolic (taupū Si) e akiaki ana i te motuhaketanga metabonomic (taupū δj) Te pikinga ake o) ētahi rōpū metabolite motuhake me te uara parepare teitei ake, me (ii) te hekenga o te kanorau metabolome (taupū Hj) nā te huringa o te tohatoha auau metabolic ki te tohatoha tinana leptin ake. I te taumata o te metabolite kotahi, e tumanakohia ana he tohatoha Si kua whakaritea, ka whakanuia e te metabolite te uara Si e ai ki tōna uara parepare (Pikitia 1B). I runga i tēnei rārangi, ka whakamāramahia e mātou te ariā MT hei matapae ko te whakaohooho ka arahi ki (i) ngā huringa kore-aronga i roto i ngā metabolite ka hua ake te hekenga o te taupū δj, me (ii) te pikinga ake o te taupū Hj nā te pikinga ake o te koretake o te metabolic. Te matapōkere rānei, ka taea te ine mā te Shannon entropy i roto i te ahua o te kanorau whānui. Mō te hanganga metabolic, ka matapae te ariā MT i te tohatoha matapōkere o Si. I te whakaaro ko ētahi matū matū kei raro i ngā āhuatanga motuhake i raro i ngā āhuatanga motuhake, ko ētahi atu āhuatanga kāore i raro i ngā āhuatanga motuhake, ā, ko tō rātou uara parepare e whakawhirinaki ana ki te taiao, i whakaarohia hoki e mātou he tauira parepare whakauru, e tohatohahia ana a δj me Hj i roto i ngā taha e rua i te taha o te Si. Ka piki ake i ngā taha katoa, ko ētahi rōpū matū matū anake, he nui ake ō rātou uara parepare, ka tino whakanui ake i te Si, ko ētahi ka tohatoha matapōkeretia (Pikitia 1B).
Hei whakamātautau i te matapae o te ariā parepare kua tautuhia anō i runga i te tuaka o te whakaahuatanga ariā mōhiohio, i whakaarahia e mātou ngā torongū kai otaota tohunga (Ms) me te whānui (Sl) i runga i ngā rau o Nepenthes pallens (Pikitia 2A). Mā te whakamahi i te tātari MS/MS, i tangohia e mātou he 599 ngā momo MS/MS kore-tāpiri (kōnae raraunga S1) mai i ngā tangohanga methanol o te kiko rau i kohia i muri i te whāngai anuhe. Mā te whakamahi i ngā taupū RDPI, Hj, me δj hei tiro i te whirihoranga anō o ngā ihirangi mōhiohio i roto i ngā kōnae whirihoranga MS/MS ka kitea he tauira whakamere (Pikitia 2B). Ko te ia whānui, e ai ki te whakaahuatanga a te whakaahuatanga mōhiohio, i te mea ka kai tonu ngā anuhe i ngā rau, ka piki ake te nui o te whakaritenga pākia katoa i roto i te wā: 72 hāora i muri i te kai a te kai otaota, ka piki ake te RDPI. Ki te whakatauritea ki te mana whakahaere kāore i pakaru, i tino heke te Hj, nā te pikinga ake o te tohu motuhake o te kōtaha pākia, i inehia e te taupū δj. He rite tonu tēnei ia āhua ki ngā matapae o te ariā OD, engari kāore i te rite ki ngā matapae matua o te ariā MT, e whakapono ana ka whakamahia ngā huringa matapōkere (kore-aronga) o ngā taumata metabolite hei huna i te ārai (Pikitia 1B). Ahakoa he rerekē te ihirangi whakaoho ā-waha (OS) me te whanonga whāngai o ēnei kararehe kai otaota e rua, i puta mai i tā rātou whāngai tika he huringa ōrite i ngā ahunga o Hj me δj i ngā wā hauhake 24-hāora me te 72-hāora. Ko te rerekētanga anake i puta i te 72 hāora o te RDPI. Ki te whakatauritea ki te mea i puta mai i te whāngai Ms, he teitei ake te pākia whānui i puta mai i te whāngai Sl.
(A) Hoahoa whakamātautau: ka whāngaihia ngā poaka noa (S1) me ngā tohunga kai otaota (Ms) ki ngā rau kua tangohia te tote o ngā tipu ipu, engari mō ngā kai otaota whakatauira, ka whakamahia te OS o Ms (W + OSM) hei whakahaere i te werohanga o ngā tūranga rau paerewa kua whara. Ngā torongū S1 (W + OSSl) wai rānei (W + W). Ko te mana whakahaere (C) he rau kāore i pakaru. (B) Te Whakaaweawe (RDPI ki te tūtohi whakahaere), te kanorau (taupū Hj) me te taupū motuhake (taupū δj) i tatauhia mō te mahere matūkai motuhake (599 MS/MS; kōnae raraunga S1). E tohu ana ngā whetū i ngā rerekētanga nui i waenga i te whāngai tika a ngā kai otaota me te rōpū whakahaere (whakamātautau-t a te ākonga me te whakamātautau-t takirua, *P<0.05 me ***P<0.001). ns, kāore i te nui. (C) Te tātaitai whakatau wā o te matua (pouaka kikorangi, waikawa amino, waikawa waro me te huka; kōnae raraunga S2) me te whānuitanga matū motuhake (pouaka whero 443 MS/MS; kōnae raraunga S1) i muri i te maimoatanga otaota whaihanga. Ko te whitiki tae e pā ana ki te wā whakawhirinaki 95%. Ko te whetū e tohu ana i te rerekētanga nui i waenga i te maimoatanga me te mana whakahaere [tātari rerekētanga tapawhā (ANOVA), ā, whai muri ko te rerekētanga nui a Tukey (HSD) mō ngā whakataurite maha i muri i te hoc, *P<0.05, **P<0.01 Me *** P <0.001]. (D) Te motuhaketanga o ngā kauwhata marara me ngā kōtaha matū motuhake (ngā tauira maha me ngā maimoatanga rerekē).
Hei tūhura mēnā kei roto i ngā panonitanga o te taumata o ngā matūkai takitahi te whakahoutanga i puta mai i te tipu otaota i te taumata metabolome, i arotahi tuatahi mātou ki ngā matūkai i akohia i mua i roto i ngā rau o Nepenthes pallens me te ātete tipu otaota kua whakamātauhia. Ko ngā phenolic amides he hononga hydroxycinnamamide-polyamine e kohia ana i te wā o te tukanga tipu otaota o ngā pepeke, ā, e mōhiotia ana ka whakaiti i te mahi a ngā pepeke (32). I rapua e mātou ngā tohu o te MS/MS e rite ana, ā, i tuhia ō rātou kōpiko nekeneke whakaemi (Pikitia S1). Ehara i te mea miharo, ko ngā phenol derivatives kāore e whai wāhi tika ana ki te parepare ki ngā tipu otaota, pērā i te waikawa chlorogenic (CGA) me te rutin, ka heke iho i muri i te tipu otaota. He rerekē, ka taea e ngā tipu otaota te hanga i ngā phenol amides kia tino kaha. Ko te whāngai tonu o ngā tipu otaota e rua i hua ake ai te whānuitanga whakaohooho tata rite o ngā phenolamides, ā, he tino kitea tēnei tauira mō te hanganga de novo o ngā phenolamides. Ka kitea anō te āhuatanga kotahi i te wā e tūhuratia ana te ara 17-hydroxygeranyl nonanediol diterpene glycosides (17-HGL-DTGs), e whakaputa ana i te maha o ngā diterpene acyclic me ngā mahi anti-herbivore whai hua (33), ā, nā Ms Feeding me Sl i whakaoho he āhua rite tonu (Pikitia S1)).
Ko te ngoikoretanga pea o ngā whakamātautau whāngai tika i ngā kararehe kai otaota ko te rerekētanga o te tere kai rau me te wā whāngai o ngā kararehe kai otaota, e uaua ai te whakakore i ngā pānga motuhake ki ngā kararehe kai otaota i puta mai i ngā patunga me ngā kararehe kai otaota. Hei whakaoti pai ake i te motuhaketanga o ngā momo kararehe kai otaota o te urupare pākia rau i whakaohokia, i whakatauira mātou i te whāngai i ngā torongū Ms me Sl mā te whakamahi tonu i te OS hou i kohia (OSM me OSS1) ki te wero paerewa W o ngā tūranga rau ōrite. Ka kiia tēnei tikanga ko te maimoatanga W + OS, ā, ka whakataurite i te whakaoho mā te wā tika i te tīmatanga o te urupare i puta mai i te kararehe kai otaota me te kore e puta he pānga whakapoauau o ngā rerekētanga o te tere, te nui rānei o te ngaronga kiko (Pikitia 2A) (34). Mā te whakamahi i te paipa tātari me te tatau MS/MS, i tikina e mātou he 443 MS/MS spectra (kōnae raraunga S1), i taupatupatu ki ngā spectra i huihuia i mua mai i ngā whakamātautau whāngai tika. I whakaatuhia e te tātari ariā mōhiohio o tēnei huinga raraunga MS/MS ko te hōtaka anō o ngā metabolome motuhake rau mā te whakatauira i ngā kararehe kai otaota i whakaatu i ngā whakaoho motuhake OS (Pikitia 2C). Ina koa, ki te whakatauritea ki te maimoatanga OSS1, i hua ake i te OSM te whakanui ake i te motuhaketanga metabolome i te 4 hāora. He mea tika kia mōhiotia, ki te whakatauritea ki te huinga raraunga whakamātautau i whāngai tika i ngā kararehe kai otaota, ko te nekehanga pākia i whakaatuhia i roto i te wāhi rua-ahu mā te whakamahi i a Hj me δj hei taunga me te ahunga o te motuhaketanga metabolome hei whakautu ki te maimoatanga kararehe kai otaota whakatauira i roto i te wā Ka piki tonu (Pikitia 2D). I taua wā anō, i inehia e mātou te ihirangi o ngā waikawa amino, ngā waikawa waro me ngā huka (kōnae raraunga S2) hei tirotiro mēnā ko tēnei pikinga whainga i roto i te tohungatanga metabolome i ahu mai i te whirihoranga anō o te pākia waro pokapū hei whakautu ki ngā kararehe kai otaota whakatauira (Pikitia S2). Hei whakamārama pai ake i tēnei tauira, i aroturukihia anō e mātou te nekehanga kohikohinga pākia o ngā ara phenolamide me te 17-HGL-DTG i kōrerohia i mua. Ko te whakaoho motuhake a ngā kararehe kai otaota OS ka hurihia hei tauira whakarerekētanga rerekē i roto i te pākia phenolamide (Pikitia S3). Ko ngā amide phenolic kei roto ngā wāhanga coumarin me te caffeoyl ka whakaohohia e te OSS1, ko ngā OSM ia ka whakaoho i te whakaoho motuhake o ngā hononga ferulyl. Mō te ara 17-HGL-DTG, i kitea te whakaoho rerekētanga o te OS mā ngā hua malonylation me te dimalonylation o raro (Pikitia S3).
Muri iho, i ako mātou i te kirihou o te transcriptome i puta mai i te OS mā te whakamahi i te huinga raraunga microarray wā-wā, e whakaata ana i te whakamahinga o ngā OSM hei rongoā i ngā rau o ngā rau tipu rosette i roto i ngā otaota kai. Ko te kinetics tauira e taupatupatu ana ki te kinetics i whakamahia i roto i tēnei rangahau metabolomics (35). Ki te whakataurite ki te whirihoranga metabolome e tino piki ake ana te kirihou metabolic i roto i te wā, ka kite mātou i ngā pakaru whakawhiti whakawhiti i roto i ngā rau i puta mai i te Ms, kei reira te inducibility transcriptome (RDPI) me te motuhaketanga (δj) kei te 1. He nui te pikinga o ngā hāora, me te kanorau (Hj) i tēnei wā, kua tino heke te whakaaturanga o BMP1, ā, whai muri ko te whakangāwari o te motuhaketanga transcriptome (Pikitia S4). Ko ngā whānau ira metabolic (pērā i te P450, glycosyltransferase, me te BAHD acyltransferase) e whai wāhi ana ki te tukanga o te kohikohi i ngā metabolites motuhake mai i ngā waeine hanganga i ahu mai i te metabolism tuatahi, e whai ana i te tauira motuhaketanga-tiketike wawe kua whakahuatia ake nei. Hei rangahau take, i tātarihia te ara phenylalanine. I whakaū te tātaritanga he nui te OS-i puta mai i ngā ira matua i roto i te pākia o te phenolamide i roto i ngā kararehe kai otaota, ki te whakaritea ki ngā tipu kāore i kukumea, ā, he tata te tūnga o ā rātou tauira whakaaturanga. Ko te ira whakawhiti MYB8 me ngā ira hanganga PAL1, PAL2, C4H me te 4CL i te tihi o tēnei ara i whakaatu i te tīmatanga wawe o te whakawhiti. Ko ngā Acyltransferases e whai wāhi ana ki te huihuinga whakamutunga o te phenolamide, pērā i te AT1, DH29, me te CV86, e whakaatu ana i tētahi tauira pikinga roa (Pikitia S4). Ko ngā kitenga i runga ake nei e tohu ana ko te tīmatanga wawe o te motuhaketanga transcriptome me te whakarei ake i muri mai o te motuhaketanga metabolomics he aratau honohono, tera pea nā te pūnaha whakahaere tukutahi e tīmata ana i tētahi urupare parepare kaha.
Ko te whirihoranga anō i roto i te tohu homoni tipu he paparanga whakahaere e whakauru ana i ngā mōhiohio kai otaota hei whakahou i te āhua o te tinana o ngā tipu. Whai muri i te whakatauira kai otaota, i inehia e mātou te kaha whakahiato o ngā kāwai homoni tipu matua, ā, i whakaatuhia te hononga-tahi i waenganui i a rātou [tauwehenga taunga Pearson (PCC)> 0.4] (Pikitia 3A). E ai ki te tumanako, ko ngā homoni tipu e pā ana ki te hanga koiora e hono ana i roto i te whatunga hononga-tahi homoni tipu. Hei tāpiri, ka maherehia te motuhaketanga pākia (taupū Si) ki tēnei whatunga hei whakaatu i ngā homoni tipu i puta mai i ngā maimoatanga rerekē. E rua ngā wāhanga matua o te urupare motuhake kai otaota kua tuhia: ko tētahi kei roto i te rōpū JA, kei reira a JA (tōna āhua kaha koiora JA-Ile) me ētahi atu JA e whakaatu ana i te kaute Si teitei rawa atu; ko tētahi ko te ethylene (ET). I whakaatu noa a Gibberellin i te pikinga pakupaku noa iho o te motuhaketanga kai otaota, ko ētahi atu homoni tipu, pērā i te cytokinin, te auxin, me te waikawa abscisic, he iti te motuhaketanga whakaoho mō ngā kai otaota. Ki te whakatauritea ki te whakamahi i te W + W anake, ka taea te huri i te whakanui ake i te uara tihi o ngā JA derivatives mā te tono OS (W + OS) hei tohu kaha motuhake o ngā JA. Kāore i te tūmanakohia, e mōhiotia ana ko te OSM me te OSS1 me ngā ihirangi elicitor rerekē ka puta he rite tonu te kohikohinga o te JA me te JA-Ile. He rerekē ki te OSS1, ka whakaohokia motuhaketia te OSM e ngā OSM, engari kāore te OSS1 e whakanui ake i te urupare o ngā patunga turanga (Pikitia 3B).
(A) Te tātari whatunga whakapuakanga-tahi i runga i te tatau PCC o te whakatauira kinetics kohikohinga homoni tipu i puta mai i ngā kararehe kai otaota. Ko te pūnga e tohu ana i tētahi homoni tipu kotahi, ā, ko te rahi o te pūnga e tohu ana i te taupū Si motuhake ki te homoni tipu i waenga i ngā maimoatanga. (B) Te kohikohinga o JA, JA-Ile me ET i roto i ngā rau i puta mai i ngā maimoatanga rerekē e tohuhia ana e ngā tae rerekē: āperekoti, W + OSM; kikorangi, W + OSSl; mangu, W + W; hina, C (mana whakahaere). Ko ngā whetū e tohu ana i ngā rerekētanga nui i waenga i te maimoatanga me te mana whakahaere (ANOVA ara-rua i whai ake ko Tukey HSD whakataurite maha post hoc, *** P <0.001). Te tātari ariā mōhiohio o (C)697 MS/MS (kōnae raraunga S1) i roto i te biosynthesis JA me te whānuitanga tirohanga ngoikore (irAOC me irCOI1) me (D)585 MS/MS (kōnae raraunga S1) i roto i te ETR1 me te tohu ET ngoikore E rua ngā maimoatanga kararehe kai otaota i whakatauira i whakaoho i ngā raina tipu me ngā tipu whakahaere waka kau (EV). E whakaatu ana ngā whetū i ngā rerekētanga nui i waenga i te maimoatanga W+OS me te mana whakahaere kāore i pakaru (ANOVA ara-rua i muri mai ko te whakataurite maha Tukey HSD post hoc, *P<0.05, **P<0.01 me ***P<0.001). (E) Ngā kauwhata marara o te whakahē marara ki te tohungatanga. Ko ngā tae e tohu ana i ngā momo momo ira kua whakarerekētia; ko ngā tohu e tohu ana i ngā tikanga maimoatanga rerekē: tapatoru, W + OSS1; tapawhā hāngai, W + OSM; porowhita C
Muri iho, ka whakamahia e mātou he momo Nepenthes kua whakarerekētia te ira (irCOI1 me te sETR1) i roto i ngā mahi matua o te hanga JA me te ET (irAOC me te irACO) me te tirohanga (irCOI1 me te sETR1) hei tātari i te pākia o ēnei homoni tipu e rua i runga i ngā kararehe kai otaota. Ko te takoha whanaunga o te hōtaka anō. E rite ana ki ngā whakamātautau o mua, i whakaūhia e mātou te whakaoho o te OS kararehe kai otaota i roto i ngā tipu kawe kau (EV) (Pikitia 3, C ki D) me te hekenga whānui o te taupū Hj i puta mai i te OSM, i te mea kua piki ake te taupū δj. He nui ake te kitea o te urupare i te urupare i puta mai i te OSS1. E whakaatu ana te kauwhata rārangi-rua e whakamahi ana i a Hj me δj hei taunga i te whakakorenga motuhake (Pikitia 3E). Ko te ia tino kitea ko roto i ngā momo kāore he tohu JA, kua tata kore rawa te kanorau metabolome me ngā huringa motuhake i puta mai i ngā kararehe kai otaota (Pikitia 3C). He rerekē, ko te tirohanga ET puku i roto i ngā tipu sETR1, ahakoa he iti iho te pānga whānui ki ngā huringa o te pākia o te kai otaota i tērā o te tohu JA, ka whakaiti i te rerekētanga o ngā tohu Hj me δj i waenga i ngā whakaohooho OSM me OSS1 (Pikitia 3D me te Pikitia S5). . E tohu ana tēnei, hei tāpiri atu ki te mahi matua o te tuku tohu JA, ka mahi anō te tuku tohu ET hei whakatikatika pai i te urupare pākia motuhake-momo o ngā kai otaota. E rite ana ki tēnei mahi whakatikatika pai, kāore he huringa i roto i te kaha whānui o te whakaoho metabolome i roto i ngā tipu sETR1. I tētahi atu taha, ki te whakatauritea ki ngā tipu sETR1, i puta mai i ngā tipu irACO ngā whānuitanga whānui o ngā huringa pākia i puta mai i ngā kai otaota, engari i whakaatu i ngā kaute Hj me δj rerekē i waenga i ngā wero OSM me OSS1 (Pikitia S5).
Hei tautuhi i ngā matū motuhake he nui ngā takoha ki te urupare motuhake a ngā kararehe kai otaota, me te whakatikatika i tā rātou whakaputanga mā roto i ngā tohu ET, i whakamahia e mātou te tikanga MS/MS hanganga kua whakawhanakehia i mua. Ka whakawhirinaki tēnei tikanga ki te tikanga whakarōpū-rua hei whakatau anō i te whānau matū mai i ngā kongakonga MS/MS [hua ira kua whakaritea (NDP)] me te kaute ōrite i runga i te ngaronga koretake (NL). Ko te huinga raraunga MS/MS i hangaia mā te tātari i ngā raina whakawhiti ET i whakaputa i te 585 MS/MS (kōnae raraunga S1), i whakatauhia mā te whakarōpū i a rātou ki roto i ngā kōwae matua e whitu o te MS/MS (M) (Pikitia 4A). Ko ētahi o ēnei kōwae he kī tonu i ngā matū motuhake kua tautuhia i mua: hei tauira, he nui ngā momo matū phenol (M1), ngā glycosides flavonoid (M2), ngā huka acyl (M3 me te M4), me te 17-HGL-DTG (M7) o M1, M2, M3, M4 me te M7. Hei tāpiritanga, ka tatauhia ngā mōhiohio motuhake mō te pākia o tētahi matūkai kotahi i roto i ia kōwae, ā, ka taea te kite i tōna tohatoha Si mā te whakaaro noa. I roto i te poto, ko ngā ira MS/MS e whakaatu ana i te tino motuhake o te tipu me te momo ira e tohuhia ana e ngā uara Si teitei, ā, e tohu ana ngā tatauranga kurtosis i te tohatoha o te huruhuru i te kokonga hiku matau. I kitea tētahi tohatoha colloid kikokore i roto i te M1, i reira ko te phenol amide i whakaatu i te wāhanga Si teitei rawa (Pikitia 4B). Ko te 17-HGL-DTG whakatipu tipu kua whakahuatia i mua ake nei i roto i te M7 i whakaatu i te kaute Si ngawari, e tohu ana i te taumata whakahaere rerekētanga ngawari i waenga i ngā momo OS e rua. He rerekē, ko te nuinga o ngā matūkai motuhake i hangaia, pērā i te rutin, te CGA, me te huka acyl, kei roto i ngā kaute Si iti rawa. Hei tūhura pai ake i te uauatanga hanganga me te tohatoha Si i waenga i ngā matūkai motuhake, i hangaia he whatunga ngota mō ia kōwae (Pikitia 4B). Ko tētahi matapae nui o te ariā OD (kua whakarāpopototia ki te Pikitia 1B) ko te whakaritenga anō o ngā matū motuhake i muri i te whakatipu otaota me arahi ki ngā huringa kotahi-ara o ngā matū he nui te uara tiaki, inā koa mā te whakanui ake i tō rātou motuhaketanga (ka rerekē ki te tohatoha matapōkere) Aratau) Te matū tiaki i matapaetia e te ariā MT. Ko te nuinga o ngā matū phenol i kohia i roto i te M1 e pā ana ki te mahi ki te hekenga o te mahi pepeke (32). I te whakataurite i ngā uara Si i roto i ngā matū M1 i waenga i ngā rau i whakaohokia me ngā rau wāhanga o ngā tipu whakahaere EV i te 24 hāora, i kite mātou he nui te pikinga o te motuhaketanga pākia o te maha o ngā matū i muri i ngā pepeke whakatipu otaota (Pikitia 4C). Ko te pikinga motuhake o te uara Si i kitea i roto i ngā phenolamide tiaki anake, engari kāore i kitea he pikinga o te uara Si i roto i ētahi atu phenol me ngā matū kāore i te mōhiotia e noho tahi ana i roto i tēnei kōwae. He tauira motuhake tēnei, e pā ana ki te ariā OD. He rite tonu ngā matapae matua mō ngā huringa pākia i puta mai i ngā tipu whakatipu otaota. Hei whakamātautau mēnā i puta mai tēnei āhuatanga motuhake o te whānuitanga phenolamide e te ET motuhake-OS, i tuhia e mātou te taupū metabolite Si, ā, i puta he uara whakaaturanga rerekē i waenga i te OSM me te OSS1 i roto i ngā momoira EV me te sETR1 (Pikitia 4D). I roto i te sETR1, i tino whakaitihia te rerekētanga i puta mai i te phenamide i waenga i te OSM me te OSS1. I whakamahia anō hoki te tikanga bi-clustering ki ngā raraunga MS/MS i kohia i roto i ngā momo he iti nei te JA hei whakatau i ngā kōwae MS/MS matua e pā ana ki te motuhaketanga metabolic e whakahaeretia ana e JA (Pikitia S6).
(A) Ko ngā hua whakarōpū o te 585 MS/MS i runga i te kongakonga tiritahi (rite NDP) me te ngaronga koretake tiritahi (rite NL) ka hua ake te ōrite o te kōwae (M) ki te whānau matū e mōhiotia ana, nā te hanganga Metabolite kāore i te mōhiotia, kāore rānei i te pai te whakapūngao. I te taha o ia kōwae, ka whakaaturia te tohatoha motuhake o te metabolite (MS/MS) (Si). (B) Whatunga ngota ngota: Ko ngā pona e tohu ana i te MS/MS me ngā taha, ngā kaute MS/MS NDP (whero) me te NL (kikorangi) (tapahanga, > 0.6). Ko te taupū motuhake metabolite kua tohua (Si) i taehia i runga i te kōwae (maui) ā, kua maherehia ki te whatunga ngota ngota (matau). (C) Kōwae M1 o te tipu EV i roto i te āhua pumau (mana) me te āhua whakaoho (kai otaota whakatauira) i te 24 hāora: te tūtohi whatunga ngota ngota (ko te uara Si te rahi o te pona, kua tohua te phenolamide parepare ki te kikorangi). (D) Ko te hoahoa whatunga ngota M1 o te rārangi whānuitanga sETR1 me te tirohanga EV me te ET ngoikore: te matū phenolic e tohuhia ana e te pona porowhita matomato, me te rerekētanga nui (uara P) i waenga i ngā maimoatanga W + OSM me W + OSS1 hei rahi o te Pona. CP, N-caffeoyl-tyrosine; CS, N-caffeoyl-spermidine; FP, N-ferulic acid ester-uric acid; FS, N-ferulyl-spermidine; CoP, N', N “-Coumarolyl-tyrosine; DCS, N', N”-dicaffeoyl-spermidine; CFS, N', N”-caffeoyl, feruloyl-spermidine; Lycium barbarum i roto i te wolfberry Son; Nick. O-AS, O-acyl sugar.
I whakawhānui ake mātou i te tātari mai i tētahi momo ira kotahi o Nepenthes kua whakaitihia ki ngā taupori tūturu, i reira kua whakaahuahia ngā huringa ā-roto-motuhake o ngā taumata JA kai otaota me ngā taumata metabolite motuhake i roto i ngā taupori tūturu (26). Whakamahia tēnei huinga raraunga hei kapi i te 43 ira-pūmua. Ko ēnei ira-pūmua he 123 ngā momo tipu mai i N. pallens. I tangohia ēnei tipu mai i ngā purapura i kohia i roto i ngā nohonga taketake rerekē i Utah, Nevada, Arizona, me California (Pikitia S7), i tatauhia e mātou te kanorau metabolome (e kiia nei ko te taumata taupori) kanorau β) me te motuhaketanga i puta mai i te OSM. E rite ana ki ngā rangahau o mua, i kite mātou i te whānuitanga o ngā huringa pākia i te taha o ngā tuaka Hj me δj, e tohu ana he rerekētanga nui o ngā ira-pūmua i roto i te kirihou o ā rātou urupare pākia ki ngā kai otaota (Pikitia S7). He rite tēnei whakahaere ki ngā kitenga o mua mō te whānuitanga hihiri o ngā huringa JA i puta mai i ngā kai otaota, ā, kua mau tonu te uara tino teitei i roto i te taupori kotahi (26, 36). Mā te whakamahi i te JA me te JA-Ile hei whakamātautau i te hononga taumata whānui i waenga i a Hj me δj, i kitea e mātou he hononga pai nui i waenga i a JA me te taupū kanorau me te motuhaketanga metabolome β (Pikitia S7). E whakaatu ana tēnei ko te rerekētanga i puta mai i ngā kararehe kai otaota i roto i te whakaoho JAs i kitea i te taumata taupori ka puta pea i ngā momo āhuahanga matua o te pūnaha i puta mai i te kōwhiringa mai i ngā kararehe kai otaota pepeke.
Kua whakaatuhia e ngā rangahau o mua he rerekē rawa ngā momo tupeka i roto i te momo me te whakawhirinakitanga whanaunga ki ngā parenga pākia i puta mai me ngā parenga pumau. E whakaponohia ana ko ēnei huringa i roto i te tuku tohu ārai-kaiota me ngā kaha parenga ka whakahaeretia e te pēhanga taupori pepeke, te huringa ora o te tipu, me ngā utu whakaputa parenga i roto i te wāhi e tipu ana tētahi momo. I akohia e mātou te ōritetanga o te whakahounga metabolome rau i puta mai i ngā kaiota o ngā momo Nicotiana e ono e noho ana i Amerika Te Raki me Amerika Te Tonga. He whanaunga tata ēnei momo ki a Nepenthes Amerika Te Raki, arā, ko Nicolas Bociflo. La, N. nicotinis, Nicotiana n. tarutaru ngoikore, Nicotiana tabacum, tupeka raina, tupeka (Nicotiana spegazzinii) me te tupeka rau tupeka (Nicotiana obtusifolia) (Pikitia 5A) (37). E ono o ēnei momo, tae atu ki te momo kua tautuhia pai ko N., he tipu ā-tau o te clade petunia, ā, ko obtusifolia N. he tipu ā-tau o te clade tuahine Trigonophyllae (38). I muri mai, i whakahaerehia te W + W, W + OSM me te W + OSS1 ki ēnei momo e whitu hei ako i te whakaritenga anō o te pākia o te whāngai pepeke i runga i te taumata momo.
(A) He rākau whakapapa irahiko i runga i te tūponotanga nui [mō te hanganga glutamine karihi (38)] me te tohatoha whenua o ngā momo Nicotiana e whitu e tata ana (ngā tae rerekē) (37). (B) He kauwhata marara o te kanorau motuhake mō ngā kōtaha pākia o ngā momo Nicotiana e whitu (939 MS/MS; kōnae raraunga S1). I te taumata momo, he hononga kino te kanorau metabolome ki te taumata o te tohungatanga. Ko te tātaritanga o te hononga taumata-momo i waenga i te kanorau pākia me te tohungatanga me te kohikohinga JA e whakaatuhia ana i te Pikitia 2. S9. Tae, ngā momo rerekē; tapatoru, W + OSS1; tapawhā hāngai, W + OSM; (C) Ko te Nicotiana JA me ngā hihiri JA-Ile kua whakarōpūtia kia rite ki te kaha whakaoho OS (ANOVA ara-rua me te Tukey HSD whakatauritenga muri-maha, * P <0.05, ** P <0.01 me te * ** Hei whakataurite i a W + OS me W + W, P <0.001). Kauwhata pouaka o (D) te kanorau me (E) te motuhaketanga o ia momo i muri i te whakatauira i te kararehe kai otaota me te methyl JA (MeJA). E tohu ana te whetū i te rerekētanga nui i waenga i te W + OS me te W + W, te lanolin rānei me te W (Lan + W) me te Lan rānei me te MeJA (Lan + MeJa) me te mana whakahaere Lan (tātaritanga rerekētanga rua-ara, ā, whai muri ko te whakataurite maha HSD a Tukey i muri i te hoc, *P<0.05, **P<0.01 me te ***P<0.001).
Mā te whakamahi i te tikanga takirua takirua, i tautuhia e mātou e 9 ngā kōwae o te 939 MS/MS (kōnae raraunga S1). He rerekē te hanganga o te MS/MS i whirihorahia anō e ngā maimoatanga rerekē i waenga i ngā kōwae rerekē i waenga i ngā momo (Pikitia S8). Mā te whakaata i a Hj (e kiia nei ko te γ-diversity taumata-momo) me δj e whakaatu ana ka huihui ngā momo rerekē ki ngā rōpū tino rerekē i roto i te wāhi pākia, i te nuinga o te wā ka kitea ake te wehenga taumata-momo i te whakaohooho. Haunga a N. linear me N. obliquus, he whānui te whānuitanga o ngā pānga whakaohooho e whakaatuhia ana e rātou (Pikitia 5B). He rerekē, ko ngā momo pēnei i a N. purpurea me N. obtusifolia he iti ake te kitea o te urupare pākia ki te maimoatanga, engari he rerekē ake te metabolome. Ko te tohatoha motuhake o te urupare pākia i whakaohokia i hua ake he hononga kino nui i waenga i te motuhaketanga me te kanorau gamma (PCC = -0.46, P = 4.9×10-8). He hononga pai ngā panoni i puta mai i te OS i roto i ngā taumata JA ki te motuhaketanga metabolome, ā, he hononga kino ki te kanorau gamma metabolic i whakaatuhia e ia momo (Pikitia 5B me te Pikitia S9). He mea tika kia mōhio ko ngā momo e kīia ana ko ngā momo "tohu urupare" i roto i te Pikitia 5C, pērā i te Nepenthes nematodes, te Nepenthes nepenthes, te Nepenthes acute, me te Nepenthes attenuated, i puta ai ngā tohu nui i te 30 meneti. Ko ngā pakarutanga JA me te JA-Ile motuhake OS tata nei, ko ētahi atu huakita i kiia ko te "tohu kore urupare", pērā i te Nepenthes mills, te Nepenthes powdery me te N. obtusifolia e whakaatu ana i te JA-Ile Edge anake me te kore he OS motuhake (Pikitia 5C). I te taumata metabolic, e ai ki te kōrero i runga ake nei, mō ngā Nepenthes kua whakaitihia, i whakaatu ngā matū urupare-tohu i te OS motuhake me te whakanui ake i te δj, me te whakaiti i te Hj. Kāore i kitea tēnei pānga priming motuhake OS i roto i ngā momo kua whakarōpūtia hei momo tohu kore-tauhohe (Pikitia 5, D me te E). He maha ake te tohatoha o ngā matū whai hua motuhake ki te OS i waenga i ngā momo e aro ana ki te tohu, ā, ka huihui ēnei tautau tohu ki ngā momo he ngoikore ake ngā urupare tohu, ko ngā momo he ngoikore ake ngā urupare tohu e whakaatu ana i te iti ake o te whakawhirinakitanga (Pikitia S8). E whakaatu ana tēnei hua ko te whakaohooho motuhake ki te OS o ngā JA me te whirihoranga anō motuhake ki te OS o te metabolome o raro e honoa ana i te taumata momo.
Muri iho, i whakamahia e mātou he whakapiri rauropi kei roto ko te methyl JA (MeJA) hei rongoā i ngā tipu hei tirotiro mēnā kei te herea ēnei momo hononga e te wātea o te JA e whakamahia ana e te JA o waho, ka noho ki roto i te cytoplasm o ngā tipu. Ko te whakakorenga tere ko te JA. I kitea e mātou te ia o te huringa āta haere mai i ngā momo e aro ana ki te tohu ki ngā momo kāore e aro ki te tohu i puta mai i te tuku tonu o te JA (Pikitia 5, D me E). I roto i te poto, i kaha te hōtaka a te maimoatanga MeJA i ngā metabolomes o ngā nematodes raina, N. obliquus, N. aquaticus, N. pallens, me N. mikimotoi, i hua ake ai he pikinga nui o te δj me te hekenga o te Hj. Ko te N. purpurea i whakaatu noa i te pikinga o te δj, engari kāore a Hj. N. obtusifolia, kua whakaaturia i mua he iti rawa ngā taumata o ngā JA, he kino hoki te urupare ki te maimoatanga MeJA e pā ana ki te whirihoranga metabolome. E whakaatu ana ēnei hua he herenga ā-tinana te whakaputanga JA, te whakawhiti tohu rānei i roto i ngā momo kāore e aro ki te tohu. Hei whakamātautau i tēnei whakapae, i ako mātou i ngā momo e whā (N. pallens, N. mills, N. pink me N. microphylla) i whakaohokia e W + W, W + OSMs me W + OSS1 Transcriptome (39). E rite ana ki te tauira o te hanga metabolome, he pai te wehewehe o ngā momo i roto i te wāhi transcriptome, i roto i ēnei ko N. attenuated i whakaatu i te RDPI OS-induced teitei rawa atu, ko N. gracilis te iti rawa (Pikitia 6A). Heoi, i kitea ko te kanorau transcriptome i whakaohokia e N. oblonga te mea iti rawa i roto i ngā momo e whā, he rerekē ki te kanorau metabonomic teitei rawa atu o N. oblonga i whakaaturia i mua i roto i ngā momo e whitu. Kua whakaatuhia e ngā rangahau o mua ko tētahi huinga ira e pā ana ki ngā tohu parepare wawe, tae atu ki ngā tohu JA, e whakamārama ana i te motuhaketanga o ngā urupare parepare wawe i whakaohokia e ngā elicitors e pā ana ki ngā otaota i roto i ngā momo Nicotiana (39). Mā te whakataurite i ngā ara tohu JA i waenga i ēnei momo e whā i kitea he tauira whakamere (Pikitia 6B). Ko te nuinga o ngā ira i roto i tēnei ara, pērā i a AOC, OPR3, ACX me COI1, i whakaatu i ngā taumata teitei o te whakaoho i roto i ēnei momo e whā. Heoi, ko tētahi ira matua, ko JAR4, e huri ana i a JA ki roto i tōna āhua koiora kaha o ngā tuhinga kua kohia mō te JA-Ile, ā, he tino iti tōna taumata whakaputa, inā koa i roto i a N. mills, Nepenthes pieris me N. microphylla. Hei tāpiri, ko te whakaputanga anake o tētahi atu ira AOS kāore i kitea i roto i a N. bifidum. Ko ēnei huringa o te whakaaturanga ira pea te take o ngā momo āhua tino nui i puta mai i te iti o te whakaputanga JA i roto i ngā momo tohu mate pukupuku me te whakaoho o N. gracilis.
(A) Te tātaritanga ariā mōhiohio o te hōtaka anō o ngā urupare whakawhiti tuatahi o ngā momo tupeka e whā e pā tata ana i tauirahia i te 30 meneti i muri i te whakaurunga otaota. Ka tatauhia te RDPI mā te whakataurite i ngā rau i whakaohokia e te OS otaota me te mana whakahaere patunga. Ka tohu ngā tae i ngā momo rerekē, ā, ka tohu ngā tohu i ngā tikanga maimoatanga rerekē. (B) Te tātaritanga o te whakaaturanga ira i roto i ngā ara tohu JA i waenga i ngā momo e whā. Ka whakaaturia te ara JA kua whakangawaritia i te taha o te kauwhata pouaka. Ka tohu ngā tae rerekē i ngā tikanga tukatuka rerekē. E tohu ana te whetū he rerekētanga nui i waenga i te maimoatanga W + OS me te mana whakahaere W + W (mō te whakamātautau-t a te Tauira mō ngā rerekētanga takirua, *P<0.05, **P<0.01 me ***P<0.001). OPDA, waikawa 12-oxophytodienoic; OPC-8: waikawa 0,3-oxo-2(2′(Z)-pentenyl)-cyclopentane-1-octanoic.
I te wāhanga whakamutunga, i ako mātou me pēhea te whakarerekētanga motuhake o te metabolome o ngā momo tipu rerekē ki ngā momo pepeke ka taea te ātete ki ngā kararehe kai otaota. I whakanuia e ngā rangahau o mua te momo Nicotiana. He tino rerekē te ātete ki a Ms me ngā torongū (40). I konei, i ako mātou i te hononga i waenga i tēnei tauira me tō rātou kirihou pākia. Mā te whakamahi i ngā momo tupeka e whā i runga ake nei, me te whakamātautau i te hononga i waenga i te kanorau me te motuhaketanga o te metabolome i puta mai i ngā kararehe kai otaota me te ātete o ngā tipu ki a Ms me Sl, i kitea e mātou he hononga pai te ātete, te kanorau me te motuhaketanga ki te Sl whānui, engari he ngoikore te hononga i waenga i te ātete ki ngā wāhine tohunga me te motuhaketanga, ā, kāore te hononga ki te kanorau i te nui (Pikitia S10). Mō te ātete S1, ko te N. chinensis me te N. gracilis kua ngoikore, i whakaaturia i mua he taumata whakawhiti tohu JA me te kirihou metabolome, he tino rerekē ngā urupare ki te whakaoho kararehe kai otaota, ā, i whakaatu hoki rātou i te ātete teitei rite tonu. Ira tangata.
I roto i ngā tau ono tekau kua pahure ake nei, kua whakaratohia e te ariā parepare tipu he anga ariā, e ahu mai ana i ngā kairangahau kua matapae i te maha o ngā whanaketanga me ngā mahi a ngā matū matū motuhake o te tipu. Ko te nuinga o ēnei ariā kāore e whai i te tukanga noa o ngā whakatau kaha (41). Ka whakaarohia e rātou ngā matapae matua (3) i te taumata tātari kotahi. Ina taea e te whakamātautau i ngā matapae matua te tātari i ngā ariā motuhake, ka tautokona tēnei, engari ka whakakahoretia ētahi atu (42). Engari, ka whakatakoto te ariā hou i ngā matapae i ngā taumata tātari rerekē, ā, ka tāpirihia he paparanga hou o ngā whakaaro whakaahua (42). Heoi, ko ngā ariā e rua i whakaarohia i te taumata mahi, MT me OD, ka taea te whakamārama ngāwari hei matapae nui mō ngā huringa matū motuhake i puta mai i ngā kararehe kai otaota: E whakapono ana te ariā OD he tino aronga ngā huringa i roto i te "wāhi" matū motuhake. E whakapono ana te ariā MT ka kore e aronga ēnei huringa, ā, ka tū matapōkeretia ki te wāhi matū, ā, he nui te uara o ngā matū matū tiaki. Kua whakamatautauria ngā whakamātautau o mua o ngā matapae OD me MT mā te whakamahi i tētahi huinga whaiti o ngā matū "tiaki" a priori. Ko ēnei whakamātautau e aro ana ki te metabolite ka ārai i te kaha ki te tātari i te whānuitanga me te ara o te whirihoranga anō o te metabolome i te wā o te whakatipu otaota, ā, kāore e whakaaetia te whakamātautau i roto i tētahi anga tatauranga ōrite kia hiahiatia he matapae matua ka taea te whakaaro whānui. Te ine i ngā huringa i roto i te metabolome tipu. I konei, i whakamahia e mātou te hangarau auaha i roto i te metabolomics i runga i te MS rorohiko, ā, i whakahaerehia he tātaritanga MS wetewete i roto i te moni whānui o ngā whakaahuatanga ariā mōhiohio hei whakamātautau i te rerekētanga i waenga i ngā mea e rua i whakaarohia i te taumata metabolomics o te ao. Ko te matapae matua o tēnei ariā. Kua whakamahia te ariā mōhiohio i roto i ngā mara maha, inā koa i roto i te horopaki o te koiora koiora me te rangahau rere matūkai (43). Heoi, e ai ki tā mātou e mōhio ana, koinei te tono tuatahi i whakamahia hei whakaahua i te wāhi mōhiohio metabolic o ngā tipu me te whakaoti rapanga taiao e pā ana ki ngā huringa metabolic rangitahi hei whakautu ki ngā tohu taiao. Ina koa, ko te kaha o tēnei tikanga kei roto i tōna kaha ki te whakataurite i ngā tauira i roto, i waenga hoki i ngā momo tipu hei tirotiro i te whanaketanga o ngā otaota mai i ngā momo rerekē ki ngā tauira macroevolutionary i waenga i ngā momo i ngā taumata whanaketanga rerekē. Metabolism.
Ka hurihia e te tātaritanga wāhanga matua (PCA) tētahi huinga raraunga maha-taurangi ki tētahi wāhi whakaiti inenga kia taea ai te whakamārama i te ia matua o ngā raraunga, nō reira ka whakamahia hei tikanga tūhura hei tātari i te huinga raraunga, pērā i te metabolome wetewete. Heoi, ka ngaro tetahi wāhanga o ngā ihirangi mōhiohio i roto i te huinga raraunga i te whakaiti inenga, ā, kāore e taea e te PCA te whakarato mōhiohio tauanga mō ngā āhuatanga e tino pā ana ki te ariā rauropi, pērā i: te āhua o te whirihoranga anō a ngā kararehe kai otaota i te kanorau i roto i ngā mara motuhake (hei tauira, te momonatanga, te tohatoha) me te nui) ngā metabolites? Ko ēhea ngā metabolites hei tohu i te āhua whakaoho o tētahi kararehe kai otaota? Mai i te tirohanga o te motuhaketanga, te kanorau me te whakaaweawe, ka wehewehea ngā ihirangi mōhiohio o te kōtaha metabolite motuhake-rau, ā, ka kitea ko te kai a ngā kararehe kai otaota ka taea te whakahohe i te pākia motuhake. I te mea ohorere, i kite mātou, e ai ki te whakaahuatanga i roto i ngā tohu ariā mōhiohio kua whakatinanahia, he nui te taupatupatu o te āhuatanga pākia i puta mai i muri i ngā whakaekenga a ngā kararehe kai otaota e rua (te tohunga whānui i te pō a Sl) me te tohunga Solanaceae a Ms. Ahakoa he tino rerekē te whanonga kai me te kukū o ā rāua whanonga. Te kaiwhakaoho waikawa ngako-waikawa amino (FAC) i roto i te OS (31). Mā te whakamahi i te OS kararehe kai otaota hei rongoā i ngā patunga werohanga paerewa, i whakaatuhia hoki e te maimoatanga kararehe kai otaota te ia o taua ia. Mā tēnei tikanga paerewa mō te whakatauira i te urupare a ngā tipu ki ngā whakaekenga a ngā kararehe kai otaota ka whakakore i ngā āhuatanga whakapoauau i puta mai i ngā huringa o te whanonga kai a ngā kararehe kai otaota, e arahi ana ki ngā taumata rerekē o te kino i ngā wā rerekē (34). Ko te FAC, e mōhiotia ana ko te tino take o te OSM, ka whakaiti i te JAS me ētahi atu urupare homoni tipu i roto i te OSS1, i te mea ka whakaitihia e te OSS1 ngā rau wā (31). Heoi, i puta i te OSS1 ngā taumata rite o te kohikohinga JA ki te OSM. Kua whakaaturia i mua ko te urupare JA i roto i te Nepenthes ngoikore he tino tairongo ki te OSM, ka taea e te FAC te pupuri i tana mahi ahakoa i whakaranua ki te 1:1000 me te wai (44). Nō reira, ki te whakatauritea ki te OSM, ahakoa he tino iti te FAC i roto i te OSS1, he rawaka hei whakaoho i te pakarutanga o te JA. Kua whakaatuhia e ngā rangahau o mua ka taea te whakamahi i ngā pūmua rite ki te porin (45) me ngā oligosaccharides (46) hei tohu ngota hei whakaoho i ngā urupare parepare tipu i roto i te OSS1. Heoi, kāore anō kia mārama mēnā ko ēnei kaiwhakaoho i roto i te OSS1 te take o te kohikohinga o te JA i kitea i roto i te rangahau o nāianei.
Ahakoa he iti noa ngā rangahau e whakaahua ana i ngā tohu rerekē o te pākia i puta mai i te whakamahinga o ngā momo kai otaota rerekē, o ngā momo JA, SA rānei o waho (waikawa salicylic) (47), kāore anō kia whakararuraruhia te whakararuraru motuhake o ngā momo kai otaota i roto i te whatunga tarutaru tipu me ōna pānga ki ngā mōhiohio whaiaro. I whakaūhia anō e tēnei tātaritanga ko te hononga whatunga homoni ā-roto me ētahi atu homoni tipu ehara i te JA te mea e hanga ana i te motuhaketanga o te whakaritenga anō o te pākia i puta mai i ngā kai otaota. Ina koa, i kitea e mātou he nui ake te ET i puta mai i te OSM i tērā i puta mai i te OSS1. He rite tonu tēnei aratau ki te nui ake o te ihirangi FAC i roto i te OSM, he āhuatanga e tika ana, e rawaka ana hoki mō te whakaoho i te pahūtanga ET (48). I roto i te horopaki o te taunekeneke i waenga i ngā tipu me ngā kai otaota, he tūpono tonu te mahi tohu a te ET ki ngā whanaketanga metabolite motuhake o te tipu, ā, he kotahi noa te rōpū matū e aro ana ki tēnei. Hei tāpiritanga, ko te nuinga o ngā rangahau kua whakamahi i te tono ā-waho o te ET, i ōna mua rānei, i ngā momo aukati rānei hei ako i te whakahaere i te ET, ā, i roto i ēnei tono matū ā-waho ka puta he maha ngā pānga taha kore-motuhake. Ki tā mātou mōhio, ko tēnei rangahau te tuatahi o ngā whakamātautau pūnaha nui o te mahi a te ET i roto i te whakamahinga o te ET hei whakaputa me te kite i ngā tipu whakawhiti-ira kua ngoikore hei whakarite i ngā mahi metabolome tipu. Ka taea e te whakaoho ET motuhake ki ngā tipu otaota te whakarerekē i te urupare metabolome. Ko te mea nui ko te whakahaere whakawhiti-ira o ngā ira ET biosynthesis (ACO) me te perception (ETR1) i whakaatu i te kohikohinga de novo motuhake ki ngā tipu otaota o ngā phenolamides. Kua whakaaturia i mua ka taea e te ET te whakatikatika i te kohikohinga nicotine i puta mai i te JA mā te whakahaere i te putrescine N-methyltransferase (49). Heoi, mai i te tirohanga miihini, kāore i te mārama me pēhea te whakatikatika a te ET i te whakaoho o te phenamide. Haunga te mahi whakawhiti tohu o te ET, ka taea hoki te whakawhiti i te rere o te pākia ki te S-adenosyl-1-methionine hei whakahaere i te haumitanga ki ngā polyaminophenol amides. Ko te S-adenosyl-1-methionine te ET me te takawaenga noa o te ara hangahanga polyamine. Me rangahau anō te tikanga e whakahaere ai te tohu ET i te taumata o te phenolamide.
Mō te wā roa, nā te nui o ngā matū motuhake kāore i te mōhiotia te hanganga, kāore i taea e te aro nui ki ngā kāwai matū motuhake te aromatawai tika i ngā huringa wā o te kanorau matū i muri i ngā taunekeneke koiora. I tēnei wā, i runga i te tātaritanga ariā mōhiohio, ko te hua matua o te whiwhinga MS/MS i runga i ngā matū kore whakaaro ko te whakaiti tonu a ngā kararehe kai otaota e kai ana, e whakatauira ana rānei i ngā kararehe kai otaota i te kanorau matū whānui o te matū rau, me te whakanui ake i tōna taumata motuhake. Ko tēnei pikinga wā poto i roto i te motuhaketanga metabolome i puta mai i ngā kararehe kai otaota e pā ana ki te pikinga tahi o te motuhaketanga transcriptome. Ko te āhuatanga e tino whai wāhi ana ki tēnei motuhaketanga metabolome nui ake (he uara Si teitei ake) ko te matū motuhake me te mahi kararehe kai kua tautuhia i mua. He rite tēnei tauira ki te matapae o te ariā OD, engari kāore te matapae o te MT e pā ana ki te matapōkeretanga o te hōtaka anō metabolome i te rite. Heoi, he rite anō hoki tēnei raraunga ki te matapae o te tauira whakauru (MT pai rawa atu; Pikitia 1B), nā te mea ko ētahi atu matū kāore i te tautuhia me ngā mahi tiaki kāore i te mōhiotia ka whai tonu pea i te tohatoha Si matapōkere.
Ko tētahi tauira whakahirahira i hopukina ake e tēnei rangahau ko te mea mai i te taumata whanaketanga-iti (te tipu kotahi me te taupori tupeka) ki tētahi tauine whanaketanga nui ake (ngā momo tupeka e pā tata ana), kei roto i te "parepare pai" ngā taumata rerekē o te whakahaere whanaketanga. He rerekētanga nui kei roto i ngā kaha o ngā kararehe kai otaota. I whakaaro motuhake a Moore et al. (20) me Kessler me Kalske (1) ki te huri i ngā taumata mahi e toru o te koiora koiora i wehewehea tuatahitia e Whittaker (50) ki ngā huringa wā-tūturu me te wā-whakaoho o te kanorau matū; kāore ēnei kaituhi i whakarāpopoto i ngā tikanga mō te kohikohi raraunga metabolome nui-tauine kāore hoki e whakamārama me pēhea te tatau i te kanorau metabolic mai i ēnei raraunga. I roto i tēnei rangahau, ko ngā whakatikatika iti ki te whakarōpūtanga mahi a Whittaker ka whakaarohia te kanorau α-metabolic hei kanorau o ngā ira MS/MS i roto i tētahi tipu, me te kanorau β-metabolic hei te metabolism intraspecific taketake o tētahi rōpū taupori. Wāhi, me te kanorau γ-metabolic hei toronga o te tātaritanga o ngā momo rite.
He mea nui te tohu JA mō te whānuitanga o ngā urupare pākia o ngā kararehe kai otaota. Heoi, kāore he whakamātautau tauanga pakari mō te takoha o te whakahaere ā-roto o te hanga koiora JA ki te kanorau metabolome, ā, kāore anō kia mārama mēnā ko te tohu JA he wāhi whānui mō te kanorau pākia i puta mai i te ahotea i runga i te tauine whanaketanga nui ake. I kite mātou ko te āhua kai otaota o te Nepenthes he kai otaota ka whakaoho i te motuhaketanga metabolome, ā, ko te rerekētanga o te motuhaketanga metabolome i roto i te taupori o ngā momo Nicotiana me ngā momo Nicotiana e whanaunga tata ana he hononga pai ki te tohu JA. Hei tāpiri, ina ngoikore te tohu JA, ka whakakorehia te motuhaketanga pākia i puta mai i tētahi momo kararehe kai otaota kotahi (Pikitia 3, C me E). Nā te mea he tauanga te nuinga o ngā huringa whānuitanga pākia o ngā taupori Nepenthes kua ngoikore noa, ko ngā huringa o te kanorau β pākia me te motuhaketanga i roto i tēnei tātaritanga ka puta pea i te nuinga o te wā nā te kaha o te whakaoho o ngā kāwai matū whai-metabolome. Ko ēnei akomanga matū te nuinga o te kōtaha metabolome, ā, ka arahi ki te hononga pai ki ngā tohu JA.
Nā te mea he tino rerekē ngā tikanga matū koiora o ngā momo tupeka e tata ana ki a ia, ka tautuhia motuhaketia ngā matū matū i roto i te taha kounga, nō reira he tātari ake. Mā te tukatuka a te ariā mōhiohio i te kōtaha matū kua hopukina e whakaatu ana ko te whakaoho kai otaota ka whakanui ake i te tauhokohoko i waenga i te kanorau gamma matū me te motuhaketanga. He mahi nui tā te tohu JA i roto i tēnei tauhokohoko. Ko te pikinga ake o te motuhaketanga metabolome e rite ana ki te matapae OD matua, ā, he hononga pai ki te tohu JA, ko te tohu JA ia he hononga kino ki te kanorau gamma matū. E tohu ana ēnei tauira ko te kaha OD o ngā tipu ka whakatauhia e te kirihou o JA, ahakoa i runga i te tauine whanaketanga moroiti, i runga rānei i te tauine whanaketanga nui ake. Ko ngā whakamātautau tono JA o waho e karo ana i ngā hapa biosynthesis JA e whakaatu ana ka taea te wehewehe i ngā momo tupeka e tata ana ki a rātou ki ngā momo e aro ana ki te tohu me ngā momo kāore e aro ki te tohu, pērā i tā rātou momo JA me te kirihou metabolome i puta mai i ngā kai otaota. Kāore e taea e ngā momo kāore e aro ki te tohu te urupare nā te kore e taea te whakaputa JA taketake, nō reira kei raro i ngā herenga ā-tinana. Tērā pea nā ngā whakarerekētanga ira o ētahi ira matua i roto i te ara tohu JA (AOS me JAR4 i roto i te N. crescens) o tēnei. E whakaatu ana tēnei hua ko ēnei tauira whanaketanga nui i waenga i ngā momo ka ahu mai pea i ngā huringa o te tirohanga ā-roto me te urupare ki ngā homoni.
Haunga te taunekeneke i waenga i ngā tipu me ngā kararehe kai otaota, ko te tūhuratanga o te kanorau pākia e pā ana ki ngā whanaketanga ariā nui katoa i roto i te rangahau o te urutau koiora ki te taiao me te whanaketanga o ngā āhuatanga matatini o te phenotypic. I te pikinga ake o te nui o ngā raraunga i kohia e ngā taputapu MS hou, ka taea e te whakamātautau whakapae mō te kanorau pākia te neke atu i ngā rerekētanga matūkai takitahi/kāwai me te mahi tātaritanga ā-ao hei whakaatu i ngā tauira ohorere. I roto i te tukanga tātaritanga tauine nui, he kupu whakarite nui ko te whakaaro ki te whakaaro i ngā mahere whai tikanga ka taea te whakamahi hei tūhura i ngā raraunga. Nō reira, ko tētahi hua nui o te whakakotahitanga o nāianei o te MS/MS metabolomics kore whakaaro me te ariā mōhiohio ko te whakarato i tētahi inenga ngāwari ka taea te whakamahi hei hanga mahere hei tirotiro i te kanorau pākia i runga i ngā tauine taxonomic rerekē. Koinei te whakaritenga taketake o tēnei tikanga. Ko te ako i te whanaketanga moroiti/macro me te rauropi hapori.
I te taumata whanaketanga whānui, ko te kaupapa matua o te ariā whanaketanga tahi o ngā tipu me ngā pepeke a Ehrlich rāua ko Raven (51) ko te matapae ko te rerekētanga o te kanorau pākia i waenga i ngā momo te take o te kanorau o ngā whakapapa tipu. Heoi, i roto i ngā tau rima tekau mai i te whakaputanga o tēnei mahi matua, he onge te whakamātautau i tēnei ariā (52). Ko te nuinga o tēnei nā ngā āhuatanga phylogenetic o ngā āhuatanga pākia rite puta noa i ngā whakapapa tipu tawhiti. Ka taea te whakamahi i te onge hei whakapūmau i ngā tikanga tātari ūnga. Ko te reremahi MS/MS o nāianei i tukatukahia e te ariā mōhiohio e ine ana i te hanganga MS/MS o ngā metabolites kāore i te mōhiotia (me te kore he kōwhiringa metabolite i mua) ā, ka huri i ēnei MS/MS ki tētahi huinga MS/MS, nō reira i roto i te pākia ngaio ka whakaritea ēnei tauira whanaketanga whānui i roto i te tauine whakarōpūtanga. Ngā tohu tatauranga māmā. He rite te tukanga ki te tātari phylogenetic, ka taea te whakamahi i te whakaritenga raupapa hei ine i te tere o te kanorau, te whanaketanga rānei o te āhua me te kore he matapae i mua.
I te taumata matū koiora, e whakaatu ana te ariā tirotiro a Firn rāua ko Jones (53) kei te pupurihia te kanorau pākia i ngā taumata rerekē hei whakarato rauemi mata hei whakahaere i ngā mahi koiora o ngā matū kore hononga, ngā matū whakakapi rānei i mua. Ka whakaratohia e ngā tikanga ariā mōhiohio he anga e taea ai te ine i ēnei whakawhiti whanaketanga motuhake-pūmua e puta ana i te wā motuhaketanga matū hei wāhanga o te tukanga tirotiro whanaketanga kua whakaarohia: te urutau kaha koiora mai i te iti o te motuhaketanga ki te teitei o te motuhaketanga Ngā matū aukati o tētahi taiao kua hoatu.
I te katoa, i ngā rā tōmua o te koiora ngota, i whakawhanakehia ngā ariā tiaki tipu nui, ā, e kiia whānuitia ana ko ngā tikanga whakapae-whakaaroaro anake te huarahi mō te ahunga whakamua pūtaiao. Ko te nuinga o tēnei nā ngā herenga hangarau o te ine i te metabolome katoa. Ahakoa he tino whai hua ngā tikanga whakapae-whakaaroaro i roto i te whiriwhiri i ētahi atu tikanga take, he iti ake tō rātou kaha ki te whakatairanga i tō tātou māramatanga ki ngā whatunga matū koiora i ngā tikanga tātai e wātea ana i roto i te pūtaiao raraunga-nui o nāianei. Nō reira, ko ngā ariā kāore e taea te matapae kei tua atu i te whānuitanga o ngā raraunga e wātea ana, nō reira kāore e taea te whakakore i te huringa tauira/whakamātautau whakapae o te ahunga whakamua i roto i te mara rangahau (4). E matapaetia ana e mātou ka taea e te reremahi tātai o te metabolomics i whakaurua mai ki konei te whakaoho anō i te hiahia ki ngā take hou (me pēhea) me te whakamutunga (he aha) o te kanorau metabolic, me te whai wāhi ki tētahi wā hou o te pūtaiao raraunga e arahina ana e te ariā. I tirotirohia anō e te wā ngā ariā nui i whakaaweawe i ngā whakatupuranga o mua.
Ko te whāngai tika i ngā kararehe kai mā te whakatipu i tētahi torongū tuarua o te tipu, i tētahi torongū Sl rānei ki runga i tētahi rau putiputi kotahi he mā te tae o tētahi tipu rōhi kotahi e puāwai ana, me te 10 ngā tāruatanga tipu mō ia tipu. I herea ngā torongū pepeke ki ngā here, ā, i kohia te kiko rau e toe ana i muri i te 24 me te 72 hāora i muri i te pangia, ā, i whakatio wawetia, kātahi ka tangohia ngā matūkai.
Whakatauirahia te maimoatanga otaota i roto i te huarahi tino tukutahi. Ko te tikanga he whakamahi i ngā wira tauira papanga hei wero i ngā rarangi tataramoa e toru i ia taha o te puku o ngā rau e toru kua whānui katoa o te tipu i te wā tipu o te tauera papanga, ā, ka pania tonutia te 1:5 Ms kua whakaranua. Mēnā rānei, whakamahia he maihao karapu hei whakauru i te S1 OS ki roto i te patunga wero. Hauhakehia, ka tukatukahia he rau e ai ki te whakaahuatanga i runga ake nei. Whakamahia te tikanga kua whakaahuatia i mua hei tango i ngā matū matū tuatahi me ngā homoni tipu (54).
Mō ngā whakamahinga JA o waho, ka rongoatia ngā rau petiole e toru o ia o ngā tipu pua rōhi e ono o ia momo ki te 20μl o te whakapiri lanolin kei roto ko te 150μg MeJA (Lan + MeJA), me te 20μl o te lanolin me te maimoatanga patunga (Lan + W), ka whakamahia rānei te 20μl o te lanolin parakore hei mana whakahaere. I hauhaketia ngā rau 72 hāora i muri i te maimoatanga, i whakatiohia ki te hauota wai, ā, i rongoatia ki te -80°C tae noa ki te whakamahinga.
E whā ngā momo ira whakawhiti JA me ET, arā, ko te irAOC (36), ko te irCOI1 (55), ko te irACO me te sETR1 (48), kua tautuhia i roto i tā mātou rōpū rangahau. I whakaatu kaha te irAOC i te hekenga o ngā taumata JA me te JA-Ile, engari kāore te irCOI1 i aro ki ngā JA. Ki te whakaritea ki te EV, i piki ake te kohikohinga JA-Ile. Waihoki, ka whakaitihia e te irACO te whakaputanga o te ET, ā, ki te whakaritea ki te EV, ka whakanuia e te sETR1, he kore aro ki te ET, te whakaputanga o te ET.
Ka whakamahia he inenga taiaho whakaahua-oro (pūoko ET wā-tūturu Sensor Sense ETD-300) hei ine i te ET, kāore he pokanga. I muri tonu i te maimoatanga, ka tapahia te haurua o ngā rau, ka whakawhitia ki tētahi ipu karāhe hiri 4-ml, ā, ka tukuna kia kohia te āputa i roto i ngā haora e 5. I te wā o te ine, ka horoia ia ipu ki te rere o te 2 rita/haora o te hau parakore mō te 8 meneti, i mua i te haere mā roto i tētahi whakakorikori i whakaratohia e Sensor Sense hei tango i te CO2 me te wai.
I whakaputaina tuatahitia ngā raraunga microarray ki (35) ā, i tiakina ki te National Center for Biotechnology Information (NCBI) Gene Expression Comprehensive Database (nama whakauru GSE30287). I tangohia ngā raraunga e pā ana ki ngā rau i puta mai i te maimoatanga W + OSMs me te mana whakahaere kāore i pakaru mō tēnei rangahau. Ko te kaha mata ko te log2. I mua i te tātaritanga tatauranga, i hurihia te turanga me te whakataurite ki tōna ōrau 75 mā te whakamahi i te kete pūmanawa R.
I tangohia ngā raraunga raupapatanga RNA taketake (RNA-seq) o ngā momo Nicotiana mai i te NCBI Short Reading Archives (SRA), ko te nama kaupapa ko PRJNA301787, i tukuna e Zhou et al. (39) ā, ka haere tonu i runga i te whakaahuatanga i (56). Ko ngā raraunga mata i tukatukahia e W + W, W + OSM me W + OSS1 e pā ana ki ngā momo Nicotiana i tīpakohia mō te tātari i roto i tēnei rangahau, ā, i tukatukahia i te huarahi e whai ake nei: Tuatahi, i hurihia ngā pānui RNA-seq mata ki te hōputu FASTQ. Ka hurihia e HISAT2 te FASTQ ki te SAM, ā, ka hurihia e SAMtools ngā kōnae SAM ki ngā kōnae BAM kua whakarōpūtia. Ka whakamahia a StringTie hei tatau i te whakaaturanga ira, ā, ko tana tikanga whakaaturanga he kongakonga mō ia mano kongakonga turanga mō ia miriona kongakonga whakawhiti raupapa.
Ko te pou chromatographic Acclaim (150 mm x 2.1 mm; rahi matūriki 2.2μm) i whakamahia i roto i te tātari me te pou tiaki 4 mm x 4 mm he mea hanga mai i te rauemi kotahi. Ko te gradient rua e whai ake nei e whakamahia ana i roto i te pūnaha Dionex UltiMate 3000 Ultra High Performance Liquid Chromatography (UHPLC): 0 ki te 0.5 meneti, isocratic 90% A [wai deionized, 0.1% (v/v) acetonitrile me te 0.05% waikawa formic], 10% B (Acetonitrile me te 0.05% waikawa formic); 0.5 ki te 23.5 meneti, ko te wāhanga gradient ko 10% A me 90% B, ia; 23.5 ki te 25 meneti, isocratic 10% A me 90% B. Ko te tere rere ko 400μl/min. Mō ngā tātaritanga MS katoa, werohia te eluent pou ki roto i tētahi tātaritanga quadrupole me te time-of-flight (qTOF) kua rite ki tētahi pūtake rehu hiko e mahi ana i te aratau ionization pai (ngaohiko capillary, 4500 V; putanga capillary 130 V; pāmahana whakamaroke 200°C; rere o te hau whakamaroke 10 rita/min).
Whakahaerehia te tātaritanga kongakonga MS / MS (e kiia nei i muri nei ko MS / MS) kāore e pā ana, e kore rānei e taea te wehewehe i ngā raraunga hei tiki i ngā mōhiohio hanganga mō te kōtaha pākia whānui ka kitea. Ko te ariā o te tikanga MS/MS kore wehewehe e whakawhirinaki ana ki te meka he matapihi wehewehe papatipu nui rawa atu tō te quadrupole [nō reira, whakaarohia ngā tohu ōwehenga papatipu-ki-utu katoa (m/z) hei kongakonga]. Mō tēnei take, nā te mea kāore i taea e te taputapu Impact II te hanga i te honga CE, he maha ngā tātaritanga motuhake i mahia mā te whakamahi i ngā uara pūngao pakaru wehewehe (CE) kua piki ake i te tukinga. I roto i te poto, tātarihia tuatahi te tauira mā te UHPLC-electrospray ionization/qTOF-MS mā te whakamahi i te aratau spectrometry papatipu kotahi (ngā tikanga pakaru iti i hangaia e te pakaru i roto i te pūtake), matawai mai i te m/z 50 ki te 1500 i te auau tāruarua o te 5 Hz. Whakamahia te hauota hei hau tukinga mō te tātaritanga MS/MS, me te mahi i ngā inenga motuhake i ngā ngaohiko pakaru wehewehe e whā e whai ake nei: 20, 30, 40, me te 50 eV. I roto i te tukanga ine, ko te whā-pū te matapihi wehenga papatipu nui rawa atu, mai i te m/z 50 ki te 1500. Ina whakatakotoria te tinana o mua m/z me te whānui wehenga ki te 200, ka whakahohehia aunoatia te awhe papatipu e te pūmanawa whakahaere o te taputapu me te 0 Da. Matawaihia mō ngā kongakonga papatipu pērā i te aratau papatipu kotahi. Whakamahia te konutai formate (50 ml isopropanol, 200 μl waikawa formic me te 1 ml 1M otinga wai NaOH) mō te whakatikatika papatipu. Mā te whakamahi i te raupapa whakatikatika tino tika a Bruker, ka whakatikatikaina te kōnae raraunga i muri i te whakahaere i te whānuitanga toharite i roto i tētahi wā kua whakaritea. Whakamahia te mahi kaweake o te pūmanawa Data Analysis v4.0 (Brook Dalton, Bremen, Tiamana) hei huri i ngā kōnae raraunga mata ki te hōputu NetCDF. Kua tiakina te huinga raraunga MS/MS ki te pātengi raraunga metabolomics tuwhera MetaboLights (www.ebi.ac.uk) me te tau whakauru. MTBLS1471.
Ka taea te whakatinana i te huihuinga MS/MS mā te tātari hononga i waenga i ngā tohu kounga MS1 me MS/MS mō te kaha tukinga iti me te kaha tukinga teitei me ngā ture hou kua whakatinanahia. Ka whakamahia te tuhinga R hei whakatinana i te tātari hononga o te tohatoha o te mua ki te hua, ā, ka whakamahia te tuhinga C# (https://github.com/MPI-DL/indiscriminant-MS-MS-assembly-pipeline) hei whakatinana i ngā ture.
Hei whakaiti i ngā hapa teka pai i puta mai i te haruru papamuri me te hononga teka i puta mai i te kitenga i ētahi āhuatanga m/z i roto i ētahi tauira torutoru noa iho, ka whakamahia e mātou te mahi "tihi kī tonu" o te mōkihi R XCMS (mō te whakatikatika haruru papamuri) Me whakamahi hei whakakapi i te kaha "NA" (tihi kāore i kitea). Ina whakamahia te mahi tihi kī, he maha tonu ngā uara kaha "0" i roto i te huinga raraunga ka pā ki te tatau hononga. Kātahi ka whakatauritea e mātou ngā hua tukatuka raraunga i whiwhihia i te wā e whakamahia ana te mahi tihi kī tonu me te wā kāore e whakamahia te mahi tihi kī tonu, ka tatau i te uara haruru papamuri i runga i te uara toharite kua whakatikatikaina, kātahi ka whakakapia ēnei uara kaha 0 ki te uara papamuri kua tatauhia. I whakaarohia anō hoki e mātou ngā āhuatanga ko te kaha e toru ngā wā neke atu i te uara papamuri, ā, i whakaarohia ko rātou he "tihi pono." Mō ngā tataunga PCC, ko ngā tohu m/z anake o te tauira mua (MS1) me ngā huinga raraunga wāhanga me te iti rawa e waru ngā tihi pono ka whakaarohia.
Mena he hononga nui te kaha o te āhuatanga kounga o mua i roto i te tauira katoa ki te kaha kua whakaitihia o te āhuatanga kounga kotahi e pāngia ana e te kaha tukinga iti, teitei rānei, ā, kāore te āhuatanga i tohua hei tihi isotope e CAMERA, ka taea te tautuhi anō. Kātahi, mā te tatau i ngā takirua hua-mua katoa i roto i te 3 hēkona (te matapihi wā pupuri whakatau tata mō te pupuri tihi), ka mahia te tātari hononga. Mēnā he iti iho te uara m/z i te uara o mua, ā, ka puta te wehewehenga MS/MS i te wāhi tauira kotahi i roto i te huinga raraunga me te mua i ahu mai ai, ka kiia he kongakonga.
I runga i ēnei ture māmā e rua, ka tangohia e mātou ngā kongakonga kua tohua me ngā uara m/z nui ake i te m/z o te mua kua tautuhia, ā, i runga anō i te tūranga tauira kei reira te mua me te kongakonga kua tohua. Ka taea hoki te tīpako i ngā āhuatanga kounga i hangaia e te maha o ngā kongakonga i roto i te pūtake i hangaia i te aratau MS1 hei mua kaitono, ā, ka puta he pūhui MS/MS tāpiri. Hei whakaiti i tēnei tāpiritanga raraunga, mēnā he nui ake te ōritetanga NDP o ngā irahiko i te 0.6, ā, nō roto rātou i te chromatogram "pcgroup" kua tohua e CAMERA, ka whakakotahi mātou i a rātou. Hei whakamutunga, ka whakakotahi mātou i ngā hua CE e whā e pā ana ki te mua me ngā kongakonga ki roto i te irahiko whakakotahi whakamutunga mā te tīpako i te tihi kaha teitei rawa atu i waenga i ngā tihi kaitono katoa me te uara m/z kotahi i ngā kaha tukinga rerekē. Ko ngā mahi tukatuka e whai ake nei e hangai ana ki te ariā o te irahiko whakakotahi, ā, ka whakaarohia ngā āhuatanga CE rerekē e hiahiatia ana hei whakanui ake i te tūponotanga o te pakaru, nā te mea ka taea anake te kite i ētahi kongakonga i raro i te kaha tukinga motuhake.
I whakamahia te RDPI (30) hei tatau i te āheinga o te kōtaha pākia. Ko te kanorau o te whānuitanga pākia (taupū Hj) i ahu mai i te nui o ngā matū mua MS/MS mā te whakamahi i te entropy Shannon o te tohatoha auau MS/MS mā te whakamahi i te whārite e whai ake nei i whakaahuahia e Martínez et al. (8). Hj = −∑i = 1mPijlog2(Pij) kei reira a Pij e rite ana ki te auau whanaunga o te MS/MS-i i roto i te tauira-j (j = 1, 2,…, m) (i = 1, 2,…, m) t).
Ko te motuhaketanga pākia (taupū Si) e tautuhia ana ko te tuakiri whakaaturanga o tētahi MS/MS e pā ana ki te auau i waenga i ngā tauira e whakaarohia ana. Ka tatauhia te motuhaketanga MS/MS hei Si = 1t (∑j = 1tPijPilog2PijPi)
Whakamahia te tātai e whai ake nei hei ine i te taupū δj motuhake-metabolome o ia tauira j, me te toharite o te motuhaketanga MS/MS δj = ∑i = 1mPijSi
Ka whakarārangihia ngā irahiko MS/MS kia takirua, ā, ka tatauhia te ōritetanga i runga i ngā kaute e rua. Tuatahi, mā te whakamahi i te NDP paerewa (e mōhiotia ana ko te tikanga taunga cosine), whakamahia te whārite e whai ake nei hei tatau i te ōritetanga wāhanga i waenga i ngā irahiko NDP = (∑iS1 & S2WS1, iWS2, i) 2∑iWS1, i2∑iWS2, i2 kei reira a S1 me S2. Mō te irahiko 1 me te irahiko 2, tae atu ki a WS1, i me WS2, ko i te tohu i te taumaha i runga i te kaha o te tihi, ko te rerekētanga o te tihi noa i waenga i ngā irahiko e rua he iti iho i te 0.01 Da. Ka tatauhia te taumaha penei: W = [kaha o te tihi] m [kounga] n, m = 0.5, n = 2, e ai ki tā MassBank.
I whakatinanahia tētahi tikanga tātai tuarua, arā, ko te tātari i te NL i tohaina i waenga i te MS/MS. Hei whakatutuki i tēnei, i whakamahia e mātou ngā rārangi NL e 52 i kitea pinepine i te wā o te tukanga wehewehe MS, me te NL tino motuhake (kōnae raraunga S1) kua whakamāramahia i mua mō te whānuitanga MS/MS o ngā matū tuarua o te momo Nepenthes kua ngoikore (9, 26). Waihangahia he whārite rua o te 1 me te 0 mō ia MS/MS, e rite ana ki te nāianei me te kore o ētahi NL. I runga i te ōrite o te tawhiti Euclidean, ka tatauhia te kaute ōrite NL mō ia takirua o ngā whārite rua NL.
Hei whakahaere i te whakarōpūtanga takirua, i whakamahia e mātou te mōkihi R DiffCoEx, e ahu mai ana i te toronga o te Weighted Gene Co-Expression Analysis (WGCNA). Mā te whakamahi i ngā matihiko whakatauranga NDP me NL o ngā ira MS/MS, i whakamahia e mātou te DiffCoEx hei tatau i te matihiko taurite. Ka mahia te whakarōpūtanga takirua mā te whakatakoto i te tawhā "cutreeDynamic" ki te tikanga = "hybrid", cutHeight = 0.9999, deepSplit = T, me te minClusterSize = 10. I tangohia te waehere pūtake R o DiffCoEx mai i te kōnae tāpiri 1 e Tesson et al. (57); Ka kitea te mōkihi pūmanawa R WGCNA e hiahiatia ana i roto i te https://horvath.genetics.ucla.edu/html/CoexpressionNetwork/Rpackages/WGCNA.
Hei whakahaere i te tātari whatunga ngota MS/MS, i tatauhia e mātou te hononga hihinga takirua i runga i ngā momo ōrite NDP me NL, ā, i whakamahia te pūmanawa Cytoscape hei whakaatu i te āhua whatunga mā te whakamahi i te tahora waro i roto i te tono toronga raupaparorohiko tahora CyFilescape yFiles.
Whakamahia te putanga R 3.0.1 hei whakahaere i te tātaritanga tatauranga mō ngā raraunga. I aromatawaihia te hiranga tatauranga mā te whakamahi i te tātaritanga rerekētanga ara-rua (ANOVA), ā, i whai ake ko te whakamātautau whai muri i te rerekētanga hiranga pono a Tukey (HSD). Hei tātari i te rerekētanga i waenga i te maimoatanga otaota me te mana whakahaere, i tātarihia te tohatoha taha-rua o ngā rōpū tauira e rua me te rerekētanga ōrite mā te whakamahi i te whakamātautau-t a Student.
Mō ngā rauemi tāpiri mō tēnei tuhinga, tirohia http://advances.sciencemag.org/cgi/content/full/6/24/eaaz0381/DC1
He tuhinga urunga-tuwhera tēnei e tohatohahia ana i raro i ngā tikanga o te Raihana Creative Commons Attribution-Non-Commercial, e āhei ai te whakamahinga, te tohatoha me te tārua i roto i tētahi momo pāpāho, mena kāore te whakamahinga whakamutunga mō te whai hua arumoni, ā, ko te whakaaro matua he tika te mahi taketake. Tohutoro.
Kia mōhio: Ko tā mātou anake he tuku mai i tō wāhitau īmēra kia mōhio ai te tangata e taunaki ana koe ki te whārangi e hiahia ana koe kia kite rātou i te īmēra, ā, ehara i te pāme. Kāore mātou e hopu i tētahi wāhitau īmēra.
Ka whakamahia tēnei pātai hei whakamātautau mēnā he manuhiri koe, hei ārai hoki i te tuku pāme aunoa.
Mā te ariā mōhiohio ka taea te whakataurite i ngā metabolome motuhake me te matapae i ngā ariā parepare whakamātautau.
Mā te ariā mōhiohio ka taea te whakataurite i ngā metabolome motuhake me te matapae i ngā ariā parepare whakamātautau.
©2021 American Association for the Advancement of Science. mana katoa. Ko AAAS te hoa o HINARI, AGORA, OARE, CHORUS, CLOCKSS, CrossRef me COUNTER. ScienceAdvances ISSN 2375-2548.